Hybrid 213 nm photodissociation of cationized Sterol lipid ions yield [M]+. Radical products for improved structural characterization using multistage tandem mass spectrometry

• Published in: Analytica chimica acta 2021-01, Vol.1141, p.100-109
• Main Authors: West, Henry, Reid, Gavin E.
• Format: Article
• Language: English
• Subjects: Animals; Cholesterol; Chromatography, High Pressure Liquid; Ions; Lipids; Oxosterol; Oxysterol; Photodissociation; Spectrometry, Mass, Electrospray Ionization; Sterol ester; Sterols; Tandem Mass Spectrometry
• ISSN: 0003-2670; 1873-4324
• DOI: 10.1016/j.aca.2020.10.013

Sterols are a class of lipid molecules that include cholesterol, oxysterols, and sterol esters. Sterol lipids play critical functional roles in mammalian biology, including the dynamic regulation of cell membrane fluidity, as precursors for the synthesis of bile acids, steroid hormones and vitamin D, as regulators of gene expression in lipid metabolism, and for cholesterol transport and storage. The most common method employed for sterol analysis is high performance liquid chromatography coupled with tandem mass spectrometry (MS/MS). However, conventional collision induced dissociation (CID) methods used for ion activation during MS/MS typically fail to provide sufficient structural information for unambiguous assignment of sterol species based on their fragmentation behaviour alone. This places a significant burden on the efficiency of the chromatographic separation methods for the effective separation of isomeric sterols. Here, toward developing an improved analysis strategy for sterol lipids, we have explored the novel use of 213 nm photodissociation MS/MS and hybrid multistage-MS/MS (i.e., MSn) data acquisition approaches for the improved structural characterization of cholesterol, representative isomeric oxysterols, and cholesteryl esters. Most notably, UVPD-MS/MS of ammoniated, lithiated and sodiated adducts of cholesterol, several representative oxysterol species, and an oxosterol lipid, are shown to give rise to abundant [M]+. radical cation products, that subsequently fragment during collision induced MS3 to yield extensive structurally informative product ions, similar to those observed by Electron Ionization, and that enable their unambiguously assignment, including isomeric differentiation of oxysterols. For cholesterol esters, a reversed hybrid collision induced-MS/MS and UVPD-MS3 approach is shown to enable assignment of the sterol backbone, and localization of the site(s) of unsaturation within esterified fatty acyl chains. [Display omitted] •Hybrid UV photodissociation and multistage tandem mass spectrometry is a novel analysis strategy for sterol lipids.•213 nm UVPD-MS/MS of cholesterol and oxysterols produces abundant [M]+. radical cation product ions.•HCD-MS3 of these products enables unambiguously structural assignment, including for differentiation of isomeric oxysterols.•HCD-MS/MS and UVPD-MS3 enables detailed structural assignment of cholesterol esters, including site(s) of unsaturation.