Validation of a reverse phase high-performance liquid chromatography method for the detection of major components and related substances in nicardipine hydrochloride injection.

A simple, feasible, isocratic elution and stable reversed-phase high performance liquid chromatography was established and verified. Chromatographic conditions were XY-C18, 4.6x150mm, 5μm column; column temperature 50oC. Mobile phase was (A) 0.025 mol/L sodium perchlorate solution, added 1ml triethylamine, and adjust the pH value to 2.0 with perchloric acid solution; (B) acetonitrile-methanol (70:30). The elution method was gradient elution; The flow rate was 1.0mL/min; the injection volume was 10μL and the wavelengths were 239nm and 258nm. The developed method had been validated according to the ICH guidelines and the system suitability, specificity, LOD, LOQ, linearity, range, accuracy, precision, durability and solution stability of the proposed method had been verified. The relationship between concentration and peak area was linear in the concentration range, with an R2>0.995. The linear relationship was good in the range of impurity concentration and the R2 was more than 0.999. The accuracy and repeatability meet the specified criteria. The method had good durability.