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In Vitro Antioxidant and Free Radical Scavenging Activity of Extracts of Rosa Damascena Flower Petals

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In Vitro Antioxidant and Free Radical Scavenging Activity of Extracts of Rosa Damascena Flower Petals Original Article In vitro Antioxidant and Free Radical Scavenging Activity of Extracts of Rosa damascena Flower Petals. Priti S. Patil1, Pratima A. Tatke*1 and Satish Y. Gabhe2 1Department of Pharmaceutical Chemistry C. U. Shah College of Pharmacy, S.N.D.T Women’s University, Juhu Campus, Santacruz (West), Mumbai 400 049, India 2Bharati Vidyapeeth Deemed University, Poona College of Pharmacy, Erandwane, Pune 411 038, India ABSTRACT Introduction: Rosa damascena mill L., well-known as Rose, has numerous pharmacological properties including antioxidant, antitussive, relaxant effect on tracheal chains, hypnotic, antibacterial and anti-HIV. Objective: The objective of the present study was to evaluate antioxidant and free radical scavenging activity of various extracts of Rose flower petals. Methods: Chloroform, acetone, ethanol, and aqueous extracts of fresh and dried Rose petals were prepared and evaluated for presence of flavonoids and phenolics. Ethanol and acetone extract showed high phenolic and flavonoid content and were selected for various in vitro free radical scavenging assays using DPPH scavenging, ABTS, peroxynitrite radicals, xanthine oxidase inhibition (XOI), superoxide scavenging and Lipid peroxidation assay. Results: All the extracts of fresh and dried Rose petals were found to Address for contain higher amount of total phenolics expressed as mg of GAE/g Correspondence of dry extract and total flavonoids expressed as mg of RE/g of dry extract. The amount of both these constituents was highest in ethanol Department of extract followed by acetone extract, aqueous extract and least in Pharmaceutical chloroform extract. The ethanol extract of fresh and dried petals Chemistry C. U. Shah showed lowest IC50 values than the acetone extract when evaluated College of Pharmacy, by DPPH, ABTS, peroxynitrite inhibition, xanthine oxidase S.N.D.T Women’s inhibition, superoxide scavenging and lipid peroxidation inhibition University, Santacruz methods indicating good free radical scavenging activity. (West), Mumbai 400 Conclusion: The present study has demonstrated that ethanol extract 049, India of fresh Rose flower petals exhibit potent antioxidant and free radical scavenging activities. E-mail: [email protected] Keywords: Rosa damascena, total phenolics, flavonoids, rutin, DPPH, ABTS. American Journal of Phytomedicine and Clinical Therapeutics www.ajpct.org Tatke et al__________________________________________________ ISSN 2321 – 2748 INTRODUCTION Free radicals are known to be the therapy of rose oil is helpful for several major cause of various chronic and allergies, migraine and headaches. degenerative diseases, including diabetes Numerous components were isolated from mellitus, inflammation, stroke, cancer, petals, flowers and hips (seed-pot) of R. coronary heart disease, and aging1,2. damascena including flavonoids, Reactive oxygen species (ROS) include free anthocyanins, glycosides, and terpenes. radicals such as .O2- (superoxide anion), Plant shows presence of kaempferol, .OH (hydroxyl radical), H2O2 (hydrogen carboxylic acid, quercetin, myrcene, and peroxide) and .O2 (singlet oxygen) which vitamin C. Flowers show presence of fatty cause cellular injuries and initiate oil, bitter principle, organic acids and peroxidation of polyunsaturated fatty acids tanning matter7.The essential oil of Rose in biological membranes3,4. The tissue injury contains more than 95% of total oil caused by ROS include protein damage, represented by eighteen compounds. β- DNA damage and oxidation of important citronellol (14.5-47.5%), geraniol (5.5- enzymes in the human cells. The 18%), nerol, kaempferol and nonadecane consequences of these events may lead to (10.5-40.5%) are the identified oils. In the occurrence of various oxidative stress addition, citrenellol (9.91%), phenyl ethyl related diseases. Recently, plants, natural alcohol (78.38%), geraniol and nonadecane foods and food derived antioxidants such as (4.35%) are also found to be present in Rose vitamins and phenolic phytochemicals have oil8. received growing attention, since they play an important role as preventive agents MATERIALS AND METHODS against damage caused due to oxidative stress5. Procurement of plant material Rosa genus, belonging to the Fresh petals of Rose flower were Rosaceae family, includes 200 species and procured from local market at Dadar, more than 18,000 cultivars. One of the most Mumbai, India and were authenticated at important Rosa species is Rosa damascena Agarkar Research Insitute, Pune, India. Mill. This plant is called damask rose because it was originally brought to Europe Chemicals and instruments from Damascus6. The