Asian Journal of Pharmacodynamics and Pharmacokinetics

Official Journal of International Quarterly Publication ISSN 1608-2281 

Original Journal Name: Asian Journal of Drug Metabolism and Pharmacokinetcs (2001-2005) 

力力 

Asian Journal of 

Pharmacodynamics and 

Pharmacokinetics 

Academic Editor-in-Chief 

Chang-Xiao Liu and Yuichi Sugiyama 

Volume Number 1 March 2009 

Hong Kong Medical Publisher

Asian Journal of Pharmacodynamics and Pharmacokinetics 

An International Quarterly Publication 

ISSN 1608-2281 NR 3880/169/00 

http://www.hktmc.com/ChineseMedia/Magazine/Medicine/ajdmpk 

Publisher President 

Kwong Cheung Ku Hong Kong Medical Publisher 24/F Yuen Long Trading Center, 99-109 Castle Peak Road, Yuen 

Long, Hong Kong, China 

Academic Editor-in-chief 

Chang-Xiao Liu Tianjin Institute of Pharmaceutical Research, 308 An-Shan West Road, Tianjin 300193, China 

Yuichi Sugiyama Department of Molecular Pharmacokinetics, The University of Tokyo, Tokyo, 113-0033, Japan 

Assistant Academic Editor-in-Chief 

Da-Fang Zhong Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 646 Songtao Road Shanghai, 

201203, China 

Guang-Hua Du Institute of Materia Medica, Chinese Academy of Medical Sciences, Beijing, 100050, China 

Qiang Zhang School of Pharmaceutical Sciences, Peking University, Beijing, 100083, China 

Publishing Editor-in-chief 

Annie Ning Editor-in-chief of Hong Kong Medical Publisher 

Hui Qing Shi Vice-editor-in-chief of Hong Kong Medical Publisher 

Members of Editorial Committee 

Henning H. Blume (Oberursel, 

GERMANY) 

Zheng-Min Chen (Tianjin, CHINA) 

Moses SS Chow (Hong Kong, CHINA) 

Guang-Hua Du (Beijing, CHINA) 

Paul Fawcett (Otago, NEW ZEALAND) 

Jack Gao (Carborough, CANADA) 

Mei-Yu Geng (QingdaoCHINA 

Patrick M. Gennissel (Paris, FRANCE) 

Kun Han (Chungbuk, KOREA) 

Guo-Zhu Han (Dalian, CHINA) 

Pei Hu (Beijing, CHINA) 

Zhuo-Han Hu (Shanghai, CHINA) 

Xi Huang (Chengdu, CHINA) 

Jun-Yan Hong (New Jersey, USA) 

Li-Ya Ju (Charenton le Pont, FRANCE) 

Devarakonda R. Krishna (Kakatiya, 

INDIA) 

June Lee (NIH, USA) 

Jian-Guo Li (Wilmington, USA) 

Lin-Lin Li (Xinjiang, CHINA) 

Zhi-Bin Lin (Beijing, CHINA) 

Terry D. Lindstrom (Indiana, USA) 

Chang-Xiao Liu (Tianjin, CHINA) 

Xiao-Dong Liu (Nanjing, CHINA) 

Ke-Xin Liu (Dalian, CHINA) 

Jian-Shi Lou (Tianjin, CHINA) 

Takeo Murakawa (Osaka, JAPAN) 

Terumichi Nakagawa (Kyoto, JAPAN) 

Charles H. Nightingale (Hartford, USA) 

Inotsume Nobuo (Hokkaido, JAPAN) 

Richard ZM Qian (Hong Kong, CHINA) 

Madhubala Rentala (New Delhi, 

INDIA) 

Jin-Xiu Ruan (Beijing, CHINA) 

Guo-Wei Sang (Beijing, CHINA) 

Xiang-Guo Shi (Boston, USA) 

Duan-Yun Si (Tianjin, CHINA) 

Chang Koo Shim (Seoul, KOREA) 

Yuichi Sugiyama (Tokyo, JAPAN) 

Ding-Feng Su (Shanghai, CHINA) 

Nikolaus Sucher (Hong Kong, CHINA) 

Yong-Da Sun (Bradford, UK) 

Hui-Qing Shi (Hong Kong, CHINA) 

Chang-Koo Shim (Seoul, KOREA) 

Guang-Ji Wang (Nanjing, CHINA) 

Hui Wang (Wuhan, CHINA) 

Ming-Wei Wang (Shanghai, CHINA) 

Zhi-Min Wang (Beijing, CHINA) 

Yi-Tao Wang (Macau, CHINA) 

Guang-Li Wei (Tianjin, CHINA) 

Chun-Fu Wu (ShenyangCHINA) 

Jinn Wu (New Jersey, USA) 

Fu-Ming Xie ( Indiana, USA) 

Guo-Wang Xu (Dalian, CHINA) 

Ming Xue (Beijing, CHINA) 

Bao-Feng Yang (HarbinCHINA) 

Zong-Hui Yuan (Wuhan, CHINA) 

Fan-Dian Zeng (Wuhan, CHINA) 

Su Zeng (Hangzhou, CHINA) 

Qiang Zhang (Beijing, CHINA) 

Jiang Zheng (SeattleUSA) 

Da-Fang Zhong (Shanghai, CHINA) 

Hong-Hao Zhou (Changsha, CHINA) 

Yao-Wei Zhu (PA, USA) 

Zhu Zhu (Beijing, CHINA)


(An International Quarterly Publication) 

Volume 9 Number 1 March 2009 

Content 

3 Ethical Guideline to Authors, Editors and Reviewers 

Special Repart 

5 Introduction to Professor Leslie Z. Benet and his Lecture in China: Predicting Drug Absorption and 

Disposition Using a Biopharmaceutics Drug Disposition Classification System 

Review Papers 

11 Hai-Yu Xu , Tie-Jun Zhang , Xue-Yu Zhu,Yu-Bo Li. Recent advance on chemical compositions and 

pharmacodynamic and pharmacokinetic studies of Rhizoma Coptidis 

27 Tie-Feng Cheng, Yong-Da Sun, Duan-Yun SiChang-Xiao Liu. Attention on research of 

pharmacology and toxicology of nanomedicines 

Research Papers 

51 Rui Zhang, Benjie Wang, Hengli Zhao, Chunmin Wei, Guiyan Yuan, Ruichen Guo. Tissue 

distribution of Curcumol in rats after intravenous injection of zedoary turmeric oil fat emulsion 

58 Sama Venkatesh, Yanadaiah JP, Zareen N, Madhava Reddy B, Ramesh M. Antinociceptive effect 

of Aerva lanata ethanolic extract in mice: A possible mechanism 

63 

Run Li, Zong-Peng Zhang, Yi-Hong Tian. Therapeutic effect and mechanism for anti-fibrosis of 

polyhydroxysilbene of Rhizoma Scirpi in hepatic fibrosis rats 

71 Xiao-Pu Nie, Wen-Yuan Gao, Pei-Gen Xiao. Changes of the adenosine content in single and mixed 

decoctions of Gualou-xiebai-baijiu Decoction 

Information and News 

10 Publication News Chinese Herbal Medicines 

26 Tianjin Centre for Drug Safety Evaluation and Research 

50 Publication News Drug Evaluation Research 

50 The 3rd Asian Pacific ISSX Meeting 

77 Successful the 2nd Asian–Pacific Regional ISSX Meeting 

78 Information for authors 

2

Ethical Guideline. Asian Journal of Pharacodynamics and Pharmacokinetics 2009;9(1):3-4 




ISSN 1608-2281 

Copyright by Hong Kong Medical Publisher 

Publisher Homepage: www.hktmc.com 

Ethical Guideline to Authors, Editors and Reviewers 

Drug products are specific goods with safety 

and effectiveness in medical health case. All of 

researchers (authors), reviewers and editors must 

abide by medical ethical obligation, and also must 

deter to ethical obligation for publication. These 

guidelines are offered as ethical behavior standards. 

We now present a set of ethical guidelines for 

persons engaged in the publication of drug research, 

specifically, for editors, authors, and reviewers. We 

believe that the guidelines offered are understood 

and subscribed to by the pharmaceutical research 

scientists. They may be helpful to those who are 

related to editors, authors, and reviewers of journal 

publication. 

Ethical Obligation of Authors 

1. An author’s key obligation is to present an 

accurate account of the research paper. 

2. An author should recognize that journal 

space is precious resource created at considerable 

cost. Therefore, an author has an obligation to use it 

widely and economically. 

3. An author should cite those publications that 

are influential in determining the nature of the 

reported study work and that will guide the reader 

to the earlier work quickly and that are essential for 

understanding the present investigation. 

4. An author is obligated to perform a literature 

search to find, and then cite, the original 

publications that describe the current research, 

closely related citation to sources should also be 

made when a non-author supplied these. 

5. An author should identify the source of all 

information quoted or offered. Information obtained 

privately, ad in conversation, should not be used in 

the author’s work without explicit permission from 

the investigators with whom the information is 

originated. 

6. The authors should reveal to the editor any 

potential conflict of interest. The authors should 

ensure that no contractual relations or proprietary 

considerations exist that would affect the 

publication of information in a submitted 

manuscript. 

7. The co-authors of a paper should be all those 

persons who have made significant scientific 

contributions to the investigation reported and share 

duty and accountability for the results. Other 

contributions should be indicated in a footnote or in 

an acknowledgments section. 

8. The author who submits a manuscript for 

publication takes the responsibility of having 

included as co-authors all persons appropriate and 

none inappropriate. 

9. The submitting author should have sent each 

living co-author a draft copy of the manuscript and 

have obtained the co-author’s assent to the 

co-authorship of it. 

10. It is improper for an author to submit 

manuscripts describe essentially the same research 

in more than one journal for primary publication, 

unless it is a resubmission of a manuscript rejected 

for or withdrawn from publication. 

Ethical Obligation of Editors 

1. An editor should respect the intellectual 

independence of authors. 

2. An editor should give unbiased 

consideration to all manuscripts submitted, and 

judge each on its merits without regard to race, 

religion, nationality, sex, seniority, or institutional 

affiliation of authors. 

3. An editor should consider manuscripts 

submitted for publication with reasonable speed. 

3

Ethical Guideline. Asian Journal of Pharacodynamics and Pharmacokinetics 2009;9(1):3-4 

4. An editor should not disclose any 

information about a manuscript under consideration 

to anyone other than those from whom professional 

advice is sought. 

5. Editorial responsibility and authority for any 

manuscript authored by an editor and submitted to 

the journal should be distributed to some other 

qualified editors of the journal. The editorial 

consideration of the manuscript in any way or form 

by the author-editor would constitute a conflict of 

interest, and is therefore improper. 

6. The sole responsibility for acceptance or 

rejection of a manuscript rests with the editor. 

Responsible and prudent exercise of the duty 

normally requires that editor seek advice from 

reviewers, chosen for their expertise and good 

judgment, as to the quality and reliability of 

manuscript submitted for publication. 

7. The editor and members of the editor’s staff 

should not disclose any information about a 

manuscript under consideration to anyone other 

than those from whom professional advice is sought. 

The editor and members of the editor’s staff may 

disclose or publish manuscript titles and author 

names of papers that have been accepted for 

publication. 

8. Unpublished information, or interpretations 

disclosed in a submitted manuscript should not be 

used in an editor’s own research without the consent 

of author. 

9. An author may request that the editor not 

choose certain reviewers in consideration of a 

manuscript. 

10. When a manuscript is closely related to the 

current or past research of an editor as to create a 

conflict of interest, the editor should arrange some 

other qualified person to take editorial responsibility 

for that manuscript. 

Ethical Obligation of Reviewers 

1As the reviewing of manuscripts is an 

essential step in journal publication, therefore in the 

operation of the reviewers, every reviewer has an 

obligation to do a fair share of reviewing. 

2. A reviewer of a manuscript should judge 

objectively the quality of the manuscript, of its 

experimental and theoretical work, of its 

interpretations and its exposition, in accordance 

with high scientific and literary standards. A 

reviewer should respect the intellectual 

independence of the authors. 

3. A chosen reviewer who feels inadequately 

qualified to judge the research reported in a 

manuscript should return it to editor. 

4. A reviewer should explain and support their 

judgment adequately so that editors and authors 

may understand the basis of their comments. 

5. A reviewer should treat a manuscript sent for 

review as a confidential document. It should neither 

be shown to nor be discussed with other expect, in 

special cases, to persons from whom specific advice 

may be sought. In that event, the identifications of 

those consulted should be disclosed to the editor. 

6. A reviewer should act promptly, submitting 

a report in a timely manner. If a reviewer receives a 

manuscript at a time when circumstances preclude 

prompt attention to it, the unreviewed manuscript 

should be returned immediately to the editor. 

7. A reviewer should be alert to failure of 

authors to cite relevant work by other persons, 

bearing in mind that compliantly that the reviewer 

own research was insufficiently cited may seem 

self-serving. A reviewer should call the editor’s 

attention any substantial similarity between the 

manuscript under consideration and any published 

paper and any manuscript submitted concurrently to 

other journals. 

8. A reviewer should be sensitive to the 

appearance of a conflict of interest when the 

manuscript under review is closely related to the 

reviewer’s work in progress or published. If in 

doubt, the reviewer should return the manuscript 

promptly without review, advising the editor of the 

conflict of interest or bias. 

9. A reviewer should not evaluate a manuscript 

authored or co-authored by a person with whom the 

reviewer has a personal or professional connection 

if the relationship would bias judgment of the 

manuscript. 

10. A reviewer should not use or disclose 

unpublished information, arguments, or 

interpretations contained in a manuscript under 

consideration. 

11. The review of submitted manuscript may 

sometimes justify criticism, even severe criticism, 

from a reviewer. 

4

Special Report. Asian Journal of Pharmacodynamics and Pharmacokinetics 2009; 9(1):5-9 




ISSN 1608-2281 



Special report 

Introduction to Professor Leslie Z. Benet and his Lecture in China: 

Predicting Drug Absorption and Disposition Using a Biopharmaceutics 

Drug Disposition Classification System 

Professor Leslie Z. Benet visited Tianjin 

Institute of Pharmaceutical Research, China on 

Wednesday January 7, 2009. During the visitation, 

Professor Benet made an academic 

lecture/presentation titled “Predicting Drug 

Absorption and Disposition Using a 

Biopharmaceutics Drug Disposition Classification 

System” to scientists from Tianjin State Laboratory 

of Pharmacokinetics and Pharmacodynamics. 

Introduction to Professor Leslie Z. Benet 

Professor Leslie Z. Benet, Ph.D., Distinguished 

Clinical Research Lecturer and chairman of the 

department of biopharmaceutical sciences 

(1978-1998), University of California, San 

Francisco (UCSF), has received international 

recognition for his work related to pharmacokinetics 

and pharmacodynamics. Dr. Benet with six 

honorary doctorates is recognized with the highest 

scientific awards of nine professional societies, 

including the Distinguished Pharmaceutical 

Scientist award of the American Association of 

Pharmaceutical Scientists, the Rawls-Palmer 

Progress in Medicine award of the American 

Society for Pharmacology and Therapeutics, the 

Higuchi Research Prize of the American 

Pharmacists Association and the Host-Madsen 

Medal of the International Pharmaceutical 

Federation (FIP). Dr. Benet formerly served as 

Chair of the Pharmacology Study Section and the 

Pharmacological Sciences Review Committee for 

the NIH, the FDA Center for Biologics (CBER) 

Peer Review Committee, the FDA Expert Panel on 

Individual Bioequivalence, the Board of 

Pharmaceutical Sciences of the FIP, the Organizing 

Committee for the Millenial World Congress of 

Pharmaceutical Sciences, the Congressionally 

mandated IOM/NRC Committee on Accelerating 

the Research, Development and Acquisition of 

Medical Countermeasures Against Biological 

Warfare Agents and as a member of the FDA 

Generic Drugs Advisory Committee and the FDA 

Science Board. He was the Founder and first 

President of the American Association of 

Pharmaceutical Scientists that now numbers more 

than 13,000 members. Dr. Benet’s research interests, 

more than 490 scientific publications, 7 books and 

11 patents are in the areas of pharmacokinetics, 

biopharmaceutics, drug delivery and 

pharmacodynamics. 

Lecture: Predicting Drug Absorption and 

Disposition Using a Biopharmaceutics Drug 

Disposition Classification System 

The Biopharmaceutics classification system 

5


The Biopharmaceutics classification system 

(BCS) has been one of the most significant 

prognostic tools created to promote product 

development in recent years. It is a scientific 

framework for classifying drug substances based on 

their aqueous solubility and intestinal permeability 

characteristics, which will substantially facilitate 

drug product selection and approval process for a 

large group of drug candidates. The goal of the BCS 

is to function as a tool for developing in vitro 

dissolution specifications for drug products that are 

predictive of their in vivo performance. 

According to the BCS, drug substances are 

classified as follows: 

Class 1: High Solubility-High Permeability: 

generally very well-absorbed compounds 

Class 2: Low Solubility-High Permeability: 

exhibit dissolution rate-limited absorption 

Class 3: High Solubility-Low Permeability: 

exhibit permeability rate-limited absorption 

Class 4: Low Solubility-Low Permeability: 

very poor oral bioavailability. 

The Class Boundaries: 

• A drug substance is considered HIGHLY 

SOLUBLE when the highest dose strength is 

soluble in250 ml water over a pH range of 1 to 

7.5. 

• A drug substance is considered HIGHLY 

PERMEABLE when the extent of absorption in 

humans is determined to be 90% of an 

administered dose, based on mass-balance or in 

comparison to an intravenous reference dose. 

• A drug product is considered to be 

RAPIDLY DISSOLVING when 85% of the 

labeled amount of drug substance dissolves within 

30 minutes using USP apparatus I or II in a volume 

of 900 ml buffer solutions. 

The pH-solubility profile of the test drug 

substance should be determined in aqueous media 

with a pH in the range of 1-7.5 using traditional 

shake-flask method as well as acid or base titration 

methods. A sufficient number of pH conditions 

should be evaluated to accurately define the 

pH-solubility profile. Concentration of the drug 

substance in selected buffers (or pH conditions) 

should be determined using a validated 

stability-indicating assay that can distinguish the 

drug substance from its degradation products. 

The permeability class of a drug substance can 

be determined in human subjects using mass 

balance, absolute BA, or intestinal perfusion 

approaches: 1. Pharmacokinetic Studies in Humans: 

a. Mass Balance Studies b. Absolute Bioavailability 

Studies; 2. Intestinal Permeability Methods: The 

following methods can be used to determine the 

permeability of a drug substance from the 

gastrointestinal tract: (1) in vivo intestinal perfusion 

studies in humans; (2) in vivo or in situ intestinal 

perfusion studies using suitable animal models; (3) 

in vitro permeation studies using excised human or 

animal intestinal tissues; or (4) in vitro permeation 

studies across a monolayer of cultured epithelial 

cells; 3. Instability in the Gastrointestinal Tract: 

determining the extent of absorption in humans 

based on mass balance studies using total 

radioactivity in urine does not take into 

consideration the extent of degradation of a drug in 

the gastrointestinal fluid prior to intestinal 

membrane permeation. 

Dissolution testing should be carried out in 

USP Apparatus I at 100 rpm or Apparatus II at 50 

rpm using 900 mL of the following dissolution 

media: (1) 0.1 N HCl or Simulated Gastric Fluid 

USP without enzymes; (2) a pH 4.5 buffer; and (3) a 

pH 6.8 buffer or Simulated Intestinal Fluid USP 

without enzymes. For capsules and tablets with 

gelatin coating, Simulated Gastric and Intestinal 

Fluids USP (with enzymes) can be used. When 

comparing the test and reference products, 

dissolution profiles should be compared using a 

similarity factor (f 2 ). The similarity factor is a 

logarithmic reciprocal square root transformation of 

the sum of squared error and is a measurement of 

the similarity in the percent (%) of dissolution 

between the two curves. Two dissolution profiles 

are considered similar when the f 2 value is 50. 

By understanding the relationship between a 

drug’s absorption, solubility, and dissolution 

characteristics, it is possible to define situations 

when in vitro dissolution data can provide a 

surrogate for in vivo bioequivalence 

assessments,that is to find under which 

circumstances dissolution testing can be prognostic 

for in vivo performance. 

6


The Biopharmaceutics Drug Disposition 

Classification System (BDDCS) 

The development of the BCS was a major step 

in bringing rational science to regulation, allowing 

waivers of in vivo bioavailability and 

bioequivalence testing (“biowaiver”) of immediate 

release dosage forms for high-solubility, 

highpermeability drugs when such drug products 

also exhibited rapid dissolution. This application of 

science yielded a decrease in the regulatory burden. 

When the BCS was first developed there was only a 

nascent understanding of the importance of drug 

transporters to bioavailability. However, as pointed 

out here, for Class 1 compounds neither efflux nor 

absorptive transporters should influence oral 

bioavailability, and meal effects on Fextent should 

be negligible. 

The Biopharmaceutics Drug Disposition 

Classification System (BDDCS) replaces the 

permeability criteria with the major route of 

elimination because of the belief that it is easier and 

less ambiguous to determine the assignment of 

BDDCS for marketed drugs based on the extent of 

metabolism than using permeability (i.e. extent of 

absorption) in BCS assignments. Designation of the 

major route of drug elimination as part or instead of 

the permeability criteria would reduce the 

regulatory burden for many more Class 1 

compounds, would eliminate the ambiguity and 

difficulty in determining 90% (or 85%) absorption 

for Classes 1 and 2 compounds, and would allow 

predictability of absorption and disposition 

characteristics of drugs in all four BDDCS. 

BDDCS Class compounds would then be 

designated as: 

Class 1: high solubility, extensive metabolism. 

Waiver of in vivo bioequivalence studies for 

BDDCS Class 1 drugs would still require rapid 

dissolution. 

Class 2: poor solubility, extensive metabolism 

Class 3: high solubility, poor metabolism 

Class 4: low solubility, poor metabolism. 

When initially proposed, “extensive 

metabolism” was defined as ≥50% metabolism of 

an oral dose in vivo in humans. Further 

consideration of this parameter designation led to 

the realization that there are very few 

drugs/compounds that are intermediately 

metabolized. It is now proposed that the 

definition of “extensive metabolism” be pushed 

to ≥70% metabolism of an oral dose in vivo in 

humans while the “poor metabolism” be defined 

as ≥50% of the dose be excreted unchanged. Use 

of the more stringent BDDCS metabolism criteria 

versus the BCS permeability characterization 

resulted in only 10 compounds requiring 

reclassification and allowed for the inclusion of 

an additional 38 drugs/compounds. 

Predicting Drug Absorption and Disposition 

The BDDCS was developed to allow prediction 

of in vivo pharmacokinetic performance of drug 

products. It may be useful in predicting overall drug 

disposition, including routes of drug elimination 

and the effects of efflux and absorptive transporters 

on oral drug absorption; when transporter-enzyme 

interplay will yield clinically significant effects (e.g., 

low bioavailability and drug-drug interactions); the 

direction, mechanism, and importance of food 

effects; and transporter effects on postabsorption 

systemic drug concentrations following oral and 

intravenous dosing. These predictions are supported 

by a series of studies investigating the effect of 

transporter inhibition and induction on drug 

metabolism. 

Major Routes of Drug Elimination: 

Class 1 and Class 2: compounds are eliminated 

primarily via metabolism; 

Class 3 and Class 4: compounds are primarily 

eliminated unchanged into the urine and bile. 

For the 130 drugs/compounds gathered from 

the literature, only 13 of the substances do not have 

readily accessible, critically evaluated 

pharmacokinetic parameters. If you know the 

intestinal absorption (or more likely a surrogate as 

Caco-2 permeability) of an NME, you can predict 

whether the major route of elimination of the NME 

will be metabolism. 

Note that the permeability parameter does not 

predict the ability for the NME to enter the liver/ 

hepatocytes (since a number of non-metabolized 

Classes 3 & 4 compounds will be excreted in the 

bile), but rather the access to the metabolic enzymes 

within the hepatocytes. 

Oral Dosing and the Predictability of 

7


Transporter Effects: 

Since extent of metabolism correctly predicts 

high vs low intestinal permeability for at least 33 of 

35 drugs , where human permeability 

measurements exist. Benet and co-workers propose 

the following: “We recommend that regulatory 

agencies add the extent of drug metabolism (i.e., 

90% metabolized) as an alternate method for the 

extent of drug absorption (i.e., 90% absorbed) in 

defining Class 1 drugs suitable for a waiver of in 

vivo studies of bioequivalence.” 

Class 1: highly soluble, high permeability, 

extensively metabolized drugs 

• Transporter effects will be minimal in the 

intestine and the liver. 

• Even compounds like verapamil that can 

be shown in certain cellular systems 

(MDR1-MDCK) to be a substrate of P-gp will 

exhibit no clinically significant P-gp substrate 

effects in the gut and liver. 

• A major proposition (and probably the 

primary advance in knowledge) of BDDCS is that 

Class 1 drugs are not substrates for transporters in 

the intestine and liver. (but not at the BBB and the 

kidney) 

Class 2: poorly soluble, highly permeable, 

extensively metabolized drugs 

• Efflux transporter effects will be 

important in the intestine and the liver. 

• In the intestine efflux transporter 

enzyme (CYP 3A4 and UGTs) interplay can 

markedly affect oral bioavailability. 

• In the liver the efflux transporter-enzyme 

interplay will yield counteractive effects to that seen 

in the intestine. 

• Uptake transporters can be important for 

the liver but not the intestine. 

• Following oral dosing, major significant 

interactions will occur for Class 2 drugs that are 

substrates for intestinal enzymes (e.g. CYP3A, 

UGTs) and efflux transporters (e.g. P-gp, MRP2, 

BCRP). Since concomitant inhibition of the 

intestinal enzyme and the efflux transporter both 

lead to less gut metabolism that synergistically 

increase systemic AUC. It is not surprising that 

drugs removed from the market due to drug-drug 

interactions predominate for orally dosed drugs that 

are substrates for CYP3A and P-gp. 

Class 3: highly soluble, low permeability, poorly 

metabolized drugs. 

• Uptake transporters will be important for 

intestinal absorption and liver entry for these poor 

permeability drugs. 

• However, once these poorly permeable 

drugs get into the enterocyte or the hepatocyte 

efflux transporter effects can occur. 

Class 4: poorly soluble, low permeability, 

poorly metabolized drugs 

• Oral bioavailability is minimal and 

transporter effects could be relevant. 

Food Effects (High-Fat Meals): 

It is well-known that food can influence drug 

bioavailability, both increasing and decreasing the 

extent of availability (Fextent) and the rate of 

availability. 

Class 1: High-fat meals will have no significant 

effect on F extent for Class 1 compounds. 

• Because complete absorption may be 

expected for high solubility/high permeability 

compounds, and as noted previously, no transporter 

drug interactions would be expected for Class 1 

compounds. 

• However, high-fat meals may delay 

stomach emptying and therefore cause an increase 

in peak time. 

Class 2: High-fat meals will increase F extent for 

Class 2 compounds 

• Due to inhibition of efflux transporters in 

the intestine and additional solubilization of drug in 

the intestinal lumen (e.g., micelle formation). 

• Peak time could decrease due to 

inhibition of efflux cycling or increase due to 

slowing of stomach emptying; 

• Formulation changes that markedly 

increase the solubility of Class 2 compounds will 

decrease or eliminate the high-fat meal effects for 

these drugs. 

Class 3: High-fat meals will decrease F extent for 

Class 3 compounds 

• Due to inhibition of uptake transporters 

in the intestine. 

Class 4: For Class 4 compounds, it is difficult to 

predict what will occur. 

Postabsorption Effects and Intravenous 

8


Dosing: 

For intravenous dosing, drug concentrations at 

the eliminating organ will always be relatively low 

due to the diluting effects of volume of distribution, 

as compared to concentrations of drug in the 

intestine. Therefore, saturation of transporters (and 

enzymes) will be minimal, if at all, and solubility 

considerations will be unimportant when 

measurable systemic concentrations of the drug are 

achieved. 

• High extraction ratio drugs, where 

clearance approaches blood flow, are mainly limited 

to Class 1 compounds. 

• Post intestinal absorption and following 

intravenous dosing, both uptake and efflux 

transporters can be important determinants of the 

disposition for Classes 2, 3, and 4 compounds. 

• Biliary secretion of parent drug can be an 

important component of disposition for Classes 3 

and 4 compounds. 

• Renal elimination of Classes 3 and 4 

compounds can be affected by both uptake and 

efflux transporters. 

Potential Drug-Drug Interactions Predicted 

by BDDCS: 

Class 1: Only metabolic in the intestine and 

liver 

• Drug-drug interactions for Class 1 

compounds will be primarily metabolic, with 

transporter-enzyme interplay only becoming 

important for those drugs where high permeability 

is a result of rapid transporter uptake rather than 

high Log P. 

