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Method Validation Report
Version
1
Page
21 of 49
Document Title
Validation Report for the Determination of Thiamin HCl, Riboflavin, Niacin
(Nicotinic Acid and Nicotinamide), Pantothenic acid, Vitamin B6
(Pyridoxine, Pyridoxal and Pyridoxamine) and Biotin in Infant Formula and
Adult Nutritionals by LC-MS/MS
Recovery: Each matrix was prepared by spiking with different concentrations of
standard at
0%, approx. 50%, and approx. 100%
over spikes based on theoretical values.
Analyze each spike level on 3 independent runs. Obtain percentage of recovery, considering
un-spiked sample as the base level. All individual recovery results should be within 90% to
110%.
Table 22: Accuracy of Thiamine HCl in Stage 3 Growing up milk formula
powder
Over
Spike
Replicates
Thiamine HCl (mcg/100g)
Un-
spiked
Result
Spiked
Result
Resulting
Spiked
Conc
Theoretical
Spiked
Conc (C)
%
Recovery
Acceptable
(A)
(B)
(A+C)
B/(A+C)
Y/N
0%
Replicate 1 886.56
Mean
Replicate 2 881.60
885.08
Replicate 3 887.07
50%
Replicate 1
1214.38 1232.77
347.70
98.51
Y
Replicate 2
1216.51 1232.77
347.70
98.68
Y
Replicate 3
1215.52 1232.77
347.70
98.60
Y
100%
Replicate 1
1539.51 1580.47
695.39
97.41
Y
Replicate 2
1560.18 1580.47
695.39
98.72
Y
Replicate 3
1602.68 1580.47
695.39
101.41
Y
Acceptance Criteria: All individual recovery results should be within 90% to 110%.
BVIT-01 (SLV)
FOR ERP/WORKING GROUP USE ONLY
DO NOT DISTRIBUTE
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