Research Journal of Medicinal Plants in Ayurveda Volume 1 Issue 2 Physico-chemical, Phytochemical and Spectroscopic Characteristic Studies on Bauhinia variegataLinn. Thamizh Selvam N1, Sudhakar D2 1Assistant Director, 2Director (Institute) Biochemistry, Department of Biochemistry and Pathology, National Ayurveda Research Institute for Panchakarma, (CCRAS, Ministry of AYUSH, Govt. of India), Cheruthuruthy, Thrissur, Kerala, India *Corresponding Author Email Id: [email protected] ABSTRACT Bauhinia variegata Linn is a medium sized deciduous tree grown in tropical regions. Even though the plant is generally used for ornamental purpose; it has wide range of medicinal values. B. variegata is used in treatment of bronchitis, leprosy and other ailments by the traditional healers. The present study scientifically documents the proximate composition, physico-chemical characteristics, phyto-chemical analysis and spectroscopic profiling of ethanolic extract, methanolic extract and aqueous extract of plant leaf. The data of this study will be highly supportive for further research on isolation of active components from B. variegata and evaluating their biopotency. Key words: Bauhinia variegata, Kachnar, Spectroscopy, Phytochemical Studies, Proximate Analysis. INTRODUCTION Bauhinia variegate Linn. commonly B. variegata has been reported for its known as ‘Mountain ebony’ is a medium various pharmacological activities. The sized, deciduous tree under family ethanolic and aqueous extracts of root have Leguminosae (Caesalpinioideae) [1]. It is been reported for hypolipidemic activity native to Southeast Asia mostly grown in and antioxidant activity [7]. The tropical regions and seen throughout India. antibacterial studies showed that the plant Leaves are broader, deeply cordate with is more effective against various gram- two leaflets, connate for about two-thirds positive and gram-negative bacteria. The up, leaflets are ovate, rounded at apex, 10- plant has also been reported for anti- 15cm long, pubescent beneath when inflammatory effect, hepatoprotective young. effect and immunomodulatory effect on experimental animals [8]. As the plant is Flowers are very showy and bloom when grown different parts of India from leaves start falling. The fruit is 15-30 cm Himalayan region to deccan region, there long, containing 10-15 seeds and this plant are various reports documented for their reproduces naturally through seeds. All physicochemical characteristics and parts of the plant including leaf, stem, proximate composition. flower, seed, root are used in traditional medicine system [2-5]. The plant is used The present study scientifically documents in the treatment of bronchitis, leprosy and the proximate composition, physico- tumours by the traditional healers. The chemical, phytochemical and stem bark of B. variegata is used as spectroscopic characteristics of plant astringent, tonic, anthelmintic and collected from Western ghat region in antidiabetic [6]. Kerala, India. CR Journals (Page 1-7) 2021. All Rights Reserved Page 1 Research Journal of Medicinal Plants in Ayurveda Volume 1 Issue 2 TAXONOMY [9] suitable temperature using rotovac Kingdom - Plantae equipment. The solvent free extract was Sub Division - Spermatophyta aliquoted and stored in refrigerated Division - Magnoliophyta condition for ready to use form. Class - Magnoliopsida Sub Classs - Rosidae Physicochemical Characteristics [11,12] Order - Fabales Physico-chemical analysis such as Family - Caesalpiniaceae moisture content, total ash, water soluble Genus - Bauhinia ash, acid insoluble ash, water soluble Species - variegata extractive and alcohol soluble extractive Botanical name -Bauhinia variegata were carried out as per the standard protocols developed and standardized at VERNACULAR NAMES NARIP was used. Mountain ebony- English Kachnar- Hindi Phytochemical Analysis [13,15] Mandaram- Malayalam The phytochemical analysis was carried out as per the standard protocols that has MATERIALS AND METHODS been standardised in NARIP. It comprised Plant collection and authentication of various tests including Froth’s test, The fresh plant materials were collected Libermann Burchard test, Fehling’s test, from the Western ghat regions of Palakkad Salkowski test, Dragendorff’s test, Keller District, Kerala, India. The collected plant Killani test and Ellagic acid test. materials were authenticated by the taxonomist, from Kerala Forest Research Spectrophotometric Analysis [16,17] Institute, (Research Institute under Spectrophotometric characteristics were Government of Kerala), Thrissur. The analysed for understanding the basic plant material was processed and chemical profiling of study plant and to herbarium was prepared as per the compare the proposition of major standard