Pharmacognostic and phytochemical investigation of leaves and ...

Research article 

Int J Pharm Biomed Sci 2012, 3(1), 13-19 

ISSN No: 0976-5263 

Research Drops 

PharmaInterScience Publishers 

Pharmacognostic and phytochemical investigation of leaves and 

flowers of Quisqualis indica Linn. 

V.A.Bairagi 1 *, P.R.Shinde 1 , 

K.L.Senthikumar 2 , N.Sandu 2 

1 KBHSS Trust’s Institute of Pharmacy, 

Bhaygaon road Malegaon- 423105, Dist- 

Nasik, Maharashtra, India 

2 Padmavati College of Pharmacy, 

Periyanahalli, Dharmapuri -635205, India. 

*Correspondence: 

Mr. V.A.bairagi 

Tel: +91 9422255266 

Fax: +91 2554250976 

E-mail: vinodbairagi@yahoo.com 

Quisqualis indica Linn. (Family Combretaceae) is commonly known as 

Rangoon creeper, traditionally used in cardiovascular disorder, fever, 

immunomodulatory. The fresh leaf and flower was studied for pharmaconostic 

evaluations, including examination of morphological and microscopic characters, 

determination of leaf constants, ash values and extractive values. The 

morphological studies revealed that the leaf is in dark green color with 

characteristic odour and slight bitter taste and the flowers are color varies in colors 

from white to pink to red. Flowers starts white and with age turn pink and then red 

with fragrant odour. The shape of Quisqualis indica leaves is as elliptical acuminate 

with entire margin, cordate base and its flowers grow in clusters and it blossoms 

year-round. Powder study of leaves revealed the presence of covering trichomes, 

annular xylem vessel, calcium oxalate crystals and anomocytic stomata and flowers 

revealed the presence glandular trichome, covering trichomes, oil glands, xylem 

vessels. Physicochemical parameters like ash value, extractive value and 

phytochemical screening with different reagents showed the presence of 

fluorescence compounds, steroids, triterpenoids, phenols, tannins and flavonoids. 

The phytochemical test and TLC study revealed the presence of slight amount of 

glycosides, tannins, flavonoids, terpenoids in petroleum extract of leaves and 

flower. And flavonoids, glycosides, tannins, terpenoids in methanolic extract of 

leaves and flower. 

Key words: Quisqualis indica, Pharmacognostic evaluation, Physico-chemical 

character, Phytochemical studies, TLC. 

Received: 28 Mar 2012 / Revised: 30 Mar 2012 / Accepted: 30 Mar 2012 / Online publication: 04 Apr 2012 

1. INTRODUCTION 

In the last few decades there has been an exponential 

growth in the field of herbal medicine. It is getting popularize 

in developing and developed countries owing to its natural 

origin and lesser side effects. In olden times, vaidyas used to 

treat patients on individual basis, and prepared drugs 

according to the requirement of the patients. But the scene 

has been changed now; herbal medicines are being 

manufactured on a large scale in mechanical units, where 

manufacturers are facing many problems such as availability 

of good quality raw material, authentication of raw material, 

availability of standards, proper standardization methodology 

of drugs and formulations, quality control parameters and etc 

[1,3]. 

Quisqualis indica Linn (Combreteceae) is a strong 

climber, ligneous vine that can reach from 2.5-8.0 meters 

(Fig.1). It is commonly known as Rangoon creeper. It is 

indigenous in Africa, Indo Malaysian region and cultivated 

all over India [7]. 

Fig.1: Quisqualis indica Linn. 

Flowers numerous, pendent, 7.5 cm long, 3.8 cm wide. At 

first they are white in colour then they become deep red [9]. 

The Indian traditional system of medicine, lays emphasis on 

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V.A.bairagi et al, Int J Pharm Biomed Sci 2012, 3(1), 13-19 

14 

promotion of health promotive, disease preventive and 

rejuvenation approach. The Indian traditional system of 

medicine, lays emphasis on promotion of health promotive, 

disease preventive and rejuvenation approach. Oxidative 

stress plays a major part in the development of chronic and 

degenerative ailments such as cancer, arthritis, aging, 

autoimmune disorders, cardiovascular and neurodegenerative 

diseases [11]. From the above literature, it is clear that no 

pharmacognostic work is carried out. The present study was 

therefore undertaken to investigate the pharmacognostic 

characters and phytochemical analysis of the plant. 