Class 2: Metabolic, efflux transporter and efflux 

transporter-enzyme interplay in the 

intestine. Metabolic, uptake 

transporter, efflux transporter and 

transporter-enzyme interplay in liver. 

• Drug-drug interactions are not limited to 

enzymatic processes but can frequently be mediated 

by transporter interactions and often involve 

transporter-enzyme interplay for Class 2 

compounds. 

• Following oral dosing, major significant 

interactions will occur for Class 2 drugs that are 

substrates for both intestinal enzymes (e.g., CYP3A, 

UGTs) and intestinal apical efflux transporters (e.g., 

P-glycoprotein, MRP2, BCRP). 

• The enzyme-efflux transporter interplay 

that is so important in the intestine will not be as 

significant in the liver (and the kidney) due to the 

reverse order in which drug molecules encounter 

the two proteins. 

• Inhibition of hepatic uptake transporters 

can lead to significantly increased systemic drug 

concentrations for Class 2 compounds 

Class 3 and 4: Uptake transporter, efflux 

transporter and uptake-efflux transporter interplay. 

Inhibition of hepatic and renal uptake 

transporters can lead to significant increases in the 

systemic concentration of Classes 3 and 4 

compounds. 

Conclusions 

The interplay between transporters, both influx 

and efflux, and metabolic enzymes in the intestine 

and the liver could differ depending on the drug’s 

solubility and permeability characteristics as 

reflected in the Biopharmaceutical Classification 

System (BCS). Although BCS has had a marked 

effect in decreasing the regulatory burden by 

allowing a waiver of in vivo bioequivalence studies 

for a limited number of Class 1 drugs, little 

predictive use has been made of Classes 2, 3, and 4 

in the BCS categorization. In general, BCS Classes 

1 and 2 are highly metabolized, whereas BCS 

Classes 3 and 4 drugs are primarily excreted 

unchanged via the biliary or renal routes. Therefore, 

changing the permeability component to a route of 

elimination component in a Biopharmaceutics Drug 

Disposition Classification System (BDDCS) will 

facilitate predictions, markedly expand the number 

of Class 1 drugs eligible for waiver of in vivo 

bioequivalence studies, and provide new insight. It 

may be easier to determine classification based on 

major routes of elimination than upon permeability; 

an extent of metabolism criterion for waiver of in 

vivo bioequivalence studies; and how predictive 

algorithms may be developed using only in vitro or 

in silico methods to facilitate class assignment in 

BDDCS. A further advantage of BDDCS is that a 

preliminary class assignment for NMEs may be 

obtained from a metabolism measure in human 

hepatocytes, prior to in vivo studies in humans. 

Understanding transporter-enzyme interactions in 

terms of the permeability and solubility of drug 

9


compounds offers the potential for predicting: 

1. Major routes of elimination; 

2. Transporter effects of drug absorption; 

3. Food (High Fat Meal) effects; 

4. Transporter effects on post absorption 

systemic levels and after i.v. dosing; 

5. Enzyme transporter interplay; 

6. Drug-drug interaction potential and its 

relationship to enzyme-transporter interplay; 

7. Previously unexplained effects of renal 

disease on hepatic metabolism that can result from 

accumulation of substances (toxins) in renal failure 

that modify hepatic uptake and efflux transporters; 

8. The translation of pharmacogenetic 

differences in metabolic enzymes (genetic 

polymorphisms) that do not always result in the 

expected differences in vivo. Phenotype-genotype 

discordance (as well as changes in the relationship 

as a function of disease states) may be explained by 

the effects of transporters on metabolic clearance; 

Obtaining a realistic perspective of new 

techniques (e.g., predictive ADME, QSAR, 

microdosing, systems biology). If validation of the 

technique primarily uses Class 1 drugs, there is no 

assurance that the technique will work for NMEs. 

It may be easier to determine classification based on 

major routes of elimination than upon permeability; an 

extent of metabolism criterion for waiver of in vivo 

bioequivalence studies; and how predictive algorithms 

may be developed using only in vitro or in silico methods 

to facilitate class assignment in BDDCS. 

(Gu Y, Si DY and Liu CX) 

Publication News 

Chinese Herbal Medicines 

Chinese Herbal Medicines, an academic journal in English edition, has been approved by the State Press 

and Publication Administration in December 2008. 

Approval Journal number is CN 12-1410/R. Chinese Herbal Medicines will officially be published in 2009, 

in China. 

Sponsored by Tianjin Institute of Pharmaceutical Research and Institute of Medicinal Plants, Chinese 

Academy of Medical Sciences. Published by Chinese herbal medicines magazine. Edited by Editorial 

Committee of Chinese herbal medicines 

Chinese Herbal Medicines, an international journal, is the official publication Sponsored by Tianjin 

Institute of Pharmaceutical Research and Institute of Medicinal Plants, Chinese Academy of Medical 

Sciences. The Journal’s purposes are to provide a forum for the studies on Chinese herbal medicines, 

traditional medicines and natural products. The Journal will accept the following contributions: 

original research articles; review papers, short communications; letters to the editor; book reviews; 

conference announcements and news. The journal covers the wide range from the research to 

development and application of traditional medicines, herbal medicines and natural products, which 

including medicinal resource, phytochemical, pharmacological, toxicological, pharmacokinetic and 

therapeutic studies of active ingredients and complex formulations in experimental and clinical trials. 

Original articles in English are published. 

10

Xu HY et al. Asian Journal of Pharmacodynamics and Pharmacokinetics 2009; 9(1):11-25 




ISSN 1608-2281 



Recent advance on chemical compositions and pharmacodynamic and 

pharmacokinetic studies of Rhizoma Coptidis 

Hai-Yu Xu 1,3 , Tie-Jun Zhang 2 , Xue-Yu Zhu 2 ,Yu-Bo Li 1,3 

1 Tianjin State Key Laboratory of Pharmacidymanamics and Pharmcokinetics, Tianjin Institute of 

Pharmaceutical Research, Tianjin 300193, China 

2 

Tianjin Research Center of Modern Chinese Medicines, Tianjin Institute of Pharmaceutical Research, 

Tianjin 300193, China 

3 Tianjin University of Traditional Chinese Medicine, Tianjing, 300193, China. 

Abstract 

Key words 

Rhizoma Coptidis has long been used for the treatment of gastrointestinal disorders, pyretic 

toxicity, abdominal typhus, bacillary dysentery, Swelling of throat in traditional oriental 

medicine in china. Recent studies found that its active constituents are alkaloids comprising 

berberine, coptisine, palmaine, jatrorrhizine etc. And also Rhizoma Coptidis has been proved to 

have extensive pharmacological activities including anti-cancer, anti-microbial activity, 

hypoglycemic activity, LDL-lowering activity, anti-inflammatory potential and antioxygen and 

free radicals scavenging etc.. In order to improve its therapeutic value and reduce its side effects, 

it is necessary to study the relationship between its activity and pharmacokinetics in vitro and in 

vivo. Some studies have been showed that berberine, an important ingredient in Coptis chinensis 

Franch displays a linear pharmacokinetic phenomenon and has poor bioavailability. This article 

summarizes chemical compositions, pharmacological actions and pharmacokinetics of Rhizoma 

Coptidis. 

Rhizoma Coptidis; chemical constituents; antimicrobial activity; hypoglycemic Activity; 

anticancer action; anti-inflammatory; antioxygen; cardiovascular action; free radicals 

scavenging; pharmacokinetics 

Article history Received 22 June 2008; Accepted 30 December 2008 

Publication data Pages: 15; Tables: 0; Figures: 4; References: 58; Paper ID: 1608-2281-2009-0901011-15 

Corresponding author Professor Zhang Tie-Jun, Tianjin Institute of Pharmaceutical Research,308 An-Shan West Road, Tianjin, 

300193, China. E-mail: tiezheng4@sina.com. 

Introduction 

Herbs have been widely employed as important 

remedies all over the world [1] . Progress in science and 

technology in recent decades has been made possibly 

not only to isolate and to characterize the biologically 

active constituents of herbs, but also to evaluate their 

biological activities. Rhizoma Coptidis, the dried root 

and rhizome of “Huang-lian” named in Chinese, has 

long been used for the treatment of gastrointestinal 

disorders, pyretic toxicity, abdominal typhus, 

bacillary dysentery, Swelling of throat in traditional 

oriental medicine. Ethnopharmacologically, the 

property and taste of Rhizoma Coptidis are bitter and 

11


cold; the meridians are into heart, liver, stomach and 

large intestine. The functions are to clear heat and dry 

dampness, and to reduce fire and dispel toxins. It is 

used to eliminate heat in the heart for insomnia, 

fidget, delirium due to high fever, inflammation of 

the month and tongue, and to stop diarrhea for acute 

enteritis and dysentery. [2] It can be devided into 3 

spicies which are Coptis chinensis Franch, (Fig 1, 

www.newdruginfo.com) Coptis deltoidea C.Y. 

Cheng et Hsiao and C. teeta Wall. Rhizoma Coptidis 

(Fig 2, www.zgycsc.com) is thought as high grade 

which has good effect and little toxin, recorded in 

TCM Classic Shennong Bencaojing. 

Recent in the chemical Constituents and 

pharmacological studies,it indicate that the total 

alkaloids is thought as main active ingredients and 

berberine is the highest in the total alkaloids of 

Rhizoma Coptidis. Rhizoma Coptidis and berberine 

have been proved to have extensive pharmacological 

activities, such as antibacterial effect, antiinflammatory 

effects, antioxygen and free radicals 

scavenging ect. and new pharmacological activities, 

such as hypoglycemic and hypolipidemic effects, 

anticancer and effects on cardiovascular system etc.. 

In order to improve its therapeutic value and reduce 

its side effects, it is necessary to study the 

relationship between its activity and 

pharmacokinetics in vitro and in vivo. Some studies 

have been showed that berberine ,an important 

ingredient in Coptis chinensis Franch displays a 

linear pharmacokinetic phenomenon and have poor 

bioavailability. 

In this review paper, we collected some 

information about chemical instistuents, 

pharmacological actions and pharmacokinetics of 

Rhizoma Coptidis. The introduced information on 

Rhizoma Coptidis will help us to know 

pharmacological actions, the mechanism of TCM, 

secondary exploitation for Rhizoma Coptidis and its 

be value for studies on TCM 

Chemical Constituents 

In Rhizoma Coptidis, major chemical 

constituents are several alkaloids such as 

berberine,coptisine, palmaine, jatrorrhizine, 

worenine, epiberberine (Fig 3) which of all are 

both protoberberine and quaternary-amine alkaloid. 

the content of berberine is approximately 5 percent 

to 8 percent ,which is the most constituent of the 

total alkaloids. In addition, Rhizoma Coptidis may 

contain magnoflorine, ferulaic acid (Fig 4) and so 

on [3] . 

Fig 1. Coptis chinensis Franch 

Fig 2. Rhizoma Coptidis 

Kobayashi [4] et al had reported that inhibitors of 

topoismerase I and topoismerase II were separated 

from Rhizoma Coptidis which could duplicate, 

transcribe and recombine DNA in the cell. Aqueous 

extract of Rhizoma Coptidis may make 

cleavable-complex of DNA Topoismerase steadily in 

mammal. Both epiberberine and groenlandicine are 

active constitutents of topoismerase I mediating DNA 

cleavage and berberrubine is a specific inductor of 

topoismeras I mediating DNA cleavage. Hirano [4] et 

al had separated lariciresinol and anti-ferulaic 

acid-duplet hydroxyl-benzene-ethyl ester. 

12


In mixed decoction of Rhizoma Coptidis and 

Radix et Rhizoma Glycyrrhizae, the former’s main 

constituent may be remained, but chemical 

constituents of Radix et Rhizoma Glycyrrhizae 

disappear completely. But no new chemical 

compound could be found in the process [6] . Studing 

on content variation of chemical constituents during 

combination of Rhizoma Coptidis with Fructus 

Evodiae,the dissolution rate of jateorhizine, coptisine, 

palmatine and berberine decreased obviously. At the 

same time, the dissolution rate lineared correlation 

with the proportion of Rhizoma Coptidis. It could be 

concluded that Rhizoma Coptidis’ Alkaloids reacted 

with Fructus Evodiae’s Flavonoids [7] . 

Berberine Coptisine Palmatine 

Jatrorrhizine Worenine Epiberberine 

Fig 3. Chemical structures of berberine,coptisine, palmatine, jatrorrhizine, worenine and epiberberine 

 

Magnoflorine 

 

 

Ferulaic acid 

Fig 4. Chemical structures of magnoflorine and ferulaic acid 

Pharmacological Actions 

In Traditional Chinese Medicine, Coptidis 

Rhizoma has the effects of suppressing fever, 

dispelling dampness and deintoxication. Through 

modern pharmacological studies, it has been proved 

to have many pharmacological effects including 

antimicrobial activity, hypoglycemic activity, 

anti-inflammatory, antioxygen and free radicals 

scavenging and anticancer effects etc..Berberine, the 

major component of this herb, has also many 

pharmacological effects including inhibition of 

adipocyte differentiation, anti-cancer effects, 

anti-microbial effects, LDL-lowering effects and 

anti-inflammatory potential [8] . 

Antimicrobial Activity 

Coptis and berberine showed wide inhibitory 

actionon bafungi prorozoon and virus in vitro and in 

vivo. [2] Berberine was found to be active against a 

number of Gram-positive and Gram-negative bacteria, 

such as Stephylococcus aureus, S. hemolylicus, 

13


Salmonella typhsa, Shigella dysenterial, S. 

paradysenterial, and so on. Berberine sulfate showed 

to be bacterial to vibrio chloral and and bacteriostatic 

to Stephylococcus aureus at concentrations of 35 and 

50 µg·mL -1 , respectively. It showed bacteriadal 

activity against Stephylococcus aureus at 50-500 

µg·ml -1 in culture medium. Berberine sulfate in 

concentrations 10-25µg·ml -1 inhibited the growth of 

11 fugni. Oral administration of berberine sulfate at 

dose of 35-700 mg·kg -1 was effective in treating 

candicans infections of the intestine in mice. [9] 

Rhizoma Coptidis has strong antibacterial action 

and broad spectrum antibacterial activities, including 

Gram-positive and Gram-negative bacteria, aerobe 

and anaerobe. The biologically active constituent of 

C. chinensis extract is characterized as the 

isoquinoline alkaloids,such as berberine, 

palmatine,jateorhizine, coptisine etc.. Those 

constituents are potent inhibitors of sortase (a 

bacterial surface protein anchoring transpeptidase 

from Staphylococcus aureus ATCC 6538p), with an 

IC 50 value of 8.7 µg·mL -1 and have antibacterial 

activity against Gram-positive bacteria with a 

minimum inhibitory concentration (MIC) in the range 

of 50–400 µg·mL -1 . Among the four isoquinoline 

alkaloids tested, berberine chloride had the strongest 

inhibitory activity [10] . George Tegos [11] et al have 

reported that the activities of berberine were 

considerably against the gram-positive bacterium 

Staphylococcus aureus and Bacillus megaterium and 

that disabling of the MDRs (multidrug resistance 

pumps ) in gram-negative species leaded to a striking 

increase in antimicrobial activity. Gram-negative 

bacteria had an effective permeability barrier, 

comprised of the outer membrane, which restricts the 

penetration of amphipathic compounds, and 

multidrug resistance pumps (MDRs), which extrude 

toxins across this barrier. Direct measurement of the 

uptake of berberine, a model plant antimicrobial, 

confirmed that disabling of the MDRs strongly 

increases the level of penetration of berberine into the 

cells of gram-negative bacteria. 

Rhizoma Coptidis have long been used for the 

treatment of gastrointestinal disorders, insomnia, 

refractoriness dental ulcer, tinnitus in traditional 

oriental medicine [12] . According to the Basic Theory 

of Chinese Traditional Medicine, it has the efficacy 

of Clearing away heat, depriving evil wetness and 

purging fire for removing toxin. These pharmacologic 

actions attribute to antibacterial action which can 

cure infectious diseases. For example, Zuo-Jin-Wan, 

a Chinese medical formula including Rhizoma 

Coptidis and Fructus Evodiae, is used to cure 

gastrointestinal disorders, such as peptic ulcer, 

chronic gastritis, functional dyspepsia etc., which are 

correlated to Zuo-Jin-Wan on anti-Hp action [13] . 

Hypoglycemic Activity 

Rhizome Coptidis and its major constituent 

berberine have good hypoglycemic and 

hypolipidemic effects. The potential glucoselowering 

effect of berberine was noted when it was 

used for diarrhea in diabetic patients. In vitro and in 

vivo studies have showed its effects on 

hyperglycemia and dyslipidemia. Zhang Y [14] et al 

have reported that in the berberine group, fasting and 

postload plasma glucose decreased from 7.0 ±0.8 to 

5.6 ± 0.9 and from 12.0 ± 2.7 to 8.9 ± 2.8 mM·L -1 , 

HbA1c from 7.5 ±1.0% to 6.6 ±0.7%, triglyceride 

from 2.51 ±2.04 to 1.61±1.10 mM·L -1 , total 

cholesterol from 5.31±0.98 to 4.35±0.96 mM·L -1 , and 

low-density lipoprotein-cholesterol from 3.23 ± 0.81 

to 2.55 ± 0.77 mM·L -1 , with all parameters differing 

from placebo significantly. The glucose disposal rate 

was increased after berberine treatment. Mild to 

moderate constipation was observed in five 

participants in the berberine group. So berberine is 

effective and safe in the treatment of type 2 diabetes 

and dyslipidemia. In one clinical study, 60 patients 

with type 2 diabetes were treated with berberine for 

1–3 months, and 90% of patients showed 

improvement in their clinical symptoms [15] . 

Recently, it has been reported that activating the 

AMP-activated protein kinase (AMPK) pathway is 

the underlying mechanism for berberine improving 

insulin resistance, lowering blood sugar, and 

correcting lipid metabolism disorders [16-19] . 

Berberine can reduce body weight and lipid 

levels and improve insulin action .Berberine acutely 

activates AMPK activity, and induces a variety of 

metabolic effects consistent with AMPK 

activation. These include activation of GLUT4 

translocation; increasing phosphorylation of AMPK, 

ACC, and p38 MAPK; reducing lipid content in 

adipocytes; increasing expression of genes involved 

14


in lipid oxidation; and decreasing expression of genes 

involved in lipid synthesis [20] . 

Berberine and other AMPK agonists, such as 

metformin,have no effect on blood glucose. The 

chronic effects involving changes in gene expression, 

which likely contribute to reducing fat cell 

differentiation and increasing mitochondrial 

biogenesis, again contribute to lipid lowering, fat 

mass reducing, and insulin sensitivity improving. A 

significant reduction in the expression of lipogenic 

genes in adipose tissue following treatment with 

berberine either in vitro or in vivo. Moreover, 

berberine inhibit adipocyte differentiation probably 

by inhibiting PPAR activity which is an important 

transcriptional regulator of adipogenesis in 3T3-L1 

cells [21, 22] . 

Anticancer action 

Berberine is the anti-tumor constituent of 

Rhizome Coptidis and can inhibit different 

tumors’cell growth including colon carcinoma, liver 

tumor, carcinoma of bladder and leukemia etc.. The 

anti-tumor mechanism of berberine is different 

according to different tumors. 

Both hepatovirus and environmental pollution 

are two main causative agents of liver tumor. And 

nitrosamine is main material of environmental 

pollution, produced by product, N-methyl- 

N′-nitro-N-nitroguanidine which is larvaceous 

carcinogenic factor in stomach acid environment. 

Berberine could significantly inhibit the 

carcinogenesis induced by 20-methylcholanthrene 

(200 µg·0.1ml -1 per a mouse) or N-nitrosodiethylamine 

(NDEA; 0.02% NDEA in distilled 

water, 2.5 ml to each an animal by gavage, five days 

a week for 20 weeks) in a dose-dependent manner. 

Administration of berberine (0.5, 2.5 or 5.0 mg·kg -1 ) 

could significantly reduce the incidence of tumour in 

animals after an injection of 20-methylcholanthrene 

and increase their life span compared with the 

control [23] . At the same time, berberine also reduced 

cell proliferation and alpha-fetoprotein expression in 

human hepatoma HepG2 cells. Multidrug resistance 

transporter (pgp-170) is known to be overexpressed 

in Hep G2 cells. Lin etc. found that berberine could 

modulate the expression and function of pgp-170 in 

hepatoma cells and other hepatoma cell lines, which 

suggest that treatment of tumor cells with berberine 

may result in reduced retention of chemotherapeutic 

agents [24]. 

Of course, berberine also can inhibit colon 

carcinoma cell growth. Li etc. thinked that the 

inhibition of cell growth by huanglian was associated 

with up to 8-fold suppression of cyclin B1 protein. 

This resulted in complete inhibition of cdc2 kinase 

activity and accumulation of cells in G 2 . Therefore, 

the effect of huanglian on inhibiting tumor growth 

seems to be mediated by the selective suppression of 

cyclin B1, which results in the inhibition of cdc2 

kinase activity. Inhibition of cyclin dependent kinase 

(cdk) activity is emerging as an attractive target for 

cancer chemotherapy [25] . Fukuda etc. used a 

beta-galactosidase reporter gene system and found 

berberine, an isoquinoline alkaloid effectively 

inhibiting COX-2 transcriptional activity in colon 

cancer cells in a dose- and time-dependent manner at 

concentrations higher than 0.3 µM [26] ; Berberine also 

inhibited arylamine Nacetyltransferase (NAT) 

activity in a human colon tumor (adenocarcinoma) 

cell line in a dose-dependent manner [27] . 

In conclusion, Rhizome Coptidis can inhibit 

tumour cells growth through different mechanisms of 

action. These results indicate that traditional Chinese 

herbs may represent a new source of agents in cancer 

therapy. Furthermore,it have many advantages such 

as high safetyaffluent resourceslow prices,little 

side effects ETC. So anti-tumor action of Rhizome 

Coptidis in clinic will have broad and bright prospect. 

Anti-Inflammatory action 

Rhizome Coptidis demonstrate anti-inflammatory 

effects in various experimental models. 

Berberine, which has strong anti-inflammatory 

effects [28-30] , is a major active constituent in Rhizoma 

Coptidis. Recently studies show that berberine could 

inhibit active chronic inflammation, delayed type 

hypersensitivity and experimental ulcerative colitis 

etc.. 

The mechanism of anti-inflammatory activity 

have not yet been clarified completely. But a lot of 

studies focused on the following aspects. Firstly, 

berberine could exhibite the inhibition for the 

adhesion of leukocyte and endothelial cell. Leukocyte 

trafficking plays an important role in inflammatory 

reaction.He Yu [31] et al studied the simulative 

inflammatory reaction state in vivo through inducing 

15


human poly morphonulear leucocyte’s(PMN) and 

endothelial cell’s adhesion increasing by IL-1TNF 

and the results showed that berberine inhibited not 

only the adhesion of endothelial cell induced by IL-1 

and TNF together with PMN but also the adhesion of 

PMN induced by TNF together with endothelial cell. 

Secondly, berberine could regulate transcription 

factors related to inflammatory diseases. Bong-Hyuk 

Choi et al reported that BBR reduced the mRNA 

expression level of inflammation factors such as TNF, 

IL-6, CRP and HP. The mRNA expression level of 

HP is most significantly reduced by BBR treatments 

among these. The mRNA expression levels of other 

inflammatory factor are decreased by 39%, 17% and 

19%, respectively [32] . Thirdly,berberine could also 

inhibite inflammatory cytokine. Kuo CL et al studied 

that the anti-inflammatory mechanism of berberine is 

mediated through COX-2 regulation and found that 

berberine induced effect occurred rapidly (3 h) as a 

result of reduced COX-2 protein, but not enzyme 

activity. These anti-inflammatory effects paralleled to 

the in vivo results where berberine pretreatment of 

Wistar rat inhibited the production of exudates and 

PGE2 in carrageenan induced air pouch [33] . 

Antioxygen and Free Radicals Scavenging 

Activating oxides which is produced by 

sunshine, ultraviolet radiation, ionizing radiation, 

chemical reaction and metabolism is very important 

reason of pathologic demages,such as DNA demage, 

cancer and cell ageing. So antioxygen agents play 

very important roles in treatment with these diseases, 

especially crude drugs are in well with people 

because of strong antioxygen and low toxicity. 

Schinella G R studied the antioxidant properties of 

twenty medical herbs used in the traditional 

Mediterranean and Chinese medicine and founded 

that Coptis chinensis at a concentration of 100 

microg/ml exhibited the highest scavenging activity 

on the superoxide radical [34] . Moreover, many 

efficacies of Coptis chinensis such as checking 

diarrhea, deintoxication, anti-inflammatory etc. were 

possiblly concerned with antixoygen and free radicals 

scavening [35] . 

The aqueous extracts of Coptis chinensis 

exhibited the highest potency in inhibiting rat 

erythrocyte hemolysis and lipid peroxidation in rat 

kidney and brain homogenates and also demonstrated 

strong superoxide- and hydroxyl radical-scavenging 

activity, but exerted only a slight pro-oxidant effect [36] . 

In Coptis chinensis, berberine, jatrorrhizine, and 

magnoflorine behaved antioxidant activities and 

antiradical activities. Jatrorrhizine and magnoflorine 

showed better activities than berberine not bearing 

any readily abstractable hydrogen on its skeleton. The 

former two showed antiperoxidative efficiency in 

DOPC liposomal membrane comparable to that of an 

effective scavenger of peroxyl radicals––stobadine. It 

could be concluded that the favorable antioxidant 

features of the hydroxylated alkaloids were most 

probably ensured by the combination of reasonably 

high antiradical reactivity with high lipophilicity. [37] 

Effects on cardiovascular system 

The antiarrythmic action of Coptis and the role 

of berberine was assessed in various animal modes. 

Berberine was shown to be able to raise the VFT of 

the normal and ischemic myocardium in aneshetized 

rats. Experiments on isolated papillary muscle and 

right atrium preparations from guinea pig leart 

indicated that berberine has positive inotropic and 

negative effects. [2] 

To observe the effects of active ingredients 

from Chinese drugs for activating blood circulation 

and detoxicating, including notoginseng saponins, 

Coptis chinensis, giant knotweed rhizome and 

rhubarb, on blood lipids and inflammatory reaction of 

aortic atherosclerotic plaques in ApoE knockout mice. 

ApoE knockout mice were fed with high-fat diet for 

26 weeks, then they were randomized into 6 groups, 

the untreated model group and the test groups treated 

with various test drugs respectively. After ending the 

13 weeks of treatment, all the mice were sacrificed 

with their blood lipids detected, and their heart and 

aorta were taken out to make slices with paraffin 

embedding. Four sections from aortic root of each 

mouse were chosen to measure and calculate the 

percentage of lipid core (LC) in the total area of 

plaque (TP) and the lipid/collagen ratio (L/C) in the 

plaque by HE and Movat staining respectively, and 

the mean value of the four sections was taken for 

analysis. The expressions of granulocyte-macrophage 

colony-stimulating factor (GM-CSF) and tumor 

necrosis factor-α (TNF-α) in mice's aorta root were 

determined by immunohistochemical staining as well. 

After being treated for 13 weeks, either the 

16


percentage of LC in TP and the L/C ratio was 

significantly lower in all the test drug treated groups 

than those in the model group, respectively (P < 0.01), 

especially prominent in the group treated with giant 

knotweed rhizome. Although lowering of the two 

indexes presented in all of the groups treated by 

notoginseng saponins, Coptis chinensis and giant 

knotweed rhizome, significant difference still 

presented between giant knotweed rhizome treated 

group vs notoginseng saponins and Coptis chinensis 

treated group (P < 0.05). As for the expressions of 

GM-CSF and TNF-α, in comparing with the 

untreated model group, significant decreasing of the 

TNF-α showed only in the rhubarb treated group, 

while that of GM-CSF could be found in all the test 

drug treated groups (P < 0.05). All the four drugs 

tested in the recommended dosage can stabilize the 

vulnerable plaques in ApoE knockout mice by 

improving the constitution of plaque, among them, 

giant knotweed rhizome and rhubarb, the drugs 

possess both the actions of activating blood 

circulation and detoxicating, show more significant 

effect, and their mechanisms may be related to their 

actions in regulating lipid metabolism and inhibiting 

inflammatory reaction. [38] 

We known that vascular smooth muscle cell 

(SMC) proliferation plays an important role in the 

pathogenesis of atherosclerosis and post-angioplasty 

restenosis. Berberine is a well-known component of 

Coptis chinensis, and is capable of inhibiting SMC 

contraction and proliferation, yet the exact 

mechanism is unknown. Therefore, Liang KW et al 

investigated the effect of berberine on SMC growth 

after mechanic injury in vitro. DNA synthesis and 

cell proliferation assay were performed to show that 

berberine inhibited serum-stimulated rat aortic SMC 

growth in a concentration-dependent manner. 

