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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 16: Quantitation Limit of Pyridoxine, Pyridoxal, Pyridoxamine and Biotin Area Ratio Lowest Standard Pyridoxine Pyridoxal Pyridoxamine Biotin Replicate 1 0.13990 0.02995 0.11110 0.05324 Replicate 2 0.14440 0.03118 0.10750 0.05659 Replicate 3 0.14250 0.02984 0.11180 0.05233 Replicate 4 0.14620 0.02931 0.11180 0.05247 Replicate 5 0.14270 0.03251 0.10990 0.05551 Replicate 6 0.14390 0.03004 0.10560 0.05580 Mean 0.14327 0.03047 0.10962 0.05432 Std. Dev. 0.00212 0.00117 0.00255 0.00186 % RSD 1.48 3.84 2.32 3.42 Acceptance Criteria RSD ≤ 5% RSD ≤ 5% RSD ≤ 5% RSD ≤ 5%

%RSD Acceptable (Y/N)

Y

Y

Y

Y

Quantitation Limit (mcg/100g)

27.86

0.32

1.12

1.13

7. Range

Range of analysis that the Linearity, Accuracy and Precision are satisfied Table 17: Standard Ranges of Thiamine HCl, Riboflavin, Nicotinic acid, Nicotinamide and Pantothenic acid Linearity range: Thiamine HCl Riboflavin Nicotinic acid Nicotinamide Pantothenic acid

min (mcg/100g)

26.08

31.35

13.03

441.93

111.06

Analyte

max (mcg/100g)

469.47

564.39

117.27

3977.33

1999.09

Acceptance Criteria

Linearity, Accuracy and Precision satisfied

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