Plerixafor is a bicyclam derivative. Cyclamis the key starting material for the synthesis of Plerixafor, hence the detection and quantification of cyclam in the Plerixafor is very essential. A reverse phase HPLC method is developed and validated for the separation and quantification of cyclam content in Plerixafor. Separation was achieved on inertsil ODS -3V (250mm*4.6mm*5µm) column with gradient elution. Sodium perchlorate mono hydrate(0.01mM) with pH 2.0 and methanol (in the ration of 97:3:: v/v) was employed as mobile phase-A, whereas methanol and water (in the ratio of 90:10::v/v )was employed as mobile phase-B with run time 45min.The flow rate was 1mL/min and detection wave length was 200nm.Method validated with reference to the ICH guidelines. Method is identified as linear form the range 0.0025mg/mL to 0.01mg/mL, with injection volume 20µL.Minimum quantification level achieved as 0.0025mg/mL, where as the minimum detection level for the cyclam was 0.0005mg/mL
