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Sensitive UHPLC-MS/MS quantification method for 4ß- and 4α-hydroxycholesterol in plasma for accurate CYP3A phenotyping.
Suzuki, Yosuke; Oda, Ayako; Negami, Jun; Toyama, Daiki; Tanaka, Ryota; Ono, Hiroyuki; Ando, Tadasuke; Shin, Toshitaka; Mimata, Hiromitsu; Itoh, Hiroki; Ohno, Keiko.
Afiliação
  • Suzuki Y; Department of Medication Use Analysis and Clinical Research, Meiji Pharmaceutical University, Kiyose, Tokyo, Japan. Electronic address: y-suzuki@my-pharm.ac.jp.
  • Oda A; Department of Medication Use Analysis and Clinical Research, Meiji Pharmaceutical University, Kiyose, Tokyo, Japan.
  • Negami J; Department of Medication Use Analysis and Clinical Research, Meiji Pharmaceutical University, Kiyose, Tokyo, Japan.
  • Toyama D; Department of Medication Use Analysis and Clinical Research, Meiji Pharmaceutical University, Kiyose, Tokyo, Japan.
  • Tanaka R; Department of Clinical Pharmacy, Oita University Hospital, Yufu, Oita, Japan.
  • Ono H; Department of Clinical Pharmacy, Oita University Hospital, Yufu, Oita, Japan.
  • Ando T; Department of Urology, Oita University Faculty of Medicine, Yufu, Oita, Japan.
  • Shin T; Department of Urology, Oita University Faculty of Medicine, Yufu, Oita, Japan.
  • Mimata H; Department of Urology, Oita University Faculty of Medicine, Yufu, Oita, Japan.
  • Itoh H; Department of Clinical Pharmacy, Oita University Hospital, Yufu, Oita, Japan.
  • Ohno K; Department of Medication Use Analysis and Clinical Research, Meiji Pharmaceutical University, Kiyose, Tokyo, Japan.
J Lipid Res ; 63(3): 100184, 2022 03.
Article em En | MEDLINE | ID: mdl-35181316
4ß-Hydroxycholesterol (4ß-OHC) is formed by Cytochrome P450 (CYP)3A and has drawn attention as an endogenous phenotyping probe for CYP3A activity. However, 4ß-OHC is also increased by cholesterol autooxidation occurring in vitro due to dysregulated storage and in vivo by oxidative stress or inflammation, independent of CYP3A activity. 4α-hydroxycholesterol (4α-OHC), a stereoisomer of 4ß-OHC, is also formed via autooxidation of cholesterol, not by CYP3A, and thus may have clinical potential in reflecting the state of cholesterol autooxidation. In this study, we establish a sensitive method for simultaneous quantification of 4ß-OHC and 4α-OHC in human plasma using ultra-high performance liquid chromatography coupled to tandem mass spectrometry. Plasma samples were prepared by saponification, two-step liquid-liquid extraction, and derivatization using picolinic acid. Intense [M+H]+ signals for 4ß-OHC and 4α-OHC di-picolinyl esters were monitored using electrospray ionization. The assay fulfilled the requirements of the US Food and Drug Administration guidance for bioanalytical method validation, with a lower limit of quantification of 0.5 ng/ml for both 4ß-OHC and 4α-OHC. Apparent recovery rates from human plasma ranged from 88.2% to 101.5% for 4ß-OHC, and 91.8% to 114.9% for 4α-OHC. Additionally, matrix effects varied between 86.2% and 117.6% for 4ß-OHC and between 89.5% and 116.9% for 4α-OHC. Plasma 4ß-OHC and 4α-OHC concentrations in healthy volunteers, stage 3-5 chronic kidney disease (CKD) patients, and stage 5D CKD patients as measured by the validated assay were within the calibration ranges in all samples. We propose this novel quantification method may contribute to accurate evaluation of in vivo CYP3A activity.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Insuficiência Renal Crônica / Hidroxicolesteróis Tipo de estudo: Diagnostic_studies / Guideline Limite: Female / Humans / Male Idioma: En Revista: J Lipid Res Ano de publicação: 2022 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Insuficiência Renal Crônica / Hidroxicolesteróis Tipo de estudo: Diagnostic_studies / Guideline Limite: Female / Humans / Male Idioma: En Revista: J Lipid Res Ano de publicação: 2022 Tipo de documento: Article
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