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Multiplex LC-MS/MS for simultaneous determination of 25-hydroxyvitamin D, 24,25-dihydroxyvitamin D3, albumin, and vitamin D-binding protein with its isoforms: One-step estimation of bioavailable vitamin D and vitamin D metabolite ratio.
Ko, Dae-Hyun; Jun, Sun-Hee; Nam, Youngwon; Song, Sang H; Han, Minje; Yun, Yeo-Min; Lee, Kyunghoon; Song, Junghan.
Afiliación
  • Ko DH; Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan Medical Center, Seoul, Republic of Korea.
  • Jun SH; Department of Laboratory Medicine, Seoul National University Bundang Hospital, Gyeonggi-do, Republic of Korea.
  • Nam Y; Department of Laboratory Medicine, Seoul National University Bundang Hospital, Gyeonggi-do, Republic of Korea; Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Republic of Korea.
  • Song SH; Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Republic of Korea; Department of Laboratory Medicine, Seoul National University Hospital, Seoul, Republic of Korea.
  • Han M; Department of Laboratory Medicine, Kangdong Sacred Heart Hospital, Seoul, Republic of Korea.
  • Yun YM; Department of Laboratory Medicine, Konkuk University School of Medicine, Seoul, Republic of Korea.
  • Lee K; Department of Laboratory Medicine, Seoul National University Bundang Hospital, Gyeonggi-do, Republic of Korea; Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Republic of Korea. Electronic address: khlee59023@snubh.org.
  • Song J; Department of Laboratory Medicine, Seoul National University Bundang Hospital, Gyeonggi-do, Republic of Korea; Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Republic of Korea. Electronic address: songjhcp@snu.ac.kr.
J Steroid Biochem Mol Biol ; 206: 105796, 2021 02.
Article en En | MEDLINE | ID: mdl-33259939
Bioavailable vitamin D and vitamin D metabolite ratio (VMR) have emerged as potential novel vitamin D markers. We developed a multiplex liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to determine all elements necessary for the calculation of bioavailable vitamin D and VMR, including 25-hydroxyvitamin D [25-(OH)D] and 24,25-dihydroxyvitamin D3 [24,25-(OH)2D3], VDBP and its isoforms, and albumin. Following separate reactions of hexane extraction and trypsin digestion, serum samples were analyzed using LC-MS/MS to measure 25-(OH)D3, 25-(OH)D2, 24,25-(OH)2D3, VDBP and its isoforms, and albumin. Analytical performances were assessed. Korean (n = 229), Arab (n = 98), White (n = 99) and Black American (n = 99) samples were analyzed. Bioavailable vitamin D and VMR were calculated. All target molecules were clearly separated and accurately quantified by LC-MS/MS. Analytical performances, including imprecision, accuracy, ion suppression, limit of quantification, linearity, and comparison with existing methods were within acceptable levels. The allele frequencies of VDBP isoforms in various races resulted similar to previously known values. The levels of bioavailable vitamin D were highest in White Americans and lowest in Black Americans. We have successfully developed a multiplex LC-MS/MS-based assay method that can simultaneously perform the measurement of all parameters needed to calculate bioavailable vitamin D and VMR. Our devised method was robust and reliable in terms of analytical performances and could be applied to routine clinical samples in the future to more accurately assess vitamin D status.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Vitamina D / Proteína de Unión a Vitamina D / 24,25-Dihidroxivitamina D 3 Límite: Humans Idioma: En Revista: J Steroid Biochem Mol Biol Asunto de la revista: BIOLOGIA MOLECULAR / BIOQUIMICA Año: 2021 Tipo del documento: Article Pais de publicación: Reino Unido
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Vitamina D / Proteína de Unión a Vitamina D / 24,25-Dihidroxivitamina D 3 Límite: Humans Idioma: En Revista: J Steroid Biochem Mol Biol Asunto de la revista: BIOLOGIA MOLECULAR / BIOQUIMICA Año: 2021 Tipo del documento: Article Pais de publicación: Reino Unido