Mechanical injury with sterile pipette tip stimulated 

the regrowth of SMCs. Treatment with berberine 

prevented the regrowth and migration of SMCs into 

the denuded trauma zone. Western blot analysis 

showed that activation of the MEK 1/2 

(mitogen-activated protein kinase 1/2 ), extracellular 

signal-regulated kinase (ERK), and up-regulation of 

early growth response gene (Egr-1), c-Fos and Cyclin 

D1 were observed sequentially after mechanic injury 

in vitro. Semi-quantitative reverse-transcription PCR 

assay further confirmed the increase of Egr-1, c-Fos, 

platelet-derived growth factor (PDGF) and Cyclin D1 

expression in a transcriptional level. However, 

berberine significantly attenuated MEK/ERK 

activation and downstream target (Egr-1, c-Fos, 

Cyclin D1 and PDGF-A) expression after mechanic 

injury in vitro. Their study showed that berberine 

blocked injury-induced SMC regrowth by 

inactivation of ERK/Egr-1 signaling pathway thereby 

preventing early signaling induced by injury in vitro. 

The anti-proliferative properties of berberine may be 

useful in treating disorders due to inappropriate SMC 

growth. [39] 

Berberine was considered to be useful in 

treating some diseases of the cardiovascular system, 

such as hypertension and chronic heart failure (CHF). 

Hong Y et al investigated the inhibitory effect of 

berberine on experimental cardiac hypertrophy, 

which is regarded as a risk factor of CHF and other 

heart diseases. Forty-two male SD rats were divided 

into four groups: age-matched control, aortic banding 

model, berberine-treated group and captopril-treated 

group. Cardiac hypertrophy was induced by 

suprarenal abdominal aorta constriction. These drugs 

were orally administered for 8 weeks starting from 4 

weeks after surgery at dosage of berberine 10 mg·kg -1 

and captopril 50 mg·kg -1 . Blood pressure (BP) was 

measured four times during the period of the 

experiment, and hemodynamic parameters, cardiac 

index, cell size of left ventricular myocardium and 

total protein of left ventricular tissue were detected 8 

weeks after treatment with these drugs. The data from 

the present study showed that: The BP of the aorta 

banded rats was increased compared with those of the 

normal (P < 0.001) and the age-matched control rats 

(P< 0.001), and berberine showed no significant 

effect on it; after 8 weeks of treatment with berberine, 

the elevated left ventricular end diastolic pressure 

(LVEDP) was slightly decreased compared with the 

aortic banded rats. Meanwhile, the maximum rates of 

contraction and relaxation (± dp/dtmax) was 

increased (P < 0.05) and the time to reach the point 

of maximum rate from beginning of contraction 

(t-dp/dt) was shortened (P < 0.01), indicating that the 

functions of heart, both contraction and relaxation, 

were improved; cardiac growth was inhibited by 

treatment with berberine. Both whole heart and left 

ventricular weight were notably decreased compared 

with the banded rats (P < 0.05 and P < 0.01); and the 

17


cell size of left ventricular myocardium was 

significantly reduced (P < 0.001) and the total protein 

of left ventricular tissue was slightly down-regulated 

by treatment with berberine. These results suggest 

that berberine can improve abnormal cardiac function 

and can prevent the development of left ventricular 

hypertrophy induced by pressure-overload. This 

indicates that it may have therapeutic potential in the 

treatment of CHF. [40] 

Pharmacokinetics studies 

It has been reported that berberine is valuable 

for long-term treatment of ventricular premature 

beats (VPBs) and leads to a decrease in mortality for 

patients with congestive heart failure (CHF). In order 

to improve its therapeutic value and reduce its side 

effects, it is necessary to study the relationship 

between its activity and plasma concentration in 

patients with CHF. Patients with CHF were treated 

with conventional therapy for 2 weeks. Immediately 

after the data from a dynamic electrocardiogram 

(DCG) and left ventricular ejection fraction (LVEF) 

were obtained, 1.2 g·d -1 of oral berberine was given. 

After 2 weeks of berberine therapy, the DCG data 

and LVEF were reassessed and the plasma berberine 

concentration was measured by HPLC. Plasma 

samples were pretreated by extraction with 

chloroform. Berberine in all samples was determined 

using a mu Bondapak C 18 column, a mobile phase of 

acetonitrile: 0.02 mol·L -1 phosphoric acid (45:55, v/v), 

and a UV detector at 346 nm. The mean recovery was 

96.5%. The linear range was 40-1600 ng·mL -1 . The 

detection limit for berberine in plasma was 0. 4 ng. 

The decrease in frequency and complexity of VPBs 

and the increase in LVEF in patients with plasma 

berberine concentrations higher than 0.11 mg·L -1 

were more significant than at concentrations lower 

than 0.11 mg·L -1 (P< 0.01 vs P < 0.05). [41] 

In the study to investigate the mechanisms by 

which berberine is transported in the secretory and 

absorptive directions across Caco-2 cell monolayers, 

the basolateral-to-apical (B-A) flux was 30-fold 

greater than the apical-to-basolateral flux and 

temperature dependent (i.e., drastic decrease at 4 

degrees C compared with 37 degrees C). The above 

results suggest the involvement of a carrier-mediated 

active transport mechanism for the B-A transport of 

berberine. However, no significant concentration 

dependency for the permeability (P app ) of berberine 

was observed for B-A transport over a concentration 

range of 5-300 µM, indicating that the K m value of 

berberine for the carrier system is greater than 300 

µM. Well-documented P-glycoprotein (P-gp) 

substrates such as verapamil, daunomycin, and 

rhodamine123 inhibited the B-A flux of berberine, 

whereas tetraethylammonium and taurocholate did 

not, suggesting that P-gp is involved in the transport. 

For the case of daunomycin, the B-A flux, but not the 

apical-to-basolateral flux, was significantly increased 

after pretreatment of the cell monolayers with 

berberine. In addition, the uptake of 1 microM 

daunomycin into Caco-2 cells was decreased as a 

result of this pretreatment. These results suggest that 

the repeated administration of berberine may 

up-regulate P-gp functions in Caco-2 cells. If this 

occurs in the gastrointestinal epithelial cells, the 

repeated administration of berberine may reduce the 

gastrointestinal absorption of P-gp substrates 

including chemotherapeutic agents such as 

daunomycin. [42] 

Berberine is an important ingredient in Coptis 

chinensis Franch but has been shown to have poor 

bioavailability in the dog. The aim of this study was 

to use the P-glycoprotein (P-glycoprotein) inhibitors 

cyclosporin A, verapamil and the monoclonal 

antibody C219 in in vivo and in vitro models of 

intestinal absorption to determine the role of 

P-glycoprotein in berberine absorption. In the rat 

recirculating perfusion model, berberine absorption 

was improved 6-times by P-glycoprotein inhibitors. 

In the rat everted intestinal sac model, berberine 

serosal-to-mucosal transport was significantly 

decreased by cyclosporin A. In Ussing-type chambers, 

the rate of serosal-to-mucosal transport across rat 

ileum was 3 times greater than in the reverse 

direction and was significantly decreased by 

cyclosporin A. In Caco-2 cells, berberine uptake was 

significantly increased by P-glycoprotein inhibitors 

and by monoclonal antibody C219. P-glycoprotein 

appears to contribute to the poor intestinal absorption 

of berberine which suggests P-glycoprotein inhibitors 

could be of therapeutic value by improving its 

bioavailability. [43] 

In order to investigate the detailed 

pharmacokinetics of berberine and its mechanisms of 

18


hepatobiliary excretion, an in vivo microdialysis 

coupled with HPLC was performed. In the control 

group, rats received berberine alone; in the 

drug-treated group, 10 min before berberine 

administration, the rats were injected with 

cyclosporin A (CsA), a P-gp inhibitor; quinidine, 

both organic cation transport (OCT) and P-gp 

inhibitors; SKF-525A (proadifen), a cytochrome 

P450 inhibitor; and probenecid to inhibit the 

glucuronidation. The results indicate that berberine 

displays a linear pharmacokinetic phenomenon in the 

dosage range from 10 to 20 mg·kg -1 , since a 

proportional increase in the area under the 

concentration-time curve (AUC) of berberine was 

observed in this dosage range. Moreover, berberine 

was processed through hepatobiliary excretion 

against a concentration gradient based on the 

bile-to-blood distribution ratio (AUC bile /AUC blood ); 

the active berberine efflux might be affected by P-gp 

and OCT since coadministration of berberine and 

CsA or quinidine at the same dosage of 10 mg·kg -1 

significantly decreased the berberine amount in bile. 

In addition, berberine was metabolized in the liver 

with phase I demethylation and phase II 

glucuronidation, as identified by liquid 

chromatography/tandem mass spectrometry. Also, the 

phase I metabolism of berberine was partially 

reduced by SKF-525A treatment, but the phase II 

glucuronidation of berberine was not obviously 

affected by probenecid under the present study 

design. [44] 

In order to investigate the pharmacokinetics of 

berberine in Coptidis rhizoma extract in rat 

hippocampus and plasma, a simple and accurate 

high-performance liquid chromatography method was 

employed in this study. Berberine was determined 

using a Hypersil C 18 column with an isocratic mobile 

phase of acetonitrile-0.05 M potassium dihydrogen 

phosphate (containing 0.5% triethylamine, pH 3.0) 

and with UV detection at 236 nm. The lower limit of 

quantification for berberine in both hippocampus and 

plasma was 24 ng·ml -1 , and the lowest concentrations 

of berberine determined in rat hippocampus and 

plasma samples were 30.7 ng·mL -1 at 48 h and 38.5 

ng·mL -1 at 4 h, respectively. The calibration curve for 

berberine was linear over the concentration range 

24--6000 ng·mL -1 . At this concentration range, the 

overall recoveries (90.6--94.2%) for berberine were 

determined and the accuracy of intra- and inter-day 

assays from rat samples were less than 7% RSD. 

Following intravenous administration of C. rhizoma 

extract at a dose of 10.2 mg/kg containing 3 mg/kg 

berberine, berberine in the plasma eliminated rapidly 

t 1/2β =1.13 h). However, berberine in the hippocampus 

increased rapidly t 1/2α =0.215 h), peaked at 3.67 h with 

a concentration of 272 ng·g -1 , and had a slow 

elimination rate t 1/2β =12.0 h), which suggests that 

berberine could have a direct action on neuron and 

accumulate in the hippocampus. This study first 

showed the pharmacokinetic characteristics of 

berberine in rat hippocampus and the kinetic 

characteristics of berberine are dissimilar in the 

hippocampus and plasma. [45] 

To study the effects of berberine (BBR) on the 

blood concentration and pharmacokinetics of 

cyclosporin A (CsA) in renal-transplant recipients., a 

randomized and controlled clinical trial was carried 

out in 52 renal-transplant recipients were treated with 

CsA and 0.2 g BBR three times daily for 3 months, 

while another 52 subjects received CsA without BBR 

co-administration. Blood trough concentration of 

CsA and biochemistry indexes for hepatic and renal 

functions were determined. For the pharmacokinetic 

study, six renal-transplant recipients were included 

with a 3-mg/kg dosage of CsA twice daily before and 

after oral co-administration of 0.2 g BBR three times 

daily for 12 days. The trough blood concentrations 

and the ratios of concentration/dose of CsA in the 

BBR-treated group increased by 88.9% and 98.4%, 

respectively, compared with those at baseline (P < 

0.05). As for the BBR-free group, they rose by 64.5% 

and 69.4%, respectively, relative to those at baseline 

(P < 0.01). Nevertheless, the final blood concentrations 

and the ratios of concentration/dose of CsA 

in BBR-treated patients were still 29.3% and 27.8%, 

respectively, higher than those in BBR-free patients 

(P < 0.05). No significant effects on liver or renal 

functions were observed under co-administration of 

BBR. After co-administration of BBR in 6 patients 

for 12 days, the mean AUC of CsA was increased by 

34.5% (P < 0.05). The mean time taken to reach the 

peak blood concentration (T max ) and the mean 

half-life (t 1/2 ) of CsA were increased by 1.7 h and 2.7 

h, respectively (P < 0.05). The average percentage 

increases in the steady-state drug concentration (C ss ) 

and minimum blood concentration (C min ) were 34.5% 

19


and 88.3%, respectively (P < 0.05). In addition, the 

average percentage decrease in CL/F was 40.4% (P < 

0.05) and the peak-to-through fluctuation index was 

significantly reduced (P < 0.01). The study showed 

that BBR can markedly elevate the blood 

concentration of CsA in renal-transplant recipients in 

both clinical and pharmacokinetic studies. This 

combination may allow a reduction of the CsA 

dosage. The mechanism for this interaction is most 

likely explained by inhibition of CYP3A4 by BBR in 

the liver and/or small intestine. [46] 

A rapid and sensitive LC-MS/MS method was 

developed to simultaneously determine berberine, 

palmatine and jatrorrhizine in rat plasma. After 

mixing with the internal standard (IS) 

tetrahydropalmatine, plasma samples were pretreated 

by protein precipitation with acetonitrile-methanol 

(1:2, v/v). Chromatographic separation was carried 

out on a C18 column using a mixture of water 

(containing 0.1% formic acid) and acetonitrile (30:70, 

v/v) as mobile phase. The detection was performed 

by selected reaction monitoring (SRM) mode via 

electrospray ionization (ESI) source operating in the 

positive ionization mode. The method was linear over 

the concentration range of 1.0-250.0 ng·mL -1 for all 

components. The intra- and inter-day precision values 

were less than 14.6% and the deviations were within 

±4.0%. The fully validated LC-MS/MS method has 

been successfully applied to the pharmacokinetic 

study of berberine, palmatine and jatrorrhizine in rat 

plasma after oral administration of coptis-evodia herb 

couple. Three peaks were observed in both individual 

and mean plasma-concentration curves of berberine, 

palmatine and jatrorrhizine, which may be attributed 

to distribution re-absorption and enterohepatic 

circulation. [47] 

Berberine (Ber) and its main metabolites were 

identified and quantified using liquid chromatography/electrospray 

ionization/ion trap mass 

spectrometry. Rat plasma contained the main 

metabolites, berberrubine, thalifendine, demethyleneberberine, 

and jatrorrhizine, as free and 

glucuronide conjugates after p.o. Ber administration. 

Moreover, the original drug, the four main 

metabolites, and their glucuronide conjugates were 

all detected in liver tissues after 0.5 h and in bile 

samples 1 h after p.o. Ber administration. Therefore, 

the metabolic site seemed to be the liver, and the 

metabolites and conjugates were evidently excreted 

into the duodenum as bile. The pharmacokinetics of 

Ber and the four metabolites were determined in 

conventional and pseudo germ-free rats (treated with 

antibiotics) after p.o. administration with 40 mg·kg -1 

Ber. The AUC 0-limt and mean transit time values of 

the metabolites significantly differed between 

conventional and pseudo germ-free rats. The amounts 

of metabolites were remarkably reduced in the 

pseudo germ-free rats, whereas levels of Ber did not 

obviously differ between the two groups. The 

intestinal flora did not exert significant metabolic 

activity against Ber and its metabolites, but it played 

a significant role in the enterohepatic circulation of 

metabolites. In this sense, the liver and intestinal 

bacteria participate in the metabolism and disposition 

of Ber in vivo. [48] 

To investigate whether baicalin and berberine 

affects the transport of nimodipine (NMD) across the 

blood-brain barrier (BBB). Primary-cultured, rat 

brain microvascular endothelial cells (rBMEC) were 

used as an in vitro model of the BBB. When cells 

became confluent, the steady-state uptake of NMD by 

rBMEC with or without baicalin and berberine was 

measured. The effects of baicalin and berberine on 

the efflux of NMD from rBMEC were also studied. 

Baicalin (2-5 µg·mL -1 ) increased the uptake of NMD, 

and baicalin (10-20 µg·mL -1 ) decreased the uptake. 

The steady-state uptake of NMD was higher than that 

of control group in the presence of 0.01-1 µg·mL -1 

berberine, but was lower in the presence of 2-10 

µg·mL -1 berberine. Their results indicated that the 

bidirectional effect of baicalin and berberine on the 

uptake of NMD by rBMEC was found. Higher 

concentration showed an inhibitory effect, and lower 

concentration demonstrated an increasing effect. [49] 

Hong ZY et al [50-57] , School of Pharmacy, 

Second Military Medical University , have studied 

the pharmacokinetics of tetrahydropalmatine (THP). 

They investigated the stereoselective pharmacokinetic 

process of THP in rats. The concentrations of 

tetrahydropalmatine enantiomers in rat plasma were 

determined by coupled achiral and chiral HPLC 

method. The differences in plasma concentrations 

and pharmacokinetic parameters between the two 

enantiomers were compared by paired t-test. The 

plasma levels of l-THP were always higher than those 

of d-THP in eight rats. There was significant 

20


difference between the main pharmacokinetic 

parameters of the two enantiomers. THP showed 

significant stereoselective pharmacokinetics in rats 

after an ig dose of the racemate. [50] 

selective chiral HPLC method coupled with 

achiral column was developed and validated to 

separate and quantify THP enantiomers in dog 

plasma. Chromatography was accomplished by two 

steps: (1) racemic THP was separated from biological 

matrix and collected on a Kromasil C18 column (150 

mmx4.6 mm, 5µm) with the mobile phase 

acetonitrile-0.1% phosphoric acid solution, adjusted 

with triethylamine to pH 6.15 (47:53); (2) 

enantiomeric separation was performed on a 

Chiralcel OJ-H column (250 mmx4.6 mm, 5µm) with 

the mobile phase anhydrous ethanol. The detection 

wavelength was set at 230 nm. (+)-THP and (-)-THP 

were separated with a resolution factor (Rs) of at 

least 1.6 and a separation factor (alpha) greater than 

1.29. Linear calibration curves were obtained over 

the range of 0.025-4 µg·mL -1 in plasma for each of 

(+)-THP and (-)-THP (R 2 >0.999) with a limit of 

detection (LOD) of 0.005 µg·mL -1 and the recovery 

was greater than 88% for each enantiomer. The 

method was used to determine the pharmacokinetics 

of THP enantiomers after oral administration of 

racemic THP. The results presented herein showed 

the stereoselective disposition kinetics of THP in 

dogs and were a further contribution to the 

understanding of the kinetic behavior of THP 

analogues. [51] 

The main objective of this study was to 

determine the brain pharmacokinetics and tissue 

distribution of THP enantiomers in rats after oral 

administration of racemic THP (rac-THP). Rats (5 

animals/group/per time) were given a single oral dose 

of rac-THP and killed after different post-treatment 

times. The concentrations of THP enantiomers in 

plasma, cortex, cerebellum, diencephalon, brain stem, 

striatum and hippocampus were measured using a 

validated chiral HPLC method coupled with an 

achiral column. The pharmacokinetic profiles of the 

two enantiomers in six brain regions were 

significantly different. The peak concentrations (C max ) 

and AUC 0-infinity values of the (-)-enantiomer were 

significantly greater than the corresponding values 

for the (+)-enantiomer while the striatum contained 

the highest peak concentrations compared with the 

plasma and other brain regions. The tissue 

distribution studies also revealed significant 

differences between the two enantiomers in all tissues 

except the lung. The highest concentrations of both 

enantiomers were found in the liver. The (-)/(+)-THP 

ratios in six brain regions and other tissues were 

consistent with that observed in plasma indicating 

that the stereoselective disposition of THP in rat brain 

and other tissues reflects the situation in plasma. [52] 

THP is the active component in Rhizoma 

corydalis and the medicine Yuanhu-Baizhi (YB), 

which consists of Rhizoma corydalis and Radix 

angelicae dahuricae. The aim of this work was to 

compare pharmacokinetic features of THP 

enantiomers in rats dosed with racemic THP 

(rac-THP), Rhizoma corydalis, or YB extracts. A 

single dose of rac-THP (5 mg·kg -1 ) or extracts of 

Rhizoma corydalis and YB (both equivalent to 5 

mg·kg -1 of rac-THP) was given orally to three groups 

of Sprague-Dawley rats, respectively. Blood samples 

were collected periodically and plasma 

concentrations of THP enantiomers were determined 

using an achiral-chiral HPLC method previously 

reported, with some modifications. The C max ratio 

(-/+) of THP was 2.91, 1.38, and 1.19, and the AUC 0 

approximately infinity) ratio (-/+) of THP was 2.84, 1.50, and 

1.35 in rats after dosed with rac-THP, extracts of 

Rhizoma corydalis and YB, respectively. The mean 

AUC 0 approximately infinity and C max of (+)-THP dosed with 

YB extracts were 0.652 ± 0.30 µg·h·ml -1 and 0.148 

±0.09 µg·ml -1 , significantly higher (P < 0.05) than 

those dosed with rac-THP and Rhizoma corydalis 

extracts. The mean AUC 0 approximately infinity and T max of 

rac-THP dosed with YB extracts were 1.500 ± 0.56 

µg·h·ml -1 and 2.12 ±1.1 h, significantly higher (P < 

0.05) than those dosed with rac-THP or Rhizoma 

corydalis extracts. These findings suggested the 

stereoselectivity in pharmacokinetics of THP 

enantiomers in rats was decreased when dosed in 

plant form, while the AUC 0-approximately infinity of 

rac-THP increased when YB extracts were dosed, 

confirming the compatibility in drug combination of 

Rhizoma corydalis and Radix angelicae dahuricae. [53] 

The main objective of this study was to 

determine the brain pharmacokinetics and tissue 

distribution of THP enantiomers in rats after oral 

administration of racemic THP (rac-THP). Rats (5 

animals/group/per time) were given a single oral dose 

21


of rac-THP and killed after different post-treatment 

times. The concentrations of THP enantiomers in 

plasma, cortex, cerebellum, diencephalon, brain stem, 

striatum and hippocampus were measured using a 

validated chiral HPLC method coupled with an 

achiral column. The pharmacokinetic profiles of the 

two enantiomers in six brain regions were 

significantly different. The peak concentrations (C max ) 

and AUC 0-infinity values of the (-)-enantiomer were 

significantly greater than the corresponding values 

for the (+)-enantiomer while the striatum contained 

the highest peak concentrations compared with the 

plasma and other brain regions. The tissue 

distribution studies also revealed significant 

differences between the two enantiomers in all tissues 

except the lung. The highest concentrations of both 

enantiomers were found in the liver. The (-)/(+)-THP 

ratios in six brain regions and other tissues were 

consistent with that observed in plasma indicating 

that the stereoselective disposition of THP in rat brain 

and other tissues reflects the situation in plasma. [54] 

To investigate the stereoselective pharmacokinetic 

process of THP in rats, the concentrations of THP 

enantiomers in rat plasma were determined by 

coupled achiral and chiral HPLC method. The plasma 

levels of l-THP were always higher than those of 

d-THP in eight rats. There was significant difference 

between the main pharmacokinetic parameters of the 

two enantiomers. THP showed significant 

stereoselective pharmacokinetics in rats after an ig 

dose of the racemate. [55] 

A selective chiral HPLC method coupled with 

achiral column was developed and validated to 

separate and quantify THP-enantiomers in dog 

plasma. Chromatography was accomplished by two 

steps: (1) racemic THP was separated from biological 

matrix and collected on a Kromasil C18 column (150 

mmx4.6 mm, 5 µm) with the mobile phase 

acetonitrile-0.1% phosphoric acid solution, adjusted 

with triethylamine to pH 6.15 (47:53); (2) 

enantiomeric separation was performed on a 

Chiralcel OJ-H column (250 mmx4.6 mm, 5 µm) 

with the mobile phase anhydrous ethanol. The 

detection wavelength was set at 230 nm. (+)-THP and 

(-)-THP were separated with a resolution factor (Rs) 

of at least 1.6 and a separation factor α greater than 

1.29. Linear calibration curves were obtained over 

the range of 0.025-4 µg·mL -1 in plasma for each of 

(+)-THP and (-)-THP (R2>0.999) with a limit of 

detection (LOD) of 0.005 µg·mL -1 and the recovery 

was greater than 88% for each enantiomer. The 

relative standard deviation (RSD) and relative error 

values were less than 10% at upper and lower 

concentrations. The method was used to determine 

the pharmacokinetics of THP enantiomers after oral 

administration of racemic THP. The results presented 

herein showed the stereoselective disposition kinetics 

of THP in dogs and were a further contribution to the 

understanding of the kinetic behavior of THP 

analogues. [56] 

THP is a racemic mixture which contains 50% 

of the (+) and 50% of (-) enantiomer. The (-) 

enantiomer accounts for most of the analgesic effects. 

Plasma concentrations of THP enantiomers were 

analyzed by chiral HPLCon a Chiralcel OJ column 

with quantification by UV at 230 nm. The method 

was used to determine the pharmacokinetics of THP 

enantiomers in rats and dogs after oral administration 

of rac-THP or (-)-THP. The pharmacokinetic profiles 

of the two enantiomers after dosing with rac-THP 

were significantly different both in rats and dogs. The 

mean C max and AUC 0-infinity values in rats were 1.93 

±0.36 µg·ml -1 and 6.65 ± 2.34 µg·h·ml -1 for the (-) 

enantiomer, and 1.11 ± 0.25 µg·mL -1 and 2.03 ±0.45 

µg·h·ml -1 for the (+) enantiomer. The mean C max and 

AUC 0-infinity in dogs were 1.60 ± 0.81µg·mL -1 and 

9.88 ±2.58 µg·h·mL -1 for the (-) enantiomer, while 

0.36 ± 0.21 µg·mL -1 and 1.22 ± 0.40 µg·h·ml -1 for the 

(+) enantiomer. The rac-THP at 40 mg·kg -1 and 

(-)-THP at 20 mg/kg had very similar plasma 

concentration-time profiles, and C max , AUC 0-infinity , 

and t 1/2 of the (-) enantiomer in both rats and dogs, 

indicating that the two treatments were equivalent 

with respect to the pharmacokinetic properties of the 

(-) enantiomer. [57] 

Application of Rhizoma Coptidis 

Rhizoma Coptidis is used in traditional Chinese 

medicine for long history in China. The earliest 

record was listed in Shengnong Bencaojing. Five to 

seven year old roots or rhizomes of Coptis chinensis 

Franch, Coptis deltoidea C.Y. Cheng et Hsiao or C. 

teeta Wall are the medicinal use in in traditional 

Chinese medicine. 

22


Rhizoma Coptidis is a very useful herb in 

traditional oriental medicine. Cheng [58] did statistics 

of 13 reciping books before the Song Dynasty in 

which Chinese medicinal formulaes including 

Rhizoma Coptidis were approximate 5 percentage of 

all. There are many known formulaes having 

continued to be used until now such as Zuojinwan 

(including Rhizoma Coptidis and Fructus Evodiae, 6 

1)Fanzuojinwan (including Rhizoma Coptidis and 

Fructus Evodiae16), Coptidis Decoction for 

Detoxification(including Rhizoma Coptidis, Radix 

Scutellariae, Cortex Phellodendri and Gardenia 

jasminoides Ellis). 

Most chemical constituents of Rhizoma Coptidis 

had been found and total alkoids had been thinked as 

main active components espcially berberine. 

Pharmacological action of Coptis chinensis and 

berberine had been studied systematically.Both in 

Traditional Chinese and Western Medicine,berberine 

was widly applied to treat with infectious diseases 

including not only mainly gastrointestinal bacterial 

infection but also respiratory infection and 

dermatologic infection. Recently, new pharmacologic 

action such as anticancer, hypoglycemic activity 

improving function of the circulatory system are 

more and more paid attention to by people. With the 

development of the living standard of the people and 

aging of societal population, the incidence of 

diabetescardiovascular disease and hypertension 

were increasing and both Coptis chinensis and 

berberine have good efficacy to treat with these 

diseases. Bescause of extensive pharmacologic 

effects and affluent resource of Coptis chinensis in 

China , it is worth doing more in-depth research on 

its chemical composition and pharmacologic action 

and making full use of it. 

Acknowledgement This project was 

supported by National Key Technologies R&D 

Program of China in the Eleventh h Five-year Plan 

under Grant No. 2006BAI06A01 and 

No.2007BAI41B06 

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berberine from rhizome of Coptis chinensis Franch using 

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29. Kuo CL, Chi CW, Liu TY. The anti-inflammatory potential 

of berberine in vitro and in vivo. Cancer Lett. 2004; 

203(20):127-137. 

30. Lee DU, Kang YJ, Park MK, Lee YS, Seo HG, Kim TS, 

Kim CH, Chang KC. Effect of 13-alkyl-substituted 

berberine alkaloids on the expression of COX-II, TNF-alpha, 

iNOS, and IL-12 production in LPS-stimulated macrophages. 

Life Sci. 2003; 73(11):1401-1412. 

31. Hao Y, Qiu QY, Wu J, Wang JJ. Effects of berberine on IL-1 

or tumour necrosis factor induced polymorphonuclear 

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32. Choi BH,Ahn IS, Kim YH, Park JW, Lee SY, Hyun CK, Do 

MS . Berberine reduces the expression of adipogenic 

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33. Kuo CL, Chi CW, Liu TY . The anti-inflammatory potential 

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28(2):126-30. 

