Available online on www.ijppr.com International Journal of Pharmacognosy and Phytochemical Research 2012-13; 4(4); 199-204 ISSN: 0975-4873 Research Article Comparative Phytochemical Studies and Evaluation of Radical Scavenging Activity in Selected Jasminum Species *1Sulaiman C.T, 2Soudha V., 1Deepak M., 1Indira Balachandran 1 Phytochemistry Division, Centre for Medicinal Plants Research, Arya Vaidya Sala, Kottakkal 2Department of Chemistry, Farook College, Calicut ABSTRACT Phytochemical studies were carried out in five Jsaminum species. The chemical profiles of different species were compared using chromatographic techniques such as TLC and HPLC. The total phenolic content (TPC) and radical scavenging capacity were also evaluated. The DPPH EC50 value was found least for Jasminum Grandiflorum (7.5µg) showing its highest antioxidant activity. The EC 50 (DPPH) values vary as, 15 μg (J.angustifolium), 12 μg (J.auriculatum), and 7.5μg (J. grandiflorum). Key words: TLC, HPLC, DPPH, Total phenolics. INTRODUCTION of 3-12 together. The plant traditionally used as an Medicinal plants and plant- derived medicines are widely analgesic, antidepressant, anti inflammatory, antiseptic, used in traditional cultures all over the world and they are aphrodisiac, sedative, expectorant and tonic (uterine) becoming increasingly popular in modern society as effects. Roots are used to treat wounds and snake bites. natural alternatives to synthetic chemical (Van Wyk and The leaves and flowers have antipyretic and decongestant Michael Wink, 2009). Nearly all cultures from ancient properties. Phytochemical studies shown that the roots times have used plants as a source of medicine. The contains dotriacontanoic acid, dotriacontanol, oleanolic World Health Organization (WHO) has listed 21,000 acid, daucosterol and hesperidin (Zhang et al, 2004) and plants worldwide, reported to have medicinal uses. India leaves contain sambacosides A, E and F (Tanahashi, is the largest producer of medicinal herbs and is called the 1988) flower contains molihuaside A-E, sambaeoside A botanical garden of the world (Trivedi, 2007). (Zhang et al, 1995). Jasminum genus with about 200 species belonging to Jasminum angustifolium, distributed in south India family Oleaceae are of three types: shrub or bush form, (kerala, Karnataka) on the hills of lower elevation (Bown, vines and trees, native to tropical and warm temperate 1995) Leaves are simple ovate-lanceolate, acute, glabrous regions. Many jasminum plants prominently feature and flowers are either solitary or more usually in three. white, yellow or pink flowers with sweet fragrance and Petals are linear, obtuse and acute Hepatoprotective effect others are unscented Jasminum species is used to treat of ethanolic and chloroform extract of Jasminum many conditions such as amenorrhoea, leprosy, skin angustifolium were evaluated against carbon tetrachloride diseases and also as an analgesic, antidepressant and anti- (1ml/kg) induced hepatic damage and was evidenced by inflammatory reduction in level of alkaline phosphatase (ALP), (http://www.ehow.com/about_6325794_jasmine-plants). alkanine amino transferase (ALT), aspartate amino Jasminum grandiflorum is a scrambling sub erect transferase (AST), cholesterol, glucose, total protein and twining evergreen shrub (Anonymous, 2004), native to bilirubin concentration in blood (Joshi et al, 2008). India, France, Italy, China, Japan, Morocco and Egypt Jasminum auriculatum grows almost throughout South [Chopra et al, 1958].The leaves are opposite, entire ovate India, on dry slopes of the Western Ghats. The roots are to somewhat elliptic in shape with acuminate mucronate useful in skin diseases especially for ringworm and apex, whereas flowers are terminal and axillary cymes, flowers are fragrant, bitter, acrid, sweet, refrigerant, calyx lobes are long, linear (Cooke, 1967 & Nadkarni, astringent, cardiotonic, diuretic and depurative in nature. 1976). Roots are useful in cephalalgia, mental debility, They are useful in burning sensation, hyperdesia, ulcers, chronic constipation, flatulence, strangury, sterility, odontalgia, stomatopathy, ophthalmopathy, cardiopathy, dysmenorrhoea, amenorrhoea, ringworm, leprosy, skin urolithiasis, nephrolithiasis, strangury and dermatopathy diseases and giddiness. (Ghosh, 1984). Jasminum auriculatum leaves have been Jasminum sambac is commercially grown in India, reported to contain lupeol and jasminol (Deshpande et al, Thailand, China and Philippines. It is an evergreen vine 1967). Alcoholic and aqueous extracts of flowers of or shrub reaching up to 1-3 m. The leaves are ovate; Jasminum auriculatum showed diuretic activity by phyllotaxy is opposite or in whorls of three. The flowers increasing the total volume of urine and concentrations of blooms throughout the year and are produced in clusters potassium and sodium salts in urine and antiurolithiatic Author for correspondence: E-mail: [email protected] Sulaiman C.T et.al./ Comparative Phytochemical