ANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR THE ESTIMATION OF ACYCLOVIR TRANSFERSOMES BY RP-HPLC
Abstract
A simple, rapid and selective RP-HPLC method was developed for the estimation of acyclovir in bulk drug and (In-house) transfersomes formulation. Acyclovir (ACV) is a synthetic purine nucleoside analog derived from guanine, is the most widely used antiviral agent. It is effective in the treatment of herpes simplex virus (HSV), mainly HSV-1 and HSV-2 and varicella zoster virus. However, it has low skin permeability. The in-house transfersomes was formulated by using phospholipids, cholesterol and Tween 80 by hand shaking method. The separation was achieved on Thermo C18 analytical column (250 mm 4.6 mm i.d., 5.0 ?m) using 0.02N HCl (100% v/v) as mobile phase and at a flow rate of 1.0 ml/min. Detection was carried out using a UV detector at 240nm. The total chromatographic analysis time per sample was about 7.0min with ACV eluting at retention time of about 6.0120.3min. The method was validated for accuracy, precision, specificity, linearity and sensitivity. Validation studies demonstrated that this HPLC method is simple, specific, rapid, reliable and reproducible. The standard curve was linear over the concentration range of 5-25?g/ml with r2 close to one (0.999). The limit of detection (LOD) and limit of quantitation (LOQ) obtained for micafungin were 0.125?g/ml and 0.374?g/ml respectively. The developed and validated method was successfully applied for the quantitative analysis of ACV in pharmaceutical formulation. The high recovery and low relative standard deviation confirm the suitability of the proposed method for the determination of ACV (In-house) transfersomes formulation.