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Hsu SL. Berberine suppresses MEK/ERK-dependent Egr-1 

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40. Hong Y, Hui SC, Chan TY, Hou JY. Effect of berberine on 

regression of pressure-overload induced cardiac hypertrophy 

in rats. Am J Chin Med. 2002; 30(4):589-99. 

41. Zeng X, Zeng X. Relationship between the clinical effects of 

berberine on severe congestive heart failure and its 

concentration in plasma studied by HPLC. Biomed 

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42. Maeng HJ, Yoo HJ, Kim IW, Song IS, Chung SJ, Shim CK. 

P-glycoprotein-mediated transport of berberine across 

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43. Pan GY, Wang GJ, Liu XD, Fawcett JP, Xie YY. The 

involvement of P-glycoprotein in berberine absorption. 

Pharmacol Toxicol. 2002; 91(4):193-7. 

44. Tsai PL, Tsai TH . Hepatobiliary excretion of berberine. 

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45. Wang X, Wang R, Xing D, Su H, Ma C, Ding Y, Du L. 

Kinetic difference of berberine between hippocampus and 

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46. Wu X, Li Q, Xin H, Yu A, Zhong M. Effects of berberine on 

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47. Deng Y, Liao Q, Li S, Bi K, Pan B, Xie Z. Simultaneous 

determination of berberine, palmatine and jatrorrhizine by 

24


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Chromatogr B Analyt Technol Biomed Life Sci. 2008; 

863(2):195-205. 

48. Zuo F, Nakamura N, Akao T, Hattori M. Pharmacokinetics 

of berberine and its main metabolites in conventional and 

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pharmacokinetic study of tetrahydropalmatine enantiomers 

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pharmacokinetics and tissue distribution of 

tetrahydropalmatine enantiomers in rats after oral 

administration of the racemate. Biopharm Drug Dispos. 

2006; 27(3):111-7. 

55. Hong ZY, Fan GR, Chai YF, Wen J, Yin XP, Wu YT. 

Stereoselective pharmacokinetics of tetrahydropalmatine in 

rats. Acta Pharm Sin. 2005; 40(8):746-9. 

56. Hong ZY, Fan GR, Chai YF, Yin XP, Wen J, Wu YT. 

Chiral liquid chromatography resolution and stereoselective 

pharmacokinetic study of tetrahydropalmatine enantiomers 

in dogs. J Chromatogr B Analyt Technol Biomed Life Sci. 

2005; 826(1-2):108-13. 

57. Hong Z, Fan G, Chai Y, Yin X, Wu Y. Stereoselective 

pharmacokinetics of tetrahydropalmatine after oral 

administration of (-)-enantiomer and the racemate. Chirality. 

2005; 17(5):293-6. 

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25

Tianjin Centre for Drug Safety Evaluation and 

Research 

Good Laboratory Practice certification 

Tianjin Centre for Drug Safety Evaluation and 

Research affiliated in Tianjin Institute of Pharmaceutical 

Research has already passed the Good Laboratory Practice 

(GLP) certification by State Food and Drug Administration 

(SFDA), which has filled the gap in Tianjin municipality 

since there was no institution for GLP safety evaluation 

before. 

SFDA designated the experts who are engaged in the 

on-the-spot inspection for the Centre according to the latest 

revision of GLP certification method (Tentative Version). 

The inspection team members learnt the reports on the GLP 

construction and checked the complete soft and hard 

conditions as well as the reference materials of non-clinical 

safety evaluation for drug experiments. Based on the GLP 

standard, all the experts confirmed that the Centre has 

already performed 493 items of various experiments for 

safety evaluation among 228 novel drugs, drafted and 

carried out 744 standard operating procedures (SOP), and 

built up a perfect quality-assured system. They also agreed 

that the studies on the safety evaluation, the arrangement, 

and the operation in the Centre could be carried on more 

smoothly and better under the desirable GLP standard. 

In the light of the comments of Mr.Xianglong DAI 

and Mr.Dongliang YANG, etc., the mayors and leaders of 

Tianjin municipality, Tianjin Centre for Drug Safety 

Evaluation and Research has been built and listed in the 

first-approval item in the support of Tianjin Special Fund 

for Technical Innovation. In Tianjin, the Centre has set up 

the modern integrated pharmaceutical Industry 

(GAP-GLP-GCP-GMP), become an essential technical 

platform in the innovative system of novel drugs, and will 

make the innovatory potential of novel drugs promote 

greatly. Also it will provide a technical prop for the 

sustainable developing in modern pharmaceutical Industry 

and the establishment of China International Innovation 

Zone for Biomedicine. 

26

Cheng TF et al. Asian Journal of Pharmacodynamics and Pharmacokinetics 2009; 9(1):27-49 




ISSN 1608-2281 



Attention on research of pharmacology and toxicology of 

nanomedicines 

Tie-Feng Cheng 1, 2 , Yong-Da Sun 3,4, , Duan-Yun Si 2,3 Chang-Xiao Liu 2,3 

1 Key Laboratory for Special Functional Materials, Henan University, Kaifeng, 475001, China 

2 Research Center of New Drug Evaluation, The State Key Laboratories of Pharmacodynamics and 

Pharmacokinetics, Tianjin Institute of Pharmaceutical Research, Tianjin 300193, China 

3 

Research Center of Biological Evaluation of Nanopharmaceuticals, China National Academy of 

Nanotechnology and Engineering, Tianjin, 300457, China 

4. 

Tianjin Crystec Pharmaceutical technology Ltd, Tianjin, 300457, China 

Abstract 

Key words 

In the 21st century, nanoscience and nanotechnology obtains the world attention due to this 

revolutionary theory and technical features. Nanoscience and nanotechnology cover the theory 

and technology of physics, chemistry, medicine, material science, biomedical engineering and 

biology, therefore, they have no less contribution to science and technology as biotechnology 

and information technology. Recent years have witnessed the rapid development of China’s 

nanoscience and nanotechnology with widespread influence. It was attended by scientists of the 

world. Research, development and application of nanotechnology research in China can be 

summed up in three characteristics: the first, China government in support of sustainable 

development; the second, significant academic achievements, and the third, a clear consensus on 

sustainable development for nanoscience and nanotechnology research and development. In this 

review paper, we discussed the pharmacology and toxicology of nanomedicines, and presented 

some issues on research and development and application of nanomedicines in the future. 

Nanoscience; nanotechnology; nanomedicines; pharmacology; toxicology; China; sustainable 

development; academic achievements 

Article history Received 26 December 2008; Accepted 27 February 2009 

Publication data Pages: 23; Tables: 2; Figures: 1; References: 47; Paper ID: 1608-2281-2009-0901027-23 

Corresponding author Professor Chang-Xiao Liu, Tianjin Institute of Pharmaceutical Research, 308 An-Shan West Road, Tianjin, 

300193, China. E-mail: liuchangxiao@vip.163.com. 

Introduction 

Nanoscience and nanotechnology has attracted 

full attention of scientists around the world due to the 

breakthrough theory and technical feature. Nanoscience 

and nanotechnology, as a newly emerging leading-edge 

discipline, cover many fields such as physics, chemistry, 

medicine, material science, biomedical engineering and 

biology, therefore, the rapid development of nanoscience 

and nanotechnology has contributed more and more deep 

27


knowledge to other discipline as biotechnology and 

information technology information technology. [1] 

Recently, nanotechnology proves its diverse 

applications to take up the international market in 

areas of biomedicine, informatics, energy resource, 

astronautics, oceanography and national defense, 

and so on. These applications offer huge economic 

and technological potentialities. Current advances in 

nanoscience and nanotechnology have led to the 

development of the new field of nanomedicine, 

which includes many applications of nanomaterials 

and nanodevices for diagnostic and therapeutic 

purposes. At the Third Annual Meeting of the 

American Academy of Nanomedicine held at the 

University of California San Diego, in San Diego, 

USA, during September 7-8, 2007. The discussion 

was focused on successful translational 

nanomedicine: from bench to bedside. There were 

four keynote lectures and eight scientific 

symposiums in this meeting. The researchers and 

investigators reported the results and process of 

current nanomedicine research and approaches to 

clinical applications. The meeting provided exciting 

information for nanomedicine clinical-related 

researches and strategy for further development of 

nanomedicine research which will be benefits to 

clinical practice. [2] 

Recent years have witnessed the rapid 

development of China’s nano-science and 

technology with widespread influence. It was 

attended by scientists of the world. On January 

2008, a proposal to convene a Sino-US symposium 

on nanomedicine and nanobiology was jointly made 

by Dr. Elias Zerhouni (Director of the National 

Institutes of Health, USA), Dr. John E. Niederhuber 

(Director of the National Cancer Institute, USA and 

Dr. Samuel Wilson (Director of the National 

Institute of Environmental Health Sciences, USA). 

To integrate outstanding research forces in China 

and carry out exchanges with scholars of the world, 

in particular US, the Xiangshan Science Conference 

(the 331st Xiangshan Science Conference) on 

nanotechnology and nanomedicine for cancer 

treatment held from 21 to 24 October in Beijing, 

China. Professor Zhao YL (Chinese National 

Research Center of Nanosceince), Professor Robert 

P. Blumenthal (Center for Cancer Research 

Nanobiology ProgramNational Cancer Institute – 

Frederick, National Institutes of Health, USA, and 

Professor Michael M. Gottesman (National Cancer 

Institute, National Institutes of Health, USA), as 

co-chairmen of this meeting, lead scientists from 

China and USA to discussed nanoscience, 

nanotechnology and nanomedicines for cancer 

treatment. An emerging field that takes full 

advantage of expertise and research approaches 

from such academic disciplines as nanotechnology, 

biology, chemistry, physics, medicine, 

pharmaceutics and public health. No single 

discipline can deal with the new field characterized 

with a lot of interdisciplinary and comprehensive 

studies. It is both a topic at the cutting-edge of 

science development and an important social issue 

closely related to people’s health and environment, 

offering unlimited opportunities for innovation. The 

meeting was focused on nanomedicines and 

nanotechnology for cancer treatment, environmental 

health of nanotechnology and its safety, and the 

strategy and policy for nanotechnology 

development. Central topics: (1) Molecular Basis of 

Nanomedicine, (2) Development of “Smart” 

Nanoparticles, biomarker and targeted delivery for 

cancer therapy and imaging, (3) Nanotechnology : 

Path to the clinic promises and hurdles and (4) The 

molecular basis for engineered nanomaterial 

interactions with human health and the environment. 

The 4 topics provided exciting information for 

nanomedicine researches in basis and clinical 

research strategy for further development of 

nanomedicines. [3] 

Research and development of nanoscience 

and nanomedicines in China 

Now in China, nanoscience and 

nanotechnology become ever more consequential in 

our lives, we in the scientific community need to 

better inform and educate the public about the 

transformations this new nano era is likely to bring. 

Among the fields that have enjoyed particularly 

rapid development in China in the past decade are 

nanoscience and nanotechnology. These terms refer 

to the growing knowledge base and technical 

framework for understanding and manipulating 

matter on nanometer scale ranging from the atomic 

to the cellular. Like many other countries, we in 

28


China expect that the development of nanoscience 

and nanotechnology will greatly affect many areas 

of scientific research and industrial development, 

and many aspects of everyday life. [4] 

Research, development and application of 

nanotechnology research in China can be summed 

up in three characteristics: the first, the government 

in support of sustainable development; the second, 

significant academic achievements, and the third, a 

clear consensus on nano-innovation. 

The government in support of sustainable 

development 

When the concept of nanoscience and 

nanotechnology was first introduced in the 1980s, it 

was received favorably in China. The initial interest 

was in part stimulated by the development of new 

tools and techniques for observing materials on the 

nanoscale, especially scanning probe microscopes 

(SPMs). Soon after the concept began trickling 

through the scientific ranks, the Chinese Academy 

of Sciences (CAS), the National Natural Science 

Foundation of China (NSFC), and the State Science 

and Technology Commission (SSTC)/ the Ministry 

of Science and Technology (MOST) began funding 

nanoscience-related work and activities. China also 

has helped those who work in nanoscience and 

nanotechnology to develop their sense of being part 

of a new research and development community. 

Since 1990, for example, dozens of international 

and domestic conferences in the field have been 

held in China. These conferences addressed a wide 

range of topics in nanoscience and nanotechnology 

and attracted wide attention and public interest. In 

the 1990s, support for the development of 

nanoscience and nanotechnology increased 

substantially, largely through several major 

initiatives. In 1990, for example, SSTC launched 

the nearly decade-long "Climbing Up" project on 

nanomaterial science. In 1999, MOST started a 

national basic research project (“973” Plan) entitled 

"Nanomaterial and Nanostructure" and has been 

funding basic research on nanomaterials, such as 

nanotubes, ever since. China National High 

Technology Plan(“863” Plan), which encompasses 

many categories of technology, has included a 

series of projects for nanomaterial applications. 

From 1990 to 2005 alone, over 1200 projects were 

implemented. In addition, during this period, NSFC 

approved nearly 1000 grants for small-scale projects 

in related areas. With so much going on in 

nano-related R&D in so many different places in 

China, we created in 2000 the National Steering 

Committee for Nanoscience and Nanotechnology to 

oversee national policy and planning in these 

arenas. [4] 

Moving forward in nanoscience and 

nanotechnology requires a particularly wide 

spectrum of skills and knowledge. The demand for 

multidisciplinary research platforms with 

components assembled from academia and industry 

and that also have educational functions has become 

especially strong in recent years. According to 

incomplete statistics, more than 50 universities, 50 

institutes and over 300 industry enterprises in China 

have engaged in nanoscience and nanotechnology 

research and development, with the involvement of 

more than 3000 researchers across China. To move 

forward and become more competitive in 

nanoscience and nanotechnology, China needs to 

continue to expand its now-limited research 

infrastructure. In some areas, such as nanoscale 

devices with novel electronic and optoelectronic 

features, efforts to consolidate resources to tackle 

key technological issues are under way. Efforts 

have also been made to pursue industrial-scale 

production of nanomaterials, such as CNTs, 

polymeric nanocomposites, and nanoparticle 

materials, with the intention of opening up 

opportunities for new businesses to sprout and grow. 

The nanoscience and nanotechnology community in 

China has made remarkable advances across the 

research and development spectrum, from 

fundamental scientific research to studies into the 

potential societal implications of new 

nanotechnologies. China still has a long way to go 

to improve the overall competitiveness of its 

nanoscience and nanotechnology enterprise. [4] 

During the Ninth Five-year Plan period 

(1996-2000), the national “863” Plan supported by 

China government starts the projects of improving 

nanobiotechnology; during the Tenth Five-year 

period (2001-2005) national 863 and 973 

Plans and National Natural science made 

29


nanoscience, nanotechnology, and nanomedicine 

studies as priority subjects to support by 

government. During the Eleventh Five-year Plan 

period (2006-2010), the state is increased support 

for nanoscience and nanotechnology research, the 

annual input on billions of funds to carry out 

research, one of three subjects of 

nanopharmaceuticals are listed of the research plan. 

Significant academic achievements 

The scientific output of Chinese nanoscientists 

is becoming ever more significant. According to the 

Scientific Citation Index, CAS ranked fourth in the 

world in total number of citations among those 

institutions and universities that published more 

than 100 nanotechnology papers from 1992 to 2002. 

Another recent analysis of nanoscience productivity 

around the world ranked China at the top for the 

first 8 months of 2004. This should not give the 

Chinese research community reason to be overly 

optimistic, however. The volume of published 

papers and total number of citations is only one 

indicator of the value of research. Another is the 

impact, or the number of citations per paper. From 

2001 to 2003, the number of citations per 

nanotechnology paper published by scientists in the 

United States, Germany, Japan, and China was 

about 6.56, 4.54, 3.7, and 2.28, respectively. 

Since 2006, Chinese basic research papers on 

nanoscience and nanotechnology and total number 

of citations have become the world's second largest, 

behind only the United States. According to 

statistical data from www.cnki.net (2004-2008), 

Chinese scholars published a large number of 

nano-page research thesis in Chinese academic 

journals (as shown in Table 1). 

Table 1. Papers published in Chinese journals from 

2004 to 2008 

Year Nanoscience and 

nanotechnology 

Nanomedicine 

2004 179 11 

2005 226 22 

2006 280 41 

2007 169 18 

2008 235 7 

A clear consensus on nano-innovation 

Facing on the arduous in nanoscience and 

nanotechnology research, and the risks of 

nanopharmaceutical industry, we think in this area 

should pay attention on four-oriented development, 

according to China's national conditions. The first is 

the practice research-oriented, combining basic and 

application. The second is to set up different 

professional disciplines for the research bases and 

to strengthen the efficacy, safety, and the 

industrialization, and feasibility study of 

nanomedicines in order to ensure sustainable 

development. The third, focus on solving the 

challenging problem of the difficult implementation, 

and breakthroughs in nanoscience and 

nanotechnology. The fourth, the complexity in 

research and development of the new technologies 

requires to support with long-term development, 

and to know the risks for technological 

transformation to industrialization. 

Nanotoxilogy and Nanopharmacology 

Nanotechnology is a newly fashionable field 

but in the world of drug development it is certainly 

not new. Nanotechnology has a vital role to play in 

realizing cost-effective diagnostic, therapeutic and 

prevent tools. The applications of nanotechnology 

for treatment, diagnosis, monitoring and control of 

biological systems have recently been referred to as 

nanomedicine. The nanocarriers have been made of 

safe materials, including synthetic biodegradable 

polymers, lipids and polysaccharides. Nanomedicines 

(nanopharmaceuticals) are the convergence 

of nanotechnology and biotechnology and an 

important component of nanotechnology. 

Application of nanotechnology is just started in 

traditional Chinese medicines. 

Nanopharmaceuticals or "Nanomedicines" can 

be developed either as drug delivery systems or 

biologically active drug products. They comprise 

nanometre size scale complex systems, consisting 

of at least two components, one of which being the 

active ingredient. Drug delivery is an 

interdisciplinary area of research that aims at 

making the administration of complex new drugs 

feasible, as well as adding critical value to the drugs 

that are currently in the market. At present, one of 

30


the most attractive areas of research in drug delivery 

is the design of nanomedicines consisting of 

nanosystems that are able to deliver drugs to the 

right place, at appropriate times. The goal of the 

present article is to review the advances we have 

made in the development and characterization of 

nanosystems intended to be used as drug carriers for 

mucosal administration. These nanocarriers are able 

to protect the associated drug against degradation 

and facilitate its transport across critical and 

specific barriers. Some are further able to release 

the associated drug to the target tissue in a 

controlled manner. These nanocarriers have been 

made of safe materials, including synthetic 

biodegradable polymers, lipids and polysaccharides. 

The change in the physicochemical and structural 

properties of engineered nanosized materials with a 

decrease in size could be responsible for a number 

of material interactions that could lead to 

toxicological effects. At present, scientists must 

accept that it is still very early in the toxicological 

evaluation for nanomaterials and nanomedicines, 

and few data on the safety and toxicity. The safety 

evaluation of nanomedicines includes workforce 

exposure limits in manufacturing process, 

environment impact with general impact and to 

patients after administration and safety for human 

use, such as depends on route of administration, 

dose and dosing frequency, as well as safety in drug 

delivery relates to toxicity of drug payload. The 

biomedical evaluation of nanomedicines includes 

biodistribution, metabolic fate, Persistance of 

non-degradable systems, Specific therapeutic issues 

and immunogenicity. We must pay an attention on 

the relative issues of nanomedicines with human 

health and safety and toxicity to develop the 

evaluation methods of nanoproducts and make 

nanotechnology play a great role in the progress in 

nanotechnology and medicines and medicine 

engineering .[5] 

Evaluation on safety and toxicology of 

nanomedicines 

The toxicology of nanomedicines used in 

device manufacture should be considered during 

their entire life cycle at stages of manufacture and 

preclinical and clinical development, consumer and 

staff safety and waste management in environment. 

The development of in vitro models of testicular 

toxicity may provide important tools for 

investigating specific mechanisms of toxicity in the 

testis. Although various systems have been reported, 

their application in toxicological studies has been 

limited by the poor ability to replicate the complex 

biochemical, molecular, and functional interactions 

observed in the testis. In vitro models have been 

established, and some of them have tried to 

reproduce the complex interactions that take place 

between the different germ cells. These models are 

limited by the poor viability of freshly isolated germ 

cells. So the development of a germ-line stem cell is 

of great interest. After previous studies to develop 

an immortalized cell line [6-8] with promising 

application in the study of testis toxicity. In vivo 

system for evaluation on safety and toxicology is 

very importance. This evidence of physiologically 

significant histopathological changes clearly 

indicates the potential of these nanomaterials for 

human toxicity at realistic doses. 

Nanoscale materials are seeming application in 

direct interventions to improve public health both 

through therapeutic strategies and environmental 

remediation. Recent years have seen the emergence 

of nano-engineered drug delivery strategies. 

Approval of abraxane, a nano-formulation of taxol 

for the treatment of breast cancer, was received by 

Food and Drug Administration (FDA), USA. This 

protein nano-bead conjugated pharmaceutical has 

increased water solubility allowing for elimination 

of the toxicity associated with the solvent vehicle 

and improved therapeutic index. The benefit of 

abraxane relies on the nanoscale formulation rather 

than on the emergent properties of the 

nanomaterials as a therapeutic modality. [9] Powers et 

al pointed out that basis nanoparticle 

characterization techniques are discussed, along 

with some of the issues and implications associated 

with measuring nanoparticle properties and their 

interactions with biological systems. Recommendations 

regarding how to approach 

nanomaterial characterization include using proper 

sampling and measurement techniques, forming 

multidisciplinary teams, and making measurements 

as close to the biological action point as possible. [10] 

The science of toxicology has provided the 

31


foundation for understanding and studying the 

interactions between chemical drugs and biology. 

While the use of nanomaterials, nanomedicines/ 

nanopharmaceuticals is rather new in the 

commercial products, the philosophical basis for 

performing the toxicological evaluation of these 

products is not expected to be different form other 

chemical drugs. 

At present, scientists must accept that it is still 

very early in the toxicological evaluation for 

nanomaterials, nanomedicines/nanopharmaceuticals, 

and few data on the safety and toxicity. The basic 

tenet of study designed to develop a study system of 

toxic effects of nanomaterials, nanomedicines/ 

nanopharmaceuticals on biological systems is to 

understand the physico-chemical characteristics of 

nanomaterials, nanomedicines. Therefore, the 

approach to addressing the safety and toxicity of 

these products will best be conducted via 

multidisciplinary terms. Many traditional methods 

and approaches will likely be applicable to study of 

nanomaterials,nanomedicines/nanopharmaceuticals. 

Nanotechnology research and development is 

directed toward understanding and creating 

improved materials, devices, and systems that 

exploit these properties. In a review, Thomas et al 

reviewed that a limited subset of products that 

contain nanoscale materials, assess the available 

data for evaluating the consume exposures and 

potential hazard associated with these products, and 

discuss the capacity of US regulatory agencies to 

address the potential risks associated with these 

products. [11] Some of the potential impacts of 

dermal exposure to nanoscale materials include the 

following: (1) enhanced amount and depth of 

penetration of active ingredients in cosmetic into 

the skin resulting in increased activity, (2) 

ingredients that are chemically unstable in air and 

light (as retinal and vitamin E) may be more readily 

used in topical products following encapsulation in 

nanoparticles, and (3) and timed release of 

ingredients may become more feasible in topical 

products and could allow for improved 

effectiveness equivalent to current controlled 

release orally administered drugs. 

Table 2. The biomedical evaluation of nanomedicines 

Evaluation terms 

Evaluation contents 

Biodistribution 

Whole organism, cellular level 

Metabolic fate 

Absorption, distribution, metabolism and excretion 

Immunogenicity 

IgG/IgM production, cytokine induction 

Persistance of non-degradable systems 

Possibility of lysosomal storage disease 

Biocompatibility 

Biological environment and toxicology and adverse 

effect to patients 

Specific therapeutic issues 

Therapeutic index of nanomedicines and its delivery 

systems in drug delivery relates to toxicity pf drug 

payload 

Due to the nanotechnology combines with 

biotechnology, a newly emerging cross-disciplinary 

field nanobiotechnology, this becomes the new 

developing area. As the research and application of 

nanotechnology, studying and understanding the 

complex relationship between nanomaterials/ 

nanomedicines and biological system will show 

special important to environmental, human health 

and safety. Criticism of the use of laboratory 

animals for the safety testing of chemicals is 

increasing, in society as a whole and also in the 

scientific world. This criticism is not only limited to 

ethical concerns, but scientific considerations also 

play a significant role. It should be realized that the 

animal bioassays presently used in toxicity testing 

are model systems for the prediction of toxicity in 

humans or the environment. In the last few decades 

new technologies and new knowledge have become 

available. This development is the result of 

intensive fundamental toxicological research and the 

implementation of new methods and technologies. [12 ] 

The biomedical evaluation of nanomedicines 

includes biodistribution, metabolic fate, persistance 

of non-degradable systems, Specific therapeutic issues 

and immunogenicity (Table 2). [1] 

32


Intraperitoneal injection of [Gd@C 82 (OH) 22 ] n 

nanoparticles decreased activities of enzymes 

associated with the metabolism of reactive oxygen 

species (ROS) in the tumor-bearing mice. Several 

physiologically relevant ROS were directly 

scavenged by nanoparticles, and lipid peroxidation 

was inhibited in this study. [Gd@C 82 (OH) 22 ] n 

nanoparticles significantly reduced the electron spin 

resonance (ESR) signal of the stable 

2,2-diphenyl-1-picryhydrazyl radical measured by 

ESR spectroscopy. Like-wise, studies using ESR 

with spin-trapping demonstrated efficient 

scavenging of superoxide radical anion, hydroxyl 

radical, and singlet oxygen (1O2) by 

[Gd@C 82 (OH) 22 ] n nanoparticles. In vitro studies 

using liposomes prepared from bovine liver 

phosphatidylcholine revealed that nanoparticles also 

had a strong inhibitory effect on lipid peroxidation. 

Consistent with their ability to scavenge ROS and 

inhibit lipid peroxidation, we determined that 

[Gd@C 82 (OH) 22 ]n nanoparticles also protected cells 

subjected in vitro to oxidative stress. Studies using 

human lung adenocarcinoma cells or rat brain 

capillary endothelial cells demonstrated that 

[Gd@C 82 (OH) 22 ] n nanoparticles reduced H 2 O 2 - 

induced ROS formation and mitochondrial damage. 

[Gd@C 82 (OH) 22 ] n nanoparticles efficiently inhibited 

the growth of malignant tumors in vivo. In 

summary, the results obtained in this study reveal 

antitumor activities of [Gd@C 82 (OH) 22 ] n 

nanoparticles in vitro and in vivo. Because ROS are 

known to be implicated in the etiology of a wide 

range of human diseases, including cancer, the 

present findings demonstrate that the potent 

inhibition of [Gd@C 82 (OH) 22 ] n nanoparticles on 

tumor growth likely relates with typical capacity of 

scavenging reactive oxygen species. [13] 

Evaluation on pharmacology of 

nanomedicines 

Nanotechnology manifests itself in a wide 

range of materials that can be useful to medical 

application. Virtually all of these materials have 

been designed with chemically modifiable surfaces 

to attach a variety of legends that can turn these 

nanomaterials into biosensors, molecular-scale 

fluorescent tags, imaging agents, targeted molecular 

delivery vehicles, and other useful biological tools. 

The unprecedented freedom to design and modify 

nanomaterials to target cells, chaperone drugs, 

image biomolecular processes, sense and signal 

molecular responses to therapeutic agents, and guide 

surgical procedures is the fundamental capability 

offered by nanotechnology, which promises to 

impact drug development, medical diagnostics, and 

clinical applications profoundly (Fig 1). [14] 

Fig 1. Medical applications of nanotechnology. The 

size and tailorability of nanoparticles may lea 

to their widespread use in a variety of medical 

applications. [14] 

Engineered nanomaterials are at the leading 

edge of the rapidly developing nanosciences and are 

founding an important class of new materials with 

specific physicochemical properties different from 

bulk materials with the same compositions. The 

potential for nanomaterials is rapidly expanding 

with novel applications constantly being explored in 

different areas. The unique size-dependent 

properties of nanomaterials make them very 

attractive for pharmaceutical applications. 

Investigations of physical, chemical and biological 

properties of engineered nanomaterials have yielded 

valuable information. Cytotoxic effects of certain 

engineered nanomaterials towards malignant cells 

form the basis for one aspect of nanomedicine. It is 

inferred that size, three dimensional shape, 

hydrophobicity and electronic configurations make 

them an appealing subject in medicinal chemistry. 