guidelines. Voucher specimen is compounds among different solvent maintained in the Department of extracts. The ethanolic extract, methanolic Biochemistry and Pathology, National extract and aqueous extract of B. variegata Ayurveda Research Institute for leaf at different concentrations were used Panchakarma (NARIP), Cheruthuruthy for for spectroscopic analysis using the ready reference. concerned solvent as blank. The spectrum characteristics were measured after Extraction [10] standardizing the procedures and analysis The air-dried, powdered plant leaf material was confirmed by carrying out triplicate was extracted by various methods using analysis at 200-800 nm. different solvents. The hot solvent extraction for 8 hours was done by Soxhlet Chemicals and Reagents method using ethanol. Methanolic extract The chemicals and reagents of AR grade was prepared by cold extraction method. purchased from make of Spectrum, Nice The plant material was soaked in methanol and HiMedia were used for the present for overnight 10-12 hours and filtered study. through Whatman filter paper No. 1. The aqueous extract was prepared as per the Instruments protocol prescribed in Ayurvedic UV Spectrophotometer of Agilent- Cary pharmacopeia of India. The solvent was 60 was used for spectroscopic analysis. removed under reduced pressure at CR Journals (Page 1-7) 2021. All Rights Reserved Page 2 Research Journal of Medicinal Plants in Ayurveda Volume 1 Issue 2 Fig 1. The herbarium authentication of Bauhinia variegata Linn. RESULTS AND DISCUSSION saponins, flavonoids, terpenoids and The authentication of the plant Bauhinia carbohydrates compounds was variegata Linn has been presented in documented in all the three types of Figure 1. The proximate composition extracts (Table 2). Spectrophotometric analysis comprising of total extractive analysis showed various prominent peaks yield, total ash content, characteristics of in the spectrum of 200-800 nm ash, characteristics of solubility of extract representing the presence of compounds has been presented in the Table 1. The like alkaloids, steroids and flavonoids. shade dried leaf of B. variegata has The spectroscopy study documented the yielded 16.3% extract and the total ash data of ethanolic extract, methanolic content was found to be 10.98%. The plant extract and aqueous extract at different extract is found to have more solubility in concentration levels and the study was water than alcohol. The phytochemical confirmed by triplicate analysis (Figure 2- analysis was done for the ethanolic extract, 4 and Table 3-5). methanolic extract and aqueous extract of shade dried leaf. The presence of Table 1. Physico Chemical Characteristics of Bauhinia variegate L Shade Dried Leaf S No Proximate Compounds Bauhinia variegate L (Shade Dried Leaf) 1 Extractive Yield (%) 16.3 2 Moisture content (%) 7.061 3 Total Ash (%) 10.989 4 Water Soluble Ash (%) 3.55 5 Acid Insoluble Ash (%) 0.297 6 Water Soluble Extractive (%) 12.78 7 Alcohol Soluble Extractive (%) 5.984 CR Journals (Page 1-7) 2021. All Rights Reserved Page 3 Research Journal of Medicinal Plants in Ayurveda Volume 1 Issue 2 Table 2: Phytochemical Characteristics of Bauhinia variegata L (Shade Dried Leaf). S. Phytochemical Method Bauhinia variegata - Shade dried Leaf No Compounds Ethanolic Methanolic Aqueous Extract Extract Extract 1 Tannins Ferric Chloride Test + ++ + 2 Saponnins Froth’s Test ++ + + 3 Terpenoids Salkowsky’s Test + ++ ++ 4 Cardiac Glycosides Keller-Killani Test + ++ + Shinoda Test ++ + + 5 Flavonoids Lead acetate Test ++ ++ ++ Alkaline Reagent Test - + + Fehling ‘s Test + + + 6 Carbohydrates Molisch Test ++ ++ ++ Benedict’s Test + + + 7 Phytosterols Liebermann Burchad Test + + + 8 Proteins Biuret test - - - Xantha protein test +++ + + Dragendorff’s Test - - + 9 Alkaloids Mayer’s Test - - + Hager’s Test - + + Wagner’s Test - - + 10 Anthraquinones Test with 10% KOH - - + +++ Highly present++Medium + Low -Nil Fig 2. UV Spectroscopic analysis of Methanolic extract of B. Variegate shade dried leaf at different concentration Table 3. Comparative Data of Spectroscopic analysis of Methanolic extract of B. variegata Shade Dried Leaf at Different Concentrations Sample Name: B. variegata Leaf-Methanol Name: B. variegata Leaf-Methanol Concentration-0.125%200-800 nm Concentration-0.0625%2 200-800 nm Wavelength Absorbance Wavelength Absorbance 665.0 1.013 665.0 0.510 368.0 1.117 265.0 0.698 344.0 1.075 240.0 2.019 CR Journals (Page 1-7) 2021. All Rights Reserved Page 4 Research Journal of Medicinal Plants in Ayurveda Volume 1 Issue 2 Fig 3. UV Spectroscopic analysis of Ethanolic extract of B. variegatashade dried leaf at different concentration Table 4. Comparative Data of Spectroscopic Analysis of Ethanolic extract of B. Variegate Shade Dried