Khyade MS et al / IJRAP 2011, 2 (1) 177-180

International Journal of Research in Ayurveda & Pharmacy, 2(1), Jan-Feb 2011 177-180

Research Article Available online through www.ijrap.net ISSN 2229-3566

PHARMACOGNOSTICAL AND PHYTOCHEMICAL EVALUATION OF LEAF OF

JATROPHA GOSSYPIFOLIA L Khyade MS*1 and Vaikos NP2

1Post Graduate Department of Botany, Sangamner Nagarpalika Arts, D.J. Malpani Commerce and B.N. Sarda Science College, Sangamner-422 605 (MS), India

2Department of Botany, Dr. Babasaheb Ambedkar, Marathwada University, Aurangabad-431004 (MS) India

Received on: 24/12/2010 Revised on: 28/01/2011 Accepted on: 09/02/2011

ABSTRACT The present study deals with pharmacognostical and phytochemical evaluation of leaves of Jatropha gossypifolia L. (Euphorbiaceae). Macromorphology, microscopy (transverse section, histochemical colour reactions and quantitative microscopy) and phytochemical components were studied to establish the salient diagnostic characters. WHO recommends physicochemical determination and phytochemical evaluation for quality control of medicinal plant materials. The various morphological, microscopical, physicochemical standards developed in this study will help for botanical identification and standardization of the drug in crude form. KEYWORDS: Jatropha gossypifolia, Pharmacognostical, Phytochemical, evaluation. *Correspondence Address Dr. Khyade MS, Post Graduate Department of Botany, Sangamner Nagarpalika Arts, D.J. Malpani Commerce and B.N. Sarda Science College, Sangamner-422 605 (MS), India Email: mskhyade@rediffmail.com , drvaikos@gmail.com INTRODUCTION Jatropha gossypifolia L. (Euphorbiaceae) is a small armed shrub, growing almost all over the India1. Its leaves are used in the Indian folk medicine for treatment of number of ailments including blood purifier, a potent antiseptic, ulcers, carbuncles, eczema and in itche, on sores in the children tongue, leaves applied on swollen breasts and febrifuge1-3. Decoction of leaves is effective as purgative and stomachic, whereas leaves are employed in intermittent fevers and swollen mammae4,2

.Crushed leaves mixed with breast milk are applied on the head to cure infantile diarrhea5. Various studies have been carried out with leaves and other organs of J. gossypifolia on the antibacterial6, anti-coagulant7, anti-inflammatory, analgesic activities8, alkaloids9, flavonoid10 triterpenoid11,13 and lignan12. Insufficient data was found in the literature about the pharmacognostical characteristics of the leaves of J. gossypifolia. Hence, the present study was designed to study the detail pharmacognostic study of the leaves including macroscopy, microscopy, quantitative microscopy, determination of various physicochemical parameters, and phytochemical screening.

MATERIALS AND METHODS The leaves of Jatropha gossypifolia L. were collected from the local areas of Aurangabad district Maharashtra, and it was identified, confirmed and authenticated by comparison with an authentic specimen at Department of Botany, Dr. Babasaheb Ambedkar Marathwada University, Aurangabad (MS), India. A voucher specimen of the plant was deposited in the Departmental herbarium for the future reference. The anatomical studies were carried out as per standard methods14,15. For anatomical studies, sections of 10–12μm thick were prepared and stained with phloroglucinol-HCl (1:1). The quantitative leaf microscopy like palisade ratio, stomatal index, vein islet and vein termination number were carried out as described methods16-18. Histochemical colour reactions were performed by the standard methods19,20. The physico-chemical characters such as extractive values, ash values, loss on drying were performed as per the official standard procedures21,22. Coarsely powdered leaves were subjected to soxhlet extraction for fractionation with petroleum ether, chloroform, acetone and methanol in the increasing order

