A validated LC-MS/MS assay for quantification of 24(S)-hydroxycholesterol in plasma and cerebrospinal fluid

Rohini Sidhu, Hui Jiang, Nicole Y. Farhat, Nuria Carrillo-Carrasco, Myra Woolery, Elizabeth Ottinger, Forbes D. Porter, Jean E. Schaffer, Daniel S. Ory, Xuntian Jiang

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31 Scopus citations

Abstract

24(S)-hydroxycholesterol [24(S)-HC] is a cholesterol metabolite that is formed almost exclusively in the brain. The concentrations of 24(S)-HC in cerebrospinal fluid (CSF) and/or plasma might be a sensitive marker of altered cholesterol metabolism in the CNS. A highly sensitive 2D-LC-MS/MS assay was developed for the quantification of 24(S)-HC in human plasma and CSF. In the development of an assay for 24(S)-HC in CSF, significant nonspecific binding of 24(S)-HC was observed and resolved with the addition of 2.5% 2-hydroxypropyl-β-cyclodextrin (HP-β-CD) into CSF samples. The sample preparation consists of liquid-liquid extraction with methyl-tert-butyl ether and derivatization with nicotinic acid. Good linearity was observed in a range from 1 to 200 ng/ml and from 0.025 to 5 ng/ml, for plasma and CSF, respectively. Acceptable precision and accuracy were obtained for concentrations over the calibration curve ranges. Stability of 24(S)-HC was reported under a variety of storage conditions. This method has been successfully applied to support a National Institutes of Health-sponsored clinical trial of HP-β-CD in Niemann-Pick type C1 patients, in which 24(S)-HC is used as a pharmacodynamic biomarker.

Original languageEnglish
Pages (from-to)1222-1233
Number of pages12
JournalJournal of lipid research
Volume56
Issue number6
DOIs
StatePublished - Jun 1 2015

Keywords

  • 2-hydroxypropyl-β-cyclodextrin
  • Niemann-Pick C1 disease
  • Two-dimensional liquid chromatography-tandem mass spectrometry
  • Validation

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    Sidhu, R., Jiang, H., Farhat, N. Y., Carrillo-Carrasco, N., Woolery, M., Ottinger, E., Porter, F. D., Schaffer, J. E., Ory, D. S., & Jiang, X. (2015). A validated LC-MS/MS assay for quantification of 24(S)-hydroxycholesterol in plasma and cerebrospinal fluid. Journal of lipid research, 56(6), 1222-1233. https://doi.org/10.1194/jlr.D058487