2. MATERIALS AND METHODS 

2.1 Plant material 

Fig.2. Transverse section of Quisqualis indica leaf 

The plant material was collected from the Malegaon city 

of Dist. Nasik, Maharashtra, in the month of September- 

2009 at the time of collection, plant including leaves and 

flowers were collected. The plant was authenticated by Mr. 

Arvind S. Dhabe (Herbarium in charge), Assistant Professor 

of Department of Botany. Dr. Babasaheb Ambedkar 

Marathwada University, Aurangabad, (M.S.) India. The 

leaves and flowers part were separated from other parts, 

washed, cleaned and dried for further use. 

2.2 Pharmacognostic studies 

2.2.1 Macroscopy 

Morphological studies were done by using simple 

microscope. The shape, apex, base, margin, taste and odor of 

leaves and flowers were determined. 

2.2.2 Microscopy 

Microscopic studies were done by preparing a thin hand 

section of midrib of leaves and pedicels of flower of 

Quisqualis indica Linn. The section were cleared with chloral 

hydrate solution, stained with Phloroglucinol and 

hydrochloric acid, and mounted with glycerin. A separate 

section were prepared and stained with iodine solution for the 

identification of starch grains. Transverse sections of the leaf 

and ovary with pedicel of Quisqualis indica are shown in 

Fig.2 and Fig.3, respectively. 

Powder of the dried leaves and flower were used for the 

observation of powder microscopical characters. The 

powdered drug was separately treated with phloroglucinol 

and HCl solution, glycerin and iodine solution to determine 

the presence of lignified cells, calcium oxalate crystals, 

trichomes and starch grains [8]. The results were registered 

by botanical illustration and photos taken by means of the 

Motic digital microscope (Motic instrument Inc, Canada) 

fitted with 1/3” CCD camera imaging accessory with motic 

image 2000 image analysis software. 

Fig.3. Transverse section of ovary with pedicel of Quisqualis indica 

2.3 Micro chemical investigation/ Histochemistry 

Micro-chemical tests and testing of behaviour of specific 

reagent towards plant drug tissue was carried out. The plant 

drug tissue was subjected to various reagents and based upon 

the inference, the histological zones were estimated [6, 16]. 

2.4 Quantitative microscopy 

As a part of quantitative microscopy, stomatal number, 

stomatal index, vein islet and vein termination number were 

determined by using fresh leaves of the plant [10]. 

2.5 Physico-chemical constants 

Total ash, water soluble ash, acid insoluble ash and 

sulphated ash were determined. Alcohol and water soluble 

extractive values were determined to find out the amount of 

water and alcohol soluble components. [5, 13]. 

2.6 Behavior of leaf powder with different chemicals 

Behavior of leaf powder with different chemical reagents 

was studied to detect the presence of phytoconstituents with 

color changes under daylight by reported method [15]. 



15 

Table 1 

TLC profile for petroleum ether and methanolic extract of Quisqualis indica 

Sl. No. Extract Mobile phase Detecting reagent 

1. PEL Pet. Ether : Ethyl acetate (8:2) Liebermann-burched reagent 

2. PEF Pet. Ether : Ethyl acetate (8:2) Liebermann-burched reagent 

3. MEL 1.Tolune: Ethyl acetate: formic acid (2.5:1:1) 

2. Ethyl acetate: Methanol : Water: Toluene (10:1.5:1.3:2) 

Methanolic FeCl 3 

Scan at UV 254& natural product reagent 

5. MEF 1.Tolune: Ethyl acetate: formic acid (2.5:1:1) 

2.Ethyl acetate: Methanol: Water: Toluene (10:1.5:1.3:2) 

Methanolic FeCl 3 

Scan at UV 254 & natural product reagent 

PEL: Petroleum ether extract of leaves, PEF: Petroleum extract of flower, MEL: methanolic extract of leaves, MEF: methanolic extract of flower. 

