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Method Validation Report
Version
1
Page
7 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
Slope
(weighting: 1/x)
0.07177
0.07257
0.07348
Y-Intercept
0.00845
0.00601
0.005490
Correlation
Coefficient
0.99902
0.99964
0.99950
Acceptance
Criteria
Correlation Coefficient (r) ≥ 0.995
Acceptable Y/N
Y
Y
Y
3. Instrument Precision
Six replicates of each of the lowest, middle and highest standard concentrations were analyzed in
a single analytical run.
Table 9: Instrument Precision of Thiamine, Riboflavin and Nicotinic acid
Thiamine HCl
Riboflavin
Nicotinic acid
Area Ratio
Area Ratio
Area Ratio
Lowest
Standard
Middle
Standard
Highest
Standard
Lowest
Standard
Middle
Standard
Highest
Standard
Lowest
Standard
Middle
Standard
Highest
Standard
Replicate 1
0.1902
1.1830
3.6200
0.1419
0.9851
2.8740
0.2246
0.4955
1.3260
Replicate 2
0.1923
1.1850
3.6270
0.1420
0.9466
2.8570
0.2227
0.4991
1.3060
Replicate 3
0.1923
1.1900
3.6620
0.1408
0.9431
2.9600
0.2240
0.4913
1.3260
Replicate 4
0.1922
1.1780
3.6440
0.1405
0.9573
2.9150
0.2236
0.4924
1.3330
Replicate 5
0.1930
1.1660
3.6310
0.1418
0.9625
2.8580
0.2141
0.5011
1.2940
Replicate 6
0.1946
1.1660
3.6340
0.1416
0.9945
2.8220
0.2166
0.4954
1.3280
Mean
0.1924
1.1780
3.6363
0.1414
0.9649
2.8810
0.2209
0.4958
1.3188
Std. Dev.
0.0014
0.0101
0.0149
0.0006
0.0208
0.0491
0.0044
0.0038
0.0153
% RSD
0.74
0.85
0.41
0.44
2.15
1.70
2.01
0.76
1.16
Acceptance
Criteria
RSD ≤
5%
RSD ≤
3%
RSD ≤
3%
RSD ≤
5%
RSD ≤
3%
RSD ≤
3%
RSD ≤
5%
RSD ≤
3%
RSD ≤
3%
%RSD
Acceptable
(Y/N)
Y
Y
Y
Y
Y
Y
Y
Y
Y
BVIT-01 (SLV)
FOR ERP/WORKING GROUP USE ONLY
DO NOT DISTRIBUTE
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