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Method Validation Report
Version
1
Page
11 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 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
570.34
1022.20
173.62
5926.41
6100.02
3949.93
Replicate 2
563.83
1054.70
180.76
6116.34
6297.09
4042.17
Replicate 3
566.16
1085.24
185.48
5936.55
6122.03
3906.27
Replicate 4
554.05
1083.12
180.17
5971.24
6151.41
3908.10
Replicate 5
529.70
1016.99
177.97
5932.55
6110.52
3830.67
Replicate 6
562.17
1066.89
179.57
6056.18
6235.75
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
Y
Y
Y
Y
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.
BVIT-01 (SLV)
FOR ERP/WORKING GROUP USE ONLY
DO NOT DISTRIBUTE
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