Abstract

A STABILITY-INDICATING UPLC METHOD FOR QUANTIFICATION OF CYCLOPHOSPHAMIDE RELATED COMPOUNDS IN PHARMACEUTICAL DOSAGE FORMS

Kiran Kumar Ankalla* and Nageswara Rao Gollapalli

ABSTRACT

A specific, precise, rapid and reliable stability indicating UPLC method has been developed and validated for estimation of Cyclophosphamide related compounds in pharmaceutical dosage forms. Chromatographic separation was achieved on an Acquity UPLC BEH C18 (2.1 x 50 mm, 1.7 μm) column using gradient composition of mixer of pH 7.0 buffer as Mobile Phase A and mixture of pH 7.0 buffer and Acetonitrile in the ratio of 20:80% v/v as Mobile Phase B at a flow rate of 0.5 mL/min and analytes were monitored at 195 nm. The retention times of the Cyclophosphamide was about 4.18 and relative retention times of Cyclophosphamide Related compound A, Cyclophosphamide Related compound B and Cyclophosphamide Related compound D were about 0.91, 0.17 and 0.23 respectively. The results of specificity studies indicate that there was no interference of diluent, excipients, impurities and degradation products at retention time of analyte and it is assured that the peak response was belongs to a single component only. The detector response was linear in the range of LOQ-150% level with respect to test concentration of Cyclophosphamide, Cyclophosphamide Related compound A, Cyclophosphamide Related compound B and Cyclophosphamide Related compound D. Correlation coefficient (R2) was not less than 0.99 for Cyclophosphamide and its related compounds. The percentage recovery of Cyclophosphamide, Cyclophosphamide Related compound A, Cyclophosphamide Related compound B and Cyclophosphamide Related compound D were meeting the predetermined acceptance criteria. The developed method was validated for specificity, linearity, precision, accuracy, solution stability, ruggedness and stress degradation studies were monitored. Hence, the developed method was specific, rapid and cost-effective, and it can be used for routine analysis of Cyclophosphamide related compounds in pharmaceutical dosage forms.

Keywords: Cyclophosphamide; Cyclophosphamide Related compound A; Cyclophosphamide Related compound B; Cyclophosphamide Related compound D; UPLC.