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Int. J. Curr. Res. Biosci. Plant Biol. (2020) 7(10), 31-37 International Journal of Current Research in Biosciences and Plant Biology Volume 7 ● Number 10 (October-2020) ● ISSN: 2349-8080 (Online) Journal homepage: www.ijcrbp.com Original Research Article doi: https://doi.org/10.20546/ijcrbp.2020.710.004 Comparative evaluation of phytochemicals in leaf, stem bark and root bark of Holarrhena antidysenterica, Wrightia tomentosa and Wrightia tinctoria Pawan Kumar Ahirwar1*, S. P. Mishra2 and Pramod Kumar3 1Department of Biological Science, M.G.C.G.V.V, Chitrakoot-485 334, Satna (M.P.), India 2Department of Crop Sciences, Faculty of Agriculture, M.G.C.G.V.V., Chitrakoot-485 334, Satna (M.P.), India 3Genetics and Tree Improvement Division, T.F.R.I., Jabalpur (M.P.), India *Corresponding author; e-mail: [email protected] Article Info ABSTRACT Date of Acceptance: Various phytochemicals like alkaloids, tannins, flavonoids, carbohydrates, proteins, 15 September 2020 saponins, polyphenolic compounds and steroids etc. are found in different parts of plant. These are primary and secondary metabolites formed as byproduct of various Date of Publication: biochemical pathways and play a major role in combat and cure of various diseases. A 06 October 2020 large number of diseases which are incurable through allopathic medicines show promising response to ayurvedic medicines. Prevention of these incurable diseases Keywords achieved through use of herbal medicines. It is necessary to identify, isolate and Allopathic medicine quantify such phytochemicals which are being used in the preparation of medicines or Herbal medicine drugs formulations. The present study deals with comparative phytochemicals analysis Holarrhena in leaf, stem bark and root bark of Holarrhena antidysenterica (Roth) Wall ex. A. DC., antidysenterica Wrightia tomentosa Roem. et Schulta and Wrightia tinctoria R. Br. Phytochemicals Wrightia tomentosa Wrightia tinctoria Introduction terpenoids, alkaloids, flavonoids, etc. of which about 12,000 secondary metabolic compounds Plants are powerful resources of bio-chemicals have been isolated. These isolated secondary and thus, good source of phytomedicines since metabolites showed wide range of ancient times. Natural constituents can be pharmacological properties such as in-vitro extracted from any part of the plant like leaves, antibacterial and antifungal activities (Zahan et bark, flowers, fruits, seeds, roots, etc. which al., 2013). Their major role i s in t h e plant contain bioactive compounds. The medicinal defense system against herbivores, property of bioactive compounds is the result of microorganisms and insects. Some are involved in the combinations of secondary metabolites plant pigmentation (tannins and quinines), odour present in the plant (Shwetha et al., 2011). Plants (terpenoids) etc. and thus responsible for a have the ability to synthesize aromatic products definite physiological effect. It clearly indicates mainly secondary metabolites such as tannins, that the medicinal property of plants lie in the P. K. Ahirwar et al. (2020) / Comparative evaluation of phytochemicals in leaf, stem bark and root bark of Holarrhena 31 antidysenterica, Wrightia tomentosa and Wrightia tinctoria Int. J. Curr. Res. Biosci. Plant Biol. (2020) 7(10), 31-37 bioactive phytochemical constituents (Patel et al., different phytochemicals in selected parts of H. 2015). Many secondary metabolites of plants are antidysenterica, W. tomentosa and W. tinctoria. commercially important and are being used in a number of pharmaceutical compounds Materials and methods (Meenakshi, 2015). Phytochemical investigation of bioactive compounds from plants has led to the Collection of plant samples and powder discovery of many therapeutic drugs which preparations effectively used in protection and treatment of various diseases including cancer (Sharma et al., Leaf and stem bark of Holarrhena 2017). With this background and abundant source antidysenterica was collected from Sati Anusuiya of unique active components harbored in plants, area and root bark was collected from Bagdara the present study was conducted on leaf, stem Ghati of Gupt Godavari area of Chitrakoot forest bark and root bark of three medicinal plants viz., whereas, leaf, stem bark and root bark of Holarrhena antidysenterica (collected from two Wrightia tinctoria was collected from Bagdara locations); Wrightia tomentosa and Wrightia Ghati of Gupt Godavari area of Chitrakoot forest tinctoria. These plant species belong to the family of Satna district of Madhya Pradesh. Samples of Apocyanaceae. Holarrhena antidysenterica were also collected from Talawada area of Guna district of Madhya H. antidysentrica is a small deciduous tree with Pradesh. While the