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) purification 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 quantification of cholesterol and CEs ranged from 0.2 to 10.0 μg/ml, while OHCs and cholesterol precursors ranged from 0.01 to 0.10 μg/ml. Linearity as the correlation coefficient was higher than 0.99 with the exception of cholesteryl laurate, myristate, oleate, and linoleate (r2 > 0.98). The precision (% coefficient 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.
Highlights
Alterations of cholesterol metabolism are responsible for vasospastic angina and atherosclerosis
Cholesterol, three cholesterol precursors, and eight OHCs were eluted within 7 min, while six cholesteryl ester (CE) were eluted in the order of the number of carbons in the hydrocarbon chain: cholesteryl laurate (CE 12:0), myristate (CE 14:0), palmitate (CE 16:0), oleate (CE 18:1), linoleate (CE 18:2), and stearate (CE 18:0) (Fig. 2)
In TMS derivatization, both hydroxyl and carbonyl ketone groups were derivatized with TMS, while CEs were unaffected by TMS agents because they do not have polar groups in their chemical structures (Fig. 1, Table 1)
Summary
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) purification coupled to high-temperature gas chromatography-mass spectrometry (HTGC-MS). The limits of quantification 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. 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.—Son, H-H., J-Y. High-temperature GC-MS-based serum cholesterol signatures may reveal sex differences in vasospastic angina. The comprehensive profiling of cholesterol and related sterols, including CEs, cholesterol precursors, and OHCs, is needed to understand the physiologies of cardiovascular events
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