ISSN 2277-4289 www.gjrmi.com International, Peer reviewed, Open access, Monthly Online Journal Research Article PRELIMINARY PHYTOCHEMICAL EVALUATION OF THE GENUINE AND MARKET SAMPLES OF THE STEM BARK OF CHIRABILWA (HOLOPTELEA INTEGRIFOLIA PLANCH.) Sinimol T P 1*, Shahul Hameed A 2 1 Research Officer (Ayurveda) Incharge, Regional Research Centre of Ayurveda, Port Blair, India 2 Associate Professor, Department of Dravyagunavijnan, Govt. Ayurveda College, Kannur, Kerala, India *Corresponding author: Email: drsinitp@gmail.com; Mob: 9446519427 ABSTRACT Received: 22/3/2017; Revised: 25/08/2017; Accepted: 31/08/2017 Chirabilwa [Holoptelea integrifolia Planch.], a large deciduous tree has been used in the treatment of various diseases including Dyspepsia, Flatulence, Colic, Helminthiasis, Vomiting, Skin diseases, Leprosy, Diabetes Mellitus, Haemorrhoids and Rheumatism. As a part of a proposed clinical study, sufficient quantity of the plant drug was required to evaluate the antidyslipidemic effects. So this opened up another area of research wherein for such huge requirements the researchers have to either depend upon market samples or collect the drug directly from the wild. Hence it was planned to do the phytochemical and physicochemical evaluation to check the genuinity and purity of market sample against the genuine hand collected sample. Shade dried samples of both market and genuine samples of the drug were subjected to analysis. Qualitative analysis indicated the presence of steroids and traces of alkaloids. Comparable results were obtained which proved the genuinity of the market samples. KEYWORDS: Phytochemical, Chirabilwa, Holoptelea integrifolia Planch. Cite this article: Sinimol T P, Shahul Hameed A (2017), PRELIMINARY PHYTOCHEMICAL EVALUATION OF THE GENUINE AND MARKET SAMPLES OF THE STEM BARK OF CHIRABILWA (HOLOPTELEA INTEGRIFOLIA PLANCH.), Global J Res. Med. Plants & Indigen. Med., Volume 6 (8): 102 108
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). 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). Preliminary phytochemical evaluation is a step towards this. Plant is a biosynthetic laboratory not only for primary metabolites but also for a multitude of secondary metabolites like steroids, glycosides, tannins etc. which exert the physiological and therapeutic effects of the plant. Preliminary phytochemical screenings are done to detect the presence of these constituents in a drug. Chirabilwa [Holoptelea integrifolia (Roxb.) (Planch.)] is a large spreading almost glabrous deciduous tree attaining a height of 15 18m, trunk upto 2 metres or more in girth near the ground level, covered with grey pustular mucilaginous bark (Kirtikar, K.R. and Basu, B.D, 2005). Main pharmacological properties include anti-inflammatory, digestive, carminative, laxative, anthelmintic, depurative, revulsive and urinary astringent action. They are useful in vitiated conditions of kapha and pitta, inflammations, dyspepsia, flatulence, colic, helminthiasis, vomiting, skin diseases, leprosy, diabetes, haemorrhoids and rheumatism (Warrier, P.K. et al., 1995). Chirabilwa [Holoptelea integrifolia (Roxb.) (Planch.)] is a tree which is included in many medohara groups like Saalasaradi, Varanadi, Aragwadadi, Arkadi (various groups of herbs) groups of Susruta Samhita (Yadavji Trikamji, 2004), Asanadi, Varanadi, Aragwadadi and Arkadi groups of Ashtanga Hridaya (Srikantha murthy K.R., 2002) and Lekhaniya mahakasaya group of Caraka Samhita (Paradkar Harishastri, 2006). But no research has been conducted yet to clinically evaluate its medohara (Dyslipidemic) effect as a single drug. Hence a clinical study to evaluate the anti dyslipidaemic effect of the stem bark of Chirabilwa was taken up. As a part of a proposed clinical study, sufficient quantity of the plant drug was required to evaluate the