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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 13A: Product Matrix 2: Stage 1 Infant formula powder (milk-based) 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

570.34 563.83 566.16 554.05 529.70 562.17

1022.20 1054.70 1085.24 1083.12 1016.99 1066.89

173.62 180.76 185.48 180.17 177.97 179.57

5926.41 6116.34 5936.55 5971.24 5932.55 6056.18

6100.02 6297.09 6122.03 6151.41 6110.52 6235.75

3949.93 4042.17 3906.27 3908.10 3830.67 4070.80

%RSD

2.64

2.80

2.15

1.31

1.29

2.29

Acceptance Criteria:

RSD ≤ 5%

RSD ≤ 5%

RSD ≤ 5%

RSD ≤ 5%

RSD ≤ 5%

RSD ≤ 5%

Acceptable (Y/N) Y Remark: Niacin is sum of Nicotinic acid and Nicotinamide, and reported as Niacin. Table 13B: Product Matrix 2: Stage 1 Infant formula powder (milk-based) Result (mcg/100g) Analyte Pyridoxine Pyridoxal Pyridoxamine Vit B6 (as Pyridoxine) Biotin Replicate 1 310.77 42.15 21.61 375.18 22.94 Replicate 2 308.99 40.84 19.82 370.27 23.60 Replicate 3 318.89 42.46 21.49 383.48 22.95 Replicate 4 315.83 42.41 20.96 379.83 23.03 Replicate 5 298.75 39.81 19.93 359.08 22.17 Replicate 6 327.56 41.85 20.76 390.81 23.78 %RSD 3.12 2.53 3.65 2.93 2.47 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. Y Y Y Y Y

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