高效液相色谱法测定枸橼酸氯米芬片中枸橼酸氯米芬及其顺式异构体

Determination of clomiphene citrate and its cis-isomer in clomiphene citrate tablet by HPLC

建立高效液相色谱法测定片剂中枸橼酸氯米芬及其异构体含量,并建立异构体定量核磁测定法。色谱柱为C_(18)柱(25 cm×4.6 mm,5μm),流动相为缓冲液(1%三乙胺溶液,用磷酸调节pH值至2.5)-甲醇(30∶70),流量为1.0 mL/min,检测波长为233 nm,进样体积为50μL,采用一维核磁共振氢谱对顺式异构体进行定量分析。枸橼酸氯米芬的质量浓度在0.02~0.10 mg/mL范围内与色谱峰面积线性关系良好。加标回收率为100.4%~100.9%,测定结果的相对标准偏差为0.18%(n=9)。在异构体含量计算中引入校正因子,可显著减小测定偏差。该方法适用于枸橼酸氯米芬片的质量评价。

A method was established for the determination of clomiphene citrate and cis-isomer content in clomiphene citrate tablet by HPLC,and a quantitative nuclear magnetic resonance(qNMR)method for the determination of cis-isomer content was set up C_(18) column(25 cm×4.6 mm,5μm)was used as chromatography column,buffer(1.0%Et3N solution,phosphoric acid was used to adjust pH at 2.5)-MeOH(30:70)was used as the mobile phase with the flow rate of 1.0 mL/min,the detection wavelength was 233 nm and the injection volume was 50μL.The cis-isomer content was confirmed by 1H NMR.The chromatograph peak area of clomiphene had excellent linear dependence relation in the range of 0.02-0.10 mg/mL.The standard addition recoveries were 100.4%-100.9%with the relative standard deviation of 0.18%(n=9).The relative correction factor was introduced in cis-isomer content calculation,which could reduce result bias significantly.This method is suitable for quality evaluation of clomiphene citrate tablet.