181 / 620
Single Lab Validation Report for GOS in Infant Formula and Adult Nutritionals
15
3.15.
Trifluoroacetic acid (TFA; Thermo Scientific p/n. 28904)
4.
Eluents
4.1.
Fractionation
4.1.1.
HPLC eluent A 1000 mM NaOH:
Transfer 1900 mL Milli-Q into a plastic eluent bottle, while flushing with nitrogen.
Add immediately 104 mL 50% NaOH solution, while flushing with nitrogen. Close
the bottle with the eluent lid and homogenize.
4.1.2.
HPLC eluent B 1000 mM NaAc:
Use 82 g NaAc, dissolve and adjust to 1 liter using Mili-Q. Transfer into in a plastic
eluent bottle.
4.1.3.
HPLC Eluent D Milli-Q
4.2.
Quantification
4.2.1.
HPLC eluent A 300 mM NaOH:
Add 1900 mL Milli-Q in a plastic eluent bottle, while flushing with nitrogen. Add
immediately 30.3 mL 50% NaOH solution, while flushing with nitrogen. Close the
bottle with the eluent lid and homogenize
4.2.2.
HPLC eluent B 1000mM NaAc:
Use 82 g NaAc, dissolve and adjust to 1 liter using Mili-Q. Transfer into in a plastic
eluent bottle.
4.2.3.
HPLC Eluent D Milli-Q
5.
Reagents
5.1.
0.2 M potassium phosphate buffer pH 6.0 ± 0.1:
•
Weigh 13.60 g KH
2
PO
4
and dissolve in 500 mL Milli-Q
•
Weigh 6.96 g K
2
HPO
4
and dissolve in 200 mL Milli-Q
Prepare a buffer solution of pH 6.0 ± 0.1 by adding K
2
HPO
4
solution to the KH
2
PO
4
solution.
5.2.
Carrez reagent 1:
•
Weigh 17.2 g K
4
Fe(CN)
6
.3H
2
O
and dissolve in 100 mL Milli-Q. Store the
solution in the refrigerator.
5.3.
Carrez reagent 2:
•
Weigh 53.5 g ZnSO
4
.7H
2
O and dissolve in 100 mL Milli-Q. Store the solution in
the refrigerator.
5.4.
80% Acetonitrile/ 0.1 % TFA:
•
Transfer 80 mL 100% acetonitrile into a beaker
•
Add 20 mL Milli-Q
•
Add 0.1 mL TFA
•
Mix and store in a closed bottle
5.5.
40% Acetonitrile/ 0.05% TFA:
•
Dilute the 80% acetonitrile with 0.1% TFA with an equal part of Milli-Q
•
Mix and store in a closed bottle
5.6.
Sugar stock solutions:
GOS-02 SLV
FOR ERP USE ONLY
DO NOT DISTRIBUTE