GJRMI - Volume 4, Issue 11, November 2015 by Dr. Hari Venkatesh.K.Rajaraman

Indexing links of GJRMI GJRMI has been indexed in the Following International Databases

Google Scholar, ProQuest, DHARA online; DOAJ; Index Copernicus; NewJour; ScienceCentral; getCITED; RoMEO; Geneva Foundation for Medical Education & Research ; Catalog ebiblioteca; Ayurbhishak; Medicinal plants (Dravya Guna); Indianscience.in; Necker; Hong Kong University of Science and Technology Library; University of Zurich; University of Kansas; Western Theological Seminary; CaRLO; Mercyhurst University; University Library of Regensberg; WZB; Jadoun science; University of California, San Fransisco (UCSF Library); University of Washington; University of Saskatchewan; University of Winnipeg; Universal Impact Factor; Global Impact factor, Ulrich’s Periodicals Directory, New York Public Library, WISE, Cite factor, DRJI, Miami University Libraries, AYUSH RESEARCH PORTAL - Department of AYUSH, Ministry of Health & Family welfare, Govt. of India -

Chakradatta Ayurveda Chikitsalaya, Mysore. (Panchakarma & Netra Roga Chikitsa Kendra)

Get treated through Ayurveda, at our Hospital. (Exclusive Panchakarma Therapy available with accommodation) Address: Beside Vikram Jyothi Hospital, Temple Road, V V Mohalla, Mysore – 12, Karnataka, India.

Contact: Mobile: +919980952358, +919035087999 E- mail: raviamrita.kumar9@gmail.com

Consultant Physician: Dr. Ravi Kumar. M. (Specialized in different types of Keraliya Ayurvedic treatments especially in ENT & Eye diseases)

Arudra Ayurveda, Bangalore (A PANCHAKARMA TREATMENT CENTRE)

All types of Keraliya Ayurvedic treatments available for all the diseases) Ayurvedic Treatments in the following diseases: Eye diseases, Asthma, Skin diseases, Joint diseases, Diseases of the nervous system, Gynaecological & Obstetric diseases, Obesity, Asthma, Stress, Anxiety, Insomnia, Depression, Loss of Memory & Concentration, Piles, digestive tract diseases, Infertility etc. Address: No. 40, IInd cross, KV Pai Layout, Konanakunte, Near Silicon city school, Bangalore – 62, Karnataka, India.

Contact: Mobile: +919480748861

An International, Peer Reviewed, Open access, Monthly E-Journal

ISSN 2277 â&#x20AC;&#x201C; 4289 www.gjrmi.com Editor-in-chief Dr Hari Venkatesh K Rajaraman

Managing Editor Dr. Shwetha Hari

Administrator & Associate Editor Miss. Shyamala Rupavahini

Advisory Board Prof. Rabinarayan Acharya Dr. Dinesh Katoch Dr. S.N.Murthy Dr. Mathew Dan Mr. Tanay Bose Dr. Nagaraja T. M.

Editorial board Dr. Nithin Ujjaliya Mr. Sriram Sridharan Dr. Ashok B.K. Dr. Madhu .K.P Dr. Sushrutha .C.K Dr. Vidhya Priya Dharshini. K. R.

Honorary Members - Editorial Board Dr Farhad Mirzaei Dr. Sabarinath Subramaniam Dr. Yogitha Bali

INDEX – GJRMI - Volume 4, Issue 11, November 2015 MEDICINAL PLANTS RESEARCH Bio-Chemistry THE CURATIVE EFFECT OF ETHANOLIC EXTRACT OF JATROPHA CURCAS L. AGAINST PRACETAMOL INDUCED HEPATIC DAMAGE Mbama Uzochukwu V*, Ifeanacho MO, Onuoha SC

216–225

INDIGENOUS MEDICINE Ayurveda – Bhaishajya Kalpana PHARMACOGNOSTICAL AND PRELIMINARY PHYSICOCHEMICAL PROFILE OF ALBIZZIA LEBBECK BENTH. STEM BARK Harmeet Kaur*, CR Harisha, Galib R, PK Prajapati

226–235

COVER PAGE PHOTOGRAPHY: DR. HARI VENKATESH K R, PLANT ID – INFLORESCENCE OF MACROPTILIUM ATROPURPUREUM (DC.) URB.* OF THE FAMILY LEGUMINOSAE PLACE – OFF KANAKAPURA ROAD, BANGALORE, KARNATAKA, INDIA *BOTANICAL NAME VALIDATED FROM www.theplantlist.org AS ON 09/12/2015

Research article THE CURATIVE EFFECT OF ETHANOLIC EXTRACT OF JATROPHA CURCAS L. AGAINST PRACETAMOL INDUCED HEPATIC DAMAGE Mbama Uzochukwu V1*, Ifeanacho MO2, Onuoha SC3 1

Research student, Department of Biochemistry, University of Port Harcourt, Nigeria Lecturer, Department of Biochemistry, Universty of Port Harcourt, Nigeria *Corresponding author: E-mail: mbamauzo@gmail.com; Phone: +2347033623523. 2,3

Received: 08/07/2015; Revised: 20/10/2015; Accepted: 10/11/2015

ABSTRACT The effect of ethanolic extract of Jatropha curcas leaf against paracetamol-induced hepatotoxicity in Wistar albino rats was studied. A total of fourty two (42) rats were used for this study. The animals were grouped into six groups of seven rats each. Group one was the normal control, group two test control, group three, treatment control while group 4, 5 and 6 were treated groups with extract. The animals except those in group one received oral administration of 2000 mg/kg of paracetamol in 3% tween 20 to induce hepatotoxicity to them; and they were treated as follows; group two received only paracetamol, group three received standard drug silymarin (25 mg/kg), group 4, 5 and 6 received varying dose of Jatropha curcas extract (200 mg/kg, 400 mg/kg, 800 mg/kg) respectively. The course of hepatotoxicity were monitored and evaluated by estimation of three liver enzymes - AST, ALT, ALP; total bilirubin, total protein and albumin after 24 hours (week 0), and seven days (week 1). AST value for group 1 in week 0 (87.71 ± 1.71) was more than three folds elevated in group 2 (294.2 ± 17.71) at p ≤ 0.05. Group 3 (75.0 ± 0.95) showed a significant decrease at p ≤ 0.05 compared to group 2 (294.29 ± 17.71). Groups 4 (188.71 ± 1.44), 5(186.86 ± 0.34) and 6(166.87 ± 0.70) also showed significant decrease at p ≤ 0.05. Same trend followed for ALT and ALP activities and was observed in week 1. Significant increase were seen for protein and albumin for group 4 (6.47 ± 0.33) and (3.40 ± 0.10), group 5 (6.17 ± 0.20) and (3.00 ± 0.35), group 6 (6.33 ± 0.23) and (3.37 ± 0.34) compared to group 2 (5.73 ± 0.07) and (2.50 ± 0.17) respectively at p ≤ 0.05. The histology studies of the liver sample confirm the serum biochemical test results as the degenerated hepatocytes were gradually reversed to normal. These results suggest that ethanolic extract of Jatropha curcas leaves has a remedying effect against paracetamol-induced hepatotoxicity in Wistar albino rats. Jatropha curcas extract also showed a dose dependent effect. KEYWORDS: liver enzymes, Jatropha curcas, paracetamol, histology. ABBREVIATIONS: ALT- Alanine transaminase, AST- Aspartate transaminase, ALP- Alkaline phosphatase

