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Method Validation Report
Version
1
Page
20 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 20B: Ruggedness of Partially hydrolyzed protein infant formula powder
Concentration (mcg/100g)
Pyridoxine
Pyridoxal Pyridoxamine
Vit B6 (as
Pyridoxine) Biotin
Day 1
376.27
3.98
22.53
402.97
23.25
Day 2
354.61
3.81
22.01
380.61
21.81
Day 3
386.85
4.37
23.65
415.07
24.34
Day 4
361.07
4.03
23.49
388.78
23.48
Day 5
344.88
3.94
22.48
371.48
22.39
Day 6
367.26
3.98
23.54
394.96
24.41
Mean
365.16
4.02
22.95
392.31
23.28
% RSD
4.13
4.67
3.03
3.98
4.46
Acetance
Criteria
5%
5%
5%
5%
5%
Accetable
(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.
9. Accuracy
Analyze NIST SRM 1849a on 3 independent runs over multiple days.
Table 21: Accuracy of SRM 1849a
Result (mcg/100g)
Acceptable
Analyte
Thiamine Riboflavin Nicotinamide
Pantothenic
acid
Pyridoxine Biotin
Y/N
Expected
Value
1159-
1355
1985-
2089
9800-11800
6630-7010
1253-
1439
186-
212
Run 1
1283.78
1999.03
10280.78
6881.59
1357.95 191.47
Y
Run 2
1306.26
1990.94
10413.05
6790.94
1285.84 191.87
Y
Run 3
1309.12
2011.72
10150.15
6984.88
1309.87 199.05
Y
BVIT-01 (SLV)
FOR ERP/WORKING GROUP USE ONLY
DO NOT DISTRIBUTE
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