title: High resolution mass spectrometry based method applicable for a wide range of 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase inhibitors in blood serum including intermediates and products of the cholesterol biosynthetic pathway.
abstract: Statins belong to the major class of hypolipidemic drugs. They act as competitive inhibitors of 3-hydroxy-3-methyl-glutaryl-coenzyme A (HMG-CoA) reductase, a rate-limiting enzyme in the cholesterol biosynthetic pathway. This inhibition not only leads to the depletion of cholesterol and its fatty acid esters, but also to the depletion of the intermediates of this metabolic pathway (mainly pyrophosphates), which can play an important role in tumor proliferation. The aim of the current study was to establish a versatile multi-analyte method capable of quantitative determination of various currently-used statins, together with free cholesterol (FC), cholesterol esters (CEs), and some key intermediates of the mevalonate pathway occurring in human serum. Various methods of sample preparation were examined in order to minimize the content of potentially interfering serum proteins, and simultaneously to assure acceptable recovery of the target analytes. Following protein precipitation with 2-propanol, separation of the sample components using ultra-high performance liquid chromatography coupled with tandem high resolution mass spectrometry (U-HPLC-HRMS/MS) was performed, employing a hyphenated quadrupole Orbitrap mass analyzer. The potential of the developed method was validated on human serum samples from patients treated with statins. This versatile method possesses wide applicability, in both clinical and experimental medicine.
authors.affiliation: University of Chemistry and Technology, Department of Food Chemistry and Analysis, Technická 3, 166 28, Praha 6, Czechia.
authors.forename: Vít
authors.initials: V
authors.lastname: Kosek
authors.affiliation: University of Chemistry and Technology, Department of Food Chemistry and Analysis, Technická 3, 166 28, Praha 6, Czechia.
authors.forename: Milena
authors.initials: M
authors.lastname: Stránská
authors.affiliation: University of Chemistry and Technology, Department of Food Chemistry and Analysis, Technická 3, 166 28, Praha 6, Czechia.
authors.forename: Marie
authors.initials: M
authors.lastname: Fenclová
authors.affiliation: University of Chemistry and Technology, Department of Biochemistry and Microbiology, Technická 3, 166 28, Praha 6, Czechia.
authors.forename: Tomáš
authors.initials: T
authors.lastname: Ruml
authors.affiliation: Institute of Medical Chemistry and Laboratory Diagnostics, and 4th Department of Internal Medicine, 1st Faculty of Medicine, Charles University in Prague, U Nemocnice 2, Praha 2, 128 01, Czechia.
authors.forename: Libor
authors.initials: L
authors.lastname: Vítek
authors.affiliation: University of Chemistry and Technology, Department of Food Chemistry and Analysis, Technická 3, 166 28, Praha 6, Czechia. Electronic address: jana.hajslova@vscht.cz.
authors.forename: Jana
authors.initials: J
authors.lastname: Hajšlová
mesh_terms: D002784:Cholesterol / Q000096:biosynthesis / Q000097:blood; D002788:Cholesterol Esters / Q000097:blood; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006801:Humans; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors / Q000097:blood*; D008798:Mevalonic Acid / Q000378:metabolism; D011106:Polyisoprenyl Phosphates / Q000097:blood; D012717:Sesquiterpenes / Q000097:blood; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D002788:Cholesterol Esters; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D011106:Polyisoprenyl Phosphates; D012717:Sesquiterpenes; C004808:farnesyl pyrophosphate; D002784:Cholesterol; C002963:geranylgeranyl pyrophosphate; D008798:Mevalonic Acid
keywords: Cholesterol; High resolution mass spectrometry; Isoprenoid pyrophosphates; Serum; Statins