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

Xinxiang WANG; Masato SHIMIZU; Fujio NUMANO; Hiroshi ASAOKA; Sachiko YAMADA

doi:10.2116/analsci.13.255

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

Xinxiang WANG, Masato SHIMIZU, Fujio NUMANO, Hiroshi ASAOKA, Sachiko YAMADA

著者情報

キーワード: Fluorometric assay, fluorescent dienophile, 25-hydroxyvitamin D₃, 24,25-dihydroxyvitamin D₃, 1,25-dihydroxyvitamin D₃, human plasma

ジャーナルフリー

1997 年 13 巻 2 号 p. 255-261

DOI https://doi.org/10.2116/analsci.13.255

詳細

Published: 1997/04/10 Received: 1996/12/20 Available on J-STAGE: 2006/07/07 Accepted: 1997/01/10 Advance online publication: - Revised: -
訂正情報
訂正日: 2006/07/07 訂正理由: - 訂正箇所: 論文抄録訂正内容: Wrong : We have developed a fluorometric method for routine analyses of 25-hydroxyvitamin D^{3 (25-OHD^{3) and 24, 25- and 1, 25-dihydroxyvitamin D^{3 [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.
Right : 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.}}}

訂正日: 2006/07/07 訂正理由: - 訂正箇所: 著者情報訂正内容: Wrong : Xinxiang WANG¹⁾, Masato SHIMIZU²⁾, Fujio NUMANO¹⁾, Hiroshi ASAOKA¹⁾, Sachiko YAMADA¹⁾
Right : Xinxiang WANG¹⁾, Masato SHIMIZU¹⁾, Fujio NUMANO¹⁾, Hiroshi ASAOKA¹⁾, Sachiko YAMADA¹⁾

訂正日: 2006/07/07 訂正理由: - 訂正箇所: PDFファイル訂正内容: -

抄録

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.

責任著者(Corresponding author)

訂正情報

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