Their unique structure coupled with immense scope 

for derivatization forms a base for exciting 

developments in therapeutics. This review article 

addresses the fate of absorption, distribution, 

metabolism and excretion (ADME) of engineered 

nanoparticles in vitro and in vivo. It updates the 

distinctive methodology used for studying the 

biopharmaceutics of nanoparticles. This review 

addresses the future potential and safety concerns 

and genotoxicity of nanoparticle formulations in 

33


general. It particularly emphasizes the effects of 

nanoparticles on metabolic enzymes as well as the 

parenteral or inhalation administration routes of 

nanoparticle formulations. This paper illustrates the 

potential of nanomedicine by discussing 

biopharmaceutics of fullerene derivatives and their 

suitability for diagnostic and therapeutic purposes. 

Future direction is discussed as well. [15] 

With the rapid development of quantum dot 

(QD) technology, water-soluble QDs have the 

prospect of being used as a biological probe for 

specific diagnoses, but their biological behaviors in 

vivo are little known. Our recent in vivo studies 

concentrated on the bio-kinetics of QDs coated by 

hydroxyl group modified silica networks (the QDs 

are 21.3±2.0 nm in diameter and have maximal 

emission at 570 nm). Male ICR mice were 

intravenously given the water-soluble QDs with a 

single dose of 5 nmol/mouse. Inductively coupled 

plasma-mass spectrometry was used to measure the 

(111)Cd content to indicate the concentration of 

QDs in plasma, organs, and excretion samples 

collected at predetermined time intervals. 

Meanwhile, the distribution and aggregation state of 

QDs in tissues were also investigated by 

pathological examination and differential 

centrifugation. The plasma half-life and clearance 

of QDs were 19.8±3.2 h and 57.3±9.2 ml·h -1·kg -1 , 

respectively. The liver and kidney were the main 

target organs for QDs. The QDs metabolized in 

three paths depending on their distinct aggregated 

states in vivo. A fraction of free QDs, maintaining 

their original form, could be filtered by glomerular 

capillaries and excreted via urine as small 

molecules within five days. Most QDs bound to 

protein and aggregated into larger particles that 

were metabolized in the liver and excreted via feces 

in vivo. After five days, 8.6% of the injected dose of 

aggregated QDs still remained in hepatic tissue and 

it was difficult for this fraction to clear. [16] 

There is growing interest in developing 

tissue-specific multifunctional drug delivery 

systems with the ability to diagnose or treat several 

diseases. One class of such agents, composite 

nanodevices (CNDs), is multifunctional 

nanomaterials with several potential medical uses, 

including cancer imaging and therapy. Nanosized 

metal-dendrimer CNDs consist of poly(amidoamine) 

dendrimers (in various sizes, surface substituents, 

and net charges) and inorganic nanoparticles, 

properties of both of which can be individually 

modified and optimized. In this study we examine 

effects of size and surface charge on the behavior of 

Au-dendrimer CNDs in mouse tumor models. 

Quantitative biodistribution and excretion analyses 

including 5-nm and 22-nm positive surface, 5-nm 

and 11-nm negative surface, and a 5-nm neutral 

surface CNDs were carried out in the B16 mouse 

melanoma tumor model system. Results seen with 

the 22-nm CND in the B16 melanoma model were 

corroborated in a prostate cancer mouse tumor 

model system. Quantitative in vivo studies confirm 

the importance of charge and show for the first time 

the importance of size in affecting CND 

biodistribution and excretion. Interestingly, CNDs 

of different size and/or surface charge had high 

levels of uptake (“selective targeting”) to certain 

organs without specific targeting moieties placed on 

their surfaces. Researchers conclude that size and 

charge greatly affect biodistribution of CNDs. 

These findings have significance for the design of 

all particle-based nanodevices for medical uses. The 

observed organ selectivity may make these 

nanodevices exciting for several targeted medical 

applications. [17] 

Study on responses of Ferric oxide 

nanoparticles: 

Ferric oxide (Fe 2 O 3 ) nanoparticles are of 

considerable interest for application in 

nanotechnology related fields. However, as iron 

being a highly redox-active transition metal, the 

safety of iron nanomaterials need to be further 

studied. In this study, the size, dose and time 

dependent of Fe 2 O 3 nanoparticle on pulmonary and 

coagulation system have been studied after 

intratracheal instillation. The Fe 2 O 3 nanoparticles 

with mean diameters of 22 and 280 nm, respectively, 

were intratracheally instilled to male Sprague 

Dawley rats at low (0.8 mg·kg -1 ) and high (20 

mg/kg) doses. The toxic effects were monitored in 

the post-instilled 1, 7 and 30 days. Our results 

showed that the Fe 2 O 3 nanoparticle exposure could 

induce oxidative stress in lung. Alveolar 

macrophage (AM) over-loading of phagocytosed 

nanoparticle by high dose treatment had occurred, 

34


while the non-phagocytosed particles were found 

entering into alveolar epithelial in day 1 after 

exposure. Several inflammatory reactions including 

inflammatory and immune cells increase, clinical 

pathological changes: follicular hyperplasia, protein 

effusion, pulmonary capillary vessel hyperaemia 

and alveolar lipoproteinosis in lung were observed. 

The sustain burden of particles in AM and 

epithelium cells has caused lung emphysema and 

pro-sign of lung fibrosis. At the post-instilled day 

30, the typical coagulation parameters, prothrombin 

time (PT) and activated partial thromboplastin time 

(APTT) in blood of low dose 22 nm-Fe 2 O 3 treated 

rats were significantly longer than the controls. We 

concluded that both of the two-sized Fe 2 O 3 particle 

intratracheal exposure could induce lung injury. 

Comparing with the submicron-sized Fe 2 O 3 particle, 

the nano-sized Fe 2 O 3 particle may increase 

microvascular permeability and cell lysis in lung 

epitheliums and disturb blood coagulation 

parameters significantly. [18] 

Superparamagnetic iron oxide nanoparticles 

(SPIONs) are applied in stem cell labeling because 

of their high magnetic susceptibility as compared 

with ordinary paramagnetic species, their low 

toxicity, and their ease of magnetic manipulation. 

The present work is the study of CD133+ stem cell 

labeling by SPIONs coupled to a specific antibody 

(AC133), resulting in the antigenic labeling of the 

CD133+ stem cell, and a method was developed for 

the quantification of the SPION content per cell, 

necessary for molecular imaging optimization. Flow 

cytometry analysis established the efficiency of the 

selection process and helped determine that the 

CD133 cells selected by chromatographic affinity 

express the transmembrane glycoprotein CD133. 

The presence of antibodies coupled to the SPION, 

expressed in the cell membrane, was observed by 

transmission electron microscopy. Quantification of 

the SPION concentration in the marked cells using 

the ferromagnetic resonance technique resulted in a 

value of 1.70 × 10–13 mol iron (9.5 pg) or 7.0 × 10 6 

nanoparticles per cell (the measurement was carried 

out in a volume of 2 µL containing about 6.16 × 10 5 

pg iron, equivalent to 4.5 × 10 11 SPIONs). [19] 

Mechanisms of development of protein 

misfolding diseases 

Misfolding and self-assembly of proteins in 

nanoaggregates of different sizes and morphologies 

(nanoensembles, primarily nanofilaments and 

nanorings) is a complex phenomenon that can be 

facilitated, impeded, or prevented by interactions 

with various intracellular metabolites, intracellular 

nanomachines controlling protein folding, and 

interactions with other proteins. A fundamental 

understanding of molecular processes leading to 

misfolding and self-aggregation of proteins 

involved in various neurodegenerative diseases will 

provide important information to help identify 

appropriate therapeutic routes to control these 

processes. An elevated propensity of misfolded 

protein conformation in solution to aggregate with 

the formation of various morphologies impedes the 

use of traditional physiochemical approaches for 

studies of misfolded conformations of proteins. 

Kransnoslobodtsev et al tethered the protein 

molecules to surfaces to prevent aggregation and, 

with force spectroscopy using an atomic force 

microscopy, probed the interaction between protein 

molecules depending on their conformations. 

Research results show that formation of filamentous 

aggregates is facilitated at pH values corresponding 

to the maximum of rupture forces. They report here 

on development of a novel surface chemistry for 

anchoring of amyloid β (Aβ) peptides at their 

N-terminal moieties. The use of the site-specific 

immobilization procedure allowed us to measure the 

rupture of Aβ-Aβ contacts at the single-molecule 

level. The rupture of these contacts is accompanied 

by the extension of the peptide chain detected by a 

characteristic elastomechanical component of the 

force-distance curves. Potential applications of 

nanomechanical studies for understanding the 

mechanisms of development of protein misfolding 

diseases are discussed. [20] 

Research and Application of Carbon 

Nanotubes 

A good representative of this fast-moving field 

is the family of nanomaterials known as carbon 

nanotubes (CNTs). These all-carbon tubes are just a 

few nanometers in diameter, which makes them 

comparable in girth to DNA molecules, and come in 

either singlewalled varieties or multiwalled varieties 

with a nesting of carbon shells resembling the 

35


structure of a retractable antenna. CNTs are 

nanodevices with important potential applications in 

biomedicine such as drug and gene delivery. 

Recognition of functionalization of 

nanotubes 

Current advances in nanotechnology have led 

to the development of the new field of 

nanomedicine, which includes many applications of 

nanomaterials and nanodevices for diagnostic and 

therapeutic purposes. The same unique physical and 

chemical properties that make nanomaterials so 

attractive may be associated with their potentially 

calamitous effects on cells and tissues. The recent 

study on nanomedicine and nanotoxicology 

published by Kagan et al demonstrated that 

aspiration of single-walled CNTs elicited an 

unusual inflammatory response in the lungs of 

exposed mice with a very early switch from the 

acute inflammatory phase to fibrogenic events 

resulting in pulmonary deposition of collagen and 

elastin. This was accompanied by a characteristic 

change in the production and release of 

proinflammatory to anti-inflammatory profibrogenic 

cytokines, decline in pulmonary function, and 

enhanced susceptibility to infection. Chemically 

unmodified (nonfunctionalized) CNTs are not 

effectively recognized by macrophages. 

Functionalization of nanotubes results in their 

increased recognition by macrophages and is thus 

used for the delivery of nanoparticles to 

macrophages and other immune cells to improve the 

quality of diagnostic and imaging techniques as 

well as for enhancement of the therapeutic 

effectiveness of drugs. These observations on 

differences in recognition of nanoparticles by 

macrophages have important implications in the 

relationship between the potentially toxic health 

effects of nanomaterials and their applications in the 

field of nanomedicine. [21] Although membrane 

proteins consist of a substantial amount of the 

human genome and are the main drug targets, the 

study of cell membrane proteins in situ is 

complicated by the technical limitations. The recent 

development of atomic force microscopy (AFM) 

opens a new way to study the functions of cell 

membrane proteins in situ at the single-molecule 

level. A detailed procedure for investigation of 

angiotensin II type 1 receptor by AFM with 

functionalized tip is introduced in this article. Some 

prospective methods to improve the imaging 

resolution are also discussed. [22] 

CNTs are nanodevices with important potential 

applications in biomedicine such as drug and gene 

delivery. Brain diseases with no current therapy 

could be candidates for CNT-based therapies. Little 

is known about toxicity of CNTs and of their 

dispersion factors in the brain. Reaearchers show 

that multiwall CNTs (MWCNTs) coated with 

Pluronic F127 (PF127) surfactant can be injected in 

the mouse cerebral cortex without causing 

degeneration of the neurons surrounding the site of 

injection. They also show that, contrary to previous 

reports on lack of PF127 toxicity on cultured cell 

lines, concentrations of PF127 as low as 0.01% can 

induce apoptosis of mouse primary cortical neurons 

in vitro within 24 hours. However, the presence of 

MWCNTs can avoid PF127-induced apoptosis. 

These results suggest that PF127-coated MWCNTs 

do not induce apoptosis of cortical neurons. 

Moreover, the presence of MWCNTs can reduce 

PF127 toxicity. [23] 

Interactions of multiwalled carbon nanotubes 

(MWCNTs) with human epidermal keratinocytes 

(HEKs) were studied with respect to the effect of 

surfactant on dispersion of MWCNT aggregates and 

cytotoxicity. Our earlier studies had shown that the 

unmodified MWCNTs were localized within the 

cytoplasmic vacuoles of HEKs and elicited an 

inflammatory response. However, MWCNTs in 

solution tend to aggregate and, therefore, cells are 

exposed to large MWCNT aggregates. The purpose 

of this study was to find a surfactant that prevents 

the formation of large aggregates of MWCNTs 

without being toxic to the HEKs. HEKs were 

exposed to serial dilutions (10% to 0.1%) of L61, 

L92, and F127 Pluronic and 20 or 60 Tween for 24 

hours. HEK viability, proportional to surfactant 

concentration, ranged from 27.1% to 98.5% with 

Pluronic F127; viability with the other surfactants 

was less than 10%. Surfactants dispersed and 

reduced MWCNT aggregation in medium. 

MWCNTs at 0.4 mg·ml -1 in 5% or 1% Pluronic 

F127 were incubated with HEKs and assayed for 

interleukin 8 (IL-8). MWCNTs were cytotoxic to 

HEKs independent of surfactant exposure. In 

36


contrast, MWCNT-induced IL-8 release was 

reduced when exposed to 1% or 5% Pluronic F127 

(P


Pegylated liposomal doxorubicin is a 

formulation of doxorubicin in which the molecule 

itself is packaged in a liposome made of various 

lipids with an outer coating of polyethylene glycol. 

Liposomal technology is being used in increasing 

amounts in the therapy of a variety of cancers, 

including ovarian cancers. A reviews written by 

Green et al on the mechanistic actions of this 

formulation, the Phase II and Phase III data that 

helped define the role of pegylated liposomal 

doxorubicin in recurrent ovarian cancer, as well as a 

discussion of some of the side-effects and their 

management. [27] Pegylated liposomal doxorubicin 

is one of a new class of drug formulations. The 

doxorubicin molecules in pegylated liposomal 

doxorubicin are encapsulated in a bilayer sphere of 

lipids. This vesicle is then surrounded by a dense 

layer of polyethylene glycol (PEG), hence the name 

pegylated liposomal doxorubicin. The size of the 

liposomes, approximately 100 nm, prevents them 

from entering tissues with tight capillary junctions, 

such as the heart and gastrointestinal tract, as well 

as selectively depositing the liposome into the 

tumor. In contrast to normal vessels, the vessels of 

the tumor are tortuous, dilated, have 

morphologically abnormal endothelial cells, and are 

leaky due to large spaces between pericytes. The 

study on mechanism of action exhibited that these 

physical characteristics allow more extravasation of 

the vesicles into the tumor, thus encouraging more 

deposition of the chemotherapy agent into the tumor. 

The PEG coating on the liposome creates a 

hydrophilic layer around the liposome that buffers 

the liposome wall from the surrounding milieu. This 

decreases proteins from binding to the lipid bilayer. 

These proteins act as opsonins, attracting 

foreign particles that in turn activate the 

mononuclear phagocytic cells. This leads to break 

down of the liposome and release of the drug. 

Therefore, the PEG coating on the liposome 

increases the longevity of the liposome. Pegylated 

liposomal doxorubicin was cleared via the 

lymphatic system and returned to the circulation. In 

tumor tissue, however, there are no lymphatics. 

Therefore, when the liposome is deposited it 

remains for a longer time. This allows a higher dose 

of doxorubicin to be released in the tumor, and a 

lower dose in normal tissue. Collectively, there is 

preferential uptake and decreased clearance of the 

drug delivery system, increasing the exposure of the 

tumor to the drug. When the liposome does leave 

the intravascular compartment, in normal tissues it 

is Phase II single-agent studies In a subsequent 

Phase II study, evaluated 79 better-defined patients 

all of whom were platinum and taxane refractory. 

Eighty-five percent of the patients had received 

more than 2 prior chemotherapy regimens. These 

“doubly refractory” patients were treated with 50 

mg/m 2 of pegylated liposomal doxorubicin every 4 

weeks. Fourteen partial responses and 1 complete 

response were reported for an overall response rate 

of 16.9%. The median time to response was 15 

weeks. The median progression-free survival for all 

patients treated in this study was 19.3 weeks (range 

0.7–86 weeks). In addition, 36 patients (57%) were 

classified as having stable disease, and achieved a 

median progression-free survival of 21.9 weeks. 

This was one of the first studies to show that disease 

stabilization in recurrent ovarian cancer is of 

clinical benefit. All patients reported at least 1 

adverse event, but the majority were grade 1 or 2. 

Asthenia and palmar-plantar erythrodysesthesia 

(PPE) were seen in 41.6%. Only 1 patient 

experienced any cardiac complications, and there 

were no treatment-related deaths. This study 

demonstrated that pegylated liposomal doxorubicin 

was useful in this drug-resistant setting, and 

associated with no life-threatening toxicities. 

In China, Liang W et al research results on 

doxorubicin- containing PEG-PE micelles are an 

important contribution to nanomedicine 

development (which is called “nanoparticles carry 

chemotherapy drug deeper into solid tumors”). 

Editorial members, Dreher MR and Chilkoti A in J 

Natl Cancer Inst get a high evaluation for their 

research. Solid tumors account for more than 85% 

of cancer mortality. To obtain nutrients for growth 

and to metastasize, cancer cells in solid tumors must 

grow around existing vessels or stimulate formation 

of new blood vessels. These new vessels are 

abnormal in structure and characterized by leakage, 

tortuousness, dilation, and a haphazard pattern of 

interconnection. Tumor structure and blood flow 

hinder the treatment of solid tumors. To reach 

cancer cells in optimal quantity, a therapeutic agent 

must pass through an imperfect blood vasculature to 

38


the tumor, cross vessel walls into the interstitium 

and penetrate multiple layers of solid tumor cells. 

Recent studies have demonstrated that poor 

penetration and limited distribution of doxorubicin 

in solid tumors are the main causes of its 

inadequacy as a chemotherapeutic agent. 

Encapsulation of doxorubicin into PEG-PE micelles 

increased its accumulation and penetration in 

tumors in terms of both the percentage of cells that 

were reached by the drug and the intracellular levels 

that were attained. This increased accumulation and 

penetration can be attributed to the efficient 

internalization of the drug-containing micelles by 

the endocytotic cell uptake mechanism and 

enhanced permeability and retention of tumors with 

leaky vasculature. High intracellular retention is 

especially important because doxorubicin must be 

internalized to be effective in tumor therapy. The 

doxorubicin- containing PEG-PE micelles had 

greatly increased antitumor activity in both 

subcutaneous and lung metastatic LLC tumor 

models compared with free doxorubicin. However, 

mice treated micelle- encapsulated doxorubicin 

showed fewer signs of toxicity than those treated 

with free doxorubicin. This drug packaging 

technology may provide a new strategy for design 

of cancer therapies. [29,30] At our laboratory, studied 

nanoparticle of doxorubicin eliminate the 

accumulation in tissues of tumor-bearing mice. 

Compared with general doxorubicin preparation, 

which is a marketed product, nanoparticle micelle 

of doxorubicin has the similar pharmacokinetics in 

the tissue, and the similar concentrations in the 

tumor tissue. Howerever, the accumulation of 

doxorubicin in the heart, spleen, kidney, lung, 

tumor, muscle and skin decreased significantly after 

three intravenous injections, showing that the 

nano-micelle can accumulatew the elimilation of 

doxorubicin in most tissues. It is deduced that the 

study was effects of doxorubicin after clinical use 

may be reduced significantly. [31] 

Pegylated liposomal doxorubicin is effective 

and well tolerated in relapsed ovarian cancer. When 

compared with topotecan in a phase III randomized 

trial, pegylated liposomal doxorubicin showed 

several advantages: improved quality of life, fewer 

severe adverse events, fewer dose modifications, 

less hematologic support, and lower total cost per 

patient. In platinum-sensitive patients, pegylated 

liposomal doxorubicin also produced a survival 

advantage. Results from prospective and 

retrospective studies further demonstrate the 

improved cardiac safety of pegylated liposomal 

doxorubicin compared to conventional 

anthracyclines. Based on survival and toxicity 

advantages and a once-monthly administration 

schedule, pegylated liposomal doxorubicin is the 

first-choice nonplatinum agent for relapsed ovarian 

cancer. Pegylated liposomal doxorubicin may also 

have clinical application in combination regimens 

for platinum-sensitive ovarian cancer, as 

consolidation/maintenance therapy for ovarian 

cancer, as a component of first-line therapy for 

ovarian cancer, and in the treatment of other 

gynecologic malignancies. Future clinical trials will 

further define and maximize the role of pegylated 

liposomal doxorubicin in the treatment of ovarian 

cancer and other gynecologic malignancies. [32] 

Doxorubicin nanoparticles 

A novel hyaluronic acid-poly(ethylene 

glycol)-poly(lactide-co-glycolide) (HA-PEG-PLGA) 

copolymer was synthesized and characterized by 

infrared and nuclear magnetic resonance 

spectroscopy. The nanoparticles of doxorubicin 

(DOX)-loaded HA-PEG-PLGA were prepared and 

compared with monomethoxy (polyethylene glycol) 

(MPEG)-PLGA nanoparticles. Nanoparticles were 

prepared using drug-to-polymer ratios of 1:1 to 1:3. 

Drug-to-polymer ratio of 1:1 is considered the 

optimum formulation on the basis of low particle 

size and high entrapment efficiency. The optimized 

nanoparticles were characterized for morphology, 

particle size measurements, differential scanning 

calorimetry, x-ray diffractometer measurement, 

drug content, hemolytic toxicity, subacute toxicity, 

and in vitro DOX release. The in vitro DOX release 

study was performed at pH 7.4 using a dialysis 

membrane. HA-PEG-PLGA nanoparticles were 

able to sustain the release for up to 15 days. The 

tissue distribution studies were performed with 

DOX-loaded HA-PEG-PLGA and MPEG-PLGA 

nanoparticles after intravenous (IV) injection in 

Ehrlich ascites tumor–bearing mice. The tissue 

distribution studies showed a higher concentration 

of DOX in the tumor as compared with 

39


MPEG-PLGA nanoparticles. The in vivo tumor 

inhibition study was also performed after IV 

injection of DOX-loaded HA-PEG-PLGA 

nanoparticles up to 15 days. DOX-loaded 

HA-PEG-PLGA nanoparticles were able to deliver 

a higher amount of DOX as compared with 

MPEG-PLGA nanoparticles. The DOX-loaded 

HA-PEG-PLGA nanoparticles reduced tumor 

volume significantly as compared with 

MPEG-PLGA nanoparticles. [33] Chitosan, PCEP 

(poly{[(cholesteryl oxocarbonylamido ethyl) methyl 

bis(ethylene) ammonium iodide] ethyl phosphate}), 

and magnetic nanoparticles (MNPs) were evaluated 

for the safe delivery of genes in the eye. Prow et al 

studied ocular nanoparticle toxicity and transfection 

of the retina and retinal pigment epithelium. Rabbits 

were injected with nanoparticles either intravitreally 

(IV) or subretinally (SR) and sacrificed 7 days later. 

Eyes were grossly evaluated for retinal pigment 

epithelium abnormalities, retinal degeneration, and 

inflammation. All eyes were cryopreserved and 

sectioned for analysis of toxicity and expression of 

either enhanced green or red fluorescent proteins. 

All of the nanoparticles were able to transfect cells 

in vitro and in vivo. IV chitosan showed 

inflammation in 12/13 eyes, whereas IV PCEP and 

IV MNPs were not inflammatory and did not induce 

retinal pathology. SR PCEP was nontoxic in the 

majority of cases but yielded poor transfection, 

whereas SR MNPs were nontoxic and yielded good 

transfection. Therefore, researchers concluded that 

the best nanoparticle evaluated in vivo was the least 

toxic nanoparticle tested, the MNP. [34] 

Liposomes targeted by fusion phage 

proteins 

Targeting of nanocarriers has long been sought 

after to improve the therapeutic indices of 

anticancer drugs. Jayanna et al provide the proof 

of principle for a novel approach of nanocarrier 

targeting through their fusion with target-specific 

phage coat proteins. The source of the targeted 

phage coat proteins are landscape phage 

libraries—collections of recombinant filamentous 

phages with foreign random peptides fused to all 

4000 copies of the major coat protein. Prashanth et 

al exploit in our approach the intrinsic 

physicochemical properties of the phage major coat 

protein as a typical membrane protein. Landscape 

phage peptides specific for specific tumors can be 

obtained by affinity selection, and purified fusion 

coat proteins can be assimilated into liposomes to 

obtain specific drug-loaded nanocarriers. As a 

paradigm for inceptive experiments, a 

streptavidin-specific phage peptide selected from a 

landscape phage library was incorporated into 

100-nm liposomes. Targeting of liposomes was 

proved by their specific binding to streptavidincoated 

beads. [35] 

Drug Loading and Release From 

Biodegradable Microcapsules 

Microcapsules made of biopolymers are of 

both scientific and technological interest and have 

many potential applications in medicine, including 

their use as controlled drug delivery devices. The 

present study makes use of the electrostatic 

interaction between polycations and polyanions to 

form a multilayered microcapsule shell and also to 

control the loading and release of charged drug 

molecules inside the microcapsule. Micron-sized 

calcium carbonate (CaCO 3 ) particles were 

synthesized and integrated with chondroitin sulfate 

(CS) through a reaction between sodium carbonate 

and calcium nitrate tetrahydrate solutions suspended 

with CS macromolecules. Oppositely charged 

biopolymers were alternately deposited onto the 

synthesized particles using electrostatic 

layer-by-layer self-assembly, and glutaraldehyde 

was introduced to cross-link the multilayered shell 

structure. Microcapsules integrated with CS inside 

the multilayered shells were obtained after 

decomposition of the CaCO 3 templates. The 

integration of a matrix (i.e., CS) permitted the 

subsequent selective control of drug loading and 

release. The CS-integrated microcapsules were 

loaded with a model drug, bovine serum albumin 

labeled with fluorescein isothiocyanate (FITC-BSA), 

and it was shown that pH was an effective means of 

controlling the loading and release of FITC-BSA. 

Such CS-integrated microcapsules may be used for 

controlled localized drug delivery as biodegradable 

devices, which have advantages in reducing 

systemic side effects and increasing drug efficacy. 

[36] 

40


Amphotericin B–intercalated liposomes 

Nanotechnology in drug delivery is a rapidly 

expanding field. Nanosized liposomal preparations 

are already in use for efficient drug delivery with 

better therapeutic indices. Existing methods of 

liposome preparation are limited by problems of 

scale-up, difficulty in controlling size, and 

intercalation efficiency. Here researchers prepare 

amphotericin B–intercalated liposomes by a novel 

process where amphotericin B and purified 

phosphatidyl choline are solubilized in suitable 

solvent and precipitated in supercritical fluid carbon 

dioxide (known as a gas antisolvent technique), to 

obtain microsized particles that are subsequently 

introduced into a buffer solution. The morphology 

of liposomes was characterized through a 

phase-contrast microscope, and the particle size 

distribution studied by laser technique showed 

nanosize with a narrow range of size distribution 

(between 0.5 and 15 µm) and a higher intercalation 

efficiency. In vitro studies conducted using 

Aspergillus fumigatus (MTCC 870) strain proved to 

be efficient in the retardation of the growth of the 

organism. [37] 

Diclofenac-loaded biopolymeric nanosuspensions 

Polymeric nanoparticle suspensions (NS) were 

prepared from poly(lactide-co- glycolide) and 

poly(lactide-co-glycolide-leucine) {poly[Lac (Glc- 

Leu)]} biodegradable polymers and loaded with 

diclofenac sodium (DS), with the aim of improving 

the ocular availability of the drug. NS were 

prepared by emulsion and solvent evaporation 

technique and characterized on the basis of 

physicochemical properties, stability, and drug 

release features. The nanoparticle system showed an 

interesting size distribution suitable for ophthalmic 

application. Stability tests (as long as 6 months' 

storage at 5°C or at 25°C/60% relative humidity) or 

freeze-drying were carried out to optimize a suitable 

pharmaceutical preparation. In vitro release tests 

showed a extended-release profile of DS from the 

nanoparticles. To verify the absence of irritation 

toward the ocular structures, blank NS were applied 

to rabbit eye and a modified Draize test performed. 

Polymer nanoparticles seemed to be devoid of any 

irritant effect on cornea, iris, and conjunctiva for as 

long as 24 hours after application, thus apparently a 

suitable inert carrier for ophthalmic drug delivery. 

[38] 

Paclitaxel nanoparticles 

Karmali et al have used tumor-homing 

peptides to target abraxane, a clinically approved 

paclitaxel-albumin nanoparticle, to tumors in mice. 

The targeting was accomplished with two peptides, 

CREKA and LyP-1 (CGNKRTRGC). Fluorescein 

(FAM)-labeled CREKA-abraxane, when injected 

intravenously into mice bearing MDA-MB-435 

human cancer xenografts, accumulated in tumor 

blood vessels, forming aggregates that contained 

red blood cells and fibrin. FAM-LyP-1-abraxane 

co-localized with extravascular islands expressing 

its receptor, p32. Self-assembled mixed micelles 

carrying the homing peptide and the label on 

different subunits accumulated in the same areas of 

tumors as LyP-1-abraxane, showing that Lyp-1 can 

deliver intact nanoparticles into extravascular sites. 

