Research Article DEVELOPMENT OF RAPID AND SIMPLE HPLC METHOD FOR THE DETERMINATION OF CYCLOPHOSPAMIDE IN HUMAN SERUM

Cyclophosphamide belongs to the group of alkylating agents which bind to DNA-usually via the binding of alkyl groups.As a result, a reactive intermediate is formed which binds to DNA and causes single or double stranded DNA breaks and may crosslink the chains of DNA thus disturbing the fundamental mechanisms concerned with the cell growth, mitotic activity, differentiation and function.This study describes a simple and fast high-performance liquid chromatography method for the determination of Cyclophosphamide in human serum.Samples were collected from adult cancer patients at Mahatma Gandhi Memorial hospital(Warangal,AP.India) at various time intervals after the end of each infusion. Serum was deproteinized with the acetonitrile and the supernatant was injected into a 250×4.6 mm ODS column. Mobile phase was made up of water and acetonitrile (70:30) with a flow rate of 1.2ml/min. Ultraviolet detection was done at 197 nm and at ambient temperature. Para aminoacetophenone was used as internal standard. Cyclophospamide and internal standard retention times were 4.6 and 9.5 minutes, respectively. Results showed that reproducibility (precision) of method within a day was 2.6 to 6 percent and between days was 5.6 to 8.5 percent. The recovery of the method was between 98.8 and 97.01 percent. The quantitation limit of the method for cyclophospamide was 0.1µM. This method is suitable for quantitation of cyclophospamide after infusion of doses of this drug and has good accuracy, precision and quantitation limit.