title: A validated LC-MS/MS assay for quantification of 24(S)-hydroxycholesterol in plasma and cerebrospinal fluid.
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.
authors: Diabetic Cardiovascular Disease Center, Washington University School of Medicine, St. Louis, MO 63110.
authors: Rohini
authors: R
authors: Sidhu
authors: Diabetic Cardiovascular Disease Center, Washington University School of Medicine, St. Louis, MO 63110.
authors: Hui
authors: H
authors: Jiang
authors: Program in Developmental Endocrinology and Genetics, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892.
authors: Nicole Y
authors: NY
authors: Farhat
authors: Therapeutics for Rare and Neglected Diseases National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD 20850.
authors: Nuria
authors: N
authors: Carrillo-Carrasco
authors: Nursing Department, National Institutes of Health, Bethesda, MD 20892.
authors: Myra
authors: M
authors: Woolery
authors: Division of Preclinical Innovation, National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD 20850.
authors: Elizabeth
authors: E
authors: Ottinger
authors: Program in Developmental Endocrinology and Genetics, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892.
authors: Forbes D
authors: FD
authors: Porter
authors: Diabetic Cardiovascular Disease Center, Washington University School of Medicine, St. Louis, MO 63110.
authors: Jean E
authors: JE
authors: Schaffer
authors: Diabetic Cardiovascular Disease Center, Washington University School of Medicine, St. Louis, MO 63110.
authors: Daniel S
authors: DS
authors: Ory
authors: Diabetic Cardiovascular Disease Center, Washington University School of Medicine, St. Louis, MO 63110.
authors: Xuntian
authors: X
authors: Jiang
mesh_terms: D000073738:2-Hydroxypropyl-beta-cyclodextrin; D002490:Central Nervous System / Q000378:metabolism; D002784:Cholesterol / Q000378:metabolism; D002853:Chromatography, Liquid; D006801:Humans; D006888:Hydroxycholesterols / Q000097:blood / Q000134:cerebrospinal fluid; D052556:Niemann-Pick Disease, Type C / Q000097:blood / Q000134:cerebrospinal fluid; D053719:Tandem Mass Spectrometry; D014481:United States; D047392:beta-Cyclodextrins / Q000494:pharmacology
chemical_list: D006888:Hydroxycholesterols; D047392:beta-Cyclodextrins; D000073738:2-Hydroxypropyl-beta-cyclodextrin; D002784:Cholesterol
keywords: 2-hydroxypropyl-β-cyclodextrin; Niemann-Pick C1 disease; two-dimensional liquid chromatography-tandem mass spectrometry; validation