HPLC法测定何首乌(黑豆酒炙)饮片中游离蒽醌的含量

Determination of Free Anthraquinones in processed Polygoni Multiflori Radix by HPLC

目的:建立中药饮片何首乌(黑豆酒炙)中游离蒽醌(大黄素和大黄素甲醚)的HPLC含量测定方法。方法:采用高效液相色谱法对何首乌(黑豆酒炙)中的游离蒽醌含量进行测定。色谱柱为Agilent ZORBAX SB-C18,4.6 mm×250 mm,5μm,流动相为甲醇-0.1%磷酸(80∶20),等度洗脱,流速1.0 mL·min-1,柱温:30℃,检测波长:254 nm。结果:大黄素在7.58~151.60μg·mL-1范围内峰面积对进样量线性关系良好(r=0.9999);大黄素甲醚在3.93~78.57μg·mL-1范围内峰面积对进样量线性关系良好(r=0.9999),大黄素和大黄素甲醚的回收率分别为:95.40%~99.63%(RSD=1.60%,n=6),98.21%~100.17%(RSD=0.80%,n=6)。7批不同批次的样品测得的大黄素和大黄素甲醚含量结果分别为0.054%~0.16%。结论:本实验经方法学验证表明,所建立的方法适用于该饮片中游离蒽醌的含量测定。

Objective:To establish an HPLC method for the assay of free anthraquinones(emodin and physcion)in processed Polygoni Multiflori Radix.Method:The contents of free anthraquinones were determined by an HPLC method.The Agilent ZORBAX SB-C18(4.6 mm×250 mm,5μm)was used as stationary phase.The mobile phase was the mixture of Methanol and Phosphoric acid 0.1%(80∶20)with the flow rate of 1.0 mL·min-1.The column temperature was set at 30℃and the detection wavelength was 254 nm.Results:The linear ranges of emodin and physcion were 7.58-151.60μg·mL-1(r=0.9999)and 3.93-78.57μg·mL-1(r=0.9999)respectively.The recoveries of emodin and physcion were 95.40%-99.63%(RSD=1.60%,n=6)and 98.21%-100.17%(RSD=0.80%,n=6).The emodin and Physcion contents detected in 7 batches of samples were ranged from 0.054%to 0.16%.Conclusion:The established method can be used for the determination of free anthraquinones in processed Polygoni Multiflori Radix.