Validation of liquid chromatography-tandem mass spectrometry for mevalonate in human plasma: Incompetent effects between treated atorvastatin & its combination with olmesartan in cardiovascular patients

Abstract

A simple and sensitive LC/MS/MS method validation was developed for the quantification of Mevalonate (MVA), a endogenous compound responsible for synthesis of cholesterol in Human Plasma. The method was validated and impended for application on human to study the concentration of Mevalonate in plasma level after administration of Atorvastatin (ATVS) individually and with combination to Olmesartan (OLM). The assay procedure involved the isolation of MVA from plasma samples using solid-phase extraction, preconditioned cartridge, washed with methanol followed by 0.1 N HCI. The analytes were eluted with3x 0.5 ml of methanol and evaporated to dryness Nitrogen stream. The residue was reconstitute for LCMS/MS analysis, were chromatographic separation was carried on a HyPurity Advance, 50X4.6mm column with a mobile phase 10mM Ammonium formate ( pH=8) and Acetonitrile. The flow rate was 0.8 ml/min throughout the process. The liquid Chromatography, Agilent 1290 coupled to electrospray ion (ESI) Mass Spectrometer (MS QQQ). The developed method was validated for specificity, accuracy, precision, stability, linearity, sensitivity and recovery. The method was linear and found to be acceptable over the range of 50–1000 ng/ml. The method was successfully applied for the drug interaction study of ATVS + OLM by quantifying changes in levels of MVA in hypertensive patients. The study revealed concentration levels of MVA in ATVS + OLM compared to ATVS as single treated condition are nearly equal. This conclude that, MVA synthesize equal level of blood cholesterol on both the stage but failed to reduce BP synergistically, that associate with vascular plague formation. Key words: Mevalonate, LC/MS/MS, human plasma level, Olmesartan, Atorvastatin.