Cite this article: Mbama Uzochukwu V, Ifeanacho MO, Onuoha SC (2015), THE CURATIVE EFFECT OF ETHANOLIC EXTRACT OF JATROPHA CURCAS AGAINST PRACETAMOL INDUCED HEPATIC DAMAGE, Global J Res. Med. Plants & Indigen. Med., Volume 4(11): 216–225

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Global J Res. Med. Plants & Indigen. Med. | Volume 4, Issue 11 | November 2015 | 216–225

INTRODUCTION The wide range of functions performed by liver like, playing a major role in metabolism and other functions including glycogen storage, decomposition of red blood cells, plasma protein synthesis, hormone production and detoxification etc., has made the liver to remain a very vital organ necessary for survival. There is no way to compensate the absence of the liver for a long time in the body system, although liver dialysis can be used short time (Maton et al., 1993). This is the more reason scientists are making efforts to use materials around us; like proven medicinal plants or other plants suspected to be potentially medicinal in preventing or remedying liver damage which most a times is chemically driven and has become a world problem today. Jatropha curcas L. (physic nut) which is a perennial plant, it is drought-resistant species which is widely cultivated in the tropics as a living fence. It is a small tree or large shrub which can reach a height of up to 5m. The plant shows articulated growth, with a morphological discontinuity at each increment. Dormancy is induced by fluctuations in rainfall and temperature/ light. The branches contain latex. Normally, five roots are formed from seedlings, one central and four peripheral. Jatropha curcas has 5 to 7 shallow lobed leaves with a length and width of 6 to 15 cm, which are arranged alternatively. Inflorescence is formed terminally on branches and is complex, possessing main and co-florescence with paracladia. Botanically, it can be described as a cyme. The plant monoecious and flowers are unisexual, occasionally hermaphrodite flowers occur (Dehgan and Webster, 1979). Branches are used as chewing stick in Nigeria (Isawumi, 1978). The sap following the stem is used to arrest bleeding of wounds (Nath and Dutta, 1992). Leaves of J.curcas are extensively used in ethnomedical practice in different forms for cure of various ailment like fever, mouth infections, guinea worm sores and joint rheumatism (Irvine, 1961; Oliver-Bever, 1986). The latex of J.curcas is reported to have

antibacterial activity against Staphylococcus aureus (Thomas 1989). J. curcas leaf extract, when combined with ciprofloxacin showed an increased activities against some selected micro organisms like Klebsiella pneumonia, Staphyllococcus aureus, Bacillius subtilis, Salmonella typhi, Pseudomonas aeruginosa, and Proteus mirabilis. The plant J. curcas is referred to as “Hospital too far” in Alu Choba, Port Harcourt Rivers State in Nigeria. It is traditionally used by the people as blood booster and also in the treatment of malnutrition in children. Drug induced liver injury is responsible for 5% of all hospital admission and 50% of all acute liver failures (Friedman et al., 2003) and (McNally & Peter F, 2006). Paracetamol induced hepatotoxicity in rodents is a widely used model to evaluate hepato protective potency of new compounds or drugs. It is a powerful inducer of cytochrome P-450 and produces a highly reactive quinineimine, which combines with sulphahydral groups of proteins and cause rapid depletion of intracellular glutathione (GSH). Normally GSH contributes significantly to the intracellular antioxidant defensive system as it is a powerful consumer of superoxide singlet oxygen, and hydroxyl radicals. The breakdown of the GSH dependent antioxidant defensive system increases the intracellular flux of oxygen apoptosis (Ladda et al., 2011). The rise in serum levels of AST, ALT and ALP has been attributed to the damaged structural integrity of the liver because these are cytoplasmic in location and are released into circulation after cellular damage (Mukherjee, 2008). The present study was taken to find out the possible actions of Jatropha curcas plant leaves ethanolic extract against paracetamol induced hepato toxicity in wistar albino rats using silymarin as a standard drug. MATERIALS AND METHODS Chemicals and reagents All chemicals and reagents used were of analytical grade and obtained from Randox

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Global J Res. Med. Plants & Indigen. Med. | Volume 4, Issue 11 | November 2015 | 216–225

Laboratories Ltd, United Kingdom. Emzor pharmaceuticals Ltd, Nigeria, Ben Merko Ltd, Nigeria. Preparation of plant extract Fresh leaves of Jatropha curcas were obtained from a garden at Alu, Port Harcourt, Rivers State. The plant was identified by Dr Nwosu E. of Plant Science and Biotechnology department, University of Port Harcourt, Port Harcourt, Nigeria. Leaves were washed under running water and air dried in shade in order to

preserve the phytochemical components of the leaves. After about 10 days, the leaves were crushed using a mill. The powdered leaves were soaked in 70% ethanol in a ratio of 1g of leaf: 5ml of ethanol. The resultant mixture were allowed to stand for 72 hours at room temperature with occasional shaking, thereafter it was filtered. The filtrate was placed in a rotary evaporator to recover some of the solvent, thereafter, water bath was used to dry the filtrate to molten form according to the method employed by Wo Jo et al. (2005).

