SPIFAN Nutrients ERP Book_9-29-15

GOS-02 SLV FOR ERP USE ONLY DO NOT DISTRIBUTE

Single Lab Validation Report for GOS in Infant Formula and Adult Nutritionals

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. 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. 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 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. HPLC Eluent D Milli-Q Quantification

4.1.2.

4.1.3.

4.2.

4.2.1.

4.2.2.

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

• and dissolve in 200 mL Milli-Q Prepare a buffer solution of pH 6.0 ± 0.1 by adding K 2 HPO 4 Weigh 6.96 g 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:

15