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Universidad del Valle Comparison of DPPH Free Radical Scavenging, Ferric Reducing Antioxidant Power (FRAP), and Total Phenolic Content of Two Meriania Species (Melastomataceae) Claudia Lorena Valverde Malaver Universidad del Valle José Hipólito Isaza Martínez Universidad del Valle Ana Julia Colmenares Dulcey Universidad del Valle Received: June 9, 215 Accepted: November 5, 215 Pag. 117-124 Abstract Searching for new antioxidants used in pharmaceuticals, cosmetics or agrochemicals have increased today. Many phenolic compounds have been reported as promising for this goal. Melastomataceae is rich in these compounds. Consequently, in this research the antioxidant power for 5, mg.l -1-1 -1 The present study revealed that Mh 2.1 and Mh 2.2 from M. hernandoi have a strong antioxidant power FRS 5 = 5.71±.85, FRAP = 53±1.5 and TPC = 224±7.1 and FRS 5 = 4.22±.1, FRAP = 65 mg±1.5, TPC = 24 ±8.6 respectively, as compared with positive control, quercetin DPPH FRS 5 = 4.2±.41, FRAP = 61±1.3. M. nobilis exhibited poor antioxidant power FRS 5 = 286±1.5, FRAP = 2.1±.79 and TPC = 26±1.5. This paper showed M. hernandoi as good source of antioxidant compounds with potential usage as cosmetic, pharmaceutical or agrochemical ingredients. Keywords: Meriania hernandoi. 1 Introduction Many studies have been published about phytochemicals extracted from plants with strong antioxidant activity. Compounds exhibiting this activity which are capable Oxidative stress is associated with an increased risk of some diseases such as diabetes, Volumen 19 No. 2, diciembre 215

C. L. Valverde, A. J. Colmenares and J. H. Isaza for this goal with similar activities to vitamins. The genera belonging to the family of Melastomataceae have been reported as plants rich in phenolic compounds such as Meriania is a neotropical genus, with more than 5 species, distributed in the Antilles, and from southern Mexico to southeast Brazil. In Colombia are known around 36 species, including Meriania nobilis and Meriania hernandoi, used mainly as ornamental. Their biological activities and phytochemicals have not been previously reported; therefore this research evaluated and compared antioxidant activity by DPPH free radical scavenging Meriania nobilis and Meriania hernandoi. 2 Materials and Methods 2.1 Plants and materials Meriania nobilis Triana Colombia, on May 212 and Meriania hernandoi Reserva Civil El refugio, in Dagua, Valle del Cauca, Colombia, on November 213. Tascón of the Universidad del Valle. 2.2 Extraction room temperature for 15 minutes. The solvent was evaporated at low pressure in an IKA 2.3 DPPH Radical - Scavenging Method DPPH free radical scavenging activity was determined in triplicate by the method of -1 a 132 mg.l -1 DPPH solution prepared in methanol. After 1 hour of reaction, the absorbance 3 118

Comparison of DPPH, FRAP, and Total Phenolic of Two Meriania Species %FRS = A c A A c s x 1 Where Ac is the absorbance of DPPH radicals without sample or positive control 5-1. 2.4 Ferric Reducing Antioxidant Power Assay (FRAP) Power reducing of Fe +3 to Fe +2 by an antioxidant was determined using the method 2 mm FeCl 3-1 4 readings were taken at 6 nm immediately after and 6 min after using a Metertech, 4 4-1 dried extract. 2.5 Estimation of Total Phenolic Content (TPC) to stand at room temperature for 15 min. The absorbance was read at 7 nm. A standard -1-1 dried extract. 2.6 Statistical analysis Results were given as the mean±sd for at least three replicates for each sample. Statistical 5 values were calculated using no nonlinear regression with one phase exponential decay calculation. Volumen 19 No. 2, diciembre 215 119

C. L. Valverde, A. J. Colmenares and J. H. Isaza 3 Results species M. nobilis and M. hernandoi M. hernandoi extracts. Figure 1. Percentage of radical scavenging capacity for extracts of the species M. nobilis, M. hernandoi. Values are the average of three replicates 5 5 values were ranging from the higher at 512 mg. L -1-1 extracts of M. nobilis showed a moderate radical scavenging activity, being Mn 2.2 the better with FRS 5 equal to 287 ± 1.7 mg.l -1 ; while M. hernandoi showed strong radical -1-1 -1-1 -1 employed. 8 FRS 5 (mgl -1 ) 6 4 512 287 5 2 4.2 5.34 Quercetin Ascorbic acid Mn 2.1 Mn 2.2 Mn 2.3 Mh 2.1 Standard and extracts 5.71 4.22 14.5 Mh 2.2 Mh 2.3 Figure 2. Activities radical scavenging for extracts of the species M. nobilis, M. hernandoi and positive controls. Values are the average of three replicates. 12