Khyade MS et al / IJRAP 2011, 2 (1) 177-180

International Journal of Research in Ayurveda & Pharmacy, 2(1), Jan-Feb 2011 177-180

of polarity. All fractions were further subjected to qualitative test for identification of various phytoconstituents23-26. RESULTS AND DISCUSSION Leaves are green or more often dark purplish-red; alternate, orbicular to broadly obovate in outline, 10-15x12-16 cm, cordate at base, 3-5 fid beyond the middle; lobes obovate, acute or acuminate, ciliate with stalked glands (Fig.3B). Petioles as long as the blades, with stalked glands on the upper surface; stipules modified into decurrently rows of stalked glands (Fig. 1). In transaction view, petiole appears oval in shape with adaxial side becoming flat. The epidermis consists of tightly arranged cells, with thick cuticle outside. Glandular and non glandular trichomes occur. It is flowed by 2-3 layered collenchyma. The cortex and pith are parenchymatous. The vascular tissue is cylindrical and forming more or less zigzag structure. Xylem is surrounded by phloem outer and inner side and sclerenchyma encloses the vascular tissue. (Fig 2). Leaf is dorsiventral and amphistomatic. Adaxial epidermis has barrel shaped cells which are compactly arranged with thick cuticle. Abaxial epidermis is of comparatively smaller cells with thin cuticle. Stomata are paracytic and occur on both the surface but, lower surface has more stomata (Fig.4 a,b). Trichomes are glandular and non glandular. Mesophyll is made up of two types of cells, palisade and spongy tissue. Palisade is 1-2 layered and spongy tissue is of loosely arranged cells. Vascular bundles are bicollateral, conjoint with xylem facing upwards. Calcium oxalate crystals occur more in phloem cells as compared to cortical cells. Hypodermis in the midrib region is collenchymatous followed by parenchymatous cortex. Pith is parenchymatous. Vascular tissue forms a distinct arc, few vascular bundles occur towards adaxial side at two end of arc forming the entire structure into circular shape (Fig. 3A) The leaf microscopic characters like stomatal frequency, stomatal index, vein islet number and vein termination number were determined (Table 1).The quantitative determination of some pharmacognostic parameters are useful for setting standards for crude drugs. The vein islet and vein termination numbers and other parameters determined in the quantitative microscopy, are relatively constant for plants and can be used to differentiate closely related species. The histochemical color reactions were carried out on transverse section of the fresh leaf (Table 2). The results indicated the presence of lignin, starch, fats, saponins, tannins and calcium oxalate crystals. Histochemical localization of certain important compounds enables to

get a preliminary idea of type of compounds and their accumulation in the plant tissues. Based on this study, one can chose the organ or tissue where the required compounds are located. The physico-chemical characters such as moisture content, total ash, acid insoluble ash, acid soluble ash and extractive values in chloroform, alcohol and water of the dried leaf powder were calculated in terms of air dried sample as shown in (Table 3). The physical constants evaluation of the drug is an important parameter for in detecting adulteration or improper handling of the drugs. The moisture content of the drug is not high (4.15%), thus it discourages bacterial, fungal or yeast growth, as the general requirement for moisture content in crude drug is not more than 14 %. Equally important in the evaluation of crude drugs, is the ash value and acid insoluble ash value determination. The total ash is particularly important in the evaluation of purity of drugs, i.e. the presence or absence of foreign inorganic matter such as metallic salts or silica. Thus it appears that the plant J. gossypifolia is useful in the traditional medicine. The results of qualitative phytochemicals are presented (Table 4). Different chemical compound such as anthraquinones, flavonoids, phlobatannin, phenolics, saponins, tannins and terpenoids are detected in J.gossypifolia, which could make the plant useful in treating different ailments and having potential for providing useful drug for human use. This is because the pharmacological activity of any plant is usually traced to particular compound. The presence of tannins and other phenolics compounds, which have antiseptic properties, could explain the use of this plant for the treatment of various diseases. It has also been mentioned that the antioxidant activity of plants might be due to their phenolic compounds27. Flavonoids are a group of polyphenolic compounds with known properties which include free radical scavenging, inhibition of hydrolytic enzymes and anti-inflammatory action28. The saponin reduces the body cholesterol by preventing its reabsorption, increasing its excretion, thereby reducing blood pressure29. The present study on Pharmacognostical and Phytochemical evaluation of Jatropha gossypifolia will provide useful information for its identification. Macro, micro and physiochemical standards discussed above can be considered as the identifying parameters to substantiate and authenticate the drug, since no such data is available for the same. ACKNOWLEDGEMENT The authors wish to thank the Head, Department of Botany, Dr. Babasaheb Ambedkar Marathwada

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International Journal of Research in Ayurveda & Pharmacy, 2(1), Jan-Feb 2011 177-180

University, Aurangabad for providing the necessary laboratory facilities. REFERENCES 1. Agarwal CK. Economic Plants of India. Calcutta, India: Kailash

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10. Subramanian SS, Nagarajan S, Sulochana N. Flavonoids of the Leaves of Jatropha gosssypifolia. Phytochem 1971; 10: 1690.

