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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