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5841 