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Journal of Medicinal Plants Research Vol. 6(12), pp. 2438-2442, 30 March, 2012 Available online at http://www.academicjournals.org/JMPR DOI: 10.5897/JMPR11.1435 ISSN 1996-0875 ©2012 Academic Journals Full Length Research Paper High-performance liquid chromatography (HPLC) determination of five active ingredients in the calyces of Physalis Alkekengi L. var. franchetii (mast.) Mskino Baoli Xu, Chengguo Ju, Huijie Guan, Liang Xu, Nan Xu and Bing Wang* Liaoning University of Traditional Chinese Medicine, 77 shengming 1 Road, DD Port, Dalian, Liaoning Province, P. R., 116600, China. Accepted 11 January, 2012 A simple high-performance liquid chromatographic (HPLC) assay using the internal standard method is developed for the simultaneous determination of five active ingredients. The analyzed compounds from the calyces of Physalis alkekengi L. var. franchetii (mast.) Mskino include Luteoloside, Luteolin, Physalin A, Physalin O and Physalin P. HPLC analysis is performed on an Agilent C18 analytical column (150×4.6 mm and 5 μm) using solvent (A) acetonitrile and (B) 0.2% aqueous phosphoric acid as the mobile phase with Ultraviolet (UV) absorption at 350 and 220 nm. The flow rate was 1.0 ml/min. The calibration curves of the five active ingredients are linear (r 2 > 0.9991) over the concentration range of 10 to 120 μg/ml. The mean recoveries were 97.59 to 101.04%. The results indicate that the HPLC method developed can easily be applied to the determination of five active ingredients in the calyces of P. alkekengi L. var. franchetii (mast.) Mskino. Key words: Calyces of Physalis alkekengi L. var. franchetii (mast.) Mskino, flavonoid, physalin. INTRODUCTION The calyces of Physalis alkekengi L. var. franchetii (mast.) Mskino major recorded in the Chinese Pharmacopoeia is a well-known traditional Chinese medicine (TCM) for the treatment of sore throat and urinate inability. The content of the luteolin not less than 0.1% in the dry calyces in accordance with the provisions of the Pharmacopoeia of PRC 2010 (The Pharmacopoeia of PRC, 2010). Flavonoids and physalins (Matsuura et al., 1970; Kawai et al., 1987, 1988, 1993; Kawal et al., 1995; Kazushi and Shii, 1992) are the main ingredients in the calyces of P. alkekengi. Flavonoids and physalins are the class of anti-inflammatory compounds (Xu et al., 2009; Melissa et al., 2005; Angelica et al., 2005). The aim of this study is to develop an assay to fully evaluate the contents of flavonoids and physalins in the calyces from different locations, wilds and purchases order to exploit and utilize them reasonably. *Corresponding author. E-mail: YZBwang@lnutcm.edu.cn. Tel: +8641187586003. MATERIALS AND METHODS Plant material Dried calyces of P. alkekengi var. franchetii were collected in six provinces of China (Heilongjiang, Jilin, Liaoning, Shanxi, Shandong and Anhui) from October 5 to 15, 2010. Voucher specimens were maintained at Liaoning University of TCM, China. Reagents The stand substances of the Luteoloside bought from Dida Technology Company Limited (Guizhou, China). The stand substances of the Luteolin were bought from State Food and Drug Administration (Beijing, China). The three standard substances of physalin were isolated from the calyces of P. Alkekengi including Physalin A, Physalin O and Physalin P by Prof. Nan Xu and Prof. Liang Xu (Liaoning University of Traditional Chinese Medicine, China). The purity of all the stand substances exceeds 98%. HPLCgrade acetonitrile was purchased from Kermel Chemical Reagent Company Limited (Tianjin, China), and the water used in all experiments was purified by a Milli-Q Ultrapure Water System (Millipore, MA). All other chemicals were of analytical reagent grade purchased from Kermel Chemical Reagent Company Limited (Tianjin, China).
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Journal of Medicinal Plants Researc
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Editors Prof. Akah Peter Achunike E
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Electronic submission of manuscript
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Fees and Charges: Authors are requi
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Table of Contents: Volume 6 Number
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Figure 1. Chuanxiong Rhizoma (http:
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Table 1. Identification of main pea
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Table 3. Pharmacokinetic parameters
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active component study, many invest
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and 1185 kg ha -1 in the first site
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Table 1. Recommendation for use fer
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Sadanandan AK, Hamza S (1996c). Stu
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and it has been traced to South Ame
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Table 3. Results of phytochemical a
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(a) (b) (c) Figure 1. (a) Normal ce
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emedies in the south of Brazil. J.
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known fatty acids and terpenoids we
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Table 1. Renal function tests of ra
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Al-Radahe et al. 2271 Figure 3.nnnn
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namely disruption of the vascular e
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Zayachkivska OS, Konturek SJ, Drozd
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β-carotene were purchased from Sig
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Figure 1. Effect of ethanol concent
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Table 3. Climatic and soil factors
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content of polysaccharides. Phospha
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Serge et al. 2285 Table 1. Relative
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Percentage inhibition % of inhibiti
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Table 1. Result of phytochemical sc
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Figure 5. T. catappa Linn. (Combret
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Figure 8. A. fumigatus: Aspect of c
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Figure 12. Sensitivity of A. fumiga
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Noormi et al. 2311 Table 1. Callus
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Table 2. Contd. NoC=No callus. 4.0
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Figure 2. Contd. at 2.0 mg/L yellow
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Table 1. Biochemical parameters in
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Figure 3. Bcl-xL reactions in inter
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ACKNOWLEDGMENTS The authors would l
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Figure 3. The most widely included
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Figure 4. Lithographic print “St.
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Table 1. List of plants included in
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Table 1. Contd. Nedelcheva 2333 Cin
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Table 1. Contd. Nedelcheva 2335 Rhe
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14 7 1 15 1 35 17 16 Imported plant
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Academy of Science Publishing House
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et al., 1996; van Wyk and Gericke,
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DPPH inhibition (%) DPPH % inhibiti
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% Mortality Concentration (µg/ml)
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information on plants used for the
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Table 1. Levels of independent vari
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Arbutin (mg/g) Co-solvent Figure 1.
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Arbutin (mg/g) Extraction pressure
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Table 3. Arbuitn content of Asian p
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Arbutin (mg/g) Extraction pressure
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Arbutin (mg/g) Extraction temperatu
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Table 1. Chemical composition of es
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Darjazi 2369 Table 2. Statistical a
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UPCOMING CONFERENCES 15th European
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