Figure 1: Chart showing preparation of plant extract

Preparation of paracetamol Stock Concentration: 2000 mg of paracetamol was dissolved in 10ml of distilled water which gives a stock concentration of 200 mg/ml. Quantity Given To the Animals: Formula = volume= weight × dose ÷ concentration Where weight = weight of animal (g), dose= 2000 mg/kg, concentration= 200 mg/ml. Quantity of Extract Given To the Animals Stock Concentration: 1g of ethanolic extract of J. curcas was dissolved in 10 ml of distilled water which gives stock concentration of 100 mg/ml. actual quantity given to each

animal is calculated thus: volume = wt × dose ÷ concentration. Experimental animals Fourty-two (42) Wistar albino rats weighing 120–225g, bred in College of Veterinary Medicine, University of Nigeria, Nsukka were used. They were randomly selected and kept in six (6) groups of seven (7) rats each. Each group was kept in a separate cage. All the animals were fed with commercially produced feed obtained from Ben Merko Limited, 21 C to C Plaza, Nkponkiti, Enugu, Nigeria. Feed and water were given. The animals were subjected to 12 hours of light cycle in a properly ventilated room. The animals were allowed to acclimatize for a

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Global J Res. Med. Plants & Indigen. Med. | Volume 4, Issue 11 | November 2015 | 216–225

period of ten (10) days before experiment commenced. Experimental design The rats were divided into six (6) groups of seven (7) rats each. The groups were treated as follows: Group one (1) is the normal control and received no drug or extract treatment. Group two (2) is the test control; received oral administration of 2000 mg/kg/body wt of paracetamol each but no extract. Group three (3) is the treatment control; received 2000 mg/kg/body wt of paracetamol each, and were treated with 25 mg/kg/body wt of silymarin (as standard drug) for 7 days. Group 4, 5 and 6; the treatment 1, treatment 2, and treatment 3; received 2000 mg/kg/body wt of paracetamol and were treated with 200 mg/kg/body wt, 400mg/kg/body wt and 800 mg/kg/body wt of extract respectively for 7 days. Note: Treatment of paracetamol induced liver damage in animals with silymarin (group 3) and plant extract (group 4, 5 and 6) commenced immediately after the initial paracetamol treatment to avoid multiple death of animals. But group 2 was not given either silymarin or plant extract. Paracetamol, silymarin and plant extracts were given to the animals orally After 24 hours of paracetamol administration, blood samples were collected to determine the baseline result of the experiment. Then, three animals were randomly selected from each group and humanely sacrificed on the 8th day. Blood and liver samples were collected for analysis. Work was done with regards to ethics on animal research. Collection of samples for analysis Each animal to be sacrificed was withdrawn from its cage 24 hours after administration of

extract and blood was collected through ocular puncture (Retro orbital plexus). The fresh blood was collected into appropriately labeled heparin and EDTA sample bottles for biochemical and hematological tests respectively. The heparinized blood was centrifuged at 5000 rpm using an MSE centrifuge for 10 minutes to obtain plasma. Analyses of the plasma analytes were carried out at the College of Veterinary Medicine Laboratory, University of Nigeria, Nsukka. After collection of blood samples, the liver tissues of the rats were dissected and preserved in a sample bottle containing 10% formaldehyde. The liver samples were later processed at the histopathology laboratory of the Department of Anatomy, College of Veterinary Medicine, University of Nigeria, Nsukka. Results values were evaluated using one way analysis of variance (ANOVA). RESULTS & DISCUSSION Enzyme Activity This study as seen in tables 1–6 evaluates if the leaf extract of J. curcas has a curative effect on liver damaged by paracetamol over dose. The liver is prone to many diseases because of its strategic location and multi dimensional functions, (NDDIC, 2010). And, damage to the liver could be determined through elevated serum enzymes, specifically the transaminases, because liver damage often manifests as abnormal liver enzyme test (McNally & Peter F, 2006 and Ostapowicz et al., 2002) Damage to the liver (Hepatotoxicity) results not from paracetamol itself, but from one of its metabolites; N-acetyl-p-benzoquinoneimine (NAPQI). NAPQI depletes the liver’s natural antioxidant glutathione and directly damages cells in the liver leading to liver failure. Under normal conditions (NAPQI) is detoxified by conjugation with glutathione to form cysteine and mercapturic acid conjugates. But, in case of paracetamol over dose, the sulfate and glucuronide pathway becomes saturated and more paracetamol is shunted to cytochrome P450 system to produce NAPQI. As a result,

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Global J Res. Med. Plants & Indigen. Med. | Volume 4, Issue 11 | November 2015 | 216–225

hepatocellular supplies of glutathione become depleted as the demand for glutathione is higher than its regeneration, (Mitchell, et al., 1973). NAPQI, therefore remains in its toxic form in the liver and reacts with cellular membrane molecule, resulting in widespread hepatcyte damage and death, (acute necrosis) (Dai and Cederbaun, 1995). The values obtained during this experimental study were analysed using one way analysis of variance (ANOVA). For biochemical study, Serum Aspartate Amino traferase; AST (GOT) : At week 0, the highest value observed was that of group 2 (294.29 ± 17.71) which is the test control group. This value elevated more than three folds when compared to group 1 (88.71 ± 1.70) which is the normal rats group. Group 2 value is a clear indication of hepatocellular damage to the rats; though AST also indicate for other organs like the heart (Mumoli et al., 2006). This is in line with the works of (Ladda, et al., 2011 and Ramadoss, et al., 2012). Groups 4 (188.71 ± 1.44), 5 (186.86 ± 0.34), and 6 (166.87 ± 0.70) also showed the damaging effect of paracetamol; at the same time the remedying effects of J. curcas were obvious. Group 3 (75.00 ± 0.95) showed that silymarin is indeed a standard drug for liver treatment which agrees with the earlier observation made by Kanda et al. (2005). This trend continued for week 1. Also, J. curcas extract curative effect was seen to be at the peak in week 1 because group 5 (67.33 ± 7.68) and group 6 (64.33 ± 7.88) were of no significant difference with group 3 (69.67 ± 9.24). Serum Alanine Amino transferase ALT (GPT): the normal rats in group 1 (7.85 ± 0.46) showed the lowest value. Group 2 (128.86 ± 1.44) value elevated more than three folds compared to group 1. This is an indication of liver damage caused by the administration of paracetamol at 2000mg/kg, (Mumoli, N., et al., 2006). Also, groups 4 (132.14 ± 1.78), 5 (132.14 ± 1.78), and 6 (113.00 ± 3.69) showed

high values, but the effect of J. curcas extract in remedying the toxicity were obvious as there were significant decrease in group 6 (113.00 ± 3.69) value compared to group 2 (128.86 ± 1.44) value. This trend continued for week 1, there were significant differences recorded with groups 4(26.00 ± 4.62), 5(22.00 ± 3.46), and 6(18.00 ± 1.15) values from group 2(37.00 ± 1.73). Serum Alkaline phosphatase, ALP: though there was initial significant elevation of value when compared group 2 (29.71 ± 1.13) with group 1 (23.71 ± 0.36), it is observed that the elevation is not up to 2 folds. This confirms that the liver injury of the rats is that of hepatocellular necrosis and not cholestasis, (Benichou C, 1990). The curative effect of J. curcas were seen in groups 5 (26.14 ± 0.51) and 6 (26.85 ± 0.86) of week 0. Also, curative effects of J. curcas were seen in groups 4, 5 and 6 in subsequent week. Bilirubin: looking at week 0 values, group 1 (0.82 ± 0.07) differed significantly from group 2 (0.99 ± 0.06). Then groups 4 (0.59 ± 0.03), 5 (0.54 ± 0.00) and 6 (0.4 ± 0.03) also showed a significant decrease confirming the curative effect of J. curcas extract. This agrees with the early observation made by Kinda et al. (2005). The pattern of increase in values of bilirubin in the rats also confirm that the defect in the liver is not cholestatic; (Benichou, 1990). Protein and Albumin: protein and albumin are the major biochemical makers of the liver since albumin is the protein specifically made by the liver. The gradual increase in the values of these parameters over the period of this experimental study is a clear confirmation of the gradual cure and remedy of the diseased livers by J. curcas and silymarin. This agrees with the work of Bipin et al. (2009) and Ramdoss et al. (2012). The results of Histopathological analysis of the Liver have been depicted in Figure 2–7.

