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Method Validation Report
Version
1
Page
15 of 49
Document Title
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
Table 18: Standard Ranges of Pyridoxine, Pyridoxal, Pyridoxamine and Biotin
Linearity range:
Pyridoxine
Pyridoxal
Pyridoxamine
Biotin
Analyte
min (mcg/100g)
27.86
0.32
1.12
1.13
max
(mcg/100g)
501.47
69.73
20.25
20.37
Acceptance
Criteria
Linearity, Accuracy and Precision satisfied
8. Ruggedness
Prepare and analyze the same lot of test samples on 6 different days. Calculated RSD
should be < 5%.
Table 18A: Ruggedness of Stage 3 Growing up milk formula powder
Concentration (mcg/100g)
Thiamine
HCl
Riboflavin
Nicotinic
acid
Nicotinamide
Niacin
Pantothenic
acid
Day 1
895.15
1175.98
251.43
8473.14
8724.57
3381.43
Day 2
876.14
1195.91
259.06
8440.51
8699.56
3225.25
Day 3
877.99
1186.05
250.08
8562.13
8812.21
3449.38
Day 4
886.88
1196.86
247.62
8338.39
8586.01
3231.22
Day 5
883.28
1160.40
265.08
9012.18
9277.27
3537.07
Day 6
845.79
1141.60
250.63
8546.75
8797.39
3454.21
Mean
877.54
1176.13
253.98
8562.18
8816.17
3379.76
% RSD
1.93
1.85
2.63
2.74
2.72
3.77
Acetance
Criteria
≤
5%
≤5%
≤5%
≤5%
≤5%
≤5%
Accetable
(Y/N)
Y
Y
Y
Y
Y
Y
Remark: Niacin is sum of Nicotinic acid and Nicotinamide, and reported as Niacin.
BVIT-01 (SLV)
FOR ERP/WORKING GROUP USE ONLY
DO NOT DISTRIBUTE
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