Determination of 25-hydroxyvitamin D in human plasma using high-performance liquid chromatography--tandem mass spectrometry.
We report here the development of a precise and sensitive method to determine 25-hydroxyvitamin D (25-OH-D(2)/ -D(3)) in human plasma using high-performance liquid chromatography-tandem mass-mass spectrometry with atmospheric pressure chemical ionization (LC-APCI-MS/MS). The method involves the use of deuterated 25-OH-D(3) as an internal standard compound for 25-OH-D(2)/-D(3), 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-D(3) and 4.5 and 5.1%, respectively, for 25-OH-D(2). The average spiked recoveries from authentic compounds added to normal human plasma samples for 25-OH-D(3) and 25-OH-D(2) were 103.8 and 98.8%, respectively. Mean plasma concentrations of 25-OH-D(3) and 25-OH-D(2) 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.
Tsugawa
Naoko
N
Department of Hygienic Sciences, Kobe Pharmaceutical University, 4-19-1, Motoyamakita-machi, Higashinada-ku, Kobe 658-8558, Japan.
Suhara
Yoshitomo
Y
Kamao
Maya
M
Okano
Toshio
T
D015652:25-Hydroxyvitamin D 2 / Q000097:blood; D002112:Calcifediol / Q000097:blood; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005260:Female; D006801:Humans; D008875:Middle Aged; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014807:Vitamin D / Q000031:analogs & derivatives* / Q000097:blood
D014807:Vitamin D; D015652:25-Hydroxyvitamin D 2; C104450:25-hydroxyvitamin D; D002112:Calcifediol