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Single Laboratory Validation – AOAC First Action Method 2012.22 Vitamin C in Infant Formula and Adult/Pediatric
Nutritional Formula by UPLC-UV
Date of Issue: February 14
th
, 2013
RDLS-RD130018
E.Campos Giménez (NRC/QS, Lausanne)
Page 6
Results
Single Laboratory Validation
System
suitability test
Relative standard deviation of retention time and response between the six
standard injections were below 1 % for all analytical series.
Specificity
All the placebo samples were free from chromatographic interferences
.
AU
0.000
0.002
0.004
0.006
0.008
0.010
0.012
0.014
0.016
0.018
0.020
Minutes
0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00
AU
0.000
0.002
0.004
0.006
0.008
0.010
0.012
0.014
0.016
0.018
0.020
Minutes
0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00
Child Formula Powder
Infant Elemental Powder
Ascorbic acid
Ascorbic acid
AU
0.000
0.002
0.004
0.006
0.008
0.010
0.012
0.014
0.016
0.018
0.020
Minutes
0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00
AdultNutritional RTF High Protein
Ascorbic acid
AU
-0.002
0.000
0.002
0.004
0.006
0.008
0.010
0.012
0.014
0.016
0.018
0.020
Minutes
0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00
Infant Formula RTF
Ascorbic acid
AU
0.000
0.002
0.004
0.006
0.008
0.010
0.012
0.014
0.016
0.018
0.020
Minutes
0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00
AdultNutritional RTF High Fat
Ascorbic acid
Linearity/
Calibration Fit
Calibration curves were plotted and linearity demonstrated by R
2
> 0.9999 and
calibration errors well below 5 % for all levels used in daily operation (0.5-10
µg/mL) (Figure 2). This corresponds to 2.5–50 mg/100 g in reconstituted
samples. Although linearity up to 500 mg/100 g has been proven (data not
shown), we recommend diluting the samples if they contain more than 50
mg/100 g.
Figure 2: linearity range/calibration fit.
Continued on next page
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VitC-03/2012.22
For ERP Use Only
DO NOT DISTRIBUTE
AOAC OMB Meeting Book
316