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A simple and sensitive LC/MS/MS method validation was developed for the quantification of Mevalonate (MVA), a autocoid responsible for synthesis of cholesterol in Human Plasma and increasing Blood pressure. The method was validated and was impended for application on human subjects 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 in combination, against ATSV treatment on hypercholestremics 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 p... ... middle of paper ... ...synthesis. The analysis of urinary mevalonic acid, either in 24h collection or in a single morning sample, is an attractive method for evaluation of long and very short term changes of the rates of cholesterol synthesis [8]. The main challenge in developing and validating a method for determining MVA in human plasma was that MVA is a polar, endogenous moiety that circulates in the blood stream at nanogram levels. In most methods, the extraction of MVA from plasma was carried out using ionexchange resins in the form of mevalonolactone (MVAL) [9-10]. A simple, specific, and sufficiently sensitive liquid chromatography-tandem mass spectrometry (negative-ion electrospray ionization)methodology to determinemevalonic acid (MVA) in human plasma is described, and its application to the analysis of rat plasma MVA levels after rosuvastatin administration is demonstrated [11].
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