Details

Autor(en) / Beteiligte

• Mochizuki, Asuka
• Kodera, Yoshio
• Saito, Tatsuya
• Satoh, Mamotu
• Sogawa, Kazuyuki
• Nishimura, Motoi
• Seimiya, Masanori
• Kubota, Masayuki
• Nomura, Fumio

Titel

Preanalytical evaluation of serum 25-hydroxyvitamin D3 and 25-hydroxyvitamin D2 measurements using LC–MS/MS

Ist Teil von

• Clinica chimica acta, 2013-05, Vol.420, p.114-120

Ort / Verlag

Netherlands: Elsevier B.V

Erscheinungsjahr

2013

Quelle

MEDLINE

Beschreibungen/Notizen

• Vitamin D testing is increasing worldwide. Although immunoassays are still widely used in Japan for the measurement of serum 25-hydroxyvitamin D (25OHD) as an indicator of vitamin D status, development of a simple and high-throughput MS-based method is still needed for routine use in clinical laboratories. We designed a method using a triple quadrupole mass spectrometer equipped with a two-step separation approach that used the Aria TLX-2 HPLC system in the selected reaction monitoring mode. Analytical performance of the system and effects of various preanalytical factors were tested. High-throughput quantitative analysis of 25OHD3 and D2 at 15samples/h was achieved using 25μl of serum/plasma. Intra- and inter-assay CVs for 25OHD3 were 5% and 7%, respectively. Limit of detection for 25OHD3 was 0.31ng/ml. No significant effects were seen for clotting time, repeated freeze–thaw cycles, anti-coagulants and possible interfering substances. A good correlation (r2=0.947) was found between the present system and the DiaSorin radioimmunoassay. Serum 25OHD3 levels in apparently healthy Japanese subjects were 25.5±9.8ng/ml for men and 20.9±7.1ng/ml for women. This high-throughput LC–MS/MS 25-OHD assay has the potential to be used as a routine clinical laboratory assay for assessing vitamin D status. ► We developed a simple MS-based method for serum 25-hydroxyvitamin D measurement. ► Quantitative 25OHD3 and D2 measurements of 350samples/day could be achieved. ► The results obtained agreed well with those by the widely used radioimmunoassays. ► The effects of various pre-analytical factors such as diet on the assays were tested. ► This method has a potential to be used on a routine basis in clinical laboratory.