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Method Validation Report
Version
1
Page
10 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
4. Method Precision
For each of the matrices, prepare the same sample 6 times from a single slurry and
analyze in one analytical run. Calculated RSD should be
≤
5% for each analyte.
Table 12A: Product Matrix 1: Stage 3 Growing up milk formula powder
Result (mcg/100g)
Analyte
Thiamine
HCl
Riboflavin
Nicotinic
acid
Nicotinamide
Niacin
Pantothenic
acid
Replicate 1
863.34
1179.51
261.59
8370.41
8632.00
3354.80
Replicate 2
878.20
1203.63
254.25
8898.23
9152.48
3435.52
Replicate 3
890.34
1199.37
267.08
8733.89
9000.97
3381.34
Replicate 4
886.56
1241.75
268.85
8670.04
8938.89
3386.93
Replicate 5
881.60
1186.54
264.78
8755.92
9020.69
3374.63
Replicate 6
887.07
1154.56
258.05
8737.74
8995.79
3426.97
%RSD
1.11
2.43
2.12
2.02
1.94
0.93
Acceptance
Criteria:
RSD ≤
5%
RSD ≤
5%
RSD ≤ 5%
RSD ≤ 5%
RSD ≤
5%
RSD ≤ 5%
Acceptable (Y/N)
Y
Y
Y
Y
Y
Y
Remark: Niacin is sum of Nicotinic acid and Nicotinamide, and reported as Niacin.
Table 12B: Product Matrix 1: Stage 3 Growing up milk formula powder
Result (mcg/100g)
Analyte
Pyridoxine
Pyridoxal Pyridoxamine
Vit B6 (as
Pyridoxine)
Biotin
Replicate 1
1126.72
107.17
13.87
1249.14
13.50
Replicate 2
1185.61
118.72
14.14
1319.99
14.05
Replicate 3
1155.33
120.09
14.82
1291.77
13.94
Replicate 4
1172.37
115.92
14.70
1304.47
14.12
Replicate 5
1192.93
115.14
13.07
1322.60
13.80
Replicate 6
1146.62
116.67
14.26
1279.04
13.97
%RSD
2.15
3.91
4.48
2.14
1.60
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.
BVIT-01 (SLV)
FOR ERP/WORKING GROUP USE ONLY
DO NOT DISTRIBUTE
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