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Dual Lab Validation of a Method for the Determination of Fructans in Infant
Formula & Adult Nutritionals /
25 Jul 2016
53
A‐10 Chromatographic conditions
The analysis of the prepared samples , standards( and blanks if required)are performed on the HPAEC‐
PAD system under the following conditions:
‐ flow mobile phase 0.25 ml/min for a 2 mm ID column
‐ temperature column oven 20 °C
‐ injection volume 20 μl
‐ linear gradient elution profile according to the scheme in table 5
Table 5
Linear gradient elution profile for the chromatographic separation of the sugars
Time
(min.)
% Eluent A
1M
CH
3
COONa
% Eluent B
0.2M
NaOH
% Eluent C
H
2
O
0
0.0
7.5
92.5
13.0
00.0
7.5
92.5
14.0
00.0
25.0
75.0
20.0
00.0
25.0
75.0
21.0
30.0
40.0
30.0
28.0
30.0
40.0
30.0
30
0.0
4.0
60.0
31
0.0
7.5
92.5
43
0.0
7.5
92.5
Post column pump:
‐ flow post column sodium hydroxide solution 0.13 ml/min
‐ minimal back pressure 500 psi
Detector adjustment:
The time program of the PAD waveform is specified in table 6
VALIDATION REPORT
FOR ERP USE ONLY
DO NOT DISTRIBUTE