AOAC SPIFAN Review Team Meeting Book-July 22, 2015

BVIT-01 (SLV) FOR ERP/WORKING GROUP USE ONLY DO NOT DISTRIBUTE

Method Validation Report

21 of 49

Version

1

Page

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

Document Title

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

Thiamine HCl (mcg/100g)

Un- spiked Result

Resulting Spiked Conc

Spiked Result

% Recovery

Acceptable

Over Spike

Replicates

Theoretical Spiked Conc (C)

(A)

(B)

(A+C)

B/(A+C)

Y/N

Replicate 1 886.56

Mean

Replicate 2 881.60

885.08

0%

Replicate 3 887.07

Replicate 1

1214.38 1232.77

347.70

98.51

Y

Replicate 2

1216.51 1232.77

347.70

98.68

Y

50%

Replicate 3

1215.52 1232.77

347.70

98.60

Y

Replicate 1

1539.51 1580.47

695.39

97.41

Y

Replicate 2

1560.18 1580.47

695.39

98.72

Y

100%

Replicate 3

1602.68 1580.47

695.39

101.41

Y

Acceptance Criteria: All individual recovery results should be within 90% to 110%.

Page 34