Resumen
O objetivo deste trabalho foi otimizar a metodologia analítica para determinação de flavonóis e flavonas em hortaliças. A hidrólise foi otimizada utilizando-se Delineamento Composto Central Rotacional (DCCR) para investigar os efeitos da concentração de HCl e do tempo de hidrólise. Essa etapa foi realizada simultaneamente com a extração por metanol aquoso 50%, em refluxo a 90ºC. Foi utilizado cromatógrafo líquido Waters com coluna Nova-Pak C18 e detector de arranjo de diodos. Os compostos estudados foram miricetina (M), quercetina (Q), kaempferol (K), luteolina (L) e apigenina (A). As condições ótimas encontradas para hidrólise de cada hortaliça foram: 1,0M HCl/6 horas para espinafre e couve, 1,6M HCl/5 horas para rúcula, 1,2M HCl/2 horas para alface, 1,7M HCl/4,3 horas para salsa e 0,8M HCl/2,5 horas para cebola. O melhor gradiente para separação (CLAE) dos flavonóides das hortaliças em estudo foi constituído de metanol:água (acidificados com 0,3% de ácido fórmico) 20:80, chegando a 45:55 em 5 minutos, 48:52 em 17 minutos e voltando a 20:80 em 20 minutos. As curvas analíticas apresentaram coeficientes de correlação maiores que 0,99. Os limites de detecção foram de 0,5, 0,4, 0,5, 0,6 e 1,0μg/mL, respectivamente, para M, Q, L, K e A.Citas
1. Anderson RF, Amarasinghe C, Fisher LJ, Mak WB, Packer JE. Reduction in free-radical-induced DNAstrand breaks and base damage through fast chemicalrepair by flavonoids. Free Rad Res 2000; 33: 91-103.
2. Hertog MGL, Feskens EJM, Hollman PCH, Katan MB, Kromhout D. Dietary antioxidant flavonoids and riskof coronary heart disease: the Zutphen Elderly Study.Lancet 1993; 342: 1007-1011.
3. Hollman PCH, Hertog MGL, Katan MB. Role of dietaryflavonoids in protection against cancer and coronaryheart disease. Biochem Soc Trans 1996; 24: 785-9.
4. Hollman PCH. Evidence for health benefits of plantphenols: local or systemic effects? J Sci Food Agric2001; 81: 842-52.
5. Knekt P, Jarvinen R, Reunanen A, Maatela J. Flavonoidintake and coronary mortality in Finland: a cohort study.Brit Med J 1996; 312: 478-81.
6. Middleton Jr E, Kandaswami, C. Effects of flavonoidson imune and inflammatory functions. BiochemPharmacol 1992; 43: 1167-79.
7. Middleton Jr E, Kandaswami C. The impact of plantflavonoids on mammalian biology: implications forimmunity, inflammation and cancer. In: Harborne JB,editor. The flavonoids. London: Chapman and Hall;1994.
8. Castelluccio C, Paganga G, Melikian N, Bolwell GP,Pridham J, Sampson J et al. Antioxidant potential ofintermediates in phenylpropanoid metabolism in higherplants. Febs Lett 1995; 368: 188-92.
9. Kandaswani C, Middleton E. Free radical scavengingand antioxidant activity of plant flavonoids.Adv ExpMed Biol 1994; 366: 351-76.
10. Rice-Evans CA, Miller NJ, Bolwell GP, Bramley PM,Pridham JB. The relative antioxidant activities of plant-derived polyphenolic flavonoids. Free Rad Res 1995;22: 375-83.
11. Rice-Evans CA, Miller, NJ, Paganga G. Structure-antioxidant activity relationships of flavonoids andphenolic acids. Free Radic Biol Med 1996; 20: 933-56.
12. Freedman JE, Parker C, Li L, Perlman JA, Frei B, IvanovV et al. Select flavonoids and whole juice from purple grapes inhibit platelet function and enhance nitric oxiderelease. Circulation 2001; 103: 2792-98.
13. Tzeng SH, Ko WC, Ko FN, Teng CM. Inhibition ofplatelet aggregation by some flavonoids. Thromb Res1991; 64: 91-100.
14. Kuntz S, Wenzel U, Daniel H. Comparative analysis ofthe effects of flavonoids on proliferation, cytotoxicity,and apoptosis in human colon cancer cell lines. Eur JClin Nutr 1999; 38: 133-42.
