Determination of 25-hydroxyvitamin D in human plasma using high-performance liquid chromatography-tandem mass spectrometry

Naoko Tsugawa, Yoshitomo Suhara, Maya Kamao, Toshio Okano

Research output: Contribution to journalArticle

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Abstract

We report here the development of a precise and sensitive method to determine 25-hydroxyvitamin D (25-OH-D2/ -D3) in human plasma using high-perfomance liquid chromatography-tandem mass-mass spectrometry with atomospheric pressure chemical ionization (LC-APCI-MS/ MS). The method involves the use of deuterated 25-OH-D3 as an internal standard compound for 25-OH-D2/-D3, which was synthesized in our laboratory, and the selection of a precursor and product ion with a MS/MS multiple reaction monitoring method. The average intraassay and interassay variation values (relative standard deviation) were 5.7 and 2.5%, respectively, for 25-OH-D3 and 4.5 and 5.1%, respectively, for 25-OH-D2. The average spiked recoveries from authentic compounds added to normal human plasma samples for 25-OH-D3 and 25-OH-D2 were 103.8 and 98.8%, respectively. Mean plasma concentrations of 25-OH-D3 and 25-OH-D2 in healthy subjects were 20.5 and 0.4 ng/mL, respectively. We conclude that this novel LC-APCI-MS/MS method would be useful for the evaluation of the vitamin D status in postmenopausal women and elderly subjects and provide useful information in the diagnosis of vitamin D insufficiency/ deficiency, as well as for the treatment and prevention of osteoporosis with vitamin D.

Original languageEnglish
Pages (from-to)3001-3007
Number of pages7
JournalAnalytical Chemistry
Volume77
Issue number9
DOIs
Publication statusPublished - 2005 May 1
Externally publishedYes

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Plasma (human)
High performance liquid chromatography
Mass spectrometry
Vitamin D
hydroxide ion
25-hydroxyvitamin D
Liquid chromatography
Ionization
Ions
Plasmas
Recovery

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Determination of 25-hydroxyvitamin D in human plasma using high-performance liquid chromatography-tandem mass spectrometry. / Tsugawa, Naoko; Suhara, Yoshitomo; Kamao, Maya; Okano, Toshio.

In: Analytical Chemistry, Vol. 77, No. 9, 01.05.2005, p. 3001-3007.

Research output: Contribution to journalArticle

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abstract = "We report here the development of a precise and sensitive method to determine 25-hydroxyvitamin D (25-OH-D2/ -D3) in human plasma using high-perfomance liquid chromatography-tandem mass-mass spectrometry with atomospheric pressure chemical ionization (LC-APCI-MS/ MS). The method involves the use of deuterated 25-OH-D3 as an internal standard compound for 25-OH-D2/-D3, which was synthesized in our laboratory, and the selection of a precursor and product ion with a MS/MS multiple reaction monitoring method. The average intraassay and interassay variation values (relative standard deviation) were 5.7 and 2.5{\%}, respectively, for 25-OH-D3 and 4.5 and 5.1{\%}, respectively, for 25-OH-D2. The average spiked recoveries from authentic compounds added to normal human plasma samples for 25-OH-D3 and 25-OH-D2 were 103.8 and 98.8{\%}, respectively. Mean plasma concentrations of 25-OH-D3 and 25-OH-D2 in healthy subjects were 20.5 and 0.4 ng/mL, respectively. We conclude that this novel LC-APCI-MS/MS method would be useful for the evaluation of the vitamin D status in postmenopausal women and elderly subjects and provide useful information in the diagnosis of vitamin D insufficiency/ deficiency, as well as for the treatment and prevention of osteoporosis with vitamin D.",
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