Table 2 

Histochemical color reactions of Quisqualis indica leaf and flower powder 

Reagents Constituents Colors Histological zones 

Aniline SO 4 + H 2 SO 4 Lignin Yellow Xylem 

Phloroglucinol + HCl Lignin Pink Xylem, Sclerenchyma 

Conc. H 2 SO 4 Cellulose Green Mesophyll 

Weak Iodine solution Starch -- -- 

Millon’s reagent Proteins White Spongy parenchyma 

H 2 SO 4 Ca. Oxalate Needles 

Mesophyll, and midrib 

SbCl 3 

Steroids/ 

Triterpenoids 

/prismatic 

Reddish pink 

parenchyma 

Mesophyll 

2.7 Successive solvent extraction 

The plant material leaves and flowers were air-dried at 

room temperature for 10 days and pulverized by grinder. 

Powdered of dried leaves and flowers of Quisqualis indica 

Linn (200 g) was successively extracted with petroleum ether 

(60-800) using a soxhlet apparatus. The extracts were 

concentrated and greenish yellow semisolid mass (coded 

“PEL and PEF”) were obtained. The resulting marc were 

exhaustively extracted by same method using methanol and 

greenish brown mass (coded “MEL and MEF”) were 

obtained after concentration on a water bath The extracts 

were stored aseptically in a desiccators at room temperature 

until demanded [14]. 

2.8 Phytochemical Screening 

The crude petroleum ether extract and methanol extracts 

were screened phytochemically for the presence of its 

constituents utilizing standard methods of analyses [2, 4, 17]. 

2.9 Thin layer chromatography 

For the TLC fingerprint the petroleum ether extract and 

methanolic extract were subjected to thin layer 

chromatographic analysis, to find the presence of number of 

chemical constituents to support the chemical test (Table 1). 

Analytical TLC plates were prepared by pouring the silica gel 

G slurry on the glass plates. Drying the thin layer plates, for 

30 min in air and then in an oven at 110°C for another 30 

min. For qualitative work, spot was applied in a row along 

one side of plate, about 2 cm from edge, by using capillary 

tubes. The range of sample volume was controlled, spreading 

not more than 0.5 cm. The plate was placed in previously 

saturated TLC chamber with mobile phase. The R f values 

were compared with standard drug and colours were recorded 

[12,18]. 

3. RESULTS 

3.1 Morphological study 

Quisqualis indica Linn is evergreen (in the tropics) and 

rambunctious vine, free branching and vigorous-growing, 

needing sturdy support. Macroscopically, the leaf is simple in 

composition, opposite in arrangement, apex and base are 

acuminate & cordate, margin is entire, venation is parallel, 

shape oblong- lanceolate and average leaf size is 6.8 cm ± 0.9 

(length) and 3.4 cm ± 0.2 (breadth). Fresh leaves are pale 

green in colour, odourless and tastless. Surface of leaf are 

silky and smooth. The midrib is prominent on lower surface. 

The flowers are raceme type arrangement; texture is very 

smooth & silky. The flowers are fragrant and tubular and 

their color varies from white to pink to red average size of 

flower about 30-35 cm long. Odour is aromatic, taste is 

aromatic & sweet. Flowers starts white and with age turn 

pink and then red. Its flowers grow in clusters and it 

blossoms year-round. Its flowers open at night with five 

bright red petals and gives out a distinct perfume (Fig.1). 

3.2 Micro chemical investigation/ Histochemistry 

Observation and result pertaining to microchemical tests 

and behavior of specific reagent towards plant tissue were 



16 

a. Parenchymatous tissue b. Unicellular trichome c. Calcium oxalate 

d. Anomocytic stomata e. Xylem vessels f. Glandular trichome 

g. Epidermis h. Oil glands (in flower) i. Spiral vessels (in flower) 

Fig.4 (a-i): Showing various characteristic features of flower and leaves of Quisqualis indica 

represented in Table 2. In the histochemical analysis the 

micro chemical tests showed the presence of midrib, vascular 

bundle and xylem vessels. 

3.3 Powder analysis 

The powder microscopy shows the fragments of 

unicellular covering and glandular trichomes, phloem fibers, 

parenchyma cells, numerous xylem vessels of spiral type and 

Epidermal cells with anomocytic stomata is shown in Fig.4. 

3.4 Quantitative microscopy 

As a part of quantitative microscopy, stomatal number, 

stomatal index, vein islet and vein termination number were 

determined by using fresh leaves of the plant (Table 3). 