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12. Das R, Das B and Kashinatham A. Gossypiline, a new lignan from Jatropha gossypifolia. Nat Prod Sci 1998;4(4): 238-240.

13. Tinto WF, John MD. Triterpenoids of Jatropha gosssypifolia. J Nat Prod 1992; 55(6): 807-809.

14. WHO-Geneva, Quality control methods for medicinal plant materials. New Delhi: A.I.T.B.S. Publisher and Distributors; 2002.

15. Kokate CK, Khandelwal KR, Pawar AP, Gokhale SB. Practical Pharmacognosy. Pune, India; Nirali Prakashan; 1995.

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17. Salisbury EJ. The interpretation of soil climate and the use of stomatal frequency as an interesting index of water relation to the plant. Beih Bot Zentralb, 1932; 49: 408-20.

18. Levin FA. The taxonomic value of the vein islet areas based upon a study of the genera Berosma, Cassia, Erythroxylon and Digitalis. Journal of Pharmamcy and Pharmacology, 1929; 2: 17-43.

19. Johansen DA. Plant Microtechnique. Tata McGraw Hill Publishing Company Ltd., New Delhi, India; 1940.

20. Guerin HP, Delaveau PG, Paris RR. Bullein de La Societe Botanique de France, 1971; 118: 29 – 36.

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Table 1: Quantitative microscopy Parameters Range Mean Palisade ratio 4-7 6.12

Stomatal frequency Upper 5-9 6.75 Lower 9-15 12.88

Stomatal index Upper 6.84-9.57 7.84 Lower 12.63-18.18 15.26

Vein islet number 20-27 25.1 Vein termination 14-23 20.8

Table 2: Histochemical colour reactions

REAGENTS

CONSTITUENTS COLOUR HISTOLOGICAL ZONE DEGREE OF

INTENSITY Aniline SO4 + H2SO4 Lignin Yellow Xy. Scl. +++ Weak Iodine solution Starch Blue Mesophyll ++ Sudan III / IV Fats Pink / Red Mesophyll, M.cor ++ Dragendroffs reagent Alkaloids -- -- -- Ba (OH)2 + K2Cr2O7 + CaCl2 Saponins Yellow Mesophyll + Fecl3 Tannins Blue green M. cor. ++ Vanillin + HCl Flavonoids -- -- -- AgNO3 + H2O2 Crystals Black M.cor. ++

$ Note= M. cor. - Midrib cortex; Xy. - Xylem; Scl. - Sclerenchyma; M.col- Midrib collenchyma.

Khyade MS et al / IJRAP 2011, 2 (1) 177-180

International Journal of Research in Ayurveda & Pharmacy, 2(1), Jan-Feb 2011 177-180

Table 3: Physical evaluation (%) Moisture content:

4.15

Extractive values: a) Chloroform 8.0

b) Alcohol 4.96 c) Water 29.2

Ash values: a) Total ash 15.8

b) A.I.A. 1.05 c) A.S.A 14.75

Table 4: Qualitative chemical analysis

Chemical constituents Observation Acubins / Iridoids

--

Alkaloids a) Dragendorff’s reagent -- b) Mayer’s reagent -- c) Wagner’s reagent -- Anthraquinone ++ Cardiac glycoside -- Coumarins -- Flavonoids ++ Leucoanthocyanins -- Phlobatannin +++ Simple phenolics + Steroids -- Saponins +++ Tannins Test – a true tannin ++ Test – b pseudotannin ++ Terpenoid ++

Fig. 1 A flowering twig with leaves

Fig 2: T. S. of Petiole [Col- Collenchyma; CO- Cortex; VB- vascular bundles]

A B

Fig 3: A T. S. of Leaf [Epi- Epidermis; Pal- Palisade cells; S- spongy cells] and B- a part of leaf margin

Fig 4: Epidermal structures [a- Adaxial, b- Abaxial epidermis]

Source of support: Nil, Conflict of interest: None Declared