A simultaneous quantitative method for vitamins A, D and E in human serum using liquid chromatography-tandem mass spectrometry

Al Bahrani, A, Rotarou, V, Roche, P and Greaves, R 2016, 'A simultaneous quantitative method for vitamins A, D and E in human serum using liquid chromatography-tandem mass spectrometry', Steroid Biochemistry and Molecular Biology, vol. 159, pp. 41-53.


Document type: Journal Article
Collection: Journal Articles

Title A simultaneous quantitative method for vitamins A, D and E in human serum using liquid chromatography-tandem mass spectrometry
Author(s) Al Bahrani, A
Rotarou, V
Roche, P
Greaves, R
Year 2016
Journal name Steroid Biochemistry and Molecular Biology
Volume number 159
Start page 41
End page 53
Total pages 13
Publisher Elsevier Ltd
Abstract Non-classical roles of fat-soluble vitamins (FSVs) in many pathologies including cancer have been identified. There is also evidence of hormonal interactions between two of these vitamins, A and D. As a result of this enhanced clinical association with disease, translational clinical research and laboratory requests for FSV measurement has significantly increased. However there are still gaps in the analytical methods available for the measurement of these vitamins. This study aimed to develop a method for simultaneous quantification of 25-hydroxyvitamin-D2 (25-OHD2), 25-hydroxyvitamin-D3 (25-OHD3) and its 3-epimer (epi-25-OHD3), retinol and α-tocopherol in human serum using liquid chromatography-tandem mass spectrometry (LC-MS/MS).The procedure was developed and validated across two LC-MS/MS platforms, using commercial calibrators referenced to certified reference materials, controls, and deuterated internal standards. The samples were prepared by liquid-liquid extraction prior to injection and LC separation (using a Pursuit-PFP column) on two Agilent MS/MS systems (6410 and 6490) in electrospray ionisation positive mode with multiple reaction monitoring.Identification and quantification of 25-OHD3 from its 3-epimer as well as 25-OHD2, retinol and α-tocopherol were achieved. The dynamic ranges were 4-160 nmol/L for 25-OHD2 and epi-25-OHD3, 4-200 nmol/L for 25-OHD3, 0.1-4.0 μmol/L for retinol and 4-70 μmol/L forα-tocopherol with correlation (r2) of 0.997-0.998. Based on participation in an external quality assurance program, the overall performance of the simultaneous methods were: imprecision (CV%) and inaccuracy (average bias) 3.0% and 3.2 nmol/L, respectively, for 25-OHD3; 5.0% and 0.04 μmol/L, respectively, for retinol; and 4.7% and 0.2 μmol/L, respectively, for α-tocopherol.In summary, two simple LC-MS/MS methods were successfully developed and validated for the simultaneous quantification of the three vit
Subject Clinical Chemistry (diagnostics)
Endocrinology
Keyword(s) Cholecalciferol
Retinol
Tocopherol
Fat-soluble vitamins
Mass spectrometry
DOI - identifier 10.1016/j.jsbmb.2016.02.019
Copyright notice © 2016 Elsevier Ltd
ISSN 0960-0760
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