Table 3 

Data representing values of quantitative microscopical studies of leaf 

Parameter 

Values 

Stomatal number 0.28-0.32 

Stomatal index 15.12-20.32 

Vein islet no. 2-5 

Vein termination no. 3-6 

Phloem fiber 

Length:6.76-78.21 

Width :1.06-1.45 

Calcium oxalate crystals 

Length:1.6-3.2 

Width :1.2-1.6 

Starch grains 1.56-8.0 

Trichomes: 

3.5 Physical constant 

Length:10.62-45.54 

Width :1.56-2.56 

The physical constant evaluation of the drugs is an 

important parameter in detecting adulteration or improper 

handling of drugs. The total ash is particularly important in 



17 

the evaluation of purity of drugs, i.e. the ash values, 

extractive values and moisture content of leaves and flowers 

were determined. The results are depicted in Table 4. 

Table 4 

Data representing physiological parameter 

Sl. No. Parameters % w/w (leaf) %w/w (flowers) 

1 Ash value 

Total ash 

Acid insoluble 

Water soluble 

Sulphated ash 

2 Extractive values 

Water soluble 

Alcohol soluble 

7 

1.5 

3.5 

5.67 

5 

1 

3 

3.45 

18 

11 

14.12 9.1 

3 Moisture content (LOD) 12 10 

Table 5 

Behavior of Ficus hispida leaf powder with different chemical reagents 

Regents Color/ppt Constituents 

Picric acid Slight ppt. Alkaloids present 

Conc. H 2 SO 4 Reddish brown Steroids/triterpenoids 

present 

Aq. FeCl 3 Bluish black ppt Tannins present 

Iodine solution No change Starch absent 

Ammonia No change Anthroquinone 

glycosides absent 

Spot test Stains observed Fixed oils present 

Aq. AgNO 3 Precipitation Proteins present 

Aq. NaOH Yellow Flavonoids present 

Mg – Hcl Magenta Flavonoids present 

Aq. Lead acetate White ppt Tannins present 

Liberman Burckhardt’s 

test 

Reddish green 

Steroids and tannins are 

present 

Table 6 

Phytochemical screening of Quisqualis indica linn. extracts 

Sl. No. Chemical test PEL PEF MEL MEF 

1 Alkaloids 

i.Dragendorff’s 

ii.Wagners 

2 Amino acids and proteins 

Xanthoproetic test + + ++ ++ 

3 Carbohydrates 

Molish test + + ++ ++ 

4 Flavonoids 

i. Shinoda test 

ii. Lead acetate test 

5 Glycoside 

i. Legal test 

ii. Baljet tet 

6 Tannins 

i. Lead acetate test 

ii. Ferric chloride test 

++ 

+ 

- 

+ 

- 

+ 

7 Phytosterol 

i. Salkowski test 

++ 

ii.Liebermann’s Burchard reaction ++ 

- 

- 

++ 

- 

- 

+ 

- 

+ 

- 

- 

++ 

++ 

Key: ++ = Highly present, + = faintly present, - = absent. 

3.6 Phytochemical investigation 

- 

+ 

++ 

++ 

- 

++ 

++ 

++ 

- 

- 

- 

+ 

++ 

++ 

- 

++ 

++ 

++ 

Successive extraction of the leaf and flower powder was 

carried out and methanolic extracts were found to give a 

maximum extractive yield. In the phytochemical tests the 

- 

- 

pet.ether extract revealed the presence of terpenoids, steroid, 

alkaloids, amino acids, and glycoside (for instance Table 5). 

Methanolic extract revealed presence of carbohydrates, 

flavonoids, vitamins, amino acids, proteins, tannins, 

glycosides and steroids. Preliminary phytochemical results of 

individual’s tests for various phytoconstituents are stated in 

Table 6. 

3.7 Thin layer chromatography 

The separation and purification of phytoconstituents of 

extract were mainly carried out using a combination of 

chromatographic techniques. The choice of techniques 

depends upon the solubility properties and volatilities of 

compound to be separated. Thin layer chromatography is a 

method of choice for separating all lipid soluble components, 

i.e. the lipids, steroids, carotenoids, flavonoids, glycosides, 

tannins, simple quinines and chlorophyll. TLC of PEL, PEF, 

MEL and MEF extracts resulted in Table 7 and Fig.5. 

