HPLC法测定依那普利氢氯噻嗪片中的马来酸依那普利有关物质

The Determination of Enalapril Related Substances in Enalapril Maleate and Hydrochlorothiazide Tablets by HPLC Method

目的:建立高效液相色谱法测定依那普利氢氯噻嗪片中马来酸依那普利有关物质含量的分析方法。方法:所用色谱柱为Waters XBridge Shield RP18 (4.6×250 mm,5μm);以0.02 mol/L磷酸二氢钠缓冲液(磷酸调至pH 2.0)作流动相A,甲醇作流动相B,梯度洗脱;流速1 mL/min;检测波长为215 nm;柱温为50℃。结果:鉴定了杂质依那普利拉和依那普利双酮,测定其校正因子,建立了依那普利拉和依那普利双酮的分析计算方法。本条件下杂质峰与主峰分离度良好,满足质量控制需要,依那普利、依那普利拉和依那普利双酮的检出限浓度分别为0.090170、0.0099342和0.091098μg/mL,并分别在0.50095~20.038,0.49671~19.868和0.5061~20.244μg/mL范围内线性关系良好。依那普利拉和依那普利双酮平均加标回收率(n=9)为95.9%和98.1%,RSD均小于5%。结论:经方法学验证,本法适用于依那普利氢氯噻嗪片中对马来酸依那普利有关物质的定量检测,本研究为复方制剂有关物质的质量控制提供了参考。

Objective:To establish an HPLC method for determination of the enalapril related substances in enalapril maleate and hydrochlorothiazide tablets.Methods:Waters XBridge Shield RP18 column(4.6×250 mm,5μm)was chosen for the analysis.The mobile phase A was 0.02 mol/L sodium dihydrogen phosphate solution(adjusted to pH 2.0 with phosphoric acid),the mobile phase B was methanol.The mobile phase was with gradient elution at a flow rate of 1.0 mL/min.The detection wavelength was 215 nm and the column temperature was 50℃.Results:Enalaprilat and enalapril diketopiperazine were identified and the correction factors were determined.The analytical method for the two impurities was also established.The resolutions of the impurities to enalapril were well under the established chromatographic condition.Detection limits of enalapril,enalaprilat and enalapril diketopiperazine were 0.090170,0.0099342,and 0.091098μg/mL,and the linearity was good over the ranges of 0.50095~20.038,0.49671~19.868,and 0.5061~20.244μg/mL,respectively.The average recoveries(n=9)of enalaprilat and enalapril diketopiperazine were 95.9%and 98.1%,with RSDs less than 5%.The HPLC method met the requirements for the quality control of enalapril maleate and hydrochlorothiazide tablets.Conclusion:The validation results show that the method can be used for quantitative analysis of impurities of enalapril in enalapril maleate and hydrochlorothiazide tablets.The research provides the reference for quality control of the related substances in compound preparations.