Untargeted, FAM-abraxane was detected in the 

form of a faint meshwork in tumor interstitium. 

LyP-1-abraxane produced a statistically highly 

significant inhibition of tumor growth compared 

with untargeted abraxane. These results show that 

nanoparticles can be effectively targeted into 

extravascular tumor tissue and that targeting can 

enhance the activity of a therapeutic nanoparticle. 

[39] 

Nano–atropine sulfate dry powder inhaler 

The work of Raisuddin Ali et al was to develop, 

characterize, and carry out a clinical trial with 

nano–atropine sulfate (nano-AS) dry powder inhaler 

(DPI), because this route may offer several 

advantages over the conventional intramuscular 

route as an emergency treatment, including ease of 

administration and more rapid bioavailability. 

Different batches of nanoparticles of AS were 

produced using variants of nanoprecipitation 

method. The influence of the process parameters, 

such as the types and quantity of solvent and 

nonsolvent, the stirring speed, the solventto-nonsolvent 

volume ratio, and the drug 

concentration, was investigated. The methodology 

resulted in optimally sized particles. Bulk properties 

of the particles made by the chosen methodology 

41


were evaluated. A clinical trial was conducted in six 

healthy individuals using a single DPI capsule 

containing 6 mg nano-AS DPI in lactose. Early 

blood bioavailability and atropinization pattern 

confirmed its value as a potential replacement to 

parenteral atropine in field conditions. The 

formulation seems to have the advantage of early 

therapeutic drug concentration in blood due to 

absorption through the lungs as well as sustained 

action due to absorption from the gut of the 

remaining portion of the drug. [40] 

Drug delivery of siRNA therapeutics 

A review by Daniela Reischl and 

Andreas Zimmer focuses on different pathways for 

siRNA delivery and summarizes recent progress 

made in the use of vector-based siRNA technology. 

Gene therapy is a promising tool for the treatment 

of human diseases that cannot be cured by rational 

therapies. The major limitation for the use of small 

interfering RNA (siRNA), both in vitro and in vivo, 

is the inability of naked siRNA to passively diffuse 

through cellular membranes due to the strong 

anionic charge of the phosphate backbone and 

consequent electrostatic repulsion from the anionic 

cell membrane surface. Therefore, the primary 

success of siRNA applications depends on suitable 

vectors to deliver therapeutic genes. Cellular 

entrance is further limited by the size of the applied 

siRNA molecule. Multiple delivery pathways, both 

viral and nonviral, have been developed to bypass 

these problems and have been successfully used to 

gain access to the intracellular environment in vitro 

and in vivo, and to induce RNA interference 

(RNAi). [41] 

Translational implications for diagnosis and 

therapy: Esophageal adenocarcinoma arises in the 

backdrop of Barrett metaplasia-dysplasia sequence, 

with the vast majority of patients presenting with 

late-stage malignancy. Mesothelin, a glycophosphatidylinositol-anchored 

protein, is aberrantly 

overexpressed on the surface of many solid cancers. 

Mesothelin expression was assessed in esophageal 

tissue microarrays encompassing the entire 

histological spectrum of Barrett-associated 

dysplasia and adenocarcinoma. Mesothelin 

expression was observed in 24/84 (29%) of invasive 

adenocarcinomas and in 5/34 (15%) lymph node 

metastases. In contrast, normal squamous and 

cardiac mucosa, as well as noninvasive Barrett 

lesions, failed to label with mesothelin. Mesothelin 

was expressed in the esophageal adenocarcinoma 

cell line JH-EsoAd1 but not in primary human 

esophageal epithelial cells. Anti-mesothelin 

antibody–conjugated CdSe/CDS/ ZnS quantum rods 

were synthesized, and confocal bioimaging 

confirmed robust binding to JH-EsoAd1 cells. 

Anti-mesothelin antibody– conjugated nanoparticles 

can be useful for the diagnosis and therapy 

of mesothelin-overexpressing esophageal adenocarcinomas. 

[42] 

Research and development of nanomedicines 

in the future 

Nanotechnology will alter our relationship with 

molecules and matter profoundly. Research on 

productive nanosystems will eventually develop 

programmable, molecular-scale systems that make 

other useful nano-structured materials and devices. 

These systems will enable a new manufacturing base 

that can produce both small and large objects 

precisely and inexpensively. Nano-risk research is 

conducted by agencies that oversee health and 

environmental regulations. Nanotechnology, dealing 

with functional structures and materials smaller than 

100nm, is emerging as a truly interdisciplinary research 

area spanning several traditional scientific disciplines. In 

keeping with the growing trend, there is a strong need for a 

platform to share original research related to applications of 

nanotechnology in biomedical fields. At the hearing, 

leaders of the Nanotechnology Environmental and 

Health Implications working group, an interagency 

panel that coordinates federal funding on health and 

environmental risks of nanotechnology, released a 

long-overdue report outlining research needed to 

buttress regulation of products in the field. There is 

far less agreement on how that money should be 

spent and coordinated. Research on Nanotech 

environmental health and safety in government 

agencies, academic institutions, and industry is being 

performed in an ad hoc fashion according to 

individual priorities. Yet the vast majority of 

nanotoxicology studies focus on those materials, 

while ignoring broad classes of other materials 

42


already on the market. [43] 

The nanomedicine research is a goal and needs a 

long-term plan, which is to quantitatively 

characterize the molecular-scale components, or 

nanomachinery, of living cells and to precisely 

control and manipulate these molecular and 

supramolecular assemblies in living cells to improve 

human health. Nanomedicine will exploit and build 

upon other research findings in nanotechnology and 

apply it to the study of molecular systems in living 

cells that contain a multitude of nanoscale structures, 

such as membrane transporters, processes such as 

self-assembly of protein–nucleic acid complexes, and 

nanomachines such as molecular motors. The 

benefits of nanomedicine include dramatically 

expanded knowledge of the human genome, a greater 

understanding of the pathophysiology of specific 

diseases at the molecular scale, more specific 

treatment of diseases, and the ability to understand 

the dynamic behavior of dysfunctional cellular 

machinery in the context of the total cell 

machinery. [44] Robert A and Freitas Jr given an 

overview of this rapidly expanding and exciting 

nanomedicine field. Over the next 5 to 10 years, 

nanomedicine will address many important medical 

problems by using nanoscale-structured materials and 

simple nanodevices that can be manufactured today. 

Many approaches to nanomedicine being pursued 

today are already close enough to fruition that it is 

fair to say that their successful development is almost 

inevitable, and their subsequent incorporation into 

valuable medical diagnostics or clinical therapeutics 

is highly likely and may occur very soon. [45] 

The science of nanomedicine exploits and builds 

upon novel research findings in nanotechnology, 

biology, and medicine; it unifies the efforts of 

scientists, engineers, and physicians determined to 

apply their latest research results to translational and 

clinical medicine by developing novel approaches 

and a better understanding of solutions to 

health-relatedissues, ultimately improving the quality 

of life. The last few years have seen unprecedented 

advances in the field of biology. The decoding of the 

human genome coupled with improving gene 

transfection technologies offer great opportunities for 

treating illnesses. In analysis and diagnosis, 

lab-on-a-chip methods have surpassed earlier ex-vivo 

and in-vivo detection methods while also aiding 

toxicology efforts. In medicine, improvements in 

targeted drug delivery, imaging, and therapy have led 

to such successful interventions in cancer therapies. 

[46] 

Although there are only a few FDA-approved 

nanopharmaceuticals on the market today, these 

formulations are already impacting medicine and 

promise to alter healthcare. Based on their ability to 

reduce time-to-market, extend the economic life of 

proprietary drugs and create additional revenue 

streams, nanopharmaceuticals should greatly impact 

medical practice and healthcare. However, if this is to 

happen effectively, there are a few key biological 

requirements for nanopharmaceuticals to fulfill: (1) 

they must exhibit stealth qualities to evade 

macrophage attack and the immune response; (2) be 

nontoxic and traceable; (3) display effective 

pharmacokinetic properties; (4) be biodegradable 

following systemic administration through any route 

(but the polymer must protect the embedded active); 

and (5) they must be selective to be effective in 

targeting specific tissue sites. Srikumaran Melethil 

(Chair and Professor of Pharmaceutical Sciences at 

the University of Findlay, Findlay, OH) discussed the 

metabolic fate of nanopharmaceuticals upon delivery 

to the human body, and presented pharmacokinetic 

data relating to numerous nanoparticulate drugs and 

highlighted the critical role of the FDA in 

nanomedicine. According to him, further knowledge 

of how the human body transports, distributes and 

clears nanoparticles via the vascular and lymphatic 

systems (i.e.,biodistribution of nanoparticles) is also 

needed to get a handle on metabolic and toxicity 

issues. Nanomedicine will eventually become an 

integral part of mainstream medicine and a standard 

in the drug industry. For example, the market impact 

of nanopharmaceuticals on the pharmaceutical and 

biotech industries is already being felt. However, for 

nanomedicine to be a viable commercial entity, 

desperately needed reforms to overhaul the PTO 

along with clearer regulatory guidelines and safety 

standards from federal agencies such as the FDA will 

be needed. 

Ethical question of nanomedicine is an important 

issue. Ginger Gruters (The President's Council on 

Bioethics, Washington, DC) presented on ethical 

considerations that are likely to play a significant role 

in nanomedicine, and stated that, as with other 

43


biomedical advances coming before it, nanomedicine 

will face significant ethical challenges as it moves 

from proof-of-concept to the clinic. Along the 

way,ethical questions regarding social justice, 

privacy and confidentiality, long-term risks and 

benefits, and human enhancement are certain to 

arise. [47] 

Appurtenances: 

Introduction to Journals on Nano 

Nature Nanotechnology 

Nature Nanotechnology is a multidisciplinary 

journal that publishes papers of the highest quality and 

significance in all areas of nanoscience and 

nanotechnology. The journal covers research into the 

design, characterization and production of structures, 

devices and systems that involve the manipulation and 

control of materials and phenomena at atomic, molecular 

and macromolecular scales. Both bottom-up and 

top-down approaches - and combinations of the two - are 

covered. Nature Nanotechnology also encourages the 

exchange of ideas between chemists, physicists, material 

scientists, biomedical researchers, engineers and other 

researchers who are active at the frontiers of this diverse 

and multidisciplinary field. Coverage extends from basic 

research in physics, chemistry and biology, including 

computational work and simulations, through to the 

development of new devices and technologies for 

applications in a wide range of industrial sectors 

(including information technology, medicine, 

manufacturing, high-performance materials, and energy 

and environmental technologies). Organic, inorganic and 

hybrid materials are all covered. Research areas covered 

in the journal include: Carbon nanotubes and fullerenes, 

Computational nanotechnology, Electronic properties and 

devices, Environmental, health and safety 

issues,Molecular machines and motors, Molecular 

self-assembly, Nanobiotechnology, Nanofluidics, 

Nanomagnetism and spintronics, Nanomaterials, 

Nanomedicine, 

Nanometrology and instrumentation, 

Nanoparticles, Nanosensors and other devices, NEMS, 

Organic–inorganic nanostructures, Photonic structures 

and devices, Quantum information, Structural properties, 

Surface patterning and imaging and Synthesis and 

processing. 

Nature Nanotechnology also publishes review 

articles, news and views, research highlights about 

important papers published in other journals, 

commentaries, book reviews, correspondence, and articles 

about the broader nanotechnology picture — funding, 

commercialization, ethical and social issues, and so on. In 

this way, the journal aims to be the voice of the 

worldwide nanoscience and nanotechnology community. 

Nature Nanotechnology offers readers and authors high 

visibility, access to a broad readership, high standards of 

copy editing and production, rigorous peer review, rapid 

publication, and independence from academic societies 

and other vested interests. 

Journal of Nanoscience and Nanotechnology 

Journal of Nanoscience and Nanotechnology (JNN) 

is an international and multidisciplinary peer-reviewed 

journal with a wide-ranging coverage, consolidating 

research activities in all areas of nanoscience and 

nanotechnology into a single and unique reference source. 

JNN is the first cross-disciplinary journal to publish 

original full research articles, rapid communications of 

important new scientific and technological findings, 

timely state-of-the-art reviews with author's photo and 

short biography, and current research news encompassing 

the fundamental and applied research in all disciplines of 

science, engineering and medicine. Topics covered in the 

journal include: Synthesis of Nanostructured and 

Nanoscale Materials, Nanofabrication and Processing of 

Nanoscale Materials and Device; Atomic and Nanoscale 

Characterization of Functional Materials and 

Bio-assemblies; Nanoprobes, Properties of Nanoscale 

Materials, Nanocatalysis; Nanocomposites, Nanoparticles, 

Nanocrystalline Materials, and Nanoclusters; 

Superlattices, Quantum Dots, Quantum Wires, Quantum 

Wells, Nanoscale Thin Films ; Fullerenes, Nanotubes, 

Nanorods, Molecular Wires, Molecular Nanotechnology; 

Supramolecules, Dendrimers, Self-Assemblies, 

Low-dimension Structures; Nanophysics, Nanoelectronics, 

Nano-Optics, Nanomagnetism and Nanodevices; Atomic 

Manipulation, Computational Nanotechnology, Molecular 

Nanoscience; Nanochips, Nanosensors and 

Nano-integration, Nanofluidics, Nanomachining; 

Structure Analysis at Atomic, Molecular, and Nanometer 

range; Nanorobotics, Nanotribology, and Novel 

Applications of Nanostructured Materials and 

Nanobiotechnology, Biochemical Assemblies, BioMEMS, 

Biomimetic Materials Nanoscale Genomics, DNA 

Sequencing, Nanomedicines, Drug Delivery, Biomedical 

Nanotechnolog. 

44


Nano 

NANO is an international peer-reviewed journal for 

nanoscience and nanotechnology that presents forefront 

fundamental research and new emerging topics. It 

features timely scientific reports of new results and 

technical breakthroughs and also contains interesting 

review articles about recent hot issues. NANO provides an 

ideal forum for presenting original reports of theoretical 

and experimental nanoscience and nanotechnology 

research. Research areas of interest include: 

nanomaterials including nano-related biomaterials, new 

phenomena and newly developed characterization tools, 

fabrication methods including by self-assembly, device 

applications, and numerical simulation, modeling, and 

theory. 

Nano Letters 

Nano Letters reports on fundamental research in all 

branches of the theory and practice of nanoscience and 

nanotechnology, providing rapid disclosure of the key 

elements of a study, publishing preliminary, experimental, 

and theoretical results on the physical, chemical, and 

biological phenomena, along with processes and 

applications of structures within the nanoscale range. 

Among the areas of interest the journal covers are: 

Synthesis and processing of organic, inorganic, and 

hybrid nanosized materials by physical, chemical, and 

biological methods; Modeling and simulation of synthetic, 

assembly, and interaction processes; Characterization of 

size-dependant properties; and Realization and 

application of novel nanostructures and nanodevices The 

Nano Letters manuscript submission process is fully 

electronic, to ensure the rapid publication of results. 

Manuscripts should be submitted via our secure Web site. 

Manuscripts submitted by hardcopy mail or by e-mail will 

not be processed. Introduction Nano Letters invites 

original reports of fundamental research in all branches of 

the theory and practice of nanoscience and 

nanotechnology. 

Journal of Nano Research 

Journal of Nano Research (J Nano R) is a 

multidisciplinary peer-reviewed journal, which publishes 

high quality work on ALL aspects of nanoscience and 

nanotechnology. Currently, it stands alone in serving the 

global “nano” community in providing up-to-date 

information on all developments and progresses being 

made in nanoscience and nanotechnology and the future 

predictions for this extraordinary technology. 

Journal of Nanotechnology Online 

The Online Journal of Nanotechnology is based on a 

free access publishing model, coupled with what is 

believed to be a unique development in the field of 

scientific publishing – the distribution of journal revenue 

between the authors, peer reviewers and site operators 

(OARS). The revenue received from the journal related 

advertising and sponsorship will be distributed according 

to the following general criteria: Authors receive a 

revenue share of 50% of the related revenue their 

contributions attract. Peer reviewers receive a revenue 

share of 20%. The site administrators receive a revenue 

share of 30%. This revenue share will apply throughout 

the on-line published life of the individual article or paper. 

The Online Journal of Nanotechnology papers will 

benefit from being hosted on the AZoNano.com website 

and database platform as they will take advantage of the 

existing AZoNano.com search tools. These search tools 

make it very easy for site visitors to locate nanotech 

information which directly relates to their research areas, 

applications and industrial sectors. 

Journal of Nano Education 

The Journal of Nano Education (JNE) is a 

peer-reviewed international journal that aims to provide 

the most complete and reliable source of information on 

current developments in nanoscale science, technology, 

engineering, and medical education. JNE publishes a 

comprehensive range of articles including topics in the 

following areas: Nanoscale science, technology, 

engineering, and medical education at the K-12, 

undergraduate and graduate levels (formal and informal, 

including public outreach and dissemination activities); 

K-12 science teacher education and professional 

development; Scientific and technological literacy/public 

understanding of nanoscale science, technology, 

engineering, and medicine; Curriculum development and 

assessment; Social and ethical issues associated with 

nanoscale science, technology, engineering, and medical 

research; Workforce preparation (professional and 

vocational); National and state science standards and their 

relationships to the goals of nanoeducation initiatives 

worldwide; Current nanoscale science, technology, 

engineering, and medical education research; Other 

pertinent areas of interest to nanoscale science, 

45


technology, engineering, and medical researchers & 

educators. JNE also will serve as a forum for commentary 

and debate on related issues. 

Journal of Computational and Theoretical 

Nanoscience 

Journal of Computational and Theoretical 

Nanoscience is an international peer-reviewed journal 

with a wide-ranging coverage, consolidates research 

activities in all aspects of computational and theoretical 

nanoscience into a single reference source. This journal 

offers scientists and engineers peer-reviewed research 

papers in all aspects of computational and theoretical 

nanoscience and nanotechnology in chemistry, physics, 

materials science, engineering and biology to publish 

original full papers and timely state-of-the-art reviews 

and short communications encompassing the fundamental 

and applied research. Topics include: Assemblers, basic 

physics, biological systems, biochemical systems, bionics, 

biophysics, CAD, carbon systems, cellular mechanisms, 

chaotic systems, circuits, clusters, cluster systems, 

complex aggregates, computer codes, crystal growth, data 

analysis, defined chain length molecules, devices, 

diffusion processes, DNA, drug design, dynamics, 

electronics, electronic properties, enzyme reactivity and 

reactions, equation of state, friction, computational 

genomics, gene technology, genetics, holistic views, 

information theory, interactions, ion channelling, kinetics, 

macromolecules, molecular interactions, large scale 

simulations, liquids, liquid crystals, luminescence, 

magnetic structures, manufacturing, many-particle 

systems, metallurgy, materials, material properties, 

mechanical models, metals, mathematical methods, 

molecule design, molecular dynamics, molecular 

mechanics, Monte Carlo simulations, multi-scale methods, 

nanomachines, nano-optics, nanorobotics, 

nanotechnology and ethics, noble gases, nonlinear optics, 

numerical algorithm, numerical procedures, oligomers, 

optoelectronics, phase transitions, phenomenological 

theory, philosophical implications and positions, photonic 

crystals, polymers, potential development, protein folding, 

quantum chemistry, quantum computers, quantum dots, 

quantum electronics and optics, quantum technology, 

replicators, RNA, semiconductors, superconductors, solid 

state physics, statistical physics, structural chemistry, 

structures, structures on surfaces, surfaces, technological 

applications, theoretical biosciences, theoretical physics, 

thermodynamics thought experiments, wear, and much 

more. 

Journal of Biomedical Nanotechnology 

Journal of Biomedical Nanotechnology (JBN) is 

being created as an international peer-reviewed periodical 

that covers applications of nanotechnology in all fields of 

life sciences. JBN publishes original full papers and 

timely state-of-the-art reviews with author's photo and 

biography, and short communications encompassing the 

fundamental and applied research aspects. To speed up 

the reviewing process, we will provide on-line refereeing 

of all articles submitted in electronic form. Authors 

receive these benefits: Electronic submission of articles, 

Fast reviewing, Rapid times to publication, No page 

charges, Free color where justified, Distinguished 

editorial board and Available in print and online 

editions. Topics include: Broadly speaking, Journal of 

Biomedical Nanotechnology covers applications of 

nanotechnology in biotechnology, medicine, biosciences, 

and all other related fields of life sciences. The coverage 

includes applications of nanotechnology in all fields of 

life sciences, all kinds of nanoscale biomaterials, 

biomimetics of biological materials and machines, 

nanoprobes, biocompatible surfaces, functional 

bioengineered materials, polypeptides, bioceramics, 

biopolymers, organic-inorganic hybrid biomaterials, 

nanocomposites, biological macromolecules, proteins, 

enzymes, kinases, phosphatases, DNA-based 

nanostructures, molecular assemblies, biomolecules, cells, 

and glycans, biochips, microarrays, biocompatibility 

aspects of materials, interactions between biomaterials, 

protein-surface, cells, tissue and organs, cellular matrix 

interaction,, artificial muscles and organs, biomembranes, 

bioseparation process, drug delivery, biopolymers for 

orthopedic and cardiovascular applications, dentistry, 

bone, bioanalysis, biosensors, molecular sensors, clinical 

diagnostic techniques, nanoparticles for drug delivery, 

dendrimers for medicine, biomedical implantation, 

biomechanics, bioinstrumentation, nanoscale physiology 

and pathology, bioinformatics, nanoscale genetics and 

genome research, gene expression, immunoassays, 

proteomics and protein-based nanostructures, sequencing 

of nucleic acid, DNA and RNA, biomarkers, 

biocomputing, instrumentation techniques for 

nanobioscience, nanoscale cellular and tissue engineering, 

nanodevices, biomedical nanoelectronics, biomedical 

microsystems, biochemistry and biophysics aspects, 

46


BioMEMS, nanofabrication, nanotubes, lab-on-a-chip, 

biological motors, biomembranes, nanofilters, biosensors, 

nanotechnologies for cell and tissues, nanofluidics, 

pharmaceutical nanotechnology, drug and gene delivery, 

therapeutic proteins, disease control, cancer therapeutics, 

diagnostic techniques, nanoscale imaging, nanoanalysis, 

spectroscopic studies using X-ray, STM, AFM, SNOM, 

systems biology, computational biology, etc., and much 

more. 

International Journal of Nanoscience (IJN) 

International Journal of Nanoscience (IJN) This 

inter-disciplinary, internationally-reviewed research 

journal covers all aspects of nanometer scale science and 

technology. Articles in any contemporary topical areas 

are sought, from basic science of nanoscale physics and 

chemistry to applications in nanodevices, quantum 

engineering and quantum computing. IJN will include 

articles in the following research areas (and other related 

areas): Properties Effected by Nanoscale Dimensions, 

Atomic Manipulation, Coupling of Properties at the 

Nanoscale; Controlled Synthesis, Fabrication and 

Processing at the Nanoscale; Nanoscale Precursors and 

Assembly, Nanostructure Arrays, Fullerenes, Carbon 

Nanotubes and Organic Nanostructures, Quantum Dots, 

Quantum Wires, Quantum Wells, Superlattices; 

Nanoelectronics, Single Electron Electronics and Devices, 

Molecular Electronics, Quantum Computing; 

Nanomechanics, Nanobiological Function and Life 

Sciences; Nanoscale Instrumentation and Characterization 

and Nano-optics, Photonic Crystals with Nanoscale 

Structural Fidelity. 

Nano Research 

With the development of modern nanotecnology 

and the inburst of various new ideas, new concepts and 

new thinking manners, more and more researchers have 

realized that nanotechnology must roots in the essences of 

international culture, with deep apperception to the 

traditional Chinese characteristics, absorbing and 

digesting the foreign consciousness, so that publishers can 

construct this journal with a high impact factor in the 

future; and find the real progress of Chinese 

nanotechnology which is recognized by the whole world. 

ACS Nano 

ACS Nano is an international forum for the 

communication of comprehensive articles on nanoscience 

and nanotechnology research at the interfaces of 

chemistry, biology, materials science, physics, and 

engineering. Moreover, the journal helps facilitate 

communication among scientists from these research 

communities in developing new research opportunities, 

advancing the field through new discoveries, and reaching 

out to scientists at all levels. ACS Nano publishes 

comprehensive articles on synthesis, assembly, 

characterization, theory, and simulation of nanostructures 

(nanomaterials and assemblies, nanodevices, and 

self-assembled structures), nanobiotechnology, 

nanofabrication, methods and tools for nanoscience and 

nanotechnology, and self- and directed-assembly. In 

addition to comprehensive, original research articles, ACS 

Nano offers thorough reviews, perspectives on 

cutting-edge research, conversations with nanoscience 

and nanotechnology thought leaders, and discussions of 

topics that provide distinctive views about the future of 

nanoscience and nanotechnology. 

Nanomedicine 

Nanomedicine: Nanotechnology, Biology, and 

Medicine (Nanomedicine: NBM) is an international, 

peer-reviewed journal. Each quarterly issue of 

Nanomedicine: NBM presents basic, clinical, and 

engineering research in the field of nanomedicine. Article 

categories include basic, diagnostic, experimental, clinical, 

engineering, pharmacologic, and toxicologic 

nanomedicine. In addition, regular features will address 

the commercialization of nanomedicine advances, ethics 

in nanomedicine, funding opportunities, and other topics 

of interest to researchers and clinicians. We invite authors 

to submit original manuscripts and review articles. The 

Journal is indexed or abstracted in PubMed/MEDLINE, 

BIOSIS Previews, EMBASE, SCOPUS, Biological 

Abstracts, Science Citation Index Expanded (SciSearch), 

Biotechnology Citation Index®, and Journal Citation 

Reports/Science Edition. 

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49

Publication News 

Drug Evaluation Research 

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Evaluation Research will officially be published in 2009, in China. Sponsored by China 

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The 3rd Asian Pacific ISSX Meeting 

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The Imperial Queen's Park Hotel| Bangkok, Thailand 

Understanding Xenobiotics for Better Drug Development and Therapy 

The 3rd Asian Pacific Regional Meeting of the International Society for the Study of 

Xenobiotics, it is our pleasure to invite you to the meeting which will be held May 10 – 12, 2009 

in Bangkok, Thailand. he theme of this meeting is Understanding Xenobiotics for Better Drug 

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50

Zhang R et al. Asian Journal of Pharmacodynamics and Pharmacokinetics 2009;9(1):51-57 




ISSN 1608-2281 



Tissue distribution of Curcumol in rats after intravenous injection 

of zedoary turmeric oil fat emulsion 

Rui Zhang 1 , Benjie Wang 1 , Hengli Zhao 2 , Chunmin Wei 1 , Guiyan Yuan 1 , Ruichen Guo 1* 

1 Institute of Clinical Pharmacology, Qilu Hospital of Shandong University, 107 Wenhua West Road, Jinan, 

250012, China 

2 Department of Pharmacy, The Second Hospital of Shandong University,247 Beiyuan Road, Jinan, 250033, 

China 

Abstract 

Key words 

Aim To determine the concentration of curcumol in different tissues and investigate the tissue 

distribution of curcumol in rats after intravenous injection of zedoary turmeric oil fat emulsion. 

Methods Wister rats were intravenously injected with a dose of 10.0 mg·kg -1 zedoary turmeric 

oil fat emulsions. The tissue samples (heart, liver, lung, kidney and brain) were collected at 

scheduled times. Curcumol was extracted with chloroform/isopropyl alcohol (95:5, v/v) from 

tissue homogenates and separated on a C 18 column with a mobile phase of acetonitrile (0.3M 

ammonium acetate containing 0.1% formic acid)/water (0.1% formic acid) (95:5, v/v). Detection 

was carried out by positive elevtrospray ionization (ESI) in multiple reactions monitoring 

(MRM) mode of 254.3→219.4 (m/z) for curcumol and 220.3→128.1 (m/z) for ornidazole (I.S.), 

respectively. Results Curcumol was distributed to liver, kidney, brain, heart, lung 10 min after 

intravenous injection and obtained the maximum concentration of 108.85±65.91, 105.19±42.92, 

92.38±17.63, 82.96±38.06, 10.01±2.97 µg·g -1 , respectively. Conclusions The method was 

successfully applied in the tissue distribution study. The results showed that the curcumol was 

markedly decreased after 0.5 h and almost eliminated at 5 h after administration. 