Studies… Fig: 1.1 TLC (UV 254) Fig: 1.1 TLC (UV 366 nm) Fig: 1.3 TLC after Derivatization activity by reducing the elevated urinary oxalate to sequential extraction using 100ml of petroleum ether, synthesis. (Bahuguna et al, 2009). chloroform, and methanol respectively for 5 hours. The final volume is made up to 50 ml to get 40-mg/ ml MATERIALS AND METHODS solution. These solutions are taken for different analyses. Extraction: Plant materials (leaf) were collected from TLC Analysis: TLC analysis of petroleum ether and 200 Herb Garden, Arya Vaidya Sala- Kottakkal and chloroform extracts was carried out on a precoated silica Page authenticated by Taxonomy division of Centre for plate (F254 Merck) using toluene: ethyl acetate as mobile Medicinal Plant Research, Arya Vaidya Sala, Kottakkal, phase in the ratio 9:1. TLC analysis of methanol extracts Kerala. The powdered samples (2g each) were subjected was carried out using toluene: ethyl acetate as mobile IJPPR, Vol-4, Issue 4, December 2012- February 2013, 199-204 Sulaiman C.T et.al./ Comparative Phytochemical Studies… Fig: 1.4 TLC of Methanol Extracts Fig: 1.5 phase in the ratio 8:2. The plates were developed up to thoroughly. After an interval of 3 min, 2 ml of 2% 9cm and visualized under UV 254 nm, UV 366 nm and in sodium carbonate solution was added and the mixture visible light after derivetizing with Anisaldehyde- was allowed to stand for 30 min with intermittent sulphuric acid (ANS) reagent. shaking. The absorbance of the mixture was measured at HPLC Analysis: HPLC profiling was done using a 550 nm using spectrophotometer (Shimadzu, Japan). Shimadzu High Performance Liquid Chromatographic Different Gallic acid standards (2, 5, 7, 10, and 15 μg/ml) system equipped with LC-10ATVP pump, SPD M10AVP were used for obtaining a standard curve (Singleton et a,l Photo Diode Array Detector in combination with 1965). The total phenolic content was expressed as Gallic CLASS-VP 6.12 SP5 integration software. The mobile acid equivalents (GAE) per gram of sample. phase used for the separation was HPLC grade 0.1% DPPH Assay: DPPH radical scavenging assay DPPH formic acid in Acetonitrile (A) and methanol (B) in a time radical scavenging activity of the leaf extracts were programming 0-10 10% A, 10-20 30% A, 20-30 50% A, determined according to the method described by Kukic 30-40 60% A and 40- 50 70 % A. The column used was et al. First, 4.0 mL of test material at different C18 – ODS (Octadecylsilane), Lichrospher RP 18e (5μm) concentrations were reacted with 0.50 mL of 1.0 mM (Merck) with a Phenomenex guard column (4mm x 2 mm DPPH solution and kept in the dark for 30 minutes, i.d: 5µm). The samples were injected using a 20 µl loop following which the absorbance was measured at 517 nm (Rheodyne Rohnet Park, CA, USA). The flow rate was against a blank sample consisting of 4.0 mL of MeOH maintained to 0.75 ml/min. and 0.50 mL DPPH solution. DPPH scavenging activity Total Phenolic Content (TPC): The assay was based on was calculated using the equation: 201 the reduction of phosphomolybdate ion of Folin- % DPPH radical scavenging activity = (Ao-As/Ao) × 100. Ciocalteu reagent by the phenolate ion of sample. A Where Ao is the absorbance of the blank sample, and As Page desired amount of plant extract, distilled water and 1 N is the absorbance of the test material. The IC50 value Folin-Ciocalteu reagent was taken into a tube and mixed represented the concentration of test material that caused IJPPR, Vol-4, Issue 4, December 2012- February 2013, 199-204 Sulaiman C.T et.al./ Comparative Phytochemical Studies… HPLC Profile of Jasminum Angustifolium 1: 254 nm, 8 nm j.angstflm j.angstflm 1800 Retention Time 3.541 1600 1400 1200 1000 4.992 mAU 800 2.603 600 5.195 400 3.733 3.904 200 1.931 4.480 6.197 6.336 6.507 6.944 7.083 7.307 7.819 8.181 8.629 9.216 9.461 9.792 11.541 13.579 16.309 17.461 17.728 18.987 20.043 0 -200 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 Minutes HPLC Profile of Jasminum Auriculatum 1: 254 nm, 8 nm 3000 j.aurcltm j.aurcltm Retention Time 2.411 2.272 2500 2000 1500 mAU 1000 2.891 500 1.899 6.443 3.339 3.765 6.123 6.795 4.213 4.352 5.163 5.440 5.760 7.584 7.893 8.896 9.856 10.773 11.328 12.896 15.179 17.739 11.947 16.181 16.875 17.899 19.381 0.928 0 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 Minutes HPLC Profile of Jasminum Flexile 1: 254 nm, 8 nm 1400 j.flxl j.flxl Retention Time2.293 1200 1000 800 mAU 600 400 3.008 200 1.920 5.013 5.397 5.781 6.283 7.349 6.592 7.584 7.829 8.619 8.896 9.419 14.805 17.387 8.331 11.200 15.840 17.685 0.555 0 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 Minutes HPLC Profile of Jasminum Grandiflorum 3000 1: 254 nm, 8 nm J.Gradiflorm04 J.Gradiflorm04 Retention Time 5.035 2500 2000 1500 mAU 1000 2.699 3.733 500 4.779 4.160 6.059 2.155 6.656 7.179 8.000 8.843 9.216 9.419 9.739 10.048 11.104 12.341 15.915 16.683 20.213 21.099 22.549 0 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 Minutes 50% scavenging activity.