Development and Validation of a Liquid Chromatographic method for the Quantification of Related Compounds in Cyclophosphamide.

A simple reverse- phase high performance liquid chromatographic method (RP-HPLC) was developed for quantification of the related compounds in Cyclophosphamide. The chromatographic separation was achieved with C18 column (250 × 4.6 mm, 5 µm particle size) with gradient elution composed of 0.02 M Potassium dihydrogen phosphate (pH-7.0) as mobile phase-A and 60:40% v/v Acetonitrile / Water as mobile phase-B at a flow rate of 0.8 mL /min and column compartment maintained at 40° C for separation. Detection was carried out at 195 nm. The correlation coefficient (≥0.99) shows the linearity response against concentration over the range of Limit of Quantification (LOQ). Precision studies showed the Relative Standard Deviation (RSD) values less than 5% for Cyclophosphamide and its related compounds. The method was substantiated with respect to specificity, precision, linearity, accuracy, limit of quantification, and robustness. The proposed method could be used for routine analysis of Cyclophosphamide formulations