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目的:建立HPLC双波长法同时测定豌豆尖中类黄酮水解产物槲皮素和山奈酚含量的方法。方法:先用甲醇-15%盐酸(4:1,v/v)混合液把豌豆尖中的类黄酮水解成槲皮素和山奈酚,后采用HPLC双波长法测定槲皮素和山奈酚的含量。GRACE Vision HT C_(18)色谱柱(250 mm×4.6mm,5μm),流动相为甲醇-0.1%冰醋酸(50:50,v/v),流速为1.0 mL/min,槲皮素检测波长为371 nm,山奈酚检测波长为366 nm,柱温为30℃。结果:槲皮素和山奈酚分别在(0.05~0.3)μg、(0.025~0.15)μg范围内线性关系良好,平均加标回收率分别为99.30%(RSD=1.74%)、98.33%(RSD=1.21%)。结论:该方法操作简便、结果可靠、重现性好,可用于豌豆尖中槲皮素和山奈酚含量的同时测定。
OBJECTIVE: To establish a method for the simultaneous determination of quercetin and kaempferol, the flavonoid hydrolyzate of pea spike by HPLC dual wavelength method. Methods: The flavonoids from peas were hydrolyzed to quercetin and kaempferol with methanol-15% hydrochloric acid (4: 1, v / v) mixture, and the contents of quercetin and kaempferol content. The mobile phase consisted of methanol-0.1% glacial acetic acid (50:50, v / v) at a flow rate of 1.0 mL / min on a GRACE Vision HT C 18 column (250 mm × 4.6 mm, 5 μm) For 371 nm, the detection wavelength of kaempferol was 366 nm, and the column temperature was 30 ℃. Results: The calibration curves of quercetin and kaempferol were linear within the range of (0.05-0.3) μg and (0.025-0.15) μg, respectively. The average recoveries were 99.30% (RSD = 1.74%) and 98.33% (RSD = 1.21%). Conclusion: The method is simple, reliable and reproducible. It can be used for the simultaneous determination of quercetin and kaempferol in peas.