目的建立去甲斑蝥素原料药含量及有关物质的反相高效液相色谱测定方法。方法色谱柱为YWG-C₁₈(4.6 mm×250 mm,5μm),流动相为甲醇-0.016 mol·L^-1磷酸二氢钾溶液(30∶70)(用磷酸调节pH值至3.1),检测波长为213 nm,流速为0.9 mL·min^-1。结果去甲斑蝥素与马来酸酐、去氢去甲斑蝥素及其降解产物均能较好的分离,且线性良好(r分别为0.999 8,0.999 9,0.999 7);检测下限分别为10.0,0.08,0.63 ng(S/N=3),平均回收率(n=9)分别为99.94%,100.43%,100.03%;含量测定方法的线性范围为0.126～4.044 g·L^-1。结论方法简便,结果准确,重复性好,适用于该产品含量及有关物质测定。

OBJECTIVE To develop a RP-HPLC method for the determination of norcantharidin and its related substances.METHODS A chromatography was carried out on a YWG-C₁₈ column(4.6 mm×250 mm,5 μm),using a mobile phase consisting of methanol-0.016 mol·L^-1 potassium dihydrogen phosphate(30∶70)(adjusted to pH 3.1 with phosphoric acid).The detection was carried out with a UV detector at 213 nm.The flow rate was 0.9 mL·min^-1.RESULTS Norcantharidin,maelic anhydride,dehydronorcantharidin and degradation products were separated.The calibration curves of norcantharidin,maelic anhydride and dehydronorcantharidin were linear(r=0.999 8,0.999 9,0.999 7).The detection limits were 10.0 ng for norcantharidin,0.08 ng for maelic anhydride and 0.63 ng for dehydronorcantharidin,respectively(S/N=3).The recoveries of norcantharidin,maelic anhydride and dehydronorcantharidin were 99.94%,100.43% and 100.03%,respectively(n=9).The norcantharidin curves were linear in the range of 0.126～4.044 g·L^-1.CONCLUSION This method is convenient,accurate and suitable for the determination of norcantharidin and related substances.