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Method Validation Report
Version
1
Page
14 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 16: Quantitation Limit of Pyridoxine, Pyridoxal, Pyridoxamine and Biotin
Area Ratio
Lowest Standard
Pyridoxine
Pyridoxal
Pyridoxamine
Biotin
Replicate 1
0.13990
0.02995
0.11110
0.05324
Replicate 2
0.14440
0.03118
0.10750
0.05659
Replicate 3
0.14250
0.02984
0.11180
0.05233
Replicate 4
0.14620
0.02931
0.11180
0.05247
Replicate 5
0.14270
0.03251
0.10990
0.05551
Replicate 6
0.14390
0.03004
0.10560
0.05580
Mean
0.14327
0.03047
0.10962
0.05432
Std. Dev.
0.00212
0.00117
0.00255
0.00186
% RSD
1.48
3.84
2.32
3.42
Acceptance Criteria
RSD ≤ 5%
RSD ≤ 5%
RSD ≤ 5%
RSD ≤ 5%
%RSD Acceptable
(Y/N)
Y
Y
Y
Y
Quantitation Limit
(mcg/100g)
27.86
0.32
1.12
1.13
7. Range
Range of analysis that the Linearity, Accuracy and Precision are satisfied
Table 17: Standard Ranges of Thiamine HCl, Riboflavin, Nicotinic acid, Nicotinamide and
Pantothenic acid
Linearity range: Thiamine
HCl
Riboflavin Nicotinic acid Nicotinamide Pantothenic
acid
Analyte
min (mcg/100g)
26.08
31.35
13.03
441.93
111.06
max
(mcg/100g)
469.47
564.39
117.27
3977.33
1999.09
Acceptance
Criteria
Linearity, Accuracy and Precision satisfied
BVIT-01 (SLV)
FOR ERP/WORKING GROUP USE ONLY
DO NOT DISTRIBUTE
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