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Method Validation Report
Version
1
Page
12 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 14A: Product Matrix 3: Partially hydrolyzed protein infant formula powder
Result (mcg/100g)
Analyte
Thiamine
HCl
Riboflavin Nicotinic
acid
Nicotinamide
Niacin
Pantothenic
acid
Replicate 1
567.49
1025.75
209.00
6990.17
7199.17
3435.83
Replicate 2
550.52
1006.88
197.47
6915.63
7113.10
3416.99
Replicate 3
578.74
1045.92
204.41
7289.48
7493.89
3595.35
Replicate 4
542.10
969.97
197.55
6916.74
7114.29
3265.82
Replicate 5
579.72
1027.81
213.29
7391.06
7604.35
3521.59
Replicate 6
595.33
1034.87
204.05
7344.39
7548.44
3529.40
%RSD
3.49
2.65
3.06
3.13
3.10
3.35
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 14B: Product Matrix 3: Partially hydrolyzed protein infant formula powder
Result (mcg/100g)
Analyte
Pyridoxine
Pyridoxal
Pyridoxamine
Vit B6 (as
Pyridoxine)
Biotin
Replicate 1
382.85
4.03
24.20
411.26
22.13
Replicate 2
351.96
3.78
23.21
379.14
21.85
Replicate 3
379.18
3.71
22.97
406.04
21.97
Replicate 4
345.64
3.73
22.07
371.62
21.94
Replicate 5
374.40
4.16
24.41
403.17
22.72
Replicate 6
369.56
3.88
23.24
396.87
22.86
%RSD
4.12
4.64
3.66
4.01
1.95
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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