15. Wenzel U, Kuntz S, Brendel MD, Daniel H. Dietaryflavone is a potent apoptosis inducer in human coloncarcinoma cells. Cancer Res 2000; 60: 3823-31.
16. Yang CS, Landau JM, Huang MT, Newmark HL.Inhibition of carcinogenesis by dietary polyphenoliccompounds. Annu Rev Nutr 2001; 21: 381-406.
17. Schroeter H, Boyd C, Spencer JPE, Williams RJ,Cadenas E, Rice-Evans C. MAPK signaling inneurodegeneration: influences of flavonoids and of nitricoxide. Neurobiol Aging 2002; 23: 861-80.
18. Crozier A, Lean MEJ, McDonald MS, Black C.Quantitative analysis of the flavonoid content ofcommercial tomatoes, onions, lettuce, and celery. J AgricFood Chem 1997; 45: 590-5.
19. Hertog MGL, Hollman PCH, Katan MB. Content ofpotentially anticarcinogenic flavonoids of 28 vegetablesand 9 fruits commonly consumed in the Netherlands. JAgric Food Chem 1992; 40: 2379-83.
20. Hertog MGL, Hollman PCH, van de Putte B. Contentof potentially anticarcinogenic flavonoids of teainfusions, wines and fruit juices. J Agric Food Chem1993; 41: 1242-6.
21. Hertog MGL, Hollman PCH, Venema DP. Optimizationof a quantitative HPLC determination of potentially anticarcinogenic flavonoids in vegetables and fruits. JAgric Food Chem 1992; 40: 1591-8.
22. McDonald M, Hughes M, Burns J, Lean MEJ,Matthews D, Crozier A. Survey of the free andconjugated myricetin and quercetin content of redwines of different geographical origins. J Agric FoodChem 1998; 46: 368-75.
23. Vries J, Janssen K, Hollman PCH, Van Staveren WA,Katan MB. Consumption of quercetin and kaempferolin free-living subjects eating a variety of diets. CancerLett 1997; 114: 141-4.
24. Merken HM, Beecher GR. Measurement offood flavonoids by high-performance liquidchromatography: A review. J Agric Food Chem 2000;48: 577-99.
25. DiCarlo G, Mascolo N, Izzo AA, Capasso F. Flavonoids:old and new aspects of a class of natural therapeutic drugs. Life Sci 1999; 65(4): 337-53.
26. Erlund I. Review of the flavonoids quercetin, hesperetin, and naringenin. Dietary sources, bioactivities, bioavailability, and epidemiology. NutrRes 2004; 24: 851-74.
27. Hermann K. On the occurrence of flavonol and flavone glycosides in vegetables. Z Lebensm Unters Forsch A1988; 186: 1-5.
28. Häkkinen SH, Kärenlampi SO, Heinonen IM, Mykkänen HM, Törrönen AR. HPLC method for screening offlavonoids and phenolic acids in berries. J Sci FoodAgric 1998; 77:543-51.
29. Nuutila AM, Kammiovirta K, Oksman-CaldenteyKM. Comparison of methods for the hydrolysis of flavonoids and phenolic acids from onion andspinach for HPLC analysis. Food Chem 2002; 76:519-25.
30. Haaland PO. Experimental design in biotechnology. New York: Marcel Dekker; 1989.
31. Box GEP, Hunter WG, Hunter JS. Statistics for experimenters: an introduction to design, data analysisand model building. New York: Wiley; 1978.
32. Rodrigues MI, Iemma AF. Planejamento de Experimentos e Otimização de Processos. Campinas: Casa do Pão Editora; 2005.
33. Caulcutt R, Boddy R. Statistic for analytical chemists. 1st ed. London: Chapman and Hall; 1983.
34. Ribani M, Bottoli CBG, Collins CH, Jardim ICSF, Melo LFC. Validação em métodos cromatográficose eletroforéticos. Quím Nova 2004; 27: 771-80.
Esta obra está bajo una licencia internacional Creative Commons Atribución 4.0.
Derechos de autor 2007 Revista del Instituto Adolfo Lutz