High-temperature GC-MS-based serum cholesterol signatures may reveal sex differences in vasospastic angina

Abstract

Alterations of cholesterol metabolism are responsible for vasospastic angina and atherosclerosis. To comprehensively evaluate cholesterol metabolism, 18 sterols, including cholesterol, 6 cholesteryl esters (CEs), 3 cholesterol precursors, and 8 hydroxycholesterols (OHCs), were simultaneously analyzed using hybrid solid-phase extraction (SPE) purifi cation coupled to high-temperature gas chromatography- mass spectrometry (HTGC-MS). Methanol-based hybrid SPE increased the selective extraction, and HTGC resulted in a good chromatographic resolution for the separation of lipophilic compounds. The limits of quantifi cation of cholesterol and CEs ranged from 0.2 to 10.0 - g/ml, while OHCs and cholesterol precursors ranged from 0.01 to 0.10 - g/ml. Linearity as the correlation coeffi cient was higher than 0.99 with the exception of cholesteryl laurate, myristate, oleate, and linoleate ( r 2 < 0.98). The precision (% coeffi cient of variation) and accuracy (% bias) ranged from 1.1 to 9.8% and from 75.9 to 125.1%, respectively. The overall recoveries of CEs ranged from 26.1 to 64.0%, and the recoveries of other sterols ranged from 83.8 to 129.3%. The cholesterol signatures showed sex differences in patients with vasospastic angina and may associate with 24-reductases. This technique can be useful for making clinical diagnoses and for an increased understanding of the pathophysiology of vasospastic angina. Copyright © 2014 by the American Society for Biochemistry and Molecular Biology, Inc.