UPLC法同时测定阿托伐他汀钙中15个杂质含量

目的建立超高效液相色谱法(UPLC)同时测定阿托伐他汀钙原料药中15个杂质含量的方法。方法采用岛津Shim-pack Velox PFPP(2.1 mm×100 mm,1.8μm)色谱柱,以3.9 g/L醋酸铵缓冲液(pH 5.0):乙腈:甲醇:四氢呋喃(无稳定剂)=67:21:6:6为流动相A,3.9 g/L醋酸铵缓冲液(pH 5.0):乙腈:甲醇:四氢呋喃(无稳定剂)=27:61:6:6为流动相B;梯度洗脱,流速0.43 ml/min,检测波长244 nm,柱温35℃,样品室温度10℃;进样量1.8μl。结果阿托伐他汀钙及各杂质峰间分离良好,在0.3~3μg/ml范围内15个杂质均呈良好的线性关系(r均>0.999,n=6),各杂质平均回收率分别为96.3%,99.1%,99.9%,102.4%,96.4%,99.8%,99.4%,104.9%,106.4%,105.4%,100.1%,98.9%,94.7%,94.4%,101.4%,且不同浓度间平行性良好。15个杂质重复性、进样精密度、中间精密度均良好,48 h内均较稳定。此法除对色谱柱要求较高外,耐用性良好。结论本方法简便、快速、分离度好、专属性强,经方法学验证可用于阿托伐他汀钙原料药杂质质量控制。

Simultaneous Determination of 15 Impurities in Atorvastatin Calcium by UPLC

Objective To establish a UPLC method for simultaneous determination of 15 impurities in atorvastatin calcium active pharmaceutical ingredient(API).Methods Analysis was performed on Shimadzu Shim-pack Velox PFPP(2.1 mm×100 mm,1.8μm)column.Gradient elution was adopted,with 3.9 g/L ammonium acetate buffer(pH 5.0):acetonitrile:methanol:tetrahydrofuran(without stabilizer)=67:21:6:6 as mobile phase A,and 3.9 g/L ammonium acetate buffer(pH 5.0):acetonitrile:methanol:tetrahydrofuran(without stabilizer)=27:61:6:6 as mobile phase B.The flow rate was 0.43 ml/min,the detection wavelength was 244 nm,the column temperature was 35℃and the sample room temperature was 10℃,and the injection volume was 1.8μl.Results Atorvastatin calcium and its impurities were well separated.The linear ranges of 15 impurities were 0.3-3μg/ml(r>0.999,n=6).The average recoveries of the impurities were 96.3%,99.1%,99.9%,102.4%,96.4%,99.8%,99.4%,104.9%,106.4%,105.4%,100.1%,98.9%,94.7%,94.4%,101.4%,respectively,and the parallelism between different concentrations was good.The repeatability,injection precision and intermediate precision of 15 impurities were all good,and the test solutions were stable within 48 h.The method showed good durability except high requirements for chromatographic column.Conclusion The method is simple,rapid,with good resolution and strong specificity,and can be used for the determination of related substances in atorvastatin calcium API.