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Global J Res. Med. Plants & Indigen. Med. | Volume 4, Issue 11 | November 2015 | 216–225

Table 1: Effect of Ethanolic Extract of J. curcas on AST Activity Parameter Groups 1 No Treatment (+ve control) 2 Test Control (-ve control) 3 Treatment Control (silymarin) 4 Treatment 1 (extract 200 mg/kg) 5 Treatment 2 (extract 400 mg/kg) 6 Treatment 3 (extract 800 mg/kg)

AST (IU/L) WKO WK 1 89.71 ± 1.70a 80.00 ± 5.77a 294.29 ± 17.71b 88.66 ± 1.76 a 75.00 ± 0.95 69.67 ± 9.2 188.71 ± 1.44c 83.33 ± 8.12 186.86 ± 0.34c 67.33 ± 7.68 c 166.87 ± 0.70 64.33 ± 7.88a

abc given in superscript expresses the statistical significance (changes) between the various groups in the weeks. Values are mean ± Standard Error of Mean of triplicate determination. Values in the same column with different superscripts are statistically significant at 95% confidence level (P ≤ 0.05).

Table 2: Effect of Ethanolic Extract of J. curcas on ALT Activity Parameter Groups 1 No Treatment 2 Test Control 3 Treatment Control 4 Treatment 1 5 Treatment 2 6 Treatment 3

ALT (IU/L) WKO 7.85 ± 0.46a 128.86 ± 1.44b 39.71 ± 1.70c 132.14 ± 1.78b 132.14 ± 1.78b 113.00 ± 3.69d

WK 1 5.33 ± 1.76a 37.00 ± 1.73b 15.00 ± 0.58c 26.00 ± 4.62d 22.00 ± 3.46cd 18.00 ± 1.15cd

Table 3: Effect of Ethanolic Extract of J. curcas on ALP Activity Parameter Groups 1 2 3 4 5 6

Normal Control Test Control Treatment Control Treatment 1 Treatment 2 Treatment 3

ALP (IU/L) WKO 23.71 ± 0.36a 29.71 ± 1.13b 29.14 ± 0.34b 29.25± 048b 26.14 ± 0.51c 26.86 ± 0.86c

WK 1 16.00 ± 2.08ab 18.00 ± 1.15a 12.00 ± 1.15bc 12.33 ± 2.02bc 12.00 ± 1.54bc 10.00 ± 1.54c

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Global J Res. Med. Plants & Indigen. Med. | Volume 4, Issue 11 | November 2015 | 216–225

Biochemical Parameters Table 4: Effect of Ethanolic Extract of J. curcas on Bilirubin Levels Parameter Groups 1 Normal Control 2 Test Control 3 Treatment Control 4 Treatment 1 5 Treatment 2 6 Treatment 3

BILIRUBIN (mg/dl) WKO WK 1 a 0.82 ± 0.07 0.88 ± 0.07a 0.99 ± 0.06b 0.74 ± 0.03ab 1.13 ± 0.02c 0.55 ± 0.03c d 0.59 ± 0.3 0.68 ± 0.09bc 0.54 ± 0.00de 0.78 ± 0.03ab 0.46 ± 0.03e 0.68 ± 0.04bc

abc given in superscript expresses the statistcal significance (changes) between the various groups in the weeks. Values are mean ± Standard Error of Mean of triplicate determination. Values in the same column with different superscripts are statistically significant at 95% confidence level (P ≤ 0.05).

Table 5: Effect of Ethanolic Extract of J. curcas on Protein Levels

1 2 3 4 5 6

Parameter Groups Normal Control Test Control Treatment Control Treatment 1 Treatment 2 Treatment 3

PROTEIN (g/dl) WKO WK 1 5.54 ± 0.03 5.13 ± 0.74a 5.41 ± 0.07 5.73 ± 0.07ab 5.57 ± 0.17 6.27 ± 0.03b 5.50 ± 0.00 6.47 ± 0.33b 5.37 ± 0.05 6.17 ± 0.20b 5.38 ± 0.03 6.33 ± 0.23b

Table 6: Effect of Ethanolic Extract of J. curcas on Albumin Levels

1 2 3 4 5 6

Parameter Groups Normal Control Test Control Treatment Control Treatment 1 Treatment 2 Treatment 3

ALBUMIN (g/dl) WKO WK 1 a 2.66 ± 0.08 2.63 ± 0.43ac 2.53 ± 0.04ab 2.50 ± 0.17a 2.36 ± 0.07b 2.47 ± 0.07a c 2.78 ± 0.09 3.40 ± 0.10bc 2.79 ± 0.09c 3.70 ± 0.35b 2.49 ± 0.05ab 3.37 ± 0.34bc

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Global J Res. Med. Plants & Indigen. Med. | Volume 4, Issue 11 | November 2015 | 216–225

Figure 2: GROUP 1(NORMAL CONTROL)

cv A Photomicrograph of liver section from control rats (group1) showing normal hepatocytes (arrows) and the central vein (CV). H and E × 40

Figure 3: GROUP 2 ( TEST CONTROL)

A histologic section of liver from rats given 2000 mg/kg/body wt pcm only showing extensive hepatocyte degeneration and necrosis whitish or pale areas(arrows). H and E × 40

Figure 4: GROUP 3 ( TREATMENT CONTROL)

vc A histologic section of liver from rats given 2000 mg/kg/body wt pcm and treated with 25 mg/kg/body wt silymarin showing focal areas of hepatocyte degeneration and venous congestion (VC). H and E × 40.

