AOAC SPIFAN Review Team Meeting Book-July 22, 2015

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

Method Validation Report

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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

Table 14A: Product Matrix 3: Partially hydrolyzed protein infant formula powder Result (mcg/100g) Analyte Thiamine HCl Riboflavin Nicotinic acid Nicotinamide Niacin

Pantothenic acid

Replicate 1 Replicate 2 Replicate 3 Replicate 4 Replicate 5 Replicate 6 Acceptance Criteria: Acceptable (Y/N) %RSD

567.49 550.52 578.74 542.10 579.72 595.33

1025.75 1006.88 1045.92 969.97 1027.81 1034.87

209.00 197.47 204.41 197.55 213.29 204.05

6990.17 6915.63 7289.48 6916.74 7391.06 7344.39

7199.17 7113.10 7493.89 7114.29 7604.35 7548.44

3435.83 3416.99 3595.35 3265.82 3521.59 3529.40

3.49

2.65

3.06

3.13

3.10

3.35

RSD ≤ 5%

RSD ≤ 5%

RSD ≤ 5%

RSD ≤ 5%

RSD ≤ 5%

RSD ≤ 5%

Y Y Remark: Niacin is sum of Nicotinic acid and Nicotinamide, and reported as Niacin. Y Y Y Y

Table 14B: Product Matrix 3: Partially hydrolyzed protein infant formula powder Result (mcg/100g) Analyte Pyridoxine Pyridoxal Pyridoxamine Vit B6 (as Pyridoxine) Biotin Replicate 1 382.85 4.03 24.20 411.26 22.13 Replicate 2 351.96 3.78 23.21 379.14 21.85 Replicate 3 379.18 3.71 22.97 406.04 21.97 Replicate 4 345.64 3.73 22.07 371.62 21.94 Replicate 5 374.40 4.16 24.41 403.17 22.72 Replicate 6 369.56 3.88 23.24 396.87 22.86 %RSD 4.12 4.64 3.66 4.01 1.95 Acceptance Criteria: RSD ≤ 5% RSD ≤ 5% RSD ≤ 5% RSD ≤ 5% RSD ≤ 5% Acceptable (Y/N) Y Y Y Y Y Remark: Vitamin B6 (as Pyridoxine) is sum of Pyridoxine, Pyridoxal and Pyridoxamine. All data have been mass corrected and expressed as Pyridoxine.

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