RP-HPLC法检查丁酸氯维地平原料药中的有关物质

Determination of Related Substances in Clevidipine Butyrate Drug Substance by RP-HPLC

目的:建立检查丁酸氯维地平原料药中有关物质含量的方法。方法:采用反相高效液相色谱法。色谱柱为Waters Symmetry ShieldTM,流动相为磷酸二氢钠水溶液与乙腈,采用梯度洗脱,流速为1.2 ml/min,检测波长为220 nm,进样量为20μl,柱温为35℃。分别采用不加及加校正因子的主成分自身对照法计算3批样品中6种及1种已知杂质(杂质B、C、D、E、F、G及H)的量。结果:7种杂质在各自的检测质量浓度范围内线性关系良好(r为0.997~1.000);回收率为94.96%~97.79%(RSD小于4.7%,n=3);丁酸氯维地平检测限为5 ng,杂质B^H的定量限依次为4、4、8、8、4、8、8 ng;3批样品中均未检出杂质B、C、D,总杂质量小于0.12%。结论:建立的方法简便、灵敏、准确,可用于丁酸氯维地平原料药中有关物质的检查。

OBJECTIVE： To develop the method for the content determination of related substances in clevidipine butyrate drug substance. METHODS： RP-HPLC method was adopted. The determination was performed on Waters Symmetry ShieldTM column with mobile phase consisted of sodium dihydrogen phosphate solution-acetonitrile （gradient elution） at the flow rate of 1.2 ml/min. The determination wavelength was set at 220 nm, and sample size was 20 μl. The column temperature was 35℃. The contents of 6 and 1 kinds of known impurities （impurity B, C, D, E, F, G and H） in 3 batches of samples were calculated by main component self-compare without or with calibration factor. RESULTS： 7 kinds of impurity from clevidipine butyrate revealed good linearity （r=0.997-1.000）; average recoveries were 94.96%-97.79% （RSD〈4.7%, n=3）. The lowest detection limit of clevidipine butyrate was 5 ng, and the limits of quantity of impurities B-H were 4, 4, 8, 8, 4, 8 and 8 ng, respectively. Impurities B, C and D were not found in 3 batches of samples, and total amount of impurity was lower than 0.12%. CONCLUSIONS： Established method is simple, sensitive and accurate, and can be used for the determination of related substances in clevidipine butyrate.