Research Article QUANTITATIVE ESTIMATION OF BERBERINE CONTENT OF BERBERIS ASIATICA FROM DIFFERENT ALTITUDE OF GARHWAL HIMALAYA ALOK MAITHANI *1 , VERSHA PARCHA 1 , AND DEEPAK KUMAR 2 1 Dept of Pharmaceutical Chemistry, Sardar Bhagwan Singh PG Institute of Biomedical Sciences and Research, Balawala, Dehradun. 2 Dept of Pharmaceutical Chemistry, Dolphin PG College of Natural Sciences, Manduwala, Dehradun. Email: alok_maithanii@rediffmail.com Received: 18 December 2013, Revised and Accepted: 16 January 2014 ABSTRACT Objective: Berberis asiatica is a popular medicinal plant of Garhwal Himalaya and its root is traditionally used for curing various ailments like jaundice, conjunctivitis, and diabetes mellitus. In the present study berberine content in B asiatica growing at seven different altitudes of Garhwal Himalayas was quantitatively determined by HPTLC analysis. Methods: Ethanolic extracts of various plant samples were analyzed with standard berberine (purified) in different concentrations 1-5µl. The peak areas of each standard and test samples were obtained from the system and a calibration graph was plotted between concentrations vs. Peak area. Results: The plant samples belonging to lowest altitude region was found to possess maximum concentration of berberine which was 2.94% however berberine content was non-linearly distributed among the other samples of higher altitudes. Keywords: Berberis asiatica, Berberine, quantitative determination, isoquinoline alkaloid, and HPTLC. Introduction: Berberise asiatica, belongs to family Berberidiacae, is a well known shrub of Garhwal Himalaya and regionally known as “Kingor”. In Ayurvedic medicinal system it is named as ‘Daruharidra” or ‘Wood Turmeric’ due to similar properties as of turmeric[1]. Berberis asiatica is reported to possess many biological activities like anti- microbial, wound healing, hepatoprotective, anti-acne, & cytotoxicity etc[2-6]. The plant yields fairly large quantity of alkaloids in which isoquinoline type alkaloids like berberine, palmatine, jetrorrhizine, and columbamine are the most studied phytoconstituents[7] (Figure 1). Thus the major pharmaceutical properties of B asiatica are attributed due to the presence of alkaloidal content of plant. Berberine is 8- substituted derivative of iso-quinoline. It exhibits significant role in management if various enteric infections, specially bacterial dysentery [8]. It is also experimentally claimed to be hepatoprotechtive, antidiabetic [9], anti-inflammatory [10] and antimicrobial [11] agent. The medicinal importance of berberine and other phytochemicals present in Berberis asiatica increases its demand in global market India consume 500tones roots of this plant alone and also Indian government exports 60 to 70 tones of roots of Berberis asiatica per year[12]. Roots of the plant are collected in fairly large quantities in Tehri Garhwal of Uttarakhand during august – September and are sold in drug market of Dehradun and Haridwar. Looking into the tremendous potential of curing disease and excellent revenue realized through the disposal of this plant, it was thought worth effective to quantitatively estimate percentage of berberine content in Berberis asiatica growing in different altitudes of Garhwal Himalaya range. Material & Methods Chemicals & Instruments All the solvents like petroleum ether, ethanol, and butanol were of analytical grade and purchased from Rankem Ltd. Standard berberine was an isolated and purified compound and received as a gift from Department of Chemistry, HNB Garhwal (A Central) Universitu, Srinagar Garhwal. HPTLC analysis was performed from Centre for Aromatic Plants (CAP), Selaqui, Dehradun. Procurement of Plant Material Roots of Berberise asiatica were collected from seven places of different altitudes, of garhwal Himalayas, ranging from 1700-5500 feet (table 1) and were identified by Dr. S.A.S. Vishwas (with a specimen no. 10127), in F.R.I. Dehradun. The samples of the plants were submitted to the herbarium of SBSPGI, Dehradun. Preparation of Extracts Finely dried roots were defatted with petroleum ether (60-80 o c) and then extracted with 80% ethanol (80 parts of ethanol and 20 parts of water). Extraction was carried out for 40 hrs, solvent was distilled off and extract was concentrated using thin film evaporator until a viscous and sticky mass obtained. The extracts of different plant materials are designate S1 – S7 in the increasing order of altitudes [13] (Table 1). HPTLC Analysis The HPTLC method provides a simple, low cost and a good statistical analysis that proves an efficient method in order to quantitatively determine berberine in ethanolic extracts of roots of Berberise asiatica of the concern altitudes. The instrument used for the estimation was Camag semi automatic sample applicator, Camag TLC scanner, CATS software for interpretation of the data. Vol 7, Suppl 1, 2014 ISSN - 0974-2441