HPLC法同时测定沙棘黄酮中槲皮素、山柰酚和异鼠李素的含量

HPLC method in determining the content of quercetin,kaempferide and isorhamnetin in Seabuckthorn flavone powder

目的:应用高效液相色谱法(HPLC)同时测定沙棘黄酮粉中槲皮素、山柰酚和异鼠李素的含量。方法:沙棘黄酮经甲醇超声提取后,加入适量盐酸75℃水浴水解,将ZORBAX SB-C18(4.6mm×250mm,5μm)柱作为固定相;甲醇(A)-0.4%磷酸水溶液(B)作为流动相,进行梯度洗脱;流速为1.0mL/min;检测波长为370nm;进样量20μL;柱温25℃。结果:槲皮素、山柰酚和异鼠李素的质量浓度分别在10.40~24.26、4.87~11.36、36.20~84.46μg/mL范围内线性关系良好,平均加样回收率分别为93.60%、98.27%、98.66%,RSD分别为0.26%、1.01%、0.21%。结论:本实验建立的HPLC方法简单易行,稳定可靠,重复性强,可用于沙棘黄酮中槲皮素、山柰酚和异鼠李素的含量测定。

Objective:To develop an HPLC method for simulaneous determination of three flavonoids such as quercetin、kaempferol and isorhamnetin in Hippophae rhamnoides.Methods:Seabuckthorn flavonoids with methanol after ultrasonic extraction,adding suitable amount of hydrochloric acid hydrolysis 75℃water bath.The optimum HPLC condition was as follows:Agilent ZORBAX SB-C18 column(4.6 mm×250 mm,5μm)as the stationary phase;mobile phase:methanol-0.4%phosphoric acid,gradient elution,detection wavelength 370 nm;column temperature 25℃;flow rate:1.0 mL/min.Results:The method had good linear relationship within the range of 10.40~24.26μg/mL for quercetin,4.87~11.36μg/mL for kaempferol,and 36.20~84.46μg/mL for isorhamnetin.The average recoveries of quercetin、kaempferol and isorhamnetin were 93.60%、98.27%and 98.66%respectively and the RSD were 0.26%、1.01%and 0.21%respectively.Conclusion:The method is simple,reliable and reproducible.It can be used for the determination of quercetin、kaempferol and isorhamnetin in flavonoids of seabuckthorn.