Comparison of DPPH, FRAP, and Total Phenolic of Two Meriania Species The antioxidant power was determined by the FRAP assay that measures the ability of the sample to reduce the Fe 3+ in a solution of TPTZ to Fe 2+. The values obtained for the samples ranged from 65 mg FeSO 4-1 ES to.28 mg FeSO 4-1 M. hernandoi extracts that showed strong antioxidant power were Mh 4-1 4-1 ES for quercetin 61 mg FeSO 4-1 ES ±1.3. Very low values of antioxidant power were obtained for M. nobilis 4-1 4-1 4 O -1 FeSO 4 7H 2 O mg(1 g DE) -1 8 61 6 53 4 2.28 2.1.57 Quercetin Mn 2.1 Mn 2.2 Mn 2.3 Mh 2.1 Standard and extracts 65 7.4 Mh 2.2 Mh 2.3 Figure 3. The antioxidant power for extracts of the species M. nobilis, M. hernandoi and positive control. Values are the average of three replicates. Total phenolic content estimation by Folin - Ciocalteu method was in the range from 11.9-1 M. hernandoi: Mh -1-1 -1 3 224 24 (AG mg)g -1 DE 2 1 12 11.9 Mn 2.1 26. 23.6 Mn 2.2 Mn 2.3 Mh 2.1 Mh 2.2 Mh 2.3 Extracts Figure 4. Estimation of total phenolic content by Folin-Ciocalteu method of the species M. nobilis and M. hernandoi. Values are the average of four replicates. Volumen 19 No. 2, diciembre 215 121

C. L. Valverde, A. J. Colmenares and J. H. Isaza 4 Discussion M. Hernandoi extracts possess a striking M. nobilis extracts the percentages were obtained FRS 5 2 5 2 2 5 assay, showing to M. hernandoi extracts, specially Mh 2.1 and Mh 2.2 as good source of antioxidant compounds from natural origin with potential usage as cosmetic, pharmaceutical or agrochemical ingredients. suggesting phenolic compounds as potential responsible for this activity. The values of FRS 5 and FRAP for the two species behaved similarly. This was checked by determining 2.423X obtained FRS 5 R 2.34X 2.423X FRS 5 8 6 4 R 2 =.9974 FRS 5 1 8 6 4 R 2 =.91 2 2 2 4 6 8 FRAP 1 2 3 TPC 3 R 2 =.995 2 TPC 1 2 4 6 8 FRAP Figure 5. Correlations between the methods used to determine antioxidant activity. FRS 5 = DPPH free radical Antioxidant Power, TPC= Total Phenolic Content. 122

Comparison of DPPH, FRAP, and Total Phenolic of Two Meriania Species Many in vitro methods are used to determine the antioxidant activity, but they do not methods with different mechanisms were used, they correlate really well, indicating that the main compounds responsible of both antioxidant activities, DPPH free radical scavenging and ferric reducing antioxidant power of M. nobilis and M. hernandoi, are phenolics. 5 Conclusions This paper showed the species M. hernandoi as good source in antioxidant compounds from of natural origin with potential usage as cosmetic, pharmaceutical or agrochemical ingredients, due to activity found by the methods in vitro free radical DPPH scavenging, of M. hernandoi are promising for future studies to isolate and elucidate the metabolites that exhibit this activity. The specie Meriania nobilis showed a poor antioxidant activity, which agree to in vitro methods employed. The in vitro method employed correlated well which allowed determining the antioxidant power of the two species studied. Acknowledgements Authors are grateful to COLCIENCIAS, Grant CT-41-211, and to Universidad del Valle Grant CI 7848 and professor Carlos Calderon, of the Reserva Civil El Refugio by providing plant material. References [1] measure of antioxidant power: The FRAP assay. Analytical Biochemistry, 239, 7-76. [2] Food Research International, 44, 212-2127. [3] of the metabolic syndrome: The oxidative stress. Nutrition, Metabolism and Cardiovascular Diseases, 2, 72-77. [4] espectrofotométrica de taninos hidrolizables y condensados en plantas melastomatáceas. Scientia Et Technica, 13, 5. [5] allied families. Phytochemistry, 15, 513-516. [6] Biochemical Systematics and Ecology, 32, 27-34. Volumen 19 No. 2, diciembre 215 123

C. L. Valverde, A. J. Colmenares and J. H. Isaza [7] Industrial Crops and Products, 51, 36-369. [8] Nono, R. N., Barboni, L., Teponno, R. B., Quassinti, L., Bramucci, M., Vitali, L. activities and phytoconstituents of extracts from the roots of Dissotis thollonii South African Journal of Botany, 93, 19-26. [9] Phytochemistry, 68, 1781-1784. [1] composition, organic acids, sugars, vitamin C and antioxidant activity in the juice of two new triploid late-season mandarins. Food Research International, 49, 462-468. [11] Experimental Physiology, 82, 291-295. [12] Neuroprotective activity of galloylated cyanogenic glucosides and hydrolysable tannins isolated from leaves of Phyllagathis rotundifolia. Fitoterapia, 83, 223-229. [13] Tarawneh, A. H., León, F., Ibrahim, M. A., Pettaway, S., McCurdy, C. R., y Cutler, Phytochemistry Letters, 7, 13-132. [14] Journal of Agricultural and Food Chemistry, 51, 6445-6451. [15] properties of ellagitannins in some plant families of the order myrtales. International Journal of Molecular Sciences, 11, 79-16. Autor s address Claudia Lorena Valverde Malaver Departamento de Química, Universidad del Valle, Cali - Colombia claudia.valverde@correounivalle.edu.co Ana Julia Colmenares Dulcey Departamento de Química, Universidad del Valle, Cali - Colombia ana.colmenares@correounivalle.edu.co José Hipólito Isaza Martínez Departamento de Química, Universidad del Valle, Cali - Colombia jose.isaza@correounivalle.edu.co 124