leaf, stem bark and root bark cream white flowers and found throughout the of Wrightia tomentosa were collected from the dry forests of India. The different parts of the Holipur area of Budhni tehsil of Sehore district of plant are used in the indigenous system of Madhya Pradesh and identity was confirmed by medicine. It is used in the treatment of dysentery, Dr. R.L.S. Sikarwar, a taxonomist of Department diarrhea, helminthic, astringent and haemostatic of Ayurveda Sadan (Research Laboratory), disorders (Ahirwar et al., 2019). Arogyadham, Deendayal Research Institute (DRI), Chitrakoot, Satna (M.P.) India. Collected W. tomentosa is an endangered medicinal tree plant samples were brought to the laboratory and placed in the category I of Red Data washed with running tap water, dried under (Nagalakshmi and Murthy, 2015) with many shade and prepared powder with the help of medicinal uses. Ethnomedically W. tomentosa is electric grinder. The powder was sieved (No. 43) widely used to treat stomach ache, tooth ache, and stored in air tight closed containers fever, hemorrhage, arthritis and snake bite separately for protection from moisture and (Srinivas et al., 2013; Ahirwar et al., 2019). contamination. Wrightia tinctoria R. Br. is a small to medium Phytochemical screening size deciduous tree, which is widely distributed all over India. In South India, it is commonly called as The powdered material weighing 2.0g was "Jaundice curative tree" traditionally (Dixit et al., extracted in 100 ml ethanol by continuous 2014). The common name of plant is Paalai and extraction for 6 hours on rotatory flask shaker called “Sweet Indrajao”. The plant are effective in and left for 18 hours for maceration. The extract stomachic, in the treatment of abdominal pain, was filtered through Whatman filter paper no. 1 anti- diarrhoeal and antihaemorrhagic and tested for various classes of active chemical (Deventhiran et al., 2016). Wrightial, constituents by using standard methods described cycloartenone, cycloeucalenol, indigotin, indirubin, in Ayurvedic Pharmacopeia of India (API) and tryptanthrin, isatin, rutin, β-sitosterol, β-amyrin, "Methods of Biochemical Analysis" (Anonymous wrigatiadione are the main isolated compounds 2010; Harborne 1984; Thimmaiah 1999). from seeds, leaves and bark of this plant (Singh et al., 2015). Reviewed on phytochemical, Phytochemicals estimation pharmacological and pharmacognostical profile of W. tinctoria and noted its anti-cancer, anti-HIV, Determination of alkaloid, flavonoid, saponins and anti-diabetic properties too (Srivastava, 2014). and crude protein by gravimetric method (in %) The aim of the present study was investigation of and total carbohydrate, soluble protein and P. K. Ahirwar et al. (2020) / Comparative evaluation of phytochemicals in leaf, stem bark and root bark of Holarrhena 32 antidysenterica, Wrightia tomentosa and Wrightia tinctoria Int. J. Curr. Res. Biosci. Plant Biol. (2020) 7(10), 31-37 tannin (mg/ml) were estimated in three replications by spectrophotometric methods in Where, B and S represent titration value of blank leaf, stem bark and root bark of three species. and sample respectively. Alkaloid was estimated by both gravimetric and spectrophotometric methods. Protein Nitrogen: Total nitrogen value multiplied with 6.25 for crude protein content Alkaloid- Harborne (1973) method was used for which also includes non-protein nitrogen. the determination of alkaloid. Total carbohydrate- Total carbohydrate or total Flavonoid- Bohm and Kocipai – Abyazan (1994) soluble sugars estimated by Anthrone method method was used for the determination of (Thimmaiah, 1999). flavonoid. Soluble protein- Lowry’s method was used for Saponins- Obadani and Ochako (2001) method the estimation of soluble protein or true protein was used for the determination of saponins with (Lowry et al., 1951; Mani et al., 2019; Sadasivam modification of petroleum ether in place of and ManickaM, 1996). diethyl ether. Tannin- Folin-Denis method was used for the Crude protein- Kel plus method was used for estimation of tannin (Sadasivam and ManickaM, estimation of crude protein (Sadasivam and 1996; Thimmaiah, 1999). Manickam 1996). Alkaloids - Alkaloids was estimated Sample preparation: Taken 0.5g powder of spectrophotometrically by Dragendorff’s reagent samples in digestion tubes and added punch of method (Nayeem et al., 2011). digestion mixture and 10 ml of sulphuric acid in all trubes. Sample was digested until green or Sample extract preparation- 10g coarsely dark green colour appeared. After cooling of powdered plant material was extracted with 25 ml tubes, sample particles attached side of the tubes 2% aqueous acetic acid at room temperature for 10 dissolved with the help of distilled water and total minute. The procedure was repeated