4. DISCUSSION 

Microscopic analysis and qualitative parameters are 

carried out on plant samples in order to establish appropriate 

data that can be used in identifying crude drugs particularly 

those supplied in powder form. Powder study of leaves 

revealed the presence of covering trichomes, annular xylem 

vessel, calcium oxalate crystals and anomocytic stomata and 

flowers revealed the presence glandular trichome, covering 

trichomes, oil glands, xylem vessels. The Moisture content, 

Total ash, acid insoluble ash, water-soluble ash values and 

sulfated ash of leaves were observed to be 8%, 9%, 12.5%, 

6.55% and 5.45% w/w respectively and flowers were 10%, 

5%, 1%, 3% and 3.5% respectively. Water-soluble extractive 

values, alcohol soluble extractive value and petroleum ether 

soluble extractive value of the leaves were observed to 

be18%, 11% and 20% w/w, respectively and flower were 

11%, 9.1% and 12%, respectively. They are standard 

pharmacognostic parameters that can be used to differentiate 

closely related plant species or varieties with similar 

constituents or pharmacological activities. The behavior of 

the powder upon treatment with different chemical reagents 

was also observed and reported. Fluorescence studies of 

powder with various reagents revealed the presence of green 

fluorescence with conc. HCl and sodium hydroxide, under 

UV light. The various qualitative chemical tests of petroleum 

ether and methanol extract indicates the presence of sterols, 

triterpenoids, flavonoids, phenols and tannins in large 

amounts whereas aromatic acids, gums, mucilage were 

totally absent in the leaf and flower extract of this plant. 

Phytoconstituents of the leaves and flower have potential of 

organic acids, phytosterols, terpenoids, flavonoids, phenolic 

compounds, tannins and reducing sugars which has to found 

possesses antioxidant, mast cells stabilizing, can act against 

allergies, ulcers, tumors, platelet aggregation, and controlling 

hypertension and immunomodulatory effects. The 



18 

Table 7 

TLC pattern of different extract of leaves and flower 

Sl .No. Extract No. of spot &Color after spraying Rf value 

1. PEL 9 spot( blue, yellow, pink, pink, violet, violet, grey, blue, violet) 0.19, 0.3, 0.33, 0.37, 0.43, 0.61, 0.72, 0.89, 0.92 respectively. 

2. PEF 8 spots(brown, orange, pink, red, violet, grey, pink, blue) 0.64, 0.54,0.45 respectively 

3. MEL 1. solvent system :3 spots (deep blue) 

2. solvent system: 9 spot (yellowish orange colour) 

1.0.25, 0.32, 0.39 respectively 

2.0.25,0.31,0.35,0.37,0.4,0.55,0.7,0.81,0.92, respectively 

4. MEF 1. mobile phase: 5 spots ( deep blue) 

2. solvent system: 10spot(yellowish orange colour) 

1. 0.25, 0.32, 0.39 respectively 

2.0.25,0.31,0.35,0.37,0.4,0.55,0.7,0.81,0.92, 0.97 respectively 

A B C D 

Fig.5. Thin layer chromatography of 

(A) PEL & PEF extracts after derivatization with Libermann burched reagent 

(B) MEL& MEF extract after derivatization with ethanolic FeCl 3. 

(C) MEL & MEF extract after derivatization with Natural product reagent 

(D) MEL& MEF extract at 254 nm 

constituents of this plant have tremendous impact on the 

health care system and may provide medical health benefits 

including the prevention and or treatment of diseases. 

5. CONCLUSION 

Since the plant, Quisqualis indica which is also used for 

the treatment of various diseases and disorders, it is important 

to standardize it for use as a drug. The pharmacognostic 

constants for the leaves and flowers of this plant, the 

diagnostic microscopic features and the numerical standards 

reported in this work could be useful for the compilation of a 

suitable monograph for its proper identification. 

6. ACKNOWLEDGEMENT 

Authors are heartily thankful to Chairman, Mr. Baliramji 

Hiray and Chief Mentor, Mr. Prasad Hiray for providing 

necessary facilities at Institute of Pharmacy, and also thanks 

Mr. Arvind S. Dhabe (Herbarium in charge), Assistant 

professor of Department of botany. Dr. Babasaheb Ambedkar 

Marathwada University, Aurangabad for identification of the 

plant. 

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