UPLC与HPLC法测定阿托伐他汀钙及其制剂中的有关物质

目的：建立测定阿托伐他汀钙有关物质及含量的超高效液相色谱法（UPLC）和高效液相色谱法（HPLC），并对方法及测定结果进行比较。方法：UPLC法，采用Acquity BEH Shield RP18色谱柱（150 mm×2.1 mm，1.7 μm），流速0.4 mL·min^-1；HPLC法，采用Symetry Shield RP18色谱柱（250 mm×4.6 mm，5 μm），流速1.5 mL·min^-1。两方法均以20 mmol·L^-1醋酸铵溶液（用冰醋酸调节pH为4.0）为流动相A，乙腈-四氢呋喃（95∶5）为流动相B，梯度洗脱，检测波长254 nm，柱温35℃。结果：UPLC法和HPLC法测定有关物质在一定范围内均呈现良好线性，r>0.999，加样回收率、精密度和样品测定结果基本无差异；UPLC检测下限略低于HPLC检测下限，主峰与非对映异构体（杂质B）分离度优于HPLC法，UPLC法测定时间为17 min，HPLC法为50 min，UPLC法溶剂使用量减少12倍。用建立的UPLC和HPLC方法对阿托伐他汀钙5个企业10批样品，阿托伐他汀钙片剂4个企业14批样品以及阿托伐他汀钙胶囊1个企业6批样品进行有关物质检查，结果无显著差异。结论：UPLC和HPLC 2种方法准确性、精密度、线性等验证试验结果以及样品测定结果基本一致；UPLC法测定阿托伐他汀钙有关物质和含量快速、灵敏度高，分离度好，更环保。

Objective: To establish UPLC and HPLC method for the determination of seven related substances in atorvastatin calcium and its preparation,and to compare the two methods as well as the determination results. Methods: A gradient UPLC system was used in the quantitative analysis of seven compounds on Acquity BEH Shield RP18 column(2.1 mm×100 mm,1.7 μm),using acetonitrile-tetrahydrofuran-ammonium acetate buffer (pH 4.0) as mobile phase,with a flow rate of 0.4 mL·min^-1.An HPLC method was conducted on Symetry Shield RP18 column(4.6 mm×250 mm,5 μm),with flow rate of 1.0 mL·min^-1 and the same mobile phase as the UPLC method.The wavelength and column temperature were set at 254 nm and 35℃,respectively in both methods.In order to compare the two methods,resolution,linearity,repeatability,recovery,limit of detection,force degradation tests,stability and the content of the seven compounds were investigated.Results: The linear relationship between the concentration and peak areas of the seven compounds were all good(r>0.999).There were no significant difference between the analytical validation (accuracy and repeatability) and impurities detection results of the two methods.UPLC shows shorter analysis time (1/3 of HPLC analysis time),less solvent(1/12),better resolution between main peak and impurity B peak,lower limits of detection.The results of two methods on related substance test for all samples have no significant differences.Conclusion: Both of the methods show almost the same results.The UPLC method is more efficient,accurate,sensitive and environmental protective.