Optimization Of A Method For Determination Of Flavonols And Flavones In Fruits By Hplc Using Statistical Design And Response Surface Analysis [otimização De Método Para Determinação De Flavonóis E Flavonas Em Frutas Por Cromatografia Líquida De Alta Eficiência Utilizando Delineamento Estatístico E Análise De Superfície De Resposta]

This work optimized the HPLC conditions for the simultaneous determination of luteolin, apigenin, myricetin, quercetin and kaempferol in aglycone form, as well defined the best conditions for hydrolysis/extraction of these flavonoids in fruits, using the statistical central composite design and response surface analysis. A reverse phase method was developed using a gradient of methanol/water acidified with 0.3% formic acid as mobile phase and a photodiode array detector. The samples were extracted with methanol/water (50:50 v/v) at 90̊C. The optimum time and HCl concentration varied for the different fruits investigated, demonstrating the necessity of optimizing these conditions for each fruit analyzed. Good recovery (87.1 to 96.3%), repeatability and linearity were obtained.31613781384+S1-S2Armstrong, B.K., Mann, J.L., Adelstein, A.M., Eskin, F., (1975) J. Chron. Dis, 28, p. 455Block, G., Patterson, B., Subar, A., (1992) Nutr. Cancer, 18, p. 1Burr, M.C., Sweetnam, P.M., (1982) Am. J. Clin. Nutr, 36, p. 673Phillips, R.L., Lemon, F.R., Beeson, W.L., Kuzma, J.W., (1978) Am. J. Clin. Nutr, 31, p. 191Verlangieri, A.J., Kapeghian, J.C., El-Dean, S., Bush, M., (1985) Med. Hypotheses, 16, p. 7Castellucio, C., Paganga, G., Melikian, N., Bolwell, G.P., Pridham, J., Sampson, J., Rice-Evans, C., (1995) FEBS Lett, 368, p. 188Hertog, M.G.L., Feskens, E.J.M., Hollman, P.C.H., Katan, M.B., Kromhout, D., (1993) Lancet, 342, p. 1007Keli, S.O., Hertog, M.G.L., Feskin, E.J.M., Kromhout, D., (1996) Arch. Int. Med, 156, p. 632Knekt, P., Jarvinen, R., Reunanen, A., Maatela, J., (1996) Br. Med. J, 312, p. 478Middleton, E., Rice-Evans, C., Miller, N.J., Bolwell, G.P., Bramley, P.M., Pridham, J.B., (1995) Free Radical Res, 22, p. 375Rimm, E.B., Katan, M.B., Ascherio, A., Stampher, M.J., Willet, W.C., (1996) Ann. Int. Med, 125, p. 384Kühnau, J., (1976) World Rev. Nutr. Diet, 24, p. 117Markham, K.R., (1989) Meth. Plant Biochem, 1, p. 197Williams, C. A.Harborne, J. B. Em The Flavonoids. Advances in Research since 1986Harborne, J.B., ed.Chapman & Hall: London, 1994Hertog, M.G.L., Hollman, P.C.H., Venema, D.P., (1992) J. Agric. Food Chem, 40, p. 1591Haaland, P.O., (1989) Experimental Design in Biotechnology, , Marcel Dekker: New YorkBox, G.E.P., Wilson, K.B., (1951) J. Roy. Statist. Soc, B13, p. 1Myers, R.H., Montgomery, D.C., (2002) Response Surface Methodology: Process and Product Optimization using Designed Experiments, , 2nd ed, Wiley: New YorkBarros Neto, B.Scarminio, I. S.Bruns, R. E.Planejamento e Otimização de Experimentos, 2a ed., Educamp: Campinas, 1995Cacace, J.E., Mazza, G., (2003) J. Food Sci, 68, p. 240Careri, M., Elviri, L., Mangia, A., Musci, M., (2000) J. Chromatogr., A, 881, p. 449Catharino, R.R., Godoy, H.T., (2001) Ciênc. Tecnol. Aliment, 21, p. 326Liyana-Pathirana, C., Shahidi, F., (2005) Food Chem, 93, p. 47Arabbi, P.R., Genovese, M.I., Lajolo, F.M., (2004) J. Agric. Food Chem, 52, p. 1124Franke, A.A., Custer, L.J., Arakaki, C., Murphy, S.P., (2004) J. Agric. Food Chem, 17, p. 1Häkkinen, S., Törrönen, R., (2000) Food Res. Int, 33, p. 517Hertog, M.G.L., Hollman, P.C.H., Katan, M.B., (1992) J. Agric. Food Chem, 40, p. 2379Shabir, G.A., (2003) J. Chromatogr., A, 987, p. 57Franke, A.A., Custer, L.J., Arakaki, C., Murphy, S.P., (2004) J. Agric. Food Chem, 17, p. 1Häkkinen, S., Törrönen, R., (2000) Food Res. Int, 33, p. 517Hertog, M.G.L., Hollman, P.C.H., Katan, M.B., (1992) J. Agric. Food Chem, 40, p. 237