HPLC双波长法测定豌豆尖中槲皮素和山奈酚的含量
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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.
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.
引文
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[2]马杰,孙勃,薛生玲,等.豌豆尖主要营养成分、生物活性物质及抗氧化能力分析[J].食品与机械,2016,(04):47-51
[3]中国疾病预防控制中心营养与食品安全所.中国食物成分表2009[M].北京:北京大学医学出版社,2009
[4]于仁文.排毒养肝的豌豆尖[J].中国老年,2016,(13):52
[5]叶春,聂开慧.对40种新鲜蔬菜中总黄酮含量的测定[J].山地农业生物学报,2000,(02):121-124
[6]J Santosa,M Herrerob,J A Mendiolab,et al.Assessment of nutritional and metabolic profiles of pea shoots:The new ready-to-eat baby-leaf vegetable[J].Food research international,2014,58:105-111
[7]中国营养学会.中国居民膳食营养素参考摄入量(2013版)[M].北京:科学出版社,2014:494-495
[8]何煜舟,汪云开,祝晨,等.山萘酚抗氧化作用研究进展[J].浙江中西医结合杂志,2012,(06):499-501
[9]陈育华,周克元,袁汉尧.山奈酚药效的研究进展[J].广东医学,2010,(08):1064-1066