Curcumol; tissue distribution; LC-MS/MS 

Article history Received 6 August 2008; Accepted 30 December 2008 

Publication data Pages: 7 ; Tables: 4; Figures: 4 ; References: 10 ; Paper ID 1608-2281-2009-0901051-07 

Corresponding author Prof. Ruichen GuoInstitute of Clinical Pharmacology, Qilu Hospital of Shandong University, 107 Wenhua 

West Road, Jinan, 250012, China Tel: +86 531 82169636; Fax: +86 531 86109975 E-mail address: 

grc7636@126.com 

Introduction 

Zedoary turmeric oil is the essential oil derived 

from the rhizome of Chinese medicinal herb 

Curcuma phaeocaulis Valeton, C. kwangsinensis S.G. 

Lee et C. F. Liang and C. wenyujin Y. H. Chen et C. 

Ling [1] . It possesses the activities of antitumor, 

antivirus, antiinflammation, antibacterial, and so on. 

Curcumol (content was about 4.12%) is an important 

ingredient with antibiosis and antiviral effects in 

zedoary turmeric oil [2-4] , which can inhibited the 

growth of Bacterium typhi, Bacterium coli, 

respiratory syncytial virus (RSV), bastard measles, 

chickenpox virus, and so on [5] . 

51


The pharmacokinetic and tissue distribution 

studies were important for clinical application. These 

studies of curcumol by isotope tracer method was 

reported [6] , it was sensitive enough to determine the 

low concentration of curcumol in biotic sample, but 

the radioactivity were the sum of 3 H labeled original 

and its metabolites, which can not exactly illustrate 

the disposition process in vivo. In order to illustrate 

the disposal process of curcumol in vivo more exactly, 

other analysis method should be developed. Several 

quantitative method of curcumol in medical material 

and preparations such as RP-HPLC [1] , infrared 

spectrophotometry [7] , thin-layer scanning method [8] 

and vapor phase chromatography [9] were reported. 

These methods were not sensitive enough to 

quantitate the curcumol in biological specimen. We 

had developed a high-performance liquid 

chromatography coupled with tandem mass 

spectrometry quantitative detection method 

(HPLC-MS/MS) to determine the curcumol in plasma 

and the method had been applied in pharmacokinetics 

study in Beagle dogs successfully [10] . In this study, 

the established method was validated in tissue 

samples and used to investigate tissue distribution of 

curcumol after intravenous injection zedoary turmeric 

oil fat emulsion in rats. The chemical structure of 

curcumol is shown in Fig 1. 

CH 2 

O 

OH 

CH 3 

CH(CH 3 ) 

Fig 1. Chemical structure of curcumol 

Materials and methods 

Chemicals and materials 

Zedoary turmeric oil fat emulsion (Lot No: 

20051213, 200mg/100ml) was supplied by the 

Research Institute of Shandong Juren Biology 

Medicinal Technology (Shandong, China). Curcumol 

standard (Lot No: 100185-200405, 99.1%) was 

obtained from National Institute for the Control of 

Pharmaceutical and Biological Products (Beijing, 

2 

China). Ornidazole (internal standard, >99.6%) was 

donated by the Institute of Wuhan Jingxi Chemical 

Production (Hubei, China). Acetonitrile (Lot No: 

044263), formic acid (Lot No: 406014), chloroform 

(Lot No: 506023) and isopropyl alcohol (Lot No: 

503027) were chromatographic grade from TEDIA 

Company (USA); ammonium acetate (Lot No: 

20050516) was analytical grade from Tianjin Tianda 

Chemistry Industry Factory (Tianjin, China). 

Animal 

Male and female Wister rats (20020 g) were 

purchased from the Laboratory Animal Center of 

Shandong University(Jinan, China) . 

Apparatus 

The LC-MS/MS system include a 1100 Series 

HPLC (Agilent company, USA) and API 4000 

MS/MS (Applied Biosystem, USA); AX-205 

Electronic Balance (METTLER TOLEDO Instrument 

Shanghai Company); XW-80A Vortex (Shanghai 

Jingke Industry Company); SORVALL Biofuge 

PRIMO centrifuge (Kendro Company,USA). 

Chromatographic Condition 

Separation was carried out on a Diamonsil TM C 18 

column (150mm×4.6mm, 5µm) (Dikma Technologies, 

China) and eluted with a mobile phase of acetonitrile 

(0.3M ammonium acetate containing 0.1% formic 

acid-water (0.1% formic acid) (95:5, v/v). The flow 

rate was 1.0 mL·min -1 . 40µL of extracted samples 

was injected for analysis. 

Mass spectrometric condition 

The ESI source was set to positive ion mode 

with the following parameters: ion source 

temperature 550 °C, spray voltage 5500 V, sheath 

gas 70 psi and auxiliary gas 40 psi, the collision 

gas was argon at a pressure of 12.5 V (curcumol) 

and 20 V (ornidazole, I.S.) for collision-induced 

dissociation (CID). Quantification was performed 

using multiple reaction monitoring (MRM) of 

254.3→219.4 (m/z) for curcumol and 

220.3→128.1 (m/z) for ornidazole (I.S.), 

respectively. 

Preparation of stock and working standard 

solutions 

52


A stock solutions of curcumol at 100 µg·mL -1 

and ornidazol at 50 µg·mL -1 were prepared by 

dissolving appropriate amounts standards in mobile 

phase, respectively. The stock solution of curcumol 

was diluted subsequently to obtain a series working 

standard solutions. The stock solution of I.S. was 

further diluted to a working concentration of 

0.05µg·mL -1 . All solutions were stored at 4 °C. 

Preparation of calibration and quality control 

Samples (QC) 

Wister rats without administration were 

sacrificed and tissues were collected. Blank tissue 

homogenate was spiked by appropriate concentration 

of working standard solution to yield final 

concentrations between 0.25-50 ng·mL -1 of curcumol 

in tissue homogenate. Quality control samples were 

prepared in the same way at three concentration of 

1.0, 10, 40ng·mL -1 . 

Sample preparation 

The tissue samples were accurately weighted 

and cut into pieces and homogenated after adding the 

appropriate amount of 0.9% sodium chloride solution 

(3 mL·g -1 tissue). 1.0 mL tissue homogenate were 

transferred to a centrifuge tube, ornidazole (15 µL, 

0.05 µg·mL -1 ) and sodium hydroxide solutions (100 

µL, 0.1M) were added and well-mixed. Then 5ml of 

chloroform / isopropyl alcohol (95:5, v/v) was added. 

The mixture was vortex-mixed for 2min and 

centrifuged at 4000 r·min -1 for 5min. The organic 

phase was moved and evaporated to dryness under 

gentle nitrogen stream in water-bath at 37 °C. The 

residues were reconstituted with 100 µL mobile phase 

and 40 µL was injected into the HPLC/MS/ MS 

system. 

Tissue distribution study 

Rats were acclimated for at least 1 week before 

experiments. Thirty Wister rats were randomized into 

six groups and intravenously injected with a single 

dose of 10.0 mg·kg -1 zedoary turmeric oil fat 

emulsions. Then five rats per group were sacrificed at 

0.17, 0.5, 1.0, 2.0, 3.0 and 5.0 h after administration 

and tissue samples (heart, liver, lung, kidney and 

brain) were collected, respectively. Tissue samples 

were rinsed individually with 0.9% sodium chloride 

solution, blotted with filter paper, and stored 

immediately at -20 °C until analysis. The 

experimental procedures were carried out in 

accordance with the Standard Operating Procedure 

for Animal Experimentation of Shandong University 

(Shandong, China). 

Results 

Method validation 

Specificity Five batches of different blank tissue 

homogenate were analyzed on HPLC-MS/MS to 

assess specificity. Typical chromatograms in kidney 

and heart were shown in Fig 2 and 3. 

A B C 

Fig 2. Chromatograms of blank kidney (A), blank kidney spiked with curcumol and I.S. 

(B), kidney sample spiked with I.S. (C) 1: ornidazole; 2: Curcumol 

53


A B C 

Fig 3. Chromatograms of blank heart (A), blank heart spiked with curcumol(B), kidney samples (C) 1: Curcumol 

Table 1. The standard curve equation of curcumol in five tissues (n=5) 

tissue regression equation linear range (ng·mL -1 ) R 2 

heart Y=4890C+1740 0.5-50.0 0.9968 

liver Y=4230C+3530 0.25-50.0 0.9990 

lung Y=8510C-1240 0.25-25.0 0.9967 

kidney Y=0.075C-0.00532 0.5-25.0 0.9988 

brain Y=3970C-729 0.5-50.0 0.9970 

Table 2. Precision and accuracy data for curcumol in tissues (n=5, mean±SD) 

Tissue 

QC 

Intra-day 

Inter-day 

( ng·mL -1 ) C( ng·mL -1 ) RSD (%) RE (%) C( ng·mL -1 ) RSD (%) RE (%) 

Heart 1.0 0.96±0.05 5.3 -4.4 0.97±0.05 5.3 -3.4 

10 9.75±0.17 1.7 -2.5 9.70±0.26 2.7 -3.0 

40 39.79±0.61 1.5 -0.5 39.56±0.71 1.8 -1.1 

Liver 0.5 0.47±0.03 6.5 -6.4 0.48±0.03 5.5 -4.3 

5.0 4.92±0.11 2.2 -1.5 4.92±0.11 2.3 -1.7 

40 39.87±0.62 1.6 -0.3 39.61±0.71 1.8 -1.0 

Lung 0.5 0.48±0.02 3.3 -4.0 0.48±0.02 4.8 -3.9 

4.0 3.91±0.10 2.7 -2.4 3.94±0.09 2.2 -1.4 

20 19.63±0.41 2.1 -1.8 19.66±0.36 1.8 -1.7 

Kidney 1.0 0.97±0.03 3.1 -2.8 0.97±0.03 3.5 -2.9 

5.0 4.95±0.06 1.3 -1.0 4.95±0.08 1.7 -1.0 

20 19.86±0.42 2.1 -0.7 19.74±0.41 2.1 -1.3 

Brain 1.0 0.97±0.04 4.3 -3.2 0.97±.04 4.6 -2.8 

5.0 4.93±0.07 1.3 -1.4 4.95±0.08 1.6 -1.0 

40 39.64±0.44 1.1 -0.9 39.72±0.87 2.2 -0.7 

54


Table 3. Stability of curcumol in liver (n=5, mean±SD) 

QC( ng·mL -1 ) C( ng·mL -1 ) RSD (%) RE (%) 

25°C,4h 0.5 0.49±0.01 2.0 -2.1 

5.0 4.93±0.06 1.2 -1.3 

40 38.77±0.69 1.8 -3.1 

freeze/thaw* 0.5 0.49±0.01 2.2 -2.6 

5.0 4.92±0.07 1.4 -1.6 

40 39.41±0.62 1.6 -1.5 

-20°C, 1day 0.5 0.48±0.01 1.9 -4.1 

5.0 4.90±0.07 1.5 -1.9 

40 38.56±0.77 2.0 -3.6 

-20°C, 7days 0.5 0.48±0.01 2.1 -3.8 

5.0 4.80±0.05 1.0 -4.1 

40 37.68±0.39 1.0 -5.8 

* After three freeze/thaw cycles at -20°C 

Calibration and LOQ Five sets of calibration 

curves were prepared over the concentration range as 

shown in Table 1. The linear regression of the curve 

for the peak area ratio (Y) versus concentration(C) 

was plotted. The R 2 values for the standard curves are 

also listed in Table 1. The calibration curve of 

curcumol in five tissues showed good linearity. All 

coefficients of correlation were between 0.9967 and 

0.9990. The limit of quantitation (LOQ) in liver and 

lung was 0.25 ng·mL -1 . For other tissues, LOQ was 

0.5 ng·mL -1 . The limit of quantitation for all samples 

was sufficient for tissue distribution study. 

Extraction recovery The extraction recovery 

was calculated at three QC levels. QC samples were 

treated as “Sample Preparation”. The extraction 

recovery was obtained by comparing the curcumol 

peak area in tissue samples with those found by direct 

injection of corresponding standard solutions. The 

mean extraction recovery for all tissues was higher 

than 42.3%. 

Precision and Accuracy The precision and 

accuracy of method were assessed by intra-day and 

inter-day RSD (relative standard deviation) and RE 

(relative error). The precision was less than 6.5% and 

the accuracy was less than 6.4%. The results were 

shown in Table 2. 

Stability The stability of curcumol in tissue 

homogenate was studied by analyzing of QC samples. 

The samples, which were placed at room temperature 

(25 °C) for 4h, freezing/thawing for three cycles, 

stored at -20 °C for 1day and 7days, were 

investigated. Results of stability in liver were shown 

in Table 3. The stabilities of curcumol in other tissues 

were investigated in the same way, the data were not 

shown. Curcumol was found to be stable (RE 

within±8.2%) at different conditions. No significant 

degradation occurred during extraction and storage 

processes. 

Tissue distribution 

Curcumol was distributed to liver, kidneys, brain, 

heart, lungs 10 min after intravenous injection and 

obtained the maximum concentration of 

108.85±65.91, 105.19±42.92, 92.38±17.63, 82.96± 

38.06, 10.01±2.97 µg·g -1 , respectively. The concentrations 

of curcumol in various tissues of rats at 

scheduled time after intravenous injection of 10 

mg·kg -1 zedoary turmeric oil fat emulsion were 

presented in Table 4 and tissues concentration-time 

column profiles of curcumol were shown in Fig 4. 

Discussion 

55


Zedoary turmeric oil fat emulsion was a new 

preparation of zedoary turmeric oil. The 

pharmacokinetic and tissue distribution results of 

animal were necessary to its clinical application. 

Zedoary turmeric oil was a multicomponent drug. We 

 

always took the active component as the target 

analyte to study its disposition process in vivo. 

Curcumol was selected in this study for its high 

content and effects. 

Table 4 . Mean tissue concentrations of curcumol in rats at scheduled time after intravenous injection of 

zedoary turmeric oil fat emulsion(µg·g -1 n=5, mean±SD 

Time Heart Liver Lung Kidney Brain 

0.17h 82.96±38.06 108.85±65.91 10.01±2.97 105.19±42.92 92.38±17.63 

0.5h 21.02±12.36 32.66±40.21 3.99±1.25 40.66±18.32 35.32±6.7 

1h 16.20±19.99 23.01±19.54 1.66±0.3 26.35±16.11 18.85±10.42 

2h 2.65±1.76 10.81±4.16 1.56±0.39 12.76±9.71 4.76±2.34 

3h 2.25±0.68 9.41±7.88 0.73±0.49 7.26±5.39 3.31±2.06 

5h 0 5.1±3.02 0.82±0.2 4.74±4.05 2.61±1.32 

ND: not detected 

120 

100 

concentration(µg/g) 

80 

60 

40 

0.17h 

0.5h 

1h 

2h 

3h 

5h 

20 

0 

Heart Liver Lung Kidney Brain 

Fig 4. Tissue distribution profiles of curcumol at scheduled time after intravenous injection 

of zedoary turmeric oil fat emulsion 

The quantitation methods of five tissues were 

different in this study. The results of specificity in 

method validation showed that no distinguish 

interference from endogenous compounds was 

observed to curcumol in all tissues, but ornidazole 

I.S. was interfered in most tissues except kidney. 

So internal standard method was selected to 

determine the curcumol in kidney, and external 

standard method was used to other tissues. 

As listed in Table 4, curcumol was detected at 

10 min after intravenous injection and undetected in 

heart or lower concentrations detected in other tissues 

at 5h after administration, which indicated that 

curcumol was distributed rapidly and no long-term 

accumulation in tissues. It can be seen from Fig 3, 

that curcumol in all tissues showed similar dynamic 

56


change, the maximum concentration was at 10 min 

and markedly decreased at 30min, then declined with 

time. The highest level of curcumol appeared in the 

liver, followed by kidney, brain, heart, and the lowest 

level of curcumol appeared in the lung. These 

implied that curcumol were mainly metabolized and 

eliminated in the liver and kidney. The higher 

concentration in the brain and heart indicated that 

curcumol can pass through the blood-brain barrier, 

which attributed to its application in cardiovascular 

and cerebrovascular disease. The lowest concentration 

in lung perhaps called in a question in the use 

of disease of respiratory system, especially 

pneumonia. 

The satisfactory outcome was obtained in 

present study, but only five main tissues of rats were 

investigated, it could not supply the distribution 

information of curcumol in vivo completely, so 

experiments including more tissues or experiments in 

other more superior animals should be studied. 

Conclusion 

The tissue distribution of curcumol after 

intravenous injection of zedoary turmeric oil fat 

emulsion was investigated in the present study. The 

established LC-MS/MS method was validated and 

applied in determination of curcumol in tissues. The 

results showed that curcumol was distributed rapidly 

and no long-term accumulation in tissues. 

References 

1. Wu YC, Liu TY, Jiang LG, Bai ZZ. Determination of 

curcumol and germacrone in zedoary turmeric oil by 

RP-HPLC. Infomation of traditional Chinese medicine 2004; 

21(4):64-65. 

2. Xia Q, Zhao KJ, Huang ZG, Zhang P, Dong TT, Li SP, Tsim 

KW. Molecular genetic and chemical assessment of 

Rhizoma Curcumae in China. J Agric Food Chem 2005; 

53(15): 6019-6026. 

3. Yang FQ, Li SP, Chen Y, Lao SC, Wang YT, Dong TT, 

Tsim KW. Identification and quantitation of eleven 

sesquiterpenes in three species of Curcuma rhizomes by 

pressurized liquid extraction and gas chromatography -mass 

spectrometry. J Pharm Biomed Anal 2005; 39(3-4):552-558. 

4. Zhou X, Li ZW, Wang DP, Liang GY, Peng BX. Study on 

fingerprint of volatile oil of Curcumol wenyujin by GC-MS. 

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5. Zhao Y, Yang R G, Luo M. Progress in pharmacological 

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6. Su CY, Liu JY, Xu HX, et al. The metabolism of 

3 H-curcumol in normal rats and tumor-bearing mice. Acta 

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7. Yang SD, Chen JM, Chen YH. Determination of curcumol 

in zedoary turmeric oil of Curcumol wenyujin. Acta Pharm 

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8. Tian SJ, Liang WF. Study of zedoary turmeric 

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57

Venkatesh S et al. Asian Journal of Pharmacodynamics and Pharmacokinetics 2009;9(1):58-62 




ISSN 1608-2281 



Antinociceptive effect of Aerva lanata ethanolic extract in mice: 

A possible mechanism 

Sama Venkatesh 1* , Yanadaiah JP 1 , Zareen N 1 , Madhava Reddy B 1 , Ramesh M 2 

1 G. Pulla Reddy College of Pharmacy, Mehdipatnam, Hyderabad-500 028, India 

2 Jubilant Innovation, Yeshwantapur, Bangalore-560 022, India 

Abstract 

Key words 

Aim Aerva lanata Juss is traditionally claimed to be useful in the treatment of urolithiasis, 

strangury, diabetes, headache and pains. In the present study, 80% aqueous ethanolic extract of 

(the dried aerial parts of) A. lanata Juss is investigated for antinociceptive activity. Methods 

Male Swiss Albino mice were used to test the antinociceptive activity on acetic acid-induced 

writhing and hot plate test, at an oral dose of 50 and 100 mg·kg -1 . The aspirin and morphine 

served as standards. Results indicate that the ethanolic extract has produced a significant 

(P


The ethanolic extract of whole plant of A. lanata was 

reported to possess nephroprotective activity and in 

the treatment of acute renal injury caused by 

nephrotoxins like cisplatin and gentamicin [13] . To the 

best of our knowledge no report is available on the 

analgesic activity of A. lanata. In view of the fact that 

the plant is used by the traditional healers to alleviate 

headache among the natives, the present study was 

undertaken to verify the claim and evaluate the 

antinociceptive property of the aerial parts of A. 

lanata with the aim of developing a natural analgesic 

agent. 

Material and Methods 

Plant Material 

Aerva lanata Juss. aerial parts were collected 

from Surampet, Karimnagar district, Andhra Pradesh, 

India and authenticated by Dr. Prabhakar Reddy, 

Taxonomist, Department of Botany, Osmania 

University, Hyderabad. A voucher specimen 

(AVL-12-2006) is being maintained in the 

Department of Pharmacognosy and Phytochemistry, 

G. Pulla Reddy College of Pharmacy, Hyderabad, 

Andhra Pradesh, India. The aerial parts were cut, air 

dried and grounded into powder. 

Preparation of Extract 

Dried aerial parts powder (650 g) was extracted 

with 80% aqueous ethanol by maceration for seven 

days. The concentrated aqueous ethanolic extract (23 

g; % yield 3.53) was tested for qualitative 

phytoconstitiuents and indicated the presence of 

alkaloids, steroids and/or triterpenoids, flavonoids 

and their glycosides and tannins 

[14] , which 

corroborated with the earlier findings. 

Animals 

Male Swiss Albino mice (25-30 g) were used 

throughout the experiment. They were maintained 

under standard environmental conditions. Animals 

had free access to feed and tap water ad libitum 

during the quarantine period. The animal 

experimentation was carried according to the 

Committee for the Purpose of Control and 

Supervision of Experimentation on Animals 

(CPCSEA) guidelines and Institutional Animal Ethics 

Committee approved all the procedures for investigating 

experimental pain in conscious animals [15] . 

Acetic acid-induced abdominal writhing test 

The Siegmund et al [16] technique modified by 

Koster et al [17] was adopted to assess the 

antinociceptive activity in pre-screened mice. Over 

night (16 h) fasted mice were divided into six groups 

of six animals each. Group-1 served as a control 

group, which received 0.5% CMC (carboxy methyl 

cellulose) in water. Group-2 and 3 received 

ethanolic extract of A. lanata at a dose of 50 and 100 

mg·kg -1 , respectively as a fine suspension in 0.5% 

CMC, orally. Group-4 animals served as positive 

control animals received acetyl salicylic acid (100 

mg·kg -1 , po). In an attempt to investigate the 

participation of opioid system in the antinociceptive 

effect of this plant, separated groups of mice were 

pretreated with non selective opioid receptor 

antagonist, naloxone (5 mg·kg -1 , ip), which was 

injected 15 min before the administration of extract 

(100 mg·kg -1 , p.o) and acetyl salicylic acid (100 

mg·kg -1 , p.o) for Group-5 and 6, respectively. After 

30 min administration, all the animals were given an 

intraperitoneal (i.p) injection of 0.6% acetic acid 

(volume of injection 0.1 ml/10 g) and number of 

writhes produced in these animals was recorded for 

30 min. 

Hot-plate test 

The method of Eddy and Leimback [18] and 

Hosseinzabeh et al [19] was employed. The 

temperature of hot plate (Eddy’s hot plate, Dolphin, 

Mumbai, India) was maintained at 55 ± 0.2°C. 

Animals were placed into the Perspex square on the 

heated surface, and the time between placement and 

licking of paws and jumping was recorded as 

response latency. Over night (16 h) fasted mice were 

divided into six groups of ten animals each. Control 

animals were treated with 0.5% CMC (Group-1), 

while morphine (5 mg·kg -1 , p.o) was used as positive 

control (Group-4). Aqueous ethanolic extract of A. 

lanata was administered orally, at dose of 50 and 100 

mg·kg -1 as a fine 0.5% CMC suspension (Group-2 

and 3, respectively). The opioid receptor antagonist 

naloxone (5 mg·kg -1 , i.p) was also tested along with 

oral administration of ethanolic extract (100 mg·kg -1 , 

Group-5) and morphine (5 mg·kg -1 , Group-6). All 

59


substances were administered 30 min before the 

beginning of experiment. The reaction time was 

measured before and at 30, 60, 90, 120 and 180 min 

after substance administration. The latency period of 

40 sec was defined as complete analysis and 

measurement was terminated if the latency exceeded 

latency period to avoid injury. 

Table 1. Effect of A. lanata ethanolic extract on the acetic acid-induced abdominal writhing in mice (Mean ± S.E.M, n = 6) 

Group 

Treatment 

Dose 

(mg·kg -1 ) 

Number of writhes 

% Inhibition 

1 Control --- 75.35 ± 1.73 --- 

2 Aq ethanolic ext. 50 38.5 ± 2.67 * 48.90 

3 Aq. ethanolic ext. 100 21.74 ± 0.89 * 71.14 

4 Acetyl salicylic acid (positive control) 100 22.72 ± 1.59 * 69.84 

5 Aq. ethanolic ext. + Naloxone (ip) 100 + 5 25.8 ± 1.11 * 65.75 

6 Acetyl salicylic acid + Naloxone (ip) 100 + 5 28.16 ± 1.07 * 62.62 

*p


Statistical Analysis 

The results were expressed as mean ± S.E.M. 

The differences between experimental groups were 

compared by one-way ANOVA (control versus 

treatment by Bonferroni’s method; using Jandal 

Scientific, Sigmastat statistical software, version 1.0) 

and were considered statistically significant when P < 

0.05. 

Results 

Effect of acetic acid- induced writhing test 

Oral administration of A. lanata ethanolic 

extract (50 and 100 mg·kg -1 ) has produced dose 

dependent significant (P


aqueous ethanolic extract of A. lanata is a potential 

analgesic agent. 

Acknowledgements The authors wish to 

thank All India Council of Technical Education, New 

Delhi, India financial support. The authors also wish 

to thank management of the college for providing 

research facilities. 

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4, India: International Book Distributors; 1996; p. 2051. 

2. Anonymous. The Wealth of India: A Dictionary of Indian 

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CSIR Publications; 1959; p. 91. 

3. Chopra RN, Nayar SL, Chopra IC, Glossary of Indian 

Medicinal Plants. India: CSIR Publications; 1956; p. 8. 

4. Zapesochnaya GG, Pervykh LN, Kurkin VA. A study of 

the herb Aerva lanata. III. Alkaloids. Chem. Nat Comp 

1991; 27: 336-40. 

5. Pervykh LN, Karasartov BS, Zapesochnaya GG. A study 

of the herb Aerva lanata. IV. 

Flavonoid 

glycosides. Chem Nat Comp 1993; 28: 509-10. 

6. Wassel GM, Ammar NM. Phytochemical study of Aerva 

lanata. Fitoterapia 1987; 58: 367. 

7. Chandra S, Shastry MS. Chemical constituents of Aerva 

lanata. Fitoterapia 1990; 61: 188. 

8. Vetrichelvan T, Jegadeesan M, Senthil MP, Murali NP, 

Sasikumar K. Diuretic and anti-inflammatory activities of 

Aerva lanata in rats. Ind J Pharm Sci 2000; 62: 300-2. 

9. Vetrichelvan T, Jegadeesan M. Anti-diabetic activity of 

alcoholic extracts of Aerva lanata Juss. in rats. J 

Ethnopharmacol 2002; 80: 103-7. 

10. Chowdhury D, Sayeed A, Islam A, Shah Alam Bhuiyan M, 

Astaq Mohal Khan GR. Antimicrobial activity and 

cytotoxicity of Aerva lanata. Fitoterapia 2002; 73: 92-4. 

11. Nevin KG, Vijayammal PL. Effect of Aerva lanata on 

solid tumor induced by DLA cells in mice. Fitoterapia 

2003; 74: 578-82. 

12. Nevin KG, Vijayammal PL. Effect of Aerva lanata against 

hepatotoxicity of carbon tetrachloride in rats. Environ 

Toxicol Pharmacol 2005; 20: 471-7. 

13. Shirwaikar A, Deepti I, Malini S. Effect of Aerva lanata on 

cisplatin and gentamicin models of acute renal failure. J 

Ethnopharmacol 2004; 90: 81-6. 

14. Kokate CK. Practical Pharmacognosy. 4 th ed, India: 

Vallabh Prakashan; 1994; p. 112-20. 

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109-10. 

16. Siegmund E, Cadmus RA, Lu G. A method for evaluating 

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17. Koster R, Andersin N, Debber EJ. Acetic acid for 

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412. 

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Dithienylbutenyl- and dithienylbutyl-amines. J Pharmacol 

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19. Hosseinzabeh H, Ramezani M, Salmami G. 

Antinociceptive, anti-inflammatory and acute toxicity 

effects of Zataria mullifora Boiss extracts in mice and rats. 

J Ethnopharmacol 2000; 73: 379-85. 