Figure 5 : GROUP 4 ( 200mg/kg/body wt J. curcas)

vc A histologic section of liver from rats given 2000 mg/kg/body wt pcm and treated with 200 mg/kg/body wt of Jatropha carcus showing mild areas of hepatocyte degeneration (arrows) and venous congestion(VC).H and E × 40

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Global J Res. Med. Plants & Indigen. Med. | Volume 4, Issue 11 | November 2015 | 216–225

Figure 6: GROUP 5 ( 400mg/kg/body wt J. curcas)

A histologic section of liver from rats given 2000 mg/kg/body wt pcm and treated with 400 mg/kg/body wt of Jatropha carcus showing mild areas of hepatocyte degeneration (arrows). H and E × 40

Figure 7 : GROUP 6 (800 mg/kg/body wt J. curcas)

vc Plate 4.6: A histologic section of liver from rats given 2000 mg/kg/body wt pcm and treated with 800 mg/kg/body wt of Jatropha carcus showing severe venous congestion(vc). See also sinusoidal congestions(arrows).H and E × 40

CONCLUSION This study shows that administration of 2000 mg/kg paracetamol to experimental rats induced acute liver damage (necrosis) as seen by the abnormal increase in serum activities of AST, ALT and reduced albumin. The study suggests that the ethanol extract of Jatropha curcas is effective in remedying liver damage as it reduces abnormally elevated biochemical

parameters. Silymarin is also confirmed by this study to be indeed a standard and effective drug in treating liver disease. Histopathology result of this study confirmed the biochemical test results as the liver tissue which reveal degeneration and lesions were gradually reversed to normal by the administration of J. curcas and silymarin. Jatropha curcas also showed a dose dependent activity.

REFERENCES Akanwariwiak, W.G., Addo-Fordjour, P., Musah, A.A. (2002). Effect of combining crude extract ethanolic extract of Jatropha curcas L. leaf and some antibiotics against some selected micro organisms. Global Journal of research on medicinal plants and indigenous medicine. Vol 1(5): 140– 148.

Benichou .C. (1990). Criteria of drug-induced liver disorders. Report of an international concensus meeting. Journal of hepatology. 11 (2):2726. Bipin, B.P., Kalpesh, G., Jean, C.P., Nema, R.K., Sharma, C.S, Abhishek, K. (2009). Hepato protective Activity of Jatropha gossipifolia against carbon tetrachloride-induced hepatic injury in rats. Asian Journal of pharmaceutical and clinical research. 2(1);50–54.

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Global J Res. Med. Plants & Indigen. Med. | Volume 4, Issue 11 | November 2015 | 216–225

Dai,Y.,

and Cederbaum, A.I (1995). Cytotoxicity of acetaminophen in human cytochrome p 450 2E1. Journal of pharmacology and experimental therapautics. 273 (3):1497505

Dehgam, B. and Webster, G.L (1979). Morphology and generic relationship of genus Jatropha (Eup-horbiceae). University of California, Botany. Page 74. Mukherjee, P.K. (2008). Quality control of herbal drugs. An approach to evaluation of botanical. Business Horizons. Pharmaceutical Publishers. Page 600– 601.

(1993). Human Biology and Health. Englewood Cliff, New Jersey, USA. McNally, Peter F. (2006). GI/ Liver secrets: with student consult Access. Saint Louis: C.V Mosby Mitchell, J.R., Jollow, D.J., Potter w.z., Gillete, J.R., Brodie, B.B. (1973). Acetaminophen-induced hepatic necrosis iv. Protective role of glutathione. The journal of pharmacology and experimental therapeutics. 187(1):211–17. Mumoli, N., Cei, M., Cosimi A. (2006). Drugrelated hepatotoxicity. New England Journal of medicine. 354(4):21913.

Friedman, S.E., Grendell, J.H., McQuid, K.R. (2003). Current diagnosis and treatment in gastro-enterology. New York: Lang Medical Books/ McGraw Hill. Page 664–679

Nath, L.K., and Dutta, S.K. (1992). Wound healing response of the proteolytic enzyme curcain. Indian journal of pharmacology. 24(2):114–115.

Isawumi, M.A. (1978). Nigeria Chewing Sticks. The Nigerian field. 43(3):111– 127

National Digestive Diseases Information Clearing House (NDDIC) (2010). Cirrhosis overview.

Kanda, S.M., Veerenda , C.Y, Bhim, C.M and Tapan, K.M. (2005). Hepatoprotective and Anti-oxi-dant Role of Berberis tinctoris Lesch Leaves on Paracetamol induced hepatic damage in rats. Iranian Journal of Pharmacology and Therapeutics 4(1):6469.

Oliver-Bever, B.E.P (1986). Medicinal plants in tropical West Africa. Cambridge University press, Cambridge. 22–27.

Ladda, P.L., Magdum, S., Naikwade, N.S. (2011). Protective Activity Of Vitex Negundo Against Paracetamol Induced Hepatotoxicty In Rats. International Journal of Pharmaceutical Research. 1(1): 19 Maton, A., Jean H., Charles, W.M, Susan, J., Maryanna, Q.M., David, L., Jill, D.W.

Source of Support: NIL

Ostapowicz, G.,Fontana, R.J., Schiodt, F.V., Larson, A., Davern, T. J., Han, S.H., McCashland, T.M; Shakil, A.O., Hay, J.E., Hynan, L., Crippin, J.S., Blei, A.T., Samuel, G., Reisch, J., Lee, W.M. (2002). Results of a prospective study of acute liver failure at 17 tertiary care centers in the United States. Annual Internal Medicine. 137(12): 947–54. Thomas, O.O. (1989). Re-examination of the anti-microbial activities of Xylopia aethiopica, Carica papaya, Ocimum gratissimum and Jatropha curcas. Fitoterapia. 60(2): 147–155. Conflict of Interest: None Declared

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Research article PHARMACOGNOSTICAL AND PRELIMINARY PHYSICOCHEMICAL PROFILE OF ALBIZZIA LEBBECK BENTH. STEM BARK Harmeet Kaur1*, CR Harisha2, Galib R3, PK Prajapati4 1,3,4

Department of Rasashastra and Bhasihajya Kalpana, Institute for Post Graduate Teaching and Research in Ayurveda, Gujarat Ayurved University, Jamnagar, Gujarat, India 2 Pharmacognosy Laboratory, Institute for Post Graduate Teaching and Research in Ayurveda, Gujarat Ayurved University, Jamnagar, Gujarat, India *Corresponding Author: E mail- harmeetkaur134@yahoo.com; Mobile: +91 7698553796

