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Titolo:
Analysis of 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors using liquid chromatography-electrospray mass spectrometry
Autore:
Park, EJ; Lee, D; Shin, YG; Lantvit, DD; van Breemen, RB; Kinghorn, AD; Pezzuto, JM;
Indirizzi:
Univ Illinois, Coll Pharm, Program Collaborat Res Pharmaceut Sci, Chicago,IL 60612 USA Univ Illinois Chicago IL USA 60612 s Pharmaceut Sci, Chicago,IL 60612 USA Univ Illinois, Coll Pharm, Dept Med Chem & Pharmacognosy, Chicago, IL 60612 USA Univ Illinois Chicago IL USA 60612 & Pharmacognosy, Chicago, IL 60612 USA
Titolo Testata:
JOURNAL OF CHROMATOGRAPHY B
fascicolo: 2, volume: 754, anno: 2001,
pagine: 327 - 332
SICI:
1387-2273(20010425)754:2<327:AO3ARI>2.0.ZU;2-V
Fonte:
ISI
Lingua:
ENG
Soggetto:
COA REDUCTASE; RAT-LIVER; IN-VITRO; LOVASTATIN; CELLS; CANCER; GROWTH; ASSAY; PHOSPHORYLATION; SIMVASTATIN;
Keywords:
3-hydroxy-3-methylglutaryl-coenzyme A reductase;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
25
Recensione:
Indirizzi per estratti:
Indirizzo: Pezzuto, JM Univ Illinois, Coll Pharm, Program Collaborat Res Pharmaceut Sci, 833 S Wood St, Chicago, IL 60612 USA Univ Illinois 833 S Wood St Chicago IL USA 60612 IL 60612 USA
Citazione:
E.J. Park et al., "Analysis of 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors using liquid chromatography-electrospray mass spectrometry", J CHROMAT B, 754(2), 2001, pp. 327-332

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 relativestandard 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 muM, respectively. These methods offer some advantages over those reported previously which employ radiolabeled substrate and products,and should be useful in searching for compounds that could lower serum cholesterol or alter cell growth and differentiation. (C) 2001 Elsevier Science B.V. All rights reserved.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 06/04/20 alle ore 08:47:11