20. Collier HO, Dinen LC, Johnson CA, Schneider C. The 

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21. Janssen PA, Niemegeers CJ, Dony JG. The inhibitory 

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62

Li R et al. Asian Journal of Pharmacodynamics and Pharmacokinetics 2009; 9(1):63-70 




ISSN 1608-2281 



Therapeutic effect and mechanism for anti-fibrosis of 

polyhydroxysilbene of Rhizoma Scirpi in hepatic fibrosis rats 

Run Li, Zong-Peng Zhang, Yi-Hong Tian 

Research Center for New Drug Evaluation, Tianjin State Key Laboratory of Pharmacokinetics and 

Pharmacodynamics, Tianjin Institute of Pharmaceutical Research, Tianjin 300193, China 

Abstract 

Key words 

Aim To study therapeutic effect and mechanism for anti-fibrosis of polyhydroxysilbene of 

Rhizoma Scirpi in hepatic fibrosis rats. Methods Fluidextract contained polyhydroxystilbene 

(PHS), The radix of Spcirpus yagara Ohwi. Was extracted with ethyl acetatethe therapeutic 

effect and mechanism on hepatic fibrosis is studied in this article. The test assay included serum 

levels of ALT, AST, TP, ALB and the levels of HA, LN, MDA, SOD and hapetic tissue 

pathomorphological examination. Results In animal experiment, study showed that the 

levels of ALT, AST in serum and the levels of HA, LN, MDA in rats liver in treatment groups is 

significant lower than that in pathologic control group(model group, group B).Compared with 

rats in pathologic control groupthe levels of SOD significant increased in the groups rats 

treated with Fluidextract or malotilate.In vitro, hepatic microsomes of rats were prepared by 

centrifugation. NADPH/Vit-C or cysteine/FeSO 4 reaction system is used to induce the 

production of MDA in rats liver microsomes. Conclusion The fluidextract suggested a better 

antioxidant effect.A dose-response relationship was found between the antioxidant effect and 

concentration of the fluidextract in the two reaction systems.The antioxidant effect may be one 

of the cardinal mechanisms in the therapeutic effect of extract of Spcirpus yagara Ohwi to 

hepatic fibrosis. 

Spcirpus yagara Ohwi, polyhydroxystilbene, Hepatic fibrosis, lipid peroxidation, Hepatic 

microsomes 

Article history Received 9 December 2006; Accepted 21 July 2008 

Publication data Pages: 8; Tables: 5 ; Figures: 7; References:18; Paper ID 1608-2281-2009-09010063-08 

Corresponding author Professor Zong-Peng Zhang, Research Center for New Drug Evaluation, Tianjin State Key 

Laboratory of Pharmacokinetics and Pharmacodynamics, Tianjin Institute of Pharmaceutical 

Research, Tianjin 300193, China. E-mail: zhang88985@163.com 

Introduction 

Hepatic fibrosis is a common pathological 

process of chronic hepatic disease, leading to the 

development of irreversible cirrhosis in patents [1-3] . 

There are various kinds of chronic liver injuries all 

over the world, causing great affection to patents. 

Therefore, searches for effective ways to inhibit 

63


fibrosis and prevent the development of cirrhosis 

are great significance [4] . Chinese traditional and 

herbal medicines, which are well known for the 

long history of prevent therapy of various diseases 

with low cost and few side effects, have particular 

potentials in the treatment of hepatic fibrosis [5-11] . 

Rhizoma Sparganii, which is a dried radix of 

Sparganium stoloniferum Buch.-Ham. (Family: 

Sparganiaceae) possesses wide pharmacological 

activities and is a commonly used traditional Chinese 

medicine for the treatment of chronic hepatitis in 

clinics of traditional medicine. Xiao and his 

co-workers reported that the Rhizoma Sparganii can 

protent hepetic cells, alleviate degeneration and 

necrosis, recover hepatic cell structure and liver 

function, and reduce the proliferation of fibrous tissue 

in the rat immunohepatic fibrosis model. Rhizoma 

Spcirpi, which is a dried radix of Spcirpus yagara 

Ohwi (family: Scirpusaceae) (Fig 1, www. 

zwy.csuft.edu.cn is also sued as a substitute for 

Rhizoma Sparganii in traditional Chinese medicine [15] . 

In chemical ingredients, the two herbs are different. 

Polyhydroxystilbene (3, 3, 4, 5-tetrahydroxystilbene), 

scirpusia A and B, veroeratrol, betulin and betulinic 

acid were found from Spciepus yagara and were not 

found from Sparganium stoloniferum. 

Polyhydroxystilbene is a main chemical ingredient of 

Spciepus yagara and its pharmacological activity on 

inhibition of lever fibrosis was not investigated. 

Fig 1. Spciepus yagara Ohwi 

In this study, the effects of Polyhydroxystilbene 

extraction from Spciepus yagara on lever 

fibrosis used an induced lever fibrosis model by 

carbon tetrachloride in rats. The antioxidant effect to 

be one of the cardinal mechanisms in the therapeutic 

effect to hepatic fibrosis was employed to examine. 


The extract and drug 

Polyhydroxystilbene fluidextract (PHS) 

extracted from plant materimals of Spcirpus yagara 

Ohwi. Rhizome with ethyl acetate is 

Yellowish-brown. Eevery gram of 

polyhydroxystibene fluidextract is equivalent to 300 

g crude materials.As a positive control drug 

malotilate tablets are purchased from Shanxi Yabao 

Pharmaceutical Co.Ltd., China. 

Animals 

Wister rats, male and female, with body weight 

from 250g to 300gsupplied by laboratory animal 

unit of Tianjin Institute of Pharmaceutical Research. 

Instruments and Reagents 

The centrifugeVITALAB Selectra 2 automatic 

biochemistry analyzer and ELISA were used in 

experiment.The biochemistry assay kits and the 

quality control serums are provided by Zhongsheng 

Beikong Bio-technology and Science Inc of china. 

The rat hyaluronic acid HAand lamininLN 

ELISA kits are manufactured by Market inc of 

U.S.A. 

Methods of animal experiment 

Rats are randomly divided into 6 groups 

(6/group), group A, B, C,D, E and F respectively.Rats 

at group BCDE and F received a single oral dose 

of 30% CCl 4 (0.2ml/100g) in sesame oil by gavage 

for 8 weeks.After 2 weeksrats at group C received a 

single oral doses of 75mg·kg -1·d -1 malotilate 

suspensions in water by gavage for 6 weeksand 

group A, D, E and F received a single oral doses of 0, 

0.15, 0.6, 2.4 mg·kg -1·d -1 polyhydroxystilbene 

fluidextract fluidextract suspensions in water/tween 

80 by gavage for 6 weeks.Rats fasted 16 h and then 

sacrificed after the last dose.At the same timesblood 

and liver sample were also collected. 

Assay methods for serum biochemiscal 

values 

The serum biochemistry values including 

alanine aminotransferase (ALT), aspartate 

aminotrasferase(AST), total protein(TP), albumin 

64


(ALB) and the values of superoxide dismutase 

(SOD)hydroxyproline (Hyp) in liver were assayed 

using automatic biochemistry analyzer with the 

methods offered by biochemistry assay kits [16] . The 

serum values of HA LN were assayed using 

ELIASA with the methods offered by ELISA 

kits.The concentration of MDA in liver was measured 

with thiobarbituric acid (TBA) method (colorimetry). 

Pathological examination 

The pathological change of hepatic slice by HE 

dyeingwere observed under the light microscope. 

The classification and statistical analysis of hepatic 

fibrosis refer to Consensus on evaluation of the 

diagnosis and efficacy of hepatic fibrosis [11] . 

Methods of anti-peroxidized experiment in 

vitro 

In vitro,hepatic microsomes of rats were 

prepared by centrifugation [12,13] . The production of 

MDA in rats liver microsomes were induced with 

NADPH/Vit-C [14] and cysteine/FeSO 4 reactive 

system [16] . The concentration of MDA in rats liver 

microsomes suggesting peroxidized levels of liver 

microsomes, were measured with thiobarbituric acid 

method (TBAcolorimetry). 

Statistical analysis 

All data were analyzed by SPSS for windows 

statistical package. Measurement data were analyzed 

by ANOVA, ranked data were analyzed by 

Kruskal-Wallis test. P values were less than 0.05 

were considered to be statistically significant. 

Results 

Animal experiment 

Effect on liver functions At the end of 

treatment period5/12 animals died in group B and D, 

3/12 animals in group C, E and F died respectively 

while no died in group A. Decrease in mortality was 

observed in group C, E and F. 

Table 1. Effect of polyhydroxystibene fluidextract on serum AST, ALT, TP and ALB values of rats (Mean± SD) 

Animal 

amountn 

ALT 

u/L 

AST 

u/L 

TP 

g/L 

ALB 

g/L 

Group A 9 31.8±9.2 ** 115.6±34.7 ** 74.1±5.5 33.8±3.5 * 

Group B 7 183.9±59.0 346.8±110.7 70.3±6.2 29.9±2.9 

Group C 9 87.6±31.2 ** 167.4±63.2 ** 71.5±4.4 30.1±2.2 

Group D 7 152.7±51.3 265.6±64.9 72.2±5.5 29.3±2.9 

Group E 8 176.5±93.8 328.6±131.1 69.6±4.9 29.7±2.9 

Group F 8 109.7±62.0 * 195.8±53.2 * 71.0±4.3 30.4±2.6 

Significant relative to group B* p


As shown in Table 1, the ALT, AST values in 

group B is significant higher than that in group A 

(P


At the concentrations of 75µg·ml -1 , and 375µ·ml -1 , the 

lipid peroxidation value was signaficantly lower than 

that of control group (p


was 1.04±0.05 in reaction system. Polyhydroxystilbene 

fluidextract completely inhibited the lipid 

peroxidation induce by cysteine and FeSO 4 in the 

system at the concentration of 5µg·ml -1 . At the 

concentrations of 5µg·ml -1 and 10 µg·ml -1 , the lipid 

peroxidation value was lower than that of control 

group (P


Discussion 

Stigma of blood stasis always appears in hepatic 

fibrosis patients. For its effect of activating blood 

circulation to dissipate blood stasis, Spcirpus yagara 

Ohwi always occurs in herbalist doctor`s 

prescriptions for treatment of the hepatic fibrosis. 

Pharmacological studies in animal models indicated, 

the pharmacological effect of Spcirpus yagara Ohwi 

is in accord with the traditional efficacy. In fact, it is 

a question that the pharmacological research of 

Spcirpus yagara Ohwi still stagnated in the research 

of its traditional effect, such as effect of activating 

blood circulation and releasing pains.From a 

literature survey ,it appears that none of the previous 

investigators is concerned with the anti-hepatic 

fibrosis effect of Spcirpus yagara Ohwi. According 

to the Chinese tridetional medical theories, stagnation 

and block of the blood vessel was the pathogenesis 

on hepatic fibrosis patients.Ameliorated the blood 

sludging very avails to hepatic fibrosis patients. 

Spcirpus yagara Ohwi can meliorate economy 

microcirculation, and can meliorate blood providing 

of liver. 

Fig 7. Effects of polyhydroxystibene fluidextract on lipid perxidation induced 

by cysteine/FeSO 4 in liver microsomes of rats 

Thereforethe therapeutic effect of Spcirpus 

yagara Ohwi on hepatic fibrosis was studied with the 

harmacological method by us in our laboratory, at the 

same time, we hope to reveal the mechanism of its. 

Experimental observation shown, 

polyhydroxystibene fluidextract which has certain 

effect on hepatic fibrosis of rats model, could 

significantly meliorate the hepatic function of model 

rats,protect hepatocyte and abate hepatocyte 

necrosis.Rats striked by CCl 4 shown obvious stigma 

of injured by lipid peroxidationthe SOD values in its 

liver decreased significantlythe content of MDA 

increased.It is generally accepted that lipid 

peroxidation is the bridge between hepatic fibrosis 

and continuate hepatitis (Liu, 2002). Thereforby 

employing reactional system of NADPH/Vit-C or 

cysteine/FeSO 4 we focused attention on the 

mechanism of therapeutic hepatic fibrosis effect of 

Spcirpus yagara Ohwi. 

As the result in above, PHS fluidextract can 

resist liver microsame injury of lipid peroxides 

completely inhibited the lipid peroxidation induced 

by Vit-c and NADPH in microsomes at the 

concentration of 5mg·ml -1 and completely inhibited 

the lipid peroxidation induced by cysteine and 

FeSO 4 in the system at the concentration of 

1.6mg·ml -1 . 

In conclusion, polyhydroxystibene fluid extract 

is an effective substantiate in treating hepatic 

fibrosis. Its mechanism might be probably related to 

the effect of anti-lipid peroxidation. 

Acknowledgments Authors are very grateful to 

Professor Liu CX for his helps. This work is completed 

partly by Guo CM, Wang JJ, Cheng XX, Hu LZhang JX 

and Wang H. This study was supported by National Basis 

Research Plan (973 Plan) No 2007CB516807. 

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peroxide.J Biol Chem 1987; 262(3):1098. 

13. Tang WX, Dan ZL, Yan HM, Wu CH, Zhang G, Liu M, Li 

Q and li SB. Experimental study of effect of ganyanping on 

fibrosis in rat livers. World J Gastroenterol 2003; 

9:1292-1295 

14. Rolf T Yasush H, Charles SL. Hepatic Microsomal 

Alcohol-oxidizing. J Biol Chem 1975; 250(18):7397. 

15. Wills ED. Lipid Peroxide Formation in Microsomes. 

Biochem.J 1969; 113:315-323. 

16. Xiao PG 2002. Rhizoma Sparganii. Modern Chinese 

material Medica 1:26-34. Beijing: Chemical Industry Press. 

17. Zhang ZP, Tian YH, Li R, Cheng XQ, Guo SM, Zhang JX, 

Wang JX, and Hu L. The comparison of the normal blood 

biochemical values of Wistar rats with different age and sex. 

Asian J Drug Metab Pharmacokinet 2004; 4(3): 215-218 

18. Andrew JF, Searl E. Stimulation of microsomal lipid 

peroxidation by iron and cyseine. Biochem J 1983; 

212:549-554. 

70

Nie XP et al. Asian Journal of Pharmacodynamics and Pharmacokinetics 2009; 9(1):71-76 




ISSN 1608-2281 



Changes of the adenosine content in single and mixed decoctions 

of Gualou-xiebai-baijiu decoction 

Xiao-Pu Nie 1 , Wen-Yuan Gao 1 *, Pei-Gen Xiao 2 

1 School of Pharmaceutical Science and Technology, Tianjin University, Tianjin 300072, China 

2 

Institute of Medicinal Plant, Chinese Academy of Medical Sciences and Peking Union Medical College, 

Beijing 100094, China 

Abstract Aim To investigate the adenosine content in single and mixed decoctions of 

Gualou-xiebai-baijiu decoction, we developed a simple and rapid High-performance liquid 

chromatographic (HPLC) method for the determination of adenosine, an active constitute to 

restrain platelet aggregation in Gualou-xiebai-baijiu decoction. Methods The decoctions were 

separated on a HiQsilC 18 column4.6mm×250mm5µmand detected by ultraviolet detection 

at a wavelength of 260nm and at the temperature of 30 °C. The use of methanol-water (10:90, 

v/v) as the mobile phase at a flow rate of 1.0 mL·min -1 enabled the baseline separation of the 

drugs free from interferences with isocratic elution. Results The content of adenosine in the 

mixed decoction is much higher than those in the single decoction which was extracted with 

20% ethanol. But it is much less than in the single decoction which was extracted with 10% and 

30% ethanol. The content of adenosine in single and mixed decoctions have exceeding 

significant difference (P


Numerous studies have shown substantial evidence 

that adenosine modulates cardiovascular control at 

the level of the nucleus tractus solitarius (NTS) [3-6] . 

In the last few years, there are more and more 

researches on the differences between the single 

decoction and the mixed decoction 

[7] .To our 

knowledge, however, there is no report on 

Gualou-xiebai-baijiu decoction in this aspect. 

Adenosine (Fig. 1) is an active constitute to inhibit 

platelet aggregation and has been separated from both 

Fructus trichosanthisi and Bulbus alliimacrostemi [8,9] . 

Moreover, Fructus trichosanthisi and Bulbus 

alliimacrostemi are often paired a basic prescription 

in many prescriptions to treat cardiovascular disease 

in traditional Chinese medicine [10] . 

Nowadays, coronary heart disease CHD is 

severely harming human health for its high 

incidence and death rate. CHD belongs to 

arthromyodynia category according to traditional 

Chinese medicine. The traditional Chinese medicine 

plays more and more important role in the treatment 

of CHD. Gualou-xiebai-baijiu decoction is a 

well-known classic preparation in traditional 

Chinese medicine. It originates from Jinkui Yaolue, 

a famous pharmaceutical book written by Zhang 

Zhong-Jing in the Han dynasty. The prescription 

consists of Fructus trichosanthisi and Bulbus 

alliimacrostemi and is extracted with wine [11] . It has 

been used to treat CHD and angina pectoris for 

several hundred years and has achieved good results 

in modern clinical practice [12] . 

HO 

N 

NH 2 

N 

H 

OH 

O 

N 

N 

OH 

Fig 1. Chemical structure of adenosine 

In this paper, we established a simple and rapid 

High-performance liquid chromatographic (HPLC) 

method for the determination of adenosine in 

Gualou-xiebai-baijiu-tang and then investigated the 

content of adenosine in single and mixed decoction 

of Gualou-xiebai-baijiu decoction. 


Chemicals and reagents 

The crude herb of Fructus trichosanthis and 

Bulbus allii macrostem were purchased from Anguo 

Meiwei Pharmaceutical Co. (Hebei, China). 

Adenosine was obtained from National Engineering 

Research Center of technique of manufacture of the 

traditional Chinese medicine solid preparation 

(Jiangxi, China). Methanol (chromatographic 

grade) was purchased from Concord Technological 

Company (Tianjin, China). Water was twice 

distilled water. Other reagents used were at least 

analytical grade. 

Equipment 

The HPLC system consists of an Agilent1100 

serials pump, solvent degasser, column oven, UV 

detector. This system was operated with a Chem 

Station software (Version: A.09.01) (Agilent, 

USA).The HPLC consists of a 5µm C18 column 

(4.6×250mm) (HiQ silC 18 , Japan) and a safeguard 

column of the same material. 

Chromatographic conditions 

The column temperature was kept constant at 

30 and the mobile phase flow rate was 

1.0mL·min -1 . All of injection volume was 10µL. 

For quantitative assay, the mobile phase was 

methanol -water (10:90, v/v) and the UV detector 

measured absorbance at 260 nm. 

Standard solutions 

Stock solution of adenosine of 19.84µg·mL -1 

was prepared in 90% methanol. Appropriate 

dilutions (v/v) of the stock solutions were made 

with 90% methanol to obtain working solutions 

from 1.984 to 19.84µg·mL -1 of adenosine. These 

solutions were used for the study of linearity, 

accuracy, repeatability and recovery. 

The linearity of the method was established by 

using 5 standard solutions for adenosine 

(concentration range of 1.984–19.84µg·mL -1 ), 

assayed in triplicate on the experiment. 

72


Sample preparation 

Mixed decoction preparation The Fructus 

Trichosanthis and Bulbus Allii Macrostemi 5.0g 

(the proportion of Fructus Trichosanthis and Bulbus 

Allii Macrostemi was 3:2), were soaked by 20% 

ethanol (40ml) for 0.5h. Then they were extracted in 

a regurgitate bath for 1 h. The decoction was 

filtered through absorbent gauze. The remainder 

slags were continually extracted in a regurgitate 

bath for 1 h by 20% ethanol (30ml). The decoction 

was filtered through absorbent gauze and merged 

with the first decoction. For determination of 

adenosine, the solution was filtered through a 

membrane (0.22µm) and then injected into HPLC. 

Single decoction preparation The Fructus 

Trichosanthis 3.0g was soaked by 20% ethanol 

(24ml) for 0.5h. Then they were extracted in a 

regurgitate bath for 1 h. The decoction was filtered 

through absorbent gauze. The remainder slags were 

continually extracted in a regurgitate bath for 1 h by 

20% ethanol (18ml). The decoction was filtered 

through absorbent gauze and merged with the first 

decoction. 

The Bulbus Allii Macrostemi 2.0g was 

extracted by the same method as Fructus 

Trichosanthis. Finally, the decoction of Fructus 

Trichosanthis and Bulbus Allii Macrostemi was 

filtered through a membrane (0.22µm) and then 

injected into HPLC. 

Data analysis 

Statistical analysis was performed using the 

repeated-measures ANOVA test, followed by t test, to 

compare different group. 

Results and discussion 

Selection of chromatographic conditions 

A good separation condition should satisfy the 

need that the analyzed peaks have baseline 

separation with adjacent peaks within a short 

analysis time as far as possible. To obtain the 

chromatograms with good separation, we tried 

various mobile phase. According to literatures [13, 14] , 

buffer salt solutions which were prepared 

complicatedly were used and damage the 

chromatographic column. So we tried the mixtures 

of acetonitrile and water (18:92, 4:96, v/v) as 

mobile phase but separation was not satisfactory. 

When the mixtures of methanol and water (15:85, 

13:87 and 12:88, v/v) were used as mobile phase, 

the analyzed peaks did not have baseline separation 

with adjacent peaks. Only when using methanol and 

water (10:90, v/v) satisfactory separation result can 

be obtained. 

Table 1. Recovery of adenosine in Gualou-xiebai-baijiu decoction 

Sample Initial content (µg) Amount added (µg) Recovery (%) Average value of recovery (%) R.S.D. (%) 

1 100.5985 100.192 99.4 

2 108.3180 108.128 102.0 

3 99.6212 103.168 99.7 

4 98.3459 98.208 98.5 

100.0 1.6 

5 102.5576 102.176 101.8 

6 100.1929 100.192 98.5 

Method validation 

Linearity The linear range for adenosine 

(1.984–19.84µ·mL -1 ) was evaluated. In linearity 

range was used 5 different standard solutions of the 

lower and upper limit concentration. Calibration 

curves were obtained by plotting the peak area 

versus adenosine concentrations (µ·mL -1 ) and 

resulted in straight lines over the concentration 

range. The linearity equations were calculated by 

using linear regression analysis, and typical 

calibration curves were defined by the following 

equations: y = 30.206x-5.3233, r=0.9997. 

It is possible to conclude that, for adenosine, 

the functions are linear in the working range. 

73


mAU 

4 

VWD1 A, Wavelength=260 nm (NIE\GX000037.D) 

19.047 

2 

0 

-2 

-4 

0 5 10 15 20 25 

A 

mi 

 


mAU 

100 

80 

60 

40 

20 

0 

-20 

0 5 10 15 20 25 

18.970 

mi 

 


mAU 

100 

80 

60 

40 

20 

0 

0 5 10 15 20 25 

 



mAU 

mAU 

18.968 

mi 

 

80 

80 

60 

40 

60 

40 

20 

0 

-20 

0 5 10 15 20 25 


mAU 

80 

60 

40 

20 

0 

18.940 

mi 

 

-20 

0 5 10 15 20 25 

18.845 

mi 

 

20 

0 

0 5 10 15 20 25 


mAU 

100 

80 

60 

40 

20 

0 

0 5 10 15 20 25 

 

18.968 

18.944 

mi 

mi 

 

 

Fig 2. HPLC chromatograms of chemical reference substance and sample 

Accuracy The accuracy of this method was 

determined by comparing the calculated 

concentration using calibration curves to the known 

concentrationR.S.D.=0.41%,n=6. 

Stability The stability study shows that the 

decoction containing adenosine is stable at room 

temperature for 12hR.S.D.=1.5%,n=6. 

Repeatability The content of adenosine in 5 

74


samples was prepared according to the method in 

2.5.1. The obtained R.S.D. values were 1.55%.The 

method is precise when working under the same 

operating conditions (e.g. same operator and 

equipment). 

Recovery The recoveries of six samples were 

investigated. Average recovery of adenosine was 

generally above 98%.The results are shown in Table 

1. 

Sample analysis 

Chromatogram Typical chromatograms of 

samples are shown in Fig.2. All samples were 

completely separated. The results presented here 

show that a HPLC method for the measurement of 

adenosine has been developed. The categories of 

compounds in Gualou-xiebai-baijiu decoction 

extracted from different solution have changed. The 

categories of compounds in Gualou-xiebai-baijiu 

decoction which extracted by same solution but 

different decoction styles are uniform on the whole. 

A chromatogram of a adenosine standard 

solution sample; B chromatogram of a mixed 

decoction sample by 10% ethanol; C chromatogram 

of a single decoction sample by 10% ethanol; D 

chromatogram of a mixed decoction sample by 20% 

ethanol; E chromatogram of a single decoction 

sample by 20% ethanol; F chromatogram of a 

mixed decoction sample by 30% ethanol; G 

chromatogram of a single decoction sample by 30% 

ethanol. 

Analysis mixed decoction samples by 

different ethanol concentrations Samples were 

prepared according to the method in 2.5.1, by 10% 

ethanol, 20% ethanol and 30% ethanol, respectively. 

Then the contents of adenosine in them were 

investigated. The results are shown in Fig.3. 

Fig 3. Results of adenosine content in Gualou-xiebai-baijiu decoction 

single decoction compared to mixed decoction which were extracted from the same solution.( ** P


ethanol had the best protection effect to the rats 

which had undergone a thirty-minute ligature of 

coronary left anterior descending branch and used 

as a model of ischemic reperfusion [15] . It is possible 

that the content of adenosine in the decoction is 

higher at 20% ethanol than in other concentration 

ethanol solutions (10% and 30%). The traditional 

Chinese medicine prescriptions often produce a 

joint action through many ways and targets. More 

active constitutes and pharmacology researches on 

the traditional Chinese medicine prescriptions 

should be done in more aspects. 

Conclusion 

A simple and rapid High-performance liquid 

chromatographic (HPLC) method for the 

determination of adenosine is developed. This 

method is achieved by methanol and reversed phase 

HPLC with ultraviolet detection. This assay method 

developed the quality control of the preparations 

including Fructus Trichosanthis and Bulbus Allii 

Macrostemi. The adenosine content in the single 

and mixed decoctions has very significant 

difference. The dissolution of adenosine isrelated 

with both the ethanol concentration and decoction. 

Active constitutes and pharmacology researches on 

the traditional Chinese medicine prescriptions,the 

influencing action of decoction style should be 

considered. 

References 

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the brainstem mediate opposite blood pressure responses. 

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constituents from Trichosanthes kirilowii. Chinese 

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9. Peng J P. Chemical constituents in Allium macrostemon 

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Gualou-xiebai-tang. Chin J Exp Tradit Med Form 1999; 

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76

Successful the 2nd Asian–Pacific Regional ISSX Meeting 

International Society for Study of 

Xenebiotics (ISSX) held the 2nd Asian–Pacifi c 

Regional Meeting at the School of Pharmacy, 

Fudan University in Shanghai, China, from May 

11–13. The meeting, co-chaired by Honghao 

Zhou, Yuichi Sugiyama and Zhuohan Hu, 

attracted a total of 517 meeting participants and 

additional accompanying persons. The four short 

courses held on Monday, May 11 had a 

combined enrollment of 398. The regional 

breakdown was 79% Asian Pacific,19% North 

American, 2% European. Eighteen countries 

were represented (Table 1). 

Table 1 Asian-Pacific Regional ISSX Meeting 

attendees by countries 

Country/ Regional Attendees 

China 239 

USA 93 

Japan 83 

South 42 

Taiwan, China 20 

UK 8 

Australia 7 

than 10 abstracts presented from 6 institutions in 

China (Table 2). 

Fig 1. News of this meeting 

340 abstracts were accepted by this meeting. 

297 abstracts were accepted as posters and 

presented during the poster sessions held on 

Monday, May 12 and Tuesday, May 13. All 

accepted abstracts have been published in a 

special supplemental issue of Drug Metabolism 

Reviews. ISSX is very appreciative of the 

extraordinary efforts of the meeting’s able 

organizing committee, led by Honghao Zhou, 

Yuichi Sugiyama, and Zhuohan Hu, as well as 

the strong support and involvement of the 

members of the ISSX Meeting 

Organizing Committee. The ISSX Web site 

will feature a full report of the scientific sessions 

of the 2nd Asian–Pacific Regional Meeting. 

News of this meeting was published in 

ISSX Newsletters 2008;28(2):6 (Fig 1). 

ISSX published a special issue of Drug 

Metabolism Reviews (IF=5.59) (Fig 2). 340 

abstracts were published in the issue. From the 

publication, we found that drug metabolism 

research is an active study filed in China. More 

Fig 2. Abstracts’ publication of this Meeting 

Table 2 More than 10 abstracts presented 

from 6 institutions 

No. Institution Paper 

number 

1 Shanghai Research Center of Drug 33 

Metabolism 

2 Shanghai Institute of liver Disease 20 

3 Tianjin Institute of Pharmaceutical 15 

Research 

4 Center South University 12 

5 China Pharmaceutical University 11 

6 Second Military Medical University 11 

77




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Koumaravelou K. Genetic polymorphism of CYP2D6 in 

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The instruction for manuscript preparation assume 

certain resources are available to the author. Although 

compliance with these instructions will increase the 

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not be discouraged in submitting manuscripts that do not 

completely comply with these instructions. If an author 

feels his/her manuscript is worthy of publication in the 

Journal, he/she should do as much as possible to comply, 

then submit it to the editor. If the editor and editorial 

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Editorial Office will assist in satisfying the publisher’s 

manuscript preparation requirements. 

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Authors will receive page proofs before publication, and 

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that the authors automatically agree to assign exclusive 

copyright to Hong Kong Medical Publisher once this 

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not be published elsewhere in any forms without written 

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