Development and validation of simplified RP-HPLC method for quantification of Darunavir in commercial tablets

Abstract

Darunavir (DRV) is a peptidic protease inhibitor class of antiretroviral drug which is regularly included in the antiretroviral therapy with several other drugs to treat Human immunodeficiency virus (HIV) patients to avoid those developing Acquired Immune Deficiency Syndrome (AIDS). In this research paper, an accurate, precise, reproducible, simple and rapid Reverse-Phase High-Performance Liquid Chromatography (RP-HPLC) method was developed and validated for quantifying DRV in the tablet dosage form (Danavir 600). The liquid chromatography was carried out on a Phenomenex C18 analytical column with dimensions of 250 × 4.6 mm, 5 µm using a mixture of acetonitrile (ACN) and water in the ratio of 30:70 v/v as mobile phase. The sample was injected in a volume of 20 µl and determined at 268 nm with a flow rate of 1 ml/min (ACN:water). The retention time of standard and sample drug was found to be 3.16 and 3.12 min respectively. The linearity in the calibration curve was achieved for DRV in the concentration range of 40–200 µg/ml and the regression coefficient (R2) was found to be 0.982. The percentage recovery of DRV was obtained in the range of 98.10–103.65% indicated the great accuracy of the current method. Reproducibility and recovery studies with % relative standard deviation (%RSD) with intra and inter-day precision was found to be less than 2%, which proven a developed method was reproducible. The limit of detection (LOD) and limit of quantification (LOQ) were found to be 0.31 and 0.96 µg/ml respectively. The method was validated as per the International Conference on Harmonization (ICH) guidelines and can be strongly adopted for routine analysis of DRV since it is the most reliable and rapid approach. Even though a lot of work has been performed for estimating DRV in dosage forms, this study was found to be under green chemistry with the rapid and economical condition. Thus, the statistical validation of the data showed that the proposed method can be applied for the estimation of DRV in commercial tablets.