Abstract

ANALYTICAL METHOD DEVELOPMENT AND VALIDATION OF PLERIXAFOR AND RELATED SUBSTANCES IN BULK FORM.

J. Anil Kumar, Saroj Kumar Sahoo* and K. Harinadha Baba

Abstract

A reverse phased HPLC method for the estimation of related substance present in Plerixafor in bulk form has been validated as per ICH guidelines. The method was developed using C18 (Kromosil 250x4.6 mm,5μm) column using the gradient program with mobile phase A (Buffer and Acetonitrile 80:20) and mobile phase B (Buffer and Acetonitrile 20:80) pH maintained at 3±0.05 and were monitored at 215 nm. The linearity level for Plerixafor was established with concentration of drug extending from 0.010 to 0.201%. The linearity level for Plerixafor impurity (H-PXFRC01 & H-PXFRC02) was established with concentration ranging from 0.005 to 0.300% & 0.007 to 0.301%. The % recovery obtained between 99.1 to 99.5% & 105.4 to 106.0%, proved that the method was accurate. The system precision results were within the limits. The low values of RSD showed that the method is precise. LOD and LOQ of method were determined, based on signal to noise method. Robustness of the proposed method was ascertained by deliberately changing the pH variation, flow rate of mobile phase and mobile phase stability. There was no significant change in the system suitability factors of Plerixafor and its impurity peak when these parameters were changed. The low values of the % RSD indicated the robustness of the method.

Keywords: HPLC method validation, Plerixafor, Acetonitrile.