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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

6. Quantitation Limit

Analyze 6 replicates of the lowest standard concentration. Calculated RSD should be <5% for each analyte. Table 15: Quantitation Limit of Thiamine HCl, Riboflavin, Nicotinic acid, Nicotinamide and Pantothenic acid Area Ratio

Lowest Standard

Nicotinic acid 0.22460 0.22270 0.22400 0.22360 0.21410 0.21660 0.22093 0.00444

Pantothenic acid

Thiamine HCl Riboflavin

Nicotinamide

Replicate 1 Replicate 2 Replicate 3 Replicate 4 Replicate 5 Replicate 6

0.18600 0.18430 0.18250 0.18250 0.18250 0.18140 0.18320 0.00166

0.12820 0.12310 0.12540 0.12430 0.12370 0.12700 0.12528 0.00199

0.27060 0.26860 0.26310 0.26900 0.27160 0.26310 0.26767 0.00370

0.18690 0.18250 0.18200 0.18650 0.18550 0.18360 0.18450 0.00209

Mean

Std. Dev.

% RSD

0.91

1.58

2.01

1.38

1.13

Acceptance Criteria %RSD Acceptable (Y/N) Quantitation Limit (mcg/100g)

RSD ≤ 5%

RSD ≤ 5%

RSD ≤ 5%

RSD ≤ 5%

RSD ≤ 5%

Y

Y

Y

Y

Y

26.08

31.35

13.03

441.93

111.06

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