antidyslipidemic effects. So this opened up another area of research wherein for such huge requirements the researchers have to either depend upon market samples or collect the drug directly from the wild. Hence it was planned to do the phytochemical and physicochemical evaluation to check the genuinity and purity of market sample against the genuine hand collected sample. Shade dried samples of both market and genuine samples of the drug were subjected to analysis. The values of this analysis were compared to prove the genuineness of the study drug. These tests are simple and quick to perform and give valuable information about the nature, genuineness and purity of a drug. Physical analysis includes parameters such as moisture content, volatile oil content etc. Physico chemical analysis includes chromatographic techniques like TLC, HPTLC etc. MATERIALS AND METHODS The genuine samples of the stem bark of Chirabilwa (Holoptelea integrifolia Planch.) were collected from the herbal garden of Pharmacognosy Unit, Govt. Ayurveda College, Trivandrum, India during January 2011. The plant identification was done by Mr. G.R. Jayakumar, Senior Research officer, Pharmacognosy Unit, Govt. Ayurveda College, Trivandrum. Market samples were collected from the local market of Trivandrum, India during January 2011. Voucher specimens were preserved in the Herbarium at the Department of Dravyagunavijnana, Ahaliya Ayurveda College, Kerala, India [Figure 1]. The collected stem barks of Chirabilwa (Holoptelea integrifolia Planch.) were washed well to remove mud and soil contaminants, dried in shade and powdered using a micropulveriser to the mesh size 60.
Figure 1:-Morphology of the stem bark and its choorna of the genuine and market samples of Chirabilwa (Holoptelea integrifolia Planch.) 1 2 3 4 1. Stem bark of H. integrifolia (genuine); 2. Stem bark of H. integrifolia (market) 3. Powder of stem bark of H. integrifolia (genuine); 4. Powder of Stem bark of H. integrifolia (market) Physico-chemical Parameters of stem bark: Physico-chemical parameters were determined as per Ayurvedic Pharmacopoeia of India (Anonymous, 2001). Moisture content, Volatile oil content, total ash value, water insoluble ash, acid insoluble ash, alcohol soluble extractive value, water soluble extractive value, fibre content and sugar content were determined. [Table 1]. The alcoholic extract of stem bark was prepared and subjected to detect the presence of various functional groups like alkaloids, phenols, flavonoids, steroids using relevant reagents (Kokate, C.K, 2005). Preliminary Phyto-chemical screening of stem bark: The alcoholic extract of stem bark was prepared and subjected to detect the presence of various functional groups like alkaloids, phenols, flavonoids, steroids using relevant reagents. [Table-2].
TLC: Solvent system used was Benzene : Ethyl acetate (8:2). Rf value of spot obtained in TLC of alcoholic extract of genuine and market samples was 0.875 [Table-3]. HPTLC Values and graphs showed comparable results. [Table 4] [Figure 2, 3, 4] RESULTS Table 1:- Results of the Physico-chemical analysis of the genuine and market samples of Holoptelea integrifolia Planch. Sl. No. Experiment Holoptelea integrifolia Genuine sample Market sample 1 Moisture content (%) 8 10 2 Volatile oil content (%) NIL NIL 3 Total ash (%) 7.97 8.95 4 Water insoluble ash (%) 5.86 5.38 5 Acid insoluble ash (%) 0.51 0.91 6 Cold water soluble extractive (%) 3.65 3.12 7 Alcohol soluble extractive (%) 5.08 4.7 8 Fibre content (%) 51.47 49.69 9 Sugar content Reducing sugar (%) Total sugar (%) Table 2:- Results of the Preliminary Phyto-chemical screening of the genuine and market samples of Holoptelea integrifolia Planch. Sl. No. Chemical constituent Genuine sample Market sample 1 Steroid + + 2 Flavanoid - - 3 Phenol - - 4 Alkaloid Table 3:- Rf value of spot obtained in TLC Solvent system Benzene : Ethyl acetate (8:2) Alcoholic extract of genuine sample Alcoholic extract of market sample Rf Detection uv/iodine Rf Detection uv/iodine 0.875 uv 0.875 uv
