Abstract
The capabilities of using particle beam (PB) liquid chromatography–mass spectrometry (LC–MS) was investigated for the analysis of cholesterol and oxycholesterols; normal-phase (NP) and reversed-phase (RP) chromatographic methods making use of narrow-bore columns were set up for the simultaneous separation of these compounds. The influence of PB interface parameters, LC conditions (solvent composition and flow-rate) and MS conditions was explored. PB mass spectra were recorded in electron impact (EI), positive-ion chemical ionization (PCI) and negative-ion chemical ionization (NCI) modes and the corresponding spectral data discussed. Better performance in terms of ion abundance were observed in EI mode both under NP and RP conditions. Using EI source and operating in selected-ion monitoring (SIM) mode, linearity, sensitivity and precision of the analysis were explored. The experimental data fitted a quadratic model in the overall range studied. Using the RP-LC–PB-MS method detection limits at the low-ng level were achieved for all the analytes; in addition, repeatability ranging from 3% for triol to 9% for cholesterol and 7-ketocholesterol at the 4 ng level was obtained.
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