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North Gujarat Life Sciences Leaflets FREE DOWNLOAD ISSN 2277-4297(Print)0976–1098(Online) EXTRACTION OF SALVADORA OLEOIDES AND ITS PERFORMANCE ALONG WITH ANTIBIOTICS FOR ANTIMICROBIAL ACTIVITIES BHARGAV DAVE1, PIYUSH VYAS1*, MADHU PATEL2 AND 3 Universal Impact NAINESH PATEL Factor 0.9285:2012; 1. 1.2210:2013 SHETH M.N SCIENCE COLLEGE, PATAN. Index Copernicus 2. NAVSARI AGRICULTURAL UNIVERSITY, SURAT. ICV 2011: 5.09, 2012: 6.42, 2013: 3. PACIFIC UNIVERSITY, UDAIPUR, RAJASTHAN. 15.8, 2014:89.16 , 2015:78.30 Corresponding author’s e-mail: [email protected] NAAS Rating 2012 : 1.3; 2013-16:2.69 ABSTRACT: 2017: 3.98 Nature has given various ways to maintain people’s health. One way is to SJIF 2012: 3.947, 2013: 4.802 use herbal medicine. Herbal medicines have been used to treat various types Infobase Index 2015:4.56 of diseases for long times. The people are more attracting towards the use of Cosmos Impact Factor herbal drugs to cure various types of diseases. For treatment of several 2015: 4.366 diseases of human beings, plant drug ‘rasayana’ has always played a vital Received on: role. According to World health organization (WHO) more than 80% of the 6th December 2017 Revised on: world population is dependent on traditional medicine for their primary th 10 December 2017 health care needs.1 Accepted on: 12th December 2017 Herbal medicine, also called botanical medicine or phytomedicine, refers to Published on: the use of a plant’s seeds, barriers, roots, leaves, bark, or flowers for 1st January 2018 medicinal purposes. Long practiced outside of conventional medicine, Volume No. herbalism is becoming more main stream as improvements in analysis and Online & Print quality control along with advances in clinical research show their value in 95 (2017) 2 Page No. the treatment and prevention of disease. 32 to 41 KEYWORDS: Extraction, Salvadora oleoids, Antibiotics, Antimicrobial Life Sciences Leaflets Activities. is a international open access print & e INTRODUCTION: journal, peer reviewed, In India, Ayurveda medicine has used many herbs such as turmeric possibly worldwide abstract as early as 4,000 BC.3,4 Earliest Sanskrit writings such as the Rig Veda, and listed, published every month with ISSN, RNI Atharva Veda are some of the earliest available documents detailing the Free- membership, medical knowledge that formed the basis of the Ayurveda system.5 Many downloads and access. other herbs and minerals used in Ayurveda were later described by ancient http://lifesciencesleaflets.petsd.org/ PEER-REVIEWED Page | 32 Life Sciences Leaflets FREE DOWNLOAD ISSN 2277-4297(Print)0976–1098(Online) Indian herbalists such as Charaka and Sushruta during the 1st millennium BC. The Sushruta Samhita attributed to Sushruta in the 6th century BC describes 700 medicinal plants, 64 preparations from mineral sources, and 57 preparations based on animal sources.6 Salvadora is a small tree with drooping branches, rarely with proper bole or exceeding a height of 20 feet and a girth of 3 feet. It is very common plant in arid tracts but becomes scarce where rainfall conditions are better. It can withstand great soil salinity. It produces new leaves during April, which on maturity become thick and leathery. The tree coppices fairly well but regenerates freely by root suckers and natural layering. It is, however, very slow growing but a dense growth is often formed around the parent plant by root suckers and some natural seedlings. The plant provides a dense shade. It is often lopped for camel and goat fodder. Fruit Small greenish white flowers are produced in March-April. The fruit is yellow and ripens in the months of May and June. It forms one of the main grazing sources for livestock owned by local farmers. It is often dried and preserved in large quantities. The seeds are spread by birds. The seedlings come up under the parent plant or under other bushes and are somewhat frost-tender.7 Common name: Bada Peelu • Hindi: Jal, Bada Peelu • Marathi: Diar, Pilu, Khabbar • Gujarati: Pilu Bada Peelu is a shrub or small tree, attaining 6-9 m height under favourable conditions. It has a short trunk, quite often twisted or bent, up to 2 m in diameter. Branches are numerous, drooping, stiff, often swollen at forks. Bark is gray or whitish-gray. Leaves are blue-green, linear-or ovate-lancelike, leathery and somewhat fleshy, dark greenish-yellow when young, gray when mature. Flowers are stalkless, greenish-white, minute in paniculate spikes, often clustered. Fruit is a spherical drupe, about 6 cm in diameter, usually yellow when ripe, dark brown or red when dry8 Plant taxonomy9 Kingdom: Plantae (unranked): Angiosperms (unranked): Eudicots (unranked): Rosids Order: Brassicales Family: Salvadoraceae Genus: Salvadora Species: S. oleiodes http://lifesciencesleaflets.petsd.org/ PEER-REVIEWED Page | 33 Life Sciences Leaflets FREE DOWNLOAD ISSN 2277-4297(Print)0976–1098(Online) MATERIALS AND METHODS: Phytochemical analysis Phytochemistry: Fruit contains sterols, beta-sitosterol and its glucosides and stigmasterol; benzylisothiocyanate, noctacosanol and