HPLC梯度洗脱法同时测定决明子中橙黄决明素、大黄素、大黄酚和大黄素甲醚的含量

The Simultaneous Measurement of Aurantio-obtusin,Emodin,Chrysophanol and Physcione in Semen Cassiae by HPLC

目的:采用HPLC梯度洗脱法同时测定决明子中橙黄决明素、大黄素、大黄酚和大黄素甲醚的含量。方法:用Kromasil 100A C_(18)色谱柱(5μm,4.6mm×150mm,迪马公司);甲醇-0.1%磷酸溶液梯度洗脱:0～15min(65:35),15～16 min(65～85:35～15),16～40 min(85:15);柱温:室温;检测波长:285nm(0～15min),254nm(15～40min);流速:1mL·min^(-1)。结果:橙黄决明素、大黄素、大黄酚和大黄素甲醚的线性范围分别为0.00038～0.00608μg(r=0.9998)、0.0029～0.0464μg(r=0.9999)、0.00105～0.0168μg(r=0.9996)、0.00044～0.00704μg(r=0.9998),平均回收率分别为99.50%(RSD=0.71%)、99.59%(RSD=1.42%)、99.31%(RSD=0.59%)、99.74%(RSD=0.91%)。结论:本方法专属性强,重复性好,可用于决明子中橙黄决明素、大黄素、大黄酚和大黄素甲醚的含量测定。

This study aimed to simultaneously measure the content of aurantio-obtusin, emodin, chrysophanol and physcione and in semen cassiae by HPLC. A Kromasil 100A C18 column （5μm, 4.6mm×150mm, DIKMA） was used for the gradient elution with a methanol-0.1% phosphonic acid solution： 0-15min （65：35）, 15-16min （65-85： 35-15）, 16- 40min （85：15）, at the flow rate of 1 mL·min^-1 at room temperature.The UV wavelength was detected at 285 nm （0- 15min） and 254nm （15-40 min）. The linear range was determined to be 0.00038-0.00608μg （r=0.9998）, 0.0029- 0.0464μg （r=0.9999）, 0.00105-0.0168μg （r=0.9996） and 0.00044-0.00704μg （r=0.9998） respectively for aurantioobtusin, emodin, ehrysophanol and physcione, with the average recovery of 99.50% （RSD=0.71%）, 99.59% （RSD= 1.42%）, 99.31% （RSD=0.59%） and 99.74% （RSD=0.91%）. The results showed that this method has good specificity and repeatability, and thus considered as a good tool for the measurement of aurantio-obtusin, emodin, chrysophanol and physeione in semen cassiae.