Figure-2: HPTLC Recording of Genuine Sample of Choorna Figure-3: HPTLC Recording of Market Sample of Choorna
Figure-4: HPTLC Comparison of Genuine And Market Sample of Choorna Table 4:-Rf values of different spots obtained in HPTLC Sl. No. of resolving Rf values bands Genuine sample Market sample 1 0.01 0.02 2 0.76 0.72 3 0.79 0.74 DISCUSSION Chirabilwa (Holoptelea integrifolia planch.) is a drug known from Samhita period, but detailed description is seen in nighantus (dictionaries for herbs). Cirivilwa was selected as the study drug because it is included in many medohara ganas (Group of drugs used in diseases of fatty tissues) by various acharyas and it possesses tikta, kasaya rasa (Bitter, Astringent taste), laghu rooksha guna (Light & rough qualities), ushna veerya (Hot in potency) and katu vipaka (Pungent transformation upon digestion). As the primary aim of this work is a clinical study, large amount of drugs were necessary. So it was collected from market. Phytochemical and Physiochemical studies of the study drug were done prior to clinical study to prove the genuinity of the market sample. In Phytochemical screening, shade dried samples of both market and genuine samples of the drug were subjected to analysis of Moisture content, Volatile oil content, Total ash, Water insoluble ash, Acid insoluble ash, Cold water soluble extractive, Alcohol soluble extractive, Fibre content, Sugar content (Reducing sugar and Total sugar). TLC and HPTLC were also carried out. Qualitative analysis indicated the presence of steroid and traces of alkaloids. Phytochemical and physiochemical studies proved the genuinity of the sample. So further clinical study was conducted confirming the identity of the drug.
CONCLUSION The physicochemical evaluation of stem bark of Chirabilwa [Holoptelea integrifolia Planch.] provided specific parameters that will be helpful in proper identification, scientific evaluation and authentication of the drug. The study proved the genuinity of the market samples. The results obtained may be referred as standard for future studies. Although the genuinity of the market sample was confirmed in this study, market samples should mandatorily be subjected to phytochemical and physico chemical analysis before conducting clinical study. REFERENCES Anonymous (2001) The Ayurvedic pharmacopoeia of India. Part 1. Volume 1. New Delhi: Ministry of Health and Family welfare. Fabio Firenzuoli, Luigi Gori (2007). Herbal medicine today: Clinical and research issues. Evidence based complementary and alternative medicine. [Online] 4(1). p. 37 40. [Accessed: 25 June 2011] Kirtikar K.R. & Basu. B.D. (2005) - Indian medicinal plants. Volume 3. New Delhi: International Book Distributors. Kokate C.K (2005). Practical pharmacognosy. 4th Ed. Delhi: Vallabh Prakashan Paradkar Harishastri. (2006) Caraka Samhita. Varanasi: Chowkhambha Krishnadas Academy. Prakash, C.P., Rakesh, K.M., Krishna, G.S. (2014). The efficacy of herbal system of medicine in the context of allopathic system in Indian central Himalaya. Journal of Herbal Medicine. [Online] 4(3). p. 147 158. [Accessed: 10 January 2011] Srikantha murthy K.R. (2002) Astanga Hridaya. Varanasi: Chowkhambha Sanskrit Sansthan. Warrier, P.K., Nambiar V.P.K., (1995) Indian medicinal plants- A compendium of 500 species. Volume 3. New Delhi: Orient Longmann. WHO (2002). WHO Traditional medicine strategy 2002 2005. Journal of drug delivery and therapeutics. [Online]. 2(4), p. 182 185. [Accessed: 20 July 2012] Yadavji Trikamji. (2004) Susruta Samhita. Varanasi : Chowkhambha Krishnadas Academy. Source of Support: NIL Conflict of Interest: None Declared