反向高效液相色谱法检测正心降脂片中丹参酮Ⅱ_(A)、葛根素和大黄素含量

Research RP-HPLC method for determination tanshinone Ⅱ_(A),puerarin and emodin in Zhengxin Jiangzhi tabets

目的建立检测正心降脂片中丹参酮ⅡA、葛根素和大黄素3种成分含量的方法。方法采用反向高效液相色谱(RPHPLC)法。取正心降脂片,除去包衣,研细,取约0.2 g,精密称定,置滤膜滤过,Lichrospher RP-C18色谱柱分离,检测波长为259 nm,流动相A为乙腈,流动相B为0.2%磷酸溶液,梯度洗脱,洗脱程序为0~6 min 12%A、6~15 min 12%~63%A、15~25 min 63%~92%A、25~27 min 91%A、27~30 min 91%~12%A、30~35 min 12%A,流速为1.0 mL/min,柱温为30℃,进样体积为10µL,二极管阵列检测器定量检测。结果葛根素、大黄素和丹参酮ⅡA分别在0.763~76.3µg/mL、0.768~76.8µg/mL和0.199~19.9µg/mL范围内,线性关系良好,平均加标回收率分别为98.5%,99.3%和99.6%,重复性相对标准偏差(RSD)分别为1.29%、2.05%和1.72%;稳定性RSD分别为2.03%、1.72%和1.33%。结论实验建立的方法具有简便、快速、准确等特点,适用于正心降脂片产品的质量控制,为进一步完善和提高该品种的质量标准提供技术参考。

Objective To establish method for determination 3 contents of tanshinoneⅡA,puerarin and emodin in Zhengxin Jiang⁃zhi tabets.Methods The coating of Zhengxin Jiangzhi Tabets samples was removed and grinded to powder.Weighing about 0.2 g pretreated samples precisely to 50 mL volumetric flask,ultrasonic treatment was performed with 95%methanol solution for 25 min.Vol‐ume the samples to 50 mL and shook it well.Samples was filtered by 0.45µm membrane filter and separated on Lichrospher RP-C18 column.Detection wavelength:259 nm.Mobile phase:acetonitrile(A)-0.2%phosphoric acid solution(B).Gradient elution procedure:0-6 min 12%A,6-15 min 12%-63%A,15-25 min 63%-92%A,25-27 min 91%A,27-30 min 91%-12%A,30-35 min 12%A.flow rate:1.0 mL/min,column temperature:30℃and injection volume was 10µL.Diode array detector was for quantitative detection.Re‑sults Puerarin,emodin and TanshinoneⅡA showed good linear relationships in following concentration range 1.526-76.3µg/mL,1.536-76.8µg/mL and 0.398-19.9µg/mL,respectively.Average recoveries were 98.5%,99.3%and 99.6%,respectively.Relative stan‐dard deviation(RSD)of repeatability(n=6)was 1.3%,1.7%and 1.1%,respectively.The stability of RSD was 2.03%,1.72%and 1.33%,respectively.Conclusion The method is simple,rapid and accurate,which is suitable for the quality control of Zhengxin Ji⁃angzhi Tabets.It also provides technical reference for further improving quality standard of this breed.