HPLC双波长法测定豌豆尖中槲皮素和山奈酚的含量  

Determination of quercetin and kaempferol in pea shoots by dual wave-length HPLC

作　　者：曹晓华[1,2] 沈旭斌 程先骄 林兆娜 陈旭[2,3] 程立立 郑金贵 

机构地区：[1]福建农林大学作物科学学院,福州350002 [2]作物遗传育种与综合利用教育部重点实验室、福建农林大学,福州350002 [3]福建农林大学食品科学学院,福州350002

出　　处：《食品科技》2017年第4期270-276,共7页Food Science and Technology

基　　金：国家科技支撑计划项目(2013BAD01B05);福建农林大学科技创新平台建设项目(PTJH13001,PTJH12015)

摘　　要：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 simultaneous determination of quercetin and kaempferol in flavonoids hydrolysate from pea shoots by dual wave-length HPLC. Methods： The flavonoids in pea shoots were hydrolyted into quercetin and kaemferol with the mixed solution of methanol-15% hydrochloric acid（4：1, v/v）, then the contents of quercetin and kaempferol were determined by dual wave-length HPLC. The sample solution was separated by using GRACE Vision HT C_（18）（250 mm×4.6 mm×5 μm） as stationary phase and a mixture of methanol and 0.1% glacial acid solution（50：50, v/v） as mobile phase at a flow rate of 1.0 mL/min. The detection wavelength of quercetin was set at 371 nm and kaempferol＇s detection wavelength was set at 366 nm, the column temperature was set at 30 ℃ and the injection volume was 10 μL. Results： Quercetin and kaempferol had good linear relationship within the range of（0.05-0.3）μg and（0.025-0.15） μg respectively. The average recovery rate of quercetin and kaempferol were 99.30%（RSD=1.74%） and 98.33%（RSD=1.21%） respectively. Conclusion： The method was simple, accurate and reproducible, which could be applied for the simultaneous determination of quercetin and kaempferol in pea shoots.

关 键 词：豌豆尖 槲皮素 山奈酚 超声提取 高效液相色谱法 双波长 

分 类 号：TS207.3[轻工技术与工程—食品科学]