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FOR EXPERT REVIEW PANEL USE ONLY
NOT FOR DISTRIBUTION
1
Expert Review Panel for Infant Formula and Adult Nutrition
Evaluation of Method: VITD-18 (2016.05)
Title:
Analysis of Vitamin D2 and Vitamin D3 by LC-MS/MS in Milk Powders, Infant
Formulas, and Adult Nutritionals
Author:
Gill, B.D
Summary of Method:
Samples are saponified at high temperature then lipid soluble components are extracted
into isooctane. A portion of the isooctane layer is transferred, washed, and an aliquot of 4-
phenyl-1,2,4-triazoline-3,5-dione is added to derivatise vitamin D to form a high molecular
mass, easily ionisable adduct. The vitamin D-adduct is then re-extracted into a small
volume of acetonitrile and analysed by reverse-phase liquid chromatography. Detection is
by triple quadrupole mass spectrometer. Stable isotope labelled vitamin D2 and vitamin
D3 internal standards are used for quantitation.
Method Scope/Applicability:
The method is applicable for vitamin D
2
or vitamin D
3
supplemented milk powders, infant formulas, and adult
nutritional products.
This method does not distinguish between contribution from pre-vitamin D and vitamin D.
and measures only an aggregate result.
General comments about the method:
1.
The method involves hot saponification followed by solvent extraction of vit D.
2.
Vitamin D is not measured directly but measured after drivatization.
3.
It does
not distinguish between contribution from pre-vitamin D and vitamin & measures only
an aggregate vitamin D2 and/or D3 value.
Method Clarity:
Satisfactory