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

Materials

1.1. 1.2. 1.3. 1.4. 1.5. 1.6. 1.7. 1.8. 1.9.

1 L, 100 mL and 50 mL glass volumetric flasks

Various beakers

2 mL safelock tubes (e.g. Eppendorf, VWR p/n. 211-2120)

Pasteur pipettes (e.g. VWR p/n. 612-1684)

Microfilters: 0.2 µm (suitable for sugar solutions e.g. GHP Acrodisc 13, VWR p/n. 516-6916)

2 mL syringe (e.g. VWR p/n. 613-3911)

HPLC vials 2 mL with split septum suitable for the ICS used

Micro inserts (e.g. VWR 548-0001)

15 mL tubes (e.g. Falcon, VWR p/n. 734-0452)

1.10. 1.11. 1.12. 1.13. 1.14.

Carbograph SPE extract clean 150 mg column (Grace Davison p/n. 210142) Carbopac PA1 column (4*250 mm) (Dionex p/n. 35391) Carbopac PA1 guard column (4*50 mm) (Dionex p/n. 43096) Carbopac PA100 column (4*250 mm) (Dionex p/n. 43055) Carbopac PA100 guard column (4*50 mm) (Dionex p/n. 043054)

2.

Apparatus

2.1. 2.2. 2.3. 2.4. 2.5.

Centrifuge for 2 mL safelock tubes (up to 13,000 rpm) (e.g. Eppendorf 5415D) Analytical balance accurate to 0.1 mg (e.g. Mettler Toledo XS205)

pH meter (e.g. Mettler Toledo Seven Multi) Vortex mixer (e.g. Minishaker, IKA MS2)

Thermo Scientific HPAEC equipment ICS-5000 or equal, equipped with:

Pump (ICS-5000 + DP)

2.5.1. 2.5.2. 2.5.3.

Thermostatic autosampler (ICS-5000 AS-AP) Detector / Chromatography module (ICS-5000 + DC)

2.5.3.1. 2.5.3.2.

ED detector PAD, with standard carbohydrate waveform for Ag/AgCl ref electrode

Au electrode for carbohydrates

2.6. 2.7. 2.8.

Speedvac (e.g. Eppendorf concentrator 5301)

Waterbath 60˚C (e.g. grant GLS 200)

Vac-elute sps-24 with pump

3.

Chemicals

3.1. 3.2. 3.3. 3.4. 3.5. 3.6. 3.7. 3.8. 3.9.

Highly purified water (Milli-Q) Sodium acetate anhydrous (C 2 H 3

NaO 2

; Thermo Scientific p/n. 059326)

Sodium hydroxide 50% (VWR p/n. BAKR3727-1000)

D(+)-galactose (Sigma p/n. G-0750) Lactose monohydrate (Sigma p/n. 61339)

Arabinose (Sigma p/n. A3256)

Helium

Carrez 1: K 4

Fe(CN) 6

.3H 2

O (VWR p/n. 1.04984.0500)

Carrez 2: ZnSO 4

.7H 2

O (VWR p/n. 1.08883.1000)

3.10. 3.11. 3.12. 3.13. 3.14.

1 M phosphate buffer pH 6.0 ± 0.1

KH 2

PO 4

(VWR p/n. 1.04871.1000) (VWR p/n. 1.05101.1000)

K 2

HPO 4

Beta galactosidase from Aspergillus oryzae (Sigma p/n. G1560 15 KU)

Acetonitrile (VWR p/n. 1000302500)

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