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

Aim: To develop a simple and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method validation for the quantification of mevalonate (MVA), an endogenous compound responsible for synthesis of cholesterol in human plasma. Settings and Design: The method was validated and impended for application on human to study the concentration of MVA in plasma level after administration of atorvastatin (ATVS) individually and with combination to olmesartan (OLM). Materials and Methods: 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 with 3 x 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, 50 x 4.6 mm column with a mobile phase 10 mM ammonium formate (pH = 8) and Acetonitrile. The flow rate was 0.8 ml/min throughout the process. The LC, Agilent 1290 coupled to electrospray ion mass spectrometer. Results: 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. Conclusion: 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.