Received: 09/10/2015; Revised: 25/10/2015; Accepted: 10/11/2015

ABSTRACT Albizzia lebbeck Benth., is one of the important herbal source, useful in many diseases . Different fractions of this plant are reported to have anti inflammatory, anti histaminic, anti microbial etc. activities. Pharmacognostical profiles of flower, leaf and heartwood of this plant are reported but the same for stem bark is not available. Considering this, an attempt has been made to evaluate pharmacognostical, phytochemical profile and to validate preliminary physicochemical profile of stem bark of the drug. Stem bark was collected in the month of February, 2013; shade dried; pulverized; sieved through 80# and preserved in an airtight glass container. Transverse section of stem bark and its powder, histochemical tests and preliminary physicochemical investigations were done to validate methods for quality control of drug and its botanical evaluation. Results showed stone cells with tannin, sclereids, rhomboidal prismatic crystals, cork cells, crystal fibre and oil globules. Preliminary physicochemical analysis revealed loss on drying (7.05%w/w), ash value (7.59%w/w), acid insoluble ash (0.68%w/w), alcohol soluble extractive (14.40%w/v) and water soluble extractive (16.30%w/v). Preliminary analysis for the presence of various functional groups revealed the presence of alkaloids, saponins, phenols and steroids. High Performance Thin Layer chromatography of alcoholic extract showed the presence of five and four spots in short UV and long UV respectively. The information generated in this study will provide relevant pharmacognostical and physicochemical data needed for proper identification, authentication and validation of pharmacopoeial characters of stem bark of Albizzia lebbeck Benth. KEYWORDS: Albizzia lebbeck, Ayurveda, Shirisha, Stem bark.

Cite this article: Harmeet Kaur, CR Harisha, Galib R, PK Prajapati (2015), PHARMACOGNOSTICAL AND PRELIMINARY PHYSICOCHEMICAL PROFILE OF ALBIZZIA LEBBECK BENTH. STEM BARK, Global J Res. Med. Plants & Indigen. Med., Volume 4(11): 226â&#x20AC;&#x201C;235

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Global J Res. Med. Plants & Indigen. Med. | Volume 4, Issue 11 | November 2015 | 226â&#x20AC;&#x201C;235

INTRODUCTION: Herbal medicine is a part of the knowledge of indigenous cultures and marginal societies across the globe which has stood the test of time (Prakash et al., 2014). During the last decade, use of traditional medicine has expanded globally and has gained popularity. Studies reveal that there are more traditional medicine providers than the allopathic providers especially in the rural areas (WHO, 2002). Development of these traditional systems of medicines with perspectives of safety, efficacy and quality control will help not only to preserve the traditional heritage but also to rationalize the use of natural products in health care (Mukherjhee et al., 2006). Majority of the world population still relies on herbal medicines to meet its health needs. It has not only continued to be used for primary health care of the poor in developing countries, but has also been used in countries where conventional medicine is predominant in the national health care systems. WHO is engaged to establish definitive guidelines for methodology of research and the appraisal of effectiveness of traditional medicine (Fabio Firenzuoli and Luigi Gori, 2007). Accurate identification and guarantee of purity through pharmacognosy and pharmaceutical chemistry measures is first step for the quality assurance and standardization of any of the herbal medicine. Albizzia lebbeck Benth., is one of such beneficial herbs. It is large erect unarmed deciduous, spreading tree common all over India, from the plains up to 900m in the Himalayas, and also in the Andamans (Wealth of India., 2006). Bark is expectorant (Tripathi et al., 1974), aphrodisiac, anti inflammatory, anti allergic (Tripathi et al., 1979), antianaphylactic (Johri et al., 1985), anti-asthmatic (Bhattathri et al., 1997), anti histaminic (Kumar et al., 2010), analgesic (Achinto et al., 2009), antioxidant (Resmi et al., 2006), immunomodulatory (Shyamlal et al., 2011.a), anticonvulsant (Kasture et al., 1996) and anti spermatogenic (Gupta et al., 2004). Pharmacognosy of heartwood (Shyamlal et al., 2011.b), flower (Shyamlal et al., 2011.c) and

leaf (Shyamlal et al., 2011.d) were reported, but was not available for stem bark. Considering these, detailed investigation of powdered stem bark of Albizzia lebbeck Benth. had been carried out using various pharmacognostical and phytochemical parameters. MATERIALS AND METHODS: Plant material: Stem bark of Albizzia lebbeck Benth. was collected from the Botanical garden of the Institute for Post Graduate Teaching & Research in Ayurveda (IPGT & RA), Gujarat Ayurved University, Jamnagar in the month of Feburary, 2013 [Figure 1]. Voucher specimen along with crude drug sample is preserved in the Pharmacognosy Lab, IPGT & RA, Gujarat Ayurved University, Jamnagar. Botanical identification was carried out by using various floras (Gamble., 1967). Macroscopic Characterization of stem bark: Macroscopic characters of stem bark were done by naked eye observations like shape, colour, taste and odour (Khandelwal, 2008). Microscopic Characterization of stem bark: For microscopical studies, free hand transverse sections of stem bark were taken and examined. Surface preparation was done and both the surfaces were observed. Powder microscopy of shade-dried powder (80#) was carried out using suitable method (Kokate, 2005.a). The powder was uniformly spread on glass slides and observed under microscope at different magnifications. For the detection of lignified tissues (stone cells, sclereids, xylem vessel, etc.), the powder was stained with phloroglucinol and hydrochloric acid and to observe the starch grains, the powder was stained with iodine solution (Trease, 1983). Photomicrographs were taken by using Carl zeiss binocular microscope attached with camera (Kokate, 2005.b). Physico-chemical Parameters of stem bark: Physico-chemical parameters were determined as per Ayurvedic Pharmacopoeia of

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Global J Res. Med. Plants & Indigen. Med. | Volume 4, Issue 11 | November 2015 | 226–235

India. Moisture content (The Ayurvedic Pharmacopoeia of India, 2001.a), total ash value (The Ayurvedic Pharmacopoeia of India, 2001.b), acid insoluble ash (The Ayurvedic Pharmacopoeia of India, 2001.c), alcohol soluble extractive value (The Ayurvedic Pharmacopoeia of India, 2001.d) and water soluble extractive value (The Ayurvedic Pharmacopoeia of India, 2001.e) were determined.

of rhytidoma and characteristic peeling of cork near the lenticels, colour of bark is buff with grayish white and blackish brown patches. Internal surface of bark is yellowish to grayish brown. Fracture in outer part of bark is laminated and inner part is fibrous and taste is astringent [Table-1, Figure 1:1–3].

Preliminary Phyto-chemical screening of stem bark:

Transverse section of stem bark of Albizzia lebbeck Benth.

The methanolic extract of stem bark was prepared and subjected to detect the presence of various functional groups like alkaloids, tannins, phenols, carbohydrates, glycosides, flavonoids, steroids, saponins by using relevant reagents. (Kokate, 2005.c) [Table-3].

The transverse sections of the mature bark are made up of alternate layers of hard and woody suberous tissues. The suberous tissues are broad and consist of several easily separable rows of thin walled rectangular cells some of which are empty, while others are filled with yellowish to brown contents. The secondary cortex or mid bark which is situated immediately beneath the rind is composed of large thin walled oblong cells. Most of them are loaded with starch grains while a number of others contain solitary prismatic crystals or yellowish brown contents which give the tissue its characteristic reddish or reddish purple colour. In this region there are numerous scattered groups of sclereids and stone cells of varies sizes and shapes.

