HPLC同时测定复方降糖方提取物中4种有效成分的含量

Simultaneous determination of four major bioactive components in extractive of compound Jiangtang by HPLC

为了建立高效液相色谱法同时测定复方降糖方提取物中4种有效成分含量的方法。采用Diamonsil C18色谱柱(4.6 mm×250 mm,5μm),流动相A为乙腈,流动相B为0.2%磷酸水溶液,梯度洗脱(0-30 min,10%A→15%A;30-50 min,15%A→25%A;50-70 min,25%A→30%A),流速1 m L/min,检测波长230 nm,柱温35℃。结果显示:在70 min内可完成对复方降糖方提取物中葛根素、芍药苷、橙皮苷和丹酚酸B 4种成分的分离测定,各成分的质量浓度分别在0.408-8.16μg/m L(r=0.9997)、2.25-45.00μg/m L(r=0.9999)、3.85-77.00μg/m L(r=0.9999)和3.675-73.50μg/m L(r=0.9999)范围内呈良好的线性关系;平均回收率分别为99.55%、102.72%、98.54%、99.09%,RSD分别为1.17%、1.33%、1.52%、1.46%;最低检测限分别0.326、1.500、0.385、0.368μg/m L。由此可知,该方法简单、准确可靠,可为含有葛根、赤芍、陈皮、丹参的成方制剂的质量控制提供参考。

To develop a HPLC method for simultaneous determination of four major active component in extractive of Compound Jiangtang. The column was Diamonsil C18 column（4.6 mm×250 mm, 5 μm）. The mobile phase A was acetonitrile and the mobile phase B was 0.2% phosphoric acid solution. The ratios of the mobile phase A were 10%→15% at 0～30 min, 15%→25% at 30-50 min, 25%→30% at 50-70 min. The flow rate was 1.0 m L/min. The detection wavelength was set at 230 nm and the column temperature was 35 ℃. The method had good linear relationship within the range of 0.408-8.16 μg/m L（r=0.9997） for puerarin, 2.25-45.00 μg/m L（r=0.9999） for paeoniflorin, 3.85-77.00 μg/m L（r=0.9999）for hesperidin and3.675-73.50 μg/m L（r=0.9999） for salvianolic acid B. The average recoveries of puerarin, paeoniflorin, hesperidin and salvianolic acid B were 99.55%, 102.72%, 98.54%, 99.09% and the RSD were 1.17%, 1.33%, 1.52%, 1.46% respectively. The detection limit were 0.326, 1.500, 0.385, 0.368 μg/m L. This method is simple, accurate and reliable. It can be reference for quality control of Chinese medicine preparations, including Radix puerariae,Paeonia lactiflora Pall,Pericarpium Citri Reticultae and Salvia miltiorrhizan Bge.