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Fonterra Co-operative Group
Page 2
Summary
An LC-MS/MS method for the determination of vitamin D
2
and vitamin D
3
in infant formula and adult
nutritionals was evaluated by single laboratory validation procedures as specified by the SPIFAN Expert
Review Panel [1]. The SPIFAN kit was used for this validation study. The method was demonstrated to be
valid and met the requirements of the vitamin D SMPR as defined by the SPIFAN Stakeholder Panel [2].
x
Acceptable linearity (vitamin D
2
linear range 0.04
–
7.3 μg hg
-1
RTF, vitamin D
3
linear range 0.06
–
11.3 μg hg
-1
RTF) was demonstrated over the range specified in the SMPR (0.12
–
5.0 μg hg
-1
RTF).
x
Acceptable repeatability (1.1
–
5.2%) was demonstrated for vitamin D
3
in samples compared to limits
specified in SMPR (
≤
11.0%).
x
Acceptable limit of detection (vitamin D
2
LOD = 0.013 μg hg
-1
RTF; vitamin D
3
LOD = 0.018 μg hg
-1
RTF) and limit of quantitation (vitamin D
2
LOQ = 0.016 μg hg
-1
RTF; vitamin D
2
LOQ = 0.028 μg hg
-1
RTF) were determined compared to limits in SMPR (LOD = 0.02 μg hg
-1
RTF, LOQ = 0.12 μg hg
-1
RTF).
x
Acceptable recovery was demonstrated (vitamin D
2
recovery = 97
–
101%; vitamin D
3
LOD = 94
–
101%) compared to limits in SMPR (90
–
110%).
x
Acceptable bias for vitamin D
3
(p-value = 0.25) demonstrated by
t
-test against certified value for
NIST 1849a.
x
Acceptable bias for vitamin D
3
(p-value = 0.09) demonstrated by
t
-test against AOAC 2002.05 as
part of method validation.
x
Method shown to be selective with a single peak in typical samples free of interferences.
x
The determination of pre-vitamin D discussed.
VitD-18 (February 2016)
FOR ERP USE ONLY
DO NOT DISTRIBUTE