The validation of a simple LC/MS/MS method for determining the level of mevalonic acid in human plasma.
A simple plasma extraction method coupled with liquid chromatography-tandem mass spectrometry (LC/MS/MS) detection was developed and validated for the analysis of endogenous mevalonic acid (MVA), a biomarker indicative of the rate of cholesterol biosynthesis, in human plasma samples. The analyte was extracted from the plasma matrix using a straightforward liquid-liquid sample preparation procedure. The extract supernatants were evaporated, reconstituted in aqueous solvent and injected into the LC/MS/MS system without further processing. The chromatographic separation was achieved on a reverse-phase high-performance liquid chromatography column. The accuracy and precision of the method was determined over the concentration range 0.25-25 ng/mL MVA from human plasma extracts in three validation batch runs. Inter-assay precision (%CV) and accuracy (%RE) of the quality control samples were ≤7.00% (at lower limit quality control) and ≤6.10%, respectively. The sensitivity and throughput of this assay was significantly improved relative to previously published methods, resulting in smaller sample requirements and shorter analysis time. Assay results from a clinical study following the oral administration of an exploratory statin demonstrate that this procedure could potentially be used in the investigation of therapies associated with hypercholesterolemia.
Department of Pharmacokinetics, Dynamics, and Metabolism, Pfizer Inc., La Jolla, CA, USA. Erick.Kindt@pfizer.com
Erick
E
Kindt
Gabriella
G
Szekely-Klepser
Scott T
ST
Fountain
D015415:Biomarkers / Q000097:blood; D002138:Calibration; D002784:Cholesterol / Q000378:metabolism; D002853:Chromatography, Liquid / Q000379:methods*; D004355:Drug Stability; D006801:Humans; D008798:Mevalonic Acid / Q000097:blood*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*
D015415:Biomarkers; D002784:Cholesterol; D008798:Mevalonic Acid