HPTLC : For High Performance Thin Layer Chromatography (HPTLC), methanol extract was prepared by taking 5 g of drug in 100 ml of methanol, it was shaken for some time; mild heat was provided to it for half an hour and then filtered on cooling. The filtrate is evaporated on water bath to approximately 20 ml and used. A CAMAG (Switzerland) HPTLC system equipped with a sample applicator Linomat V was used for application of samples. CAMAG TLC Scanner 3, Reprostar and Wincats 4.02 were used for scanning the plates. CAMAG twin through glass chamber was used for developing the plates. Pre-coated silica gel GF 254 plate was used as stationary phase. Toluene : Ethyl acetate : formic acid ( 7 : 2 : 1) v/v was used as mobile phase. After 30 minutes of chamber saturation, plate was developed, and then scanned under short UV (254 nm) and long UV (366 nm).

Microscopic characteristics:

RESULTS AND DISCUSSION: Macroscopic Characters of stem bark:

The inner bark is a fairly broad zone composed of thin walled phloem alternating with tangential strips of sclerenchyma and medullary rays. The medullary rays are mostly biseriate while uni and triseriate rays are also present. The living tissue of the bark is mainly composed of wide secondary phloem consists of groups of tangentially running sieve tissues frequently getting obliterated companion cells and phloem parenchyma alternating with phloem fibre are associated with idioblast containing prisms of calcium oxalate crystals, groups of stone cells occasionally associated with fibres.

Macroscopically, bark pieces are slightly curved. External surface is very rough due to longitudinal and transverse fissures, exfoliation

The elements of wood are arranged in series. Extended xylem fibres form the mass of the wood [Figure 1:4–8 and Figure 2:1–4].

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Global J Res. Med. Plants & Indigen. Med. | Volume 4, Issue 11 | November 2015 | 226â&#x20AC;&#x201C;235

FIGURE 1- Morphology and transverse section of stem bark and its powder

1.Whole plant; 2. Stem bark of shirisha; 3. Powder of shirisha; 4. TS of bark with cork; 5.TS showing cortical region; 6. TS with pericyclic fibres and stone cells; 7. Parenchyma cells with tannin content; 8. Cork zone with tannin content

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Global J Res. Med. Plants & Indigen. Med. | Volume 4, Issue 11 | November 2015 | 226â&#x20AC;&#x201C;235

FIGURE 2- Transverse section of stem bark and its powder

1.Parenchyma cells with simple and compound starch grains; 2. Solid cork with stone cells layers; 3. Group of stone cells; 4. Sclereids

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Global J Res. Med. Plants & Indigen. Med. | Volume 4, Issue 11 | November 2015 | 226â&#x20AC;&#x201C;235

Powder microscopy of stem bark: Organoleptic characters are shown in [Table-1]. Plenty of stone cells with tannin, parenchymatous cells, sclereids, fragment of fibres, rhomboidal prismatic crystals, brown content, oil globule, cork with tannin content,

cork cells, fibres passing through medullary rays, lignified stone cells, lignified parenchymatous cells, corky surface and crystal fibre were observed under the microscope [Figure 3:1â&#x20AC;&#x201C;8].

Table-1: Organoleptic characters of stem bark powder of Albizia lebbeck Benth. Sr. no. 1 2 3 4

Character Colour Odour Texture Taste

Observation Brownish Sweetish to aromatic Rough Sweetish astringent

FIGURE 3- Powder microscopy of stem bark of shirisha

1.Cork in surface view; 2. Oil globule; 3. Tannin content; 4. lignified stone cells; 5. stone cells with tannin content; 6. rhomboidal crystals; 7. lignified pitted parenchyma; 8. lignified fibres

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Global J Res. Med. Plants & Indigen. Med. | Volume 4, Issue 11 | November 2015 | 226–235

Physicochemical Parameters of stem bark: Results of physicochemical parameters of stem bark are provided in [Table -2] Preliminary Phytochemical Screening of stem bark: Qualitative analysis for the presence of various functional groups was

carried out in methanol soluble extractive [Table-3]. HPTLC: Alcoholic extract of stem bark showed the presence of five and four spots in short UV (254 nm) and long UV (366 nm) respectively [Table-4] [Figure 4:1–2].

Table-2: Physicochemical evaluations of stem bark of Albizia lebbeck Benth. Parameters Values obtained (%) 7.05 w/w Loss on drying 7.59 w/w Ash Value 0.68 w/w Acid insoluble ash 16.30 w/v Water soluble extractive Alcohol soluble extractive 14.40 w/v

Values in API (%)Part I, Vol.III -----Not more than 8 percent Not more than 1 percent Not less than 6 percent Not less than 12 percent

Table-3: Qualitative analysis for presence of functional groups in Albizia lebbeck Benth. stem bark Sr. no. 1 2 3 4 5

6 7

Material

Reagents

Functional groups Dragendorff’s reagent Alkaloids Dil. FeCl3 Tannins Neutral FeCl3 Phenols Benedict’s reagent Carbohydrates Conc.H2SO4 Glycosides

Methanolic extract of stem bark powder Lead acetate flavonoid Chloroform, Steroids Conc.H2SO4, distilled water Shaking in test-tube Saponins

Observation

Result

No Brown ppt. Blue brownish color Violet color Yellow ppt. color change brown colour disappears

Present Present Present Present Present

Yellow color Brown ring rinse

Present with Present

Frothing honeycomb appearance

with Present

Table-4: HPTLC of Shirisha stem bark Chromatogram No. of spots Max. Rf value 5 0.04, 0.15, 0.53, 0.78,0.98 254 nm 4 0.04, 0.23, 0.35,0.78 366 nm

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Global J Res. Med. Plants & Indigen. Med. | Volume 4, Issue 11 | November 2015 | 226â&#x20AC;&#x201C;235

FIGURE 4- HPTLC of shirisha stem bark powder

1. AT 254nm 2. AT 365nm parenchymatous cells are other identification characters of stem bark. The physico-chemical CONCLUSION: profile observed in the current study was Pharmacognostical and physicochemical complying with the standards laid down in the evaluation of stem bark of Albizzia lebbeck Ayurvedic Pharmacopoeia of India, revealing Benth. provided specific parameters that will be its genuinity. As no published reports are helpful in proper identification, scientific available on Pharmacognostical aspects of the evaluation and authentication of the drug. stem bark; the results obtained may be referred Externally bark is whitish, crackled with as standard in future studies. rhomboidal crystals of calcium oxalate, crystal fibres and tannin content that are the specific ACKNOWLEDGEMENTS: characteristic features to identify Shirisha bark Authors are thankful to Dr. VJ Shukla, and its powder. Stone cells with tannin, Head, Pharmaceutical Chemistry Lab for parenchymatous cells, sclereids, brown content, providing basic facilities during Chemical oil globule, cork cells, lignified analysis.

