Method Development and Validation for Tenofovir an Antiretroviral Drug in Plasma by LC-MS/MS Technique

Abstract

: Bioanalytical method development for Tenofovir (TFR) as an antiretroviral drug by LCMS Technique. A developed Bioanalytical analysis method for TFR can be used routinely in a commercial laboratory. All the solvents used were of HPLC grade. 4000 QTrap along with the Shimadzu LC 20AD LC System used to develop and validate the method. The LLOQ and LOQ for Tenofovir was found were 5ng/mL and 15ng/mL. The method was accurate (within ±15% of control) and precise (coefficient of variation ≤ 15%). Analytes were stable for five freeze/thaw cycles and up to 6 days at room temperature, whereas long-term at − 20°C or at − 80°C. For Precision study using QCs of the drug-85%, 100% and 115% concentration of drug chosen and the levels M1QC (75ng/mL), MQC (300ng/mL) and HQC (600ng/mL) where the % CV were observed of ≤ 15%. In a Precision and Accuracy study (inter day and intraday), the % CV obtained for Tenofovir was observed ≤ 15%. Recovery studies for extracted samples with LQC (15ng/mL), MQC (300ng/mL) and HQC (600ng/mL) were 94.51%, 91.83% and 90.91% respectively. Stability was within 15% deviation. The results of System Suitability Test for TFR and Acyclovir (ACR) are an internal standard with observed %CV ≤ 2.0%. The aim of the study was to develop a method that could be used as an alternative to the existing Tenofovir indirect method. The existing method observes separating the parent drug from the metabolite in LCMS/MS. This method is a good alternative to the indirect methods currently in use.