PMID: 11339276May 8, 2001Paper

Analysis of 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors using liquid chromatography-electrospray mass spectrometry

Journal of Chromatography. B, Biomedical Sciences and Applications
E J ParkJ M Pezzuto

Abstract

Employing high-performance liquid chromatography-electrospray mass spectrometry, we describe a new assay for monitoring 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase activity. Incubations were carried out with HMG-CoA reductase (rat liver), HMG-CoA and NADPH, and terminated by the addition of HCl. The reaction product, mevalonolactone, and internal standard, were extracted with ethyl acetate, dissolved in methanol, and analyzed by LC-MS. Using an isocratic mobile phase of 10% acetonitrile and 0.1% formic acid (flow-rate, 0.2 ml/min), the protonated molecules of mevalonolactone at m/z 131 and internal standard, beta,beta-dimethyl-gamma-(hydroxymethyl)-gamma-butyrolactone, at m/z 145, were detected using selected ion monitoring. The limit of detection was approximately 6.5 pg, and the limit of quantitation was approximately 16.3 pg. Extraction recovery was >90%. The relative standard deviations for intra- and inter-day assays were approximately 4.1+/-2.7 and 9.4+/-3.4%, respectively. Mevalonolactone was examined over a period of 3 days and found to be stable. Using this assay, lovastatin and mevastatin inhibited HMG-CoA reductase activity with IC50 values 0.24+/-0.02 and 2.16+/-0.31 microM, respectively. These methods...Continue Reading

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Citations

Jul 27, 2001·Journal of Mass Spectrometry : JMS
Mar 28, 2008·Applied Microbiology and Biotechnology·Bin ZhugeJian Zhuge
Apr 27, 2010·Journal of Lipid Research·Matteo MozzicafreddoMauro Angeletti
May 12, 2004·Analytical Biochemistry·Raphaele GerberDouglas S Clark
Jul 16, 2008·European Journal of Immunology·Magdalena KistowskaGennaro De Libero
Feb 3, 2007·Journal of Lipid Research·Akira HondaHiroshi Miyazaki
May 30, 2008·Journal of Biomolecular Screening·Marjo SimonenDaniela Gabriel
Apr 19, 2005·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Michele BuffaliniVilberto Stocchi

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