Multiple Fluorometric Assay of Three Major Vitamin D3 Metabolites (25-Hydroxyvitamin D3, 24,25-Dihydroxyvitamin D3 and 1,25-Dihydroxyvitamin D3) in Plasma and Its Application to Clinical Studies.

• WANG Xinxiang

  Institute for Medical and Dental Engineering and Third Department of

• SHIMIZU Masato

  Institute for Medical and Dental Engineering and Third Department of

• NUMANO Fujio

  Institute for Medical and Dental Engineering and Third Department of

• ASAOKA Hiroshi

  Institute for Medical and Dental Engineering and Third Department of

• YAMADA Sachiko

  Institute for Medical and Dental Engineering and Third Department of

Bibliographic Information

Other Title

• Multiple Fluorometric Assay of Three Ma

Abstract

We have developed a fluorometric method for routine analyses of 25-hydroxyvitamin D₃ (25-OHD₃) and 24, 25- and 1, 25-dihydroxyvitamin D₃ [24, 25- and 1, 25-(OH)₂D₃] in human plasma. Lipid extracts from plasma (2.5ml) were purified using a Sep-Pak silica cartridge followed by normal-phase HPLC, and each of the three metabolites was assayed in separate lines. Each metabolite fraction was treated with fluorescent dienophile (DMEQ-TAD) and quantified on reversed-phase HPLC after appropriate cleanup steps. Labeled 25-OHD₃ was quantified directly, 24, 25-(OH)₂D₃ was analyzed after cleanup by a Bond Elut PSA cartridge and 1, 25-(OH)₂D₃ was quantified after purification by a Bond Elut PSA cartridge and normal-phase HPLC. In the assay of 25-OHD₃, we demonstrated the usefulness of a non-radioactive internal standard for the fluorometric assay. The multiple fluorometric assay was successfully applied to the clinical monitoring of vitamin D₃ therapy in a vitamin D-deficient patient with metabolic bone disease.

Journal

• Analytical Sciences

  Analytical Sciences 13 (2), 255-261, 1997

  The Japan Society for Analytical Chemistry

Keywords

• Fluorometric assay
• fluorescent dienophile
• 25-hydroxyvitamin D₃
• 24,25-dihydroxyvitamin D₃
• 1,25-dihydroxyvitamin D₃
• human plasma

Details

• CRID: 1390001204257861888
• NII Article ID: 10002411174
• NII Book ID: AA10500785
• DOI: 10.2116/analsci.13.255
• COI: 1:CAS:528:DyaK2sXis12msr4%3D
• ISSN: 13482246
• ISSN: 09106340
• ISSN: http://id.crossref.org/issn/09106340
• NDL BIB ID: 4196985
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I4196985
• Web Site: https://link.springer.com/content/pdf/10.2116/analsci.13.255.pdf
• Web Site: https://link.springer.com/article/10.2116/analsci.13.255/fulltext.html
• Web Site: http://www.jstage.jst.go.jp/article/analsci1985/13/2/13_2_255/_pdf
• Web Site: https://search.jamas.or.jp/link/ui/1997144951
• Text Lang: en
• Data Source:
  • JaLC
  • NDL
  • Crossref
  • CiNii Articles
• Abstract License Flag: Disallowed