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Single Lab Validation of a Method for the Determination of β‐
Galactooligosaccharides in Infant Formula & Adult Nutritionals / Sean Austin (NRC,
Lausanne)
25 Jul 2016
CONFIDENTIAL
∙ This document may not be reproduced or disclosed to third parties without prior authorization
Page 11 / 35
2.2.7
Calibration & Calculations
Use the instrument software to plot a six point standard curve of “Instrument response for
maltotriose / Instrument response for laminaritriose” against the “concentration of maltotriose” in
the standard (in µmol/mL). Fit a linear model to the data without forcing through zero.
Use the standard curve to calculate the molar concentration (in µmol/mL) of each oligosaccharide in
the chromatogram (C
m
) without enzyme treatment, and calculate the total oligosaccharides in that
sample as follows:‐
ܥ
்ைௌ
ൌ ሺ
ܥ
ൈ
ܯ
ܹሻ ൈ ܸ ݉ ൈ 0.0001
Use the standard curve to calculate the molar concentration (in µmol/mL) of each oligosaccharide in
the chromatogram (C
m
) with enzyme treatment, and calculate the total oligosaccharides in that
sample as follows:‐
ܥ
ൌ ሺ
ܥ
ൈ
ܯ
ܹሻ ൈ ܸ ݉ ൈ 0.0001
Then calculate the GOS content of sample as follows:‐
ܥ
ீைௌ
ൌ
ܥ
்ைௌ
െ
ܥ
Where,
C
TOS
= Total concentration of oligosaccharides in the untreated sample (in g/100 g)
C
B
= Total concentration of oligosaccharides in the enzyme‐treated sample (in g/100 g)
C
GOS
= Total concentration of GOS in the sample (in g/100 g)
C
m
= molar concentration of each individual oligosaccharide in the sample (in µmol/mL)
MW = molecular weight of each individual oligosaccharide in the sample
V = volume to which the original sample weight was diluted (in mL)
m = weight of sample diluted to volume (V) (in g)
0.0001 = factor to convert result from µg/g to g/100 g.
VALIDATION REPORT
FOR ERP USE ONLY
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