Determination of serum vitamins 25-OH-D2 and 25-OH-D3 with liquid chromatography-tandem mass spectrometry using atmospheric pressure chemical ionization or electrospray source and core-shell or sub-2 μm particle columns: a comparative study.

Abstract:

OBJECTIVE:Develop a simple LC-MS-MS method for determination of 25-hydroxymetabolites of vitamin D2 and D3 in serum. DESIGN AND METHODS:0.1 mL serum was mixed with 0.3 mL deproteinizing mixture containing two deuterated internal standards. Two LC-MS-MS systems were used: Waters Premier with an ESI source and a Hypersil Gold RP 18 50 × 2.1 mm column, and a Thermo Quantum, with an APCI source and a Kinetex RP 18 core-shell 50 × 2.1 mm column. Certified reference material 972 from NIST was used for method calibration. The samples were additionally analyzed with commercial HPLC procedure. RESULTS:Baseline separation of compounds was achieved in 6 min run time. Both systems have limits of detection of 0.5 nmol/L and recovery ranging from 86% to 112%. The measurement of 25-OH-D3 gave comparable results for all procedures; the results of 25-OH-D2 measurement obtained with the HPLC often differed from those obtained with LC-MS-MS methods. CONCLUSIONS:Established procedure performed well for ESI and APCI sources and totally porous and core-shell LC columns. The use of two internal standards enhanced the accuracy of the measurements.

journal_name

Clin Biochem

journal_title

Clinical biochemistry

authors

Bogusz MJ,Al Enazi E,Tahtamoni M,Jawaad JA,Al Tufail M

doi

10.1016/j.clinbiochem.2011.08.1134

subject

Has Abstract

pub_date

2011-11-01 00:00:00

pages

1329-37

issue

16

eissn

0009-9120

issn

1873-2933

pii

S0009-9120(11)02596-3

journal_volume

44

pub_type

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