Robust and Rugged Stability-Indicating HPLC Method for the Determination of Plerixafor and Its Related Impurities in Drug Substances.

A novel, reversed-phase high-performance liquid chromatographic method was developed and validated for the determination of related substances in Plerixafor (PLX) drug substance. PLX is an immunostimulant used to mobilize hematopoietic stem cells in cancer patients. The method is efficient for estimation of all degradation and process-related impurities. The method was developed using the Phenomenex Luna L11 column using the gradient program with mobile phase A and mobile phase B, where mobile phase A consists of pH 2.0 1-heptanesulfonic acid sodium salt buffer and acetonitrile in the ratio of 80:20 (v/v) and mobile phase B consists of pH 2.0 1-heptanesulfonic acid sodium salt buffer and acetonitrile in the ratio of 20:80 (v/v). PLX and its impurities were monitored at 210 nm. The present work is describing the role of ion-pair reagent in the separation of polar compounds. PLX was subjected to various stress conditions of oxidative, acid, base, hydrolytic, thermal, humidity and photolytic degradations. The degradation products were well separated from each other and with the main peak, demonstrating the stability-indicating power of the method. The performance of the method was validated according to the present ICH guidelines for specificity, limit of detection, limit of quantification, linearity, accuracy, precision, ruggedness and robustness.