缬沙坦中2-氰基-4’-溴甲基联苯和2-氰基-4’,4’-二溴甲基联苯的高效液相色谱法测定方法的建立及评价

Establishment and Evaluation of HPLC Method for the Determination of 2-cyano-4 ’-Bromomethylbiphenyl and 2-cyano-4’, 4 ’-Dibromomethylbiphenyl in Valsartan

目的建立缬沙坦中2-氰基-4’-溴甲基联苯(杂质Ⅰ)和2-氰基-4’,4’-二溴甲基联苯(杂质Ⅱ)的高效液相色谱法(HPLC)测定方法。方法采用HPLC,C18色谱柱(推荐使用ACE EXCEL 3 C18-AR,150×46 mm),流动相为乙腈:0.1%乙酸溶液(52:48),检测波长为260nm,柱温为40℃,流速为1.0ml/min,进样量为20μl。结果杂质Ⅰ与缬沙坦和杂质Ⅱ的分离度符合要求。杂质Ⅰ和杂质Ⅱ浓度在10~500 ng/ml的范围内线性良好,杂质Ⅰ和杂质Ⅱ检测限为5 ng/ml,定量限为10 ng/ml,精密度、重复性、准确度(加样回收率)、耐用性符合要求。样品溶液室温放置和光照后稳定,对照品溶液需4℃放置。结论该方法简单、准确、重现性好、耐用性好,可用于控制缬沙坦中杂质Ⅰ、杂质Ⅱ含量。

Objective To establish a HPLC method to test content of 2-cyano 4 ’-bromine-biphenyl(impurity Ⅰ) and 2 – cyano-4’, 4 ’-dibromomethyl-biphenyl(impurityⅡ) of valsartan. Methods HPLC, C18 column was used.(ACE EXCEL 3 c18-ar, 150×46 mm is recommended to use) with the mobile phase of acetonitrile-0.1% acetic acid solution(52:48) at a flow rate of 1.0 m L/min, with the detection wavelength at 260 nm, the column temperature of 40 ℃ and the injection volume of 20 μL. Results The degree of separation of Impurity Ⅰ with valsartan and impurityⅡ conformed to requirements. The concentration for Impurity Ⅰand impurityⅡ were in the range of 10-500 ng·m L-1. The detection limit of impurityⅠand impurity Ⅱ was 5 ng/ml, quantitative limit was 10 ng/ml. Furthermore, the precision, repeatability, accuracy(sample recovery rate) and durability meet the requirements. The sample solution was stable under room temperature and light, and the control solution needed to be placed at 4 ℃. Conclusion The method is simple, accurate, reproducible and durable. It can be used to control the concentration of impurity I and II in valsartan.