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.
- Cardiovascular disease
- Hybrid solid-phase extraction-precipitation
ASJC Scopus subject areas
- Cell Biology