Simultaneous quantitative analysis of eight vitamin D analogues in milk using liquid chromatographyetandem mass spectrometry

Gomes, Fabio P., Shaw, P. Nicholas, Whitfield, Karen and Hewavitharana, Amitha K. (2015) Simultaneous quantitative analysis of eight vitamin D analogues in milk using liquid chromatographyetandem mass spectrometry. Analytica Chimica Acta, 891 211-220. doi:10.1016/j.aca.2015.08.017


Author Gomes, Fabio P.
Shaw, P. Nicholas
Whitfield, Karen
Hewavitharana, Amitha K.
Title Simultaneous quantitative analysis of eight vitamin D analogues in milk using liquid chromatographyetandem mass spectrometry
Journal name Analytica Chimica Acta   Check publisher's open access policy
ISSN 0003-2670
1873-4324
Publication date 2015-01-01
Sub-type Article (original research)
DOI 10.1016/j.aca.2015.08.017
Open Access Status Not Open Access
Volume 891
Start page 211
End page 220
Total pages 10
Place of publication Amsterdam, Netherlands
Publisher Elsevier
Collection year 2016
Language eng
Formatted abstract
Milk is an important source of nutrients for various risk populations, including infants. The accurate measurement of vitamin D in milk is necessary to provide adequate supplementation advice for risk groups and to monitor regulatory compliance. Currently used liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods are capable of measuring only four analogues of vitamin D in unfortified milk. We report here an accurate quantitative analytical method for eight analogues of vitamin D: Vitamin D2 and D3 (D2 and D3), 25-hydroxy D2 and D3, 24,25-dihydroxy D2 and D3, and 1,25-dihydroxyD2 and D3. In this study, we compared saponification and protein precipitation for the extraction of vitamin D from milk and found the latter to be more effective. We also optimised the pre-column derivatisation using 4-phenyl-l,2,4-triazoline-3,5-dione (PTAD), to achieve the highest sensitivity and accuracy for all major vitamin D forms in milk. Chromatography was optimised to reduce matrix effects such as ion-suppression, and the matrix effects were eliminated using co-eluting stable isotope labelled internal standards for the calibration of each analogue. The analogues, 25-hydroxyD3 (25(OH)D3) and its epimer (3-epi-25(OH)D3) were chromatographically resolved, to prevent over-estimation of 25(OH)D3. The method was validated and subsequently applied for the measurement of total vitamin D levels in human, cow, mare, goat and sheep milk samples. The detection limits, repeatability standard deviations, and recovery ranges were from 0.2 to 0.4 femtomols, 6.30-13.5%, and 88.2-105%, respectively.
Keyword Derivatisation
Liquid chromatography-mass spectrometry
Milk
Protein precipitation
Saponification
Vitamin D analogues
Vitamin D metabolites
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2016 Collection
School of Pharmacy Publications
 
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Created: Fri, 02 Oct 2015, 22:45:38 EST by Ms Felicity Lindberg on behalf of School of Pharmacy