A Liquid Chromatography-UV (LC-UV) Method was Developed for Quantification of Six Potential Impurities in Androstanolone Active Pharmaceutical Ingredient

Abstract

A sensitive, selective reverse phase method was developed for the quantitative determination of six potential impurities in Androstanolone active pharmaceutical ingredient. Efficient chromatographic separation was achieved on Zorbax Eclipse XDB C8 (250×4.6 mm, 5 μm) column with mobile phase containing a gradient mixture of solvent-A and solvent-B. The eluted compounds were monitored at 200 nm. All six potential impurities were identified by mass spectrophotometer and characterized by nuclear magnetic resonance. The developed method was validated as per ICH guidelines with respect to specificity, precision, linearity, quantitation limit, detection limit and accuracy. Regression coefficient value was greater than 0.99 for Androstanolone impurities. Detection limit of impurity-A, impurity-B, impurity-C, impurity-D, impurity-E and impurity-F were in the range of 0.0002-0.003% respectively. The quantitation limit of impurity-A, impurity-B, impurity-C, impurity-D, impurity-E and impurity-F were in the range of 0.003-0.013% respectively with respect to sample concentration. The accuracy of the method was established based on the recovery obtained between 92.72-106.90% for all impurities.