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Global J Res. Med. Plants & Indigen. Med. | Volume 4, Issue 11 | November 2015 | 226–235

REFERENCES: Achinto Saha, Ahmad Muniruddin (2009). The analgesic and anti-inflammatory activities of the extract of Albizia lebbeck in animal model. Pak. J. Pharm. Sci. 22(1): 74–77. Bhattathri PPN, Rao PV, Acharya MV, Bhikshapathi T, Swami JK (1997). Clinical Evaluation of Shirisha Twak Kwatha in the management of Tamaka Shwasa. Journal of Research of Ayurveda and Siddha 18 (1–2): 21–27. Fabio Firenzuoli and Luigi Gori (2007). Herbal Medicine Today: Clinical and research Issues. Evidencebased Complementary and Alternative Medicine 4(1): 37–40. Gamble JS (1967). The Flora of Presidency of Madras. Vol. 1. London: Adlard and Sons. p. 1353. Gupta RS, Kachhawa JB and Chaudhary R (2004). Antifertility effects of methanolic pod extract of Albizia lebbeck Benth. in male rats. Asian J. Androl 6(2): 155–159. Johri RK, Zutshi U, Kameshwara L, Atal C K (1985). Effect of quercetin and Albizzia saponins on rat mast cell. Indian J. Physiol Pharmacol 29(1):43–46. Kasture VS, Pal SC (1996). Anticonvulsant activity of Albizzia lebbeck leaves. Indian journal of Experimental Biology 34(1):78–80. Khandelwal KR (2008). Practical and Pharmacognosy: Techniques and Experiments. 17th ed. Pune: Nirali Prakashan. p.10–19. Kokate CK (2005.a). Practical Pharmacognosy. 4th edition. Delhi: Published by Vallabh prakashan. p. 28.

Kokate CK (2005.b). Practical Pharmacognosy. 4th edition. Delhi: Published by Vallabh prakashan. p. 22–23. Kokate CK, Purohit AP, Gokhale SB (2005.c). Pharmacognosy: CBS Publishers and distributors. p. 169. Kumar S, Bansal P, Gupta V, Sannd R, Rao MM (2010). Clinical efficacy of Albizia lebbeck stem bark decoction on Bronchial asthma. International Journal of Pharmaceutical Science and Drug Research 2(1): 48–50. Mukherhee PK, Wahile A (2006). Integrated approach towards drug development from Ayurveda and other system of medicines. J Ethanopharmacology 103: 25–35. Prakash CP, Rakesh KM, Krishna GS (2014). The efficacy of herbal system of medicine in the context of allopathic system in Indian Central Himalaya. Journal of Herbal Medicine 4(3): 147– 158. Resmi CR. Antioxidant activity of albizia lebbeck in alloxan diabetic rats (2006). Indian J Physiol Pharmacol 50(3): 297– 302. Shyamlal SY, Galib, Ravishankar, Prajapati PK, Ashok BK (2011.a). Evaluation of Immunomodulatory activity of “Shirishavaleha” - An Ayurvedic Compound formulation in albino rats. Journal of Ayurveda and Integrative Medicine 2(4): 192–196.

Shyamlal SY (2011.b). Pharmacognostical Screening and Phytochemical Evaluation of Albizia lebbeck Benth. Heartwood. Asian Journal of Biomedical and Pharmaceutical Sciences 1(5), 01–06.

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Global J Res. Med. Plants & Indigen. Med. | Volume 4, Issue 11 | November 2015 | 226–235

Shyamlal SY (2011.c). Pharmacognostical and Physico-Chemical Investigations of Albizia lebbeck Benth. Flower. International Journal of Pharmaceutical & Biological Archives 2(5):1434– 1438. Shyamlal SY (2011.d). Pharmacognostical and physicochemical investigations of Albizia Lebbeck Benth. Leaf. International Journal of Science Innovations and Discoveries 1 (2), 178– 185. The

The

Ayurvedic Pharmacopoeia of India (2001.a). Reprinted 1st ed. Part 1, Vol. I. Appendix-2(2.2.9) Govt. of India: Ministry of Health and Family Welfare. New Delhi . p. 143. Ayurvedic Pharmacopoeia of India (2001.b). Part 1, Vol. 5. Appendix2(2.2.3.) Govt. of India: Ministry of Health and Family Welfare. New Delhi. p. 213. Ayurvedic Pharmacopoeia of India (2001.c). Part 1, Vol. 5. Appendix2(2.2.4.) Govt. of India: Ministry of Health and Family Welfare. New Delhi. p.213.

The

Ayurvedic Pharmacopoeia of India (2001.d). Reprinted 1st ed. Part 1, Vol. I. Appendix-2(2.2.6) Govt. of India: Ministry of Health and Family Welfare. New Delhi. p. 143.

The

Ayurvedic Pharmacopoeia of India (2001.e). Reprinted 1st ed. Part 1, Vol. I. Appendix-2(2.2.7) Govt. of India: Ministry of Health and Family Welfare. New Delhi. p. 143.

Trease GE, Evans WC (1983). Pharmacognosy. 12th ed. U.K: Baillier Tindall Can Macmillan Publishers. p. 95–9, 512–47. Tripathi RM, Sen PC, Das PK (1979). Studies on the mechanism of action of Albizzia lebbeck, an Indian indigenous drug used in the treatment of atopic allergy. J Ethnopharmacol 1(4):385–396. Tripathi VJ, Ray AB, Das Gupta B (1974). Neutral constituents of Albizzia lebbeck. Curr Sci 43:46–8. Wealth of India: A dictionary of Indian Raw Materials and Industrial Products (2006).1st ed. Vol. I: A. New Delhi. CSIR. p.126. WHO. WHO Traditional Medicine Strategy 2002–2005 (2002). Geneva: World Health Organization.

Source of Support: NIL

Conflict of Interest: None Declared

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Call for Papers â&#x20AC;&#x201C; Vol. 5, Issue 1, January 2016 Submit your manuscripts (Research articles, Review articles, Short Communications, Letters to the Editor, Book Reviews) to Global Journal of Research on Medicinal plants & Indigenous medicine â&#x20AC;&#x201C; GJRMI Submit it online through www.gjrmi.com or mail it to submitarticle@gjrmi.com on or before December 10th 2015.

To advertise on the Flip book Cover page freely, write to chiefeditor@gjrmi.com or editorinchief@gjrmi.com Or Call - +919590574495