essential oil of Rosa 1,1-Diphenyl-2-picrylhydrazyl damascena is known for its fine perfumery (DPPH), Gallic acid, 6-Hydroxy-2, 5, 7, 8- applications and the use in cosmetic tetramethylchroman-2-carboxylic acid preparations. The use of Rosa damascena is (Trolox), 2,2’-azinobis-(3-ethylbenzo- increasing steadily worldwide for its thiazoline)-6-sulphonic acid (ABTS), medicinal properties health promoting Allopurinol, Bovine Brain Extract (BBE), benefits. Recently, the antioxidant, Bovine Serum Albumin (BSA), were antibacterial, anti-HIV and activities of its procured from Sigma Chemical Co. (USA). essential oil have been demonstrated. This Xanthine oxidase, Nitroblue tetrazolium plant is also used as a gentle laxative. Rose chloride (NBT) and xanthine were procured oil is used for healing nervous stress, from Himedia Ltd. Mumbai, India. Folin- tension, grief and depression. It helps in the Ciocalteu solution, Pyrogallol Red, reduction of thirst, curing old cough, treating potassium dihydrogen phosphate and special complaints of women, wound thiobarbituric acid, dipotassium hydrogen healing, and improving skin health. Vapour phosphate, trichloroacetic acid, anhydrous AJPCT[3][09][2015] 589-601 Tatke et al__________________________________________________ ISSN 2321 – 2748 sodium carbonate, ascorbic acid, butylated made upto 10 ml. The final mixture was hydroxytoluene (BHT) potassium persulfate, allowed to stand at room temperature for 2 h ferrous ascorbate, ethylene diamine tetra with intermittent shaking. Then the acetic acid (EDTA), were purchased from absorbance of the dark blue colour that S.D.Fine Chemicals, Mumbai, India. All developed was measured at 725 nm using other chemicals and solvents used were of UV-Vis spectrophotometer. Gallic acid was analytical grade. used as standard for preparing the The instruments used for the study calibration curve (10 μg/ml - 100 μg/ml). were UV spectrophotometer, (Jasco, V-630), The concentration of total phenolic laboratory centrifuge (Remi motors, R4C) compounds in the extracts was calculated and digital pH meter (Equip-tronics, EQ- using the following linear equation based on 610). the calibration curve: y = 0.010x + 0.01 with R2= 0.994. The total phenolic content in the Preparation of extracts plant extract was expressed as mg of gallic Fresh petals of Rose were acid equivalent per g of dry weight (mg immediately used for extraction. Some of GAE/g) of extract. All the tests were carried the fresh petals were dried, stored in air-tight out in triplicate and the results are expressed container and then used for extraction. The as mean ± SD. extraction of plant material was carried out by Soxhlet extraction method using various Quantification of total flavonoids solvents such as chloroform, acetone, The total flavonoid content of all the methanol. and water. All extracts were extracts was determined by aluminium filtered individually and further evaporated chloride colorimetric method9. 0.5 ml of to get dry extracts. The extractive values sample solution was mixed with 2 ml of were determined. After drying, crude distilled water and subsequently with 0.15 extracts were stored in desiccator till further ml of 5% NaNO2 solution. After 6 min of use. incubation, 0.15 ml of 10% AlCl3 solution was added and then allowed to stand for 6 Phytochemicals screening min, followed by adding 2 ml of 4% NaOH Extracts were qualitatively evaluated solution to the mixture. Immediately water for the presence or absence of active was added to the sample to make the final principles such as phytosterols, tannins, volume to 5 ml, the mixture was thoroughly flavonoids, saponins, alkaloids, glycoside, mixed and allowed to stand for another 15 triterpenoids and proteins. min. The absorbance was determined at wavelength 510 nm. The total flavonoid Quantification of total phenolics – by Folin- content was expressed in mg of rutin Ciocalteu method equivalent per g of extract (mg RE/g) of The total phenolic content in various extract. extracts of the plants were measured using Folin-Ciocalteu reagent9. 1 ml of extract In vitro antioxidant studies solution was added to 0.5 ml of Folin The ethanol and acetone extracts of Ciocalteau reagent and 5 ml of distilled fresh and dried Rose flower petals with high water. The mixture was incubated at room phenolic and flavonoid content were further temperature for 10 min. Then 1.5 ml of evaluated for their antioxidant and radical anhydrous sodium carbonate solution (10% scavenging potential w/v) was added and the final volume was AJPCT[3][09][2015] 589-601 Tatke et al__________________________________________________ ISSN 2321 – 2748 DPPH (1, 1, diphenyl 2-picryl hydrazyl) The % inhibition of pyrogallol red bleaching assay was determined using the formula [(A1- The free radical scavenging activity A2)/A1] X 100, where A1 is the absorbance of the extracts, based on the scavenging in
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