tetracosane; flavonoids including quercetin and rutin; thiourea derivatives and phospholipids10. Essential oil of leaves and stems of Salvadora oleoides was extracted by hydro distillation and analyzed by gas chromatography and mass spectrometry (GC-MS). Thirty five (94.0%) and twenty five (91.1%) components were identified in the leaves and stems, respectively. The results show that leaves contain high concentration of 2-methoxy-4-vinylphenol (25.4%), cis-3-hexenyl benzoate (16.8%), phytol (13.9%), n-hexadecanoic acid (6.9%), and trans-ß-damascenone (2.1%). Stems contain high concentration of 2-methoxy4-vinylphenol (21.6%), phytol (12.9%), n- hexadecanoic acid (3.6%), octacosane (7.9%), nonacosane (7.3%), 1octadecene (5.8%), heptacosane (5.9%), hexacosane (4.5%), pentacosane (3.4%), squalene (3.9%) and transß-damascenone (2.3%)11 Medical and traditional uses This tree species is having a number of proven medicinal applications and almost all parts have been found to be pharmaceutically important. Decoction of leaves is gien o the cattles to promote the expulsion of dead fetus from the uterus. The leaves were used as a cooling agent, blood purifier, laxative, expectorant and also used as purgative in horses12,13. Salvadora oleoides is extracted by using soxhlet extraction. Here different solvents like acetone, methanol and ethanol used. These extracts are combined with different Antibiotics and their combinations were applied for zone of inhibition test against different microorganism. Antimicrobial Activity : Antimicrobial activity and antifungal activity of plant extract alone, antibiotics alone and combination of both was carried out by a well diffusion method. In vitro antibacterial activity of the crude extracts was studied by the agar well diffusion method. After getting the turbidity equal to 0.5 McFarland standards, inoculums were aseptically introduced on to the surface of sterile agar plates and sterilized cotton swabs were used for even distribution of the inoculums. Wells were prepared in the agar plates using a sterile cork borer of 10.0 mm diameter. The plant extract and antibiotic drug were dissolved in DMSO to get desired concentration. The wells were filled with plant extract 25µl, 50µl and antibiotic drug (100µl). The plates are incubated at 37℃for 48 hours and then zone of inhibition was measured. In case of combination of plant extract and antibiotic, equal volume of each was added in the well and zone of inhibition was measured. The experiment was replicated two times. (Zone of Inhibition = ±1mm) Qualitative Phytochemical Analysis Preliminary qualitative analysis of all plant extract was carried out as per standard methods as follows. http://lifesciencesleaflets.petsd.org/ PEER-REVIEWED Page | 34 Life Sciences Leaflets FREE DOWNLOAD ISSN 2277-4297(Print)0976–1098(Online) 1. Test for alkaloids Extracts were dissolved in dil.HCl and filtered. A. Dragendroff’s Test: Filtrate was treated with Dragendroff’s reagent (Potassium Bismuth Iodide solution). Formation of red precipitate indicates the presence of alkaloids. B. Wagner’s Test: Filtrate was treated with Wagner’s reagent (Iodine in Potassium Iodide). Formation of brown/reddish precipitate indicates the presence of alkaloids. 2. Test for flavonoids A. Lead acetate Test: Extract was treated with 10% lead acetate solution. Yellow colour precipitate formation indicates the presence of flavonoids. B. Test with Sodium hydroxide: Extract was treated with few drops of NaOH solution. The yellow colour indicates the presence of flavonoids. 3. Test for Glycosides A. Keller-Kiiani Test: Add glacial acetic acid, one drop 5% FeCl3 and conc. H2SO4 to the extract. Formation of reddish brown color at junction of the two liquid layers and bluish green at upper layer indicates the presence of glycosides. B. Legal Test: On treatment of Extracts with sodium nitropruside in pyridine and sodium hydroxide, observed pink to blood red colour that indicates the presence of cardiac glycosides. 4. Test for Phenolics Test with Ferric Chloride: Extract was treated with few drops of ferric chloride solution and formed bluish black colour indicates the presence of phenols. 5. Test for Saponins Froth Test: The extract was diluted with 20 ml of distilled water and then shaken in a graduated cylinder for 15 minutes. Formation of 1 cm layer of foam indicates the presence of saponins. 6. Test for Tannins Test with Ferric Chloride: Extract was boiled with water and filtered it. Added FeCl3 to the filtrate and formation of brownish green or a blue-black coloration indicates the presence of tannins. http://lifesciencesleaflets.petsd.org/ PEER-REVIEWED Page | 35 Life Sciences Leaflets FREE DOWNLOAD ISSN 2277-4297(Print)0976–1098(Online) 7. Test for Terpenoids Salkowski Test: To the extract, added 2ml chloroform and 3ml Conc.H2SO4 by the wall of test tube carefully. Formation reddish brown colour at interface indicates the presence of terpenoids. RESULTS AND DISCUSSION: Table A showed phytochemical analysis of Salvadora oleoides by using acetone, methanol and ethanol extracts.
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