Journal
JOURNAL OF LIPID RESEARCH
Volume 59, Issue 6, Pages 1058-1070Publisher
ELSEVIER
DOI: 10.1194/jlr.D083246
Keywords
sterols; cholesterol/metabolism; Niemann-Pick type C; oxidized lipids; tandem mass spectrometry
Categories
Funding
- Biotechnology and Biological Sciences Research Council [BB/I001735/1, BB/N015932/1, BB/L001942/1]
- Welsh Government's A4B project
- KESS2 award from the Welsh Government
- European Social Fund
- PhD studentship from Imperial College Healthcare Charities
- European Society of Paediatric Infectious Diseases
- Medical Research Council [MR/N001427/1]
- BBSRC [BB/I001735/1, BB/L001942/1, BB/N015932/1] Funding Source: UKRI
- MRC [MR/N001427/1] Funding Source: UKRI
Ask authors/readers for more resources
7-Oxocholesterol (7-OC), 5,6-epoxycholesterol (5,6-EC), and its hydrolysis product cholestane-3 beta,5 alpha,6 beta-triol (3 beta,5 alpha,6 beta-triol) are normally minor oxysterols in human samples; however, in disease, their levels may be greatly elevated. This is the case in plasma from patients suffering from some lysosomal storage disorders, e.g., Niemann-Pick disease type C, or the inborn errors of sterol metabolism, e.g., Smith-Lemli-Opitz syndrome and cerebrotendinous xanthomatosis. A complication in the analysis of 7-OC and 5,6-EC is that they can also be formed ex vivo from cholesterol during sample handling in air, causing confusion with molecules formed in vivo. When formed endogenously, 7-OC, 5,6-EC, and 3,5,6-triol can be converted to bile acids. Here, we describe methodology based on chemical derivatization and LC/MS with multistage fragmentation (MSn) to identify the necessary intermediates in the conversion of 7-OC to 3-hydroxy-7-oxochol-5-enoic acid and 5,6-EC and 3 beta,5 alpha,6 beta-triol to 3 beta,5 alpha,6 beta-triol can be converted to bile acids. Here, we describe methodology based on chemical derivatization and LC/MS with multistage fragmentation (MS n) to identify the necessary intermediates in the conversion of 7-OC to 3. hydroxy-7-oxochol-5-enoic acid and 5,6-EC and 3 beta,5 alpha,6 beta triol to 3 beta,5 alpha,6 beta-trihydroxycholanoic acid. Identification of intermediate metabolites is facilitated by their unusual MSn fragmentation patterns. Semiquantitative measurements are possible, but absolute values await the synthesis of isotopelabeled standards.-Griffiths,
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available