Keywords
HPLC analysis
vegetables
How to Cite
Abstract
The objective of this investigation was to optimize the analytical methodology for determining flavonols and flavones in vegetables. The hydrolysis procedure was optimized using Central Composite Rotational Design (CCRD) to investigate the effects of HCl concentration and hydrolysis time. This step was carried out simultaneously with extraction with 50% aqueous methanol, and refluxing at 90°C. A Waters liquid chromatograph, with Nova-Pak C18 column and photodiode array detector, was used. The analyzed compounds were myricetin (M), quercetin (Q), kaempferol (K), luteolin (L), and apigenin (A). The optimum conditions found for hydrolysis for each vegetable were: 1.0M HCl for 6 hours for spinach and kale, 1.6M HCl for 5 hours for roquette, 1.2M HCl for 2 hours for lettuce, 1.7M HCl for 4.3 hours for parsley, and 0.8M HCL for 2.5 hours for onion. The best gradient (HPLC) for separating flavonoids from these vegetables consisted of methanol:water (acidified with 0.03% formic acid) 20:80, changing to 45:55 in 5 minutes, 48:52 in 17 minutes, returning to 20:80 in 20 minutes. The standard curves of the flavonoids had coefficients of correlation higher than 0.99. The detection limits were 0.5, 0.4, 0.5, 0.6 and 1.0μg/mL for M, Q, L, K, and A, respectively.
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