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Method Validation Report
Version
1
Page
1 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
I. BACKGROUND
Method 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.
II. TEST SAMPLES:
No.
Matrix/Test Sample
1
Reference Material SRM 1849a Infant formula
2
Stage 3 Growing up milk formula powder
3
Stage 1 Infant formula powder(milk-based)
4
Partially hydrolyzed protein infant formula powder
III. VALIDATION DATA:
1. Linearity
Table 1: Linearity of Thiamine HCl
Cncentration
Area
Response 1
Area
Response 2
Area
Response 3
(ng/ml)
Standard 1
16.3
0.1819
0.1845
0.1844
Standard 2
32.6
0.3343
0.3334
0.3325
Standard 3
65.2
0.6785
0.6860
0.6762
Standard 4
97.81
1.0010
1.0080
1.0170
Standard 5
130.41
1.3670
1.3580
1.3650
Standard 6
163.01
1.6950
1.6890
1.6880
Standard 7
195.61
2.0360
2.0610
2.0250
Standard 8
228.21
2.4280
2.4170
2.3980
Standard 9
260.82
2.7740
2.7830
2.7530
Standard 10
293.42
3.1000
3.0850
3.1010
Slope
(weighting: 1/x)
0.01049
0.0105
0.01045
Y-Intercept
0.0027
0.00331
0.00401
Correlation
Coefficient
0.99983
0.99984
0.99987
Acceptance
Criteria
Correlation Coefficient (r) ≥ 0.995
Acceptable Y/N
Y
Y
Y
BVIT-01 (SLV)
FOR ERP/WORKING GROUP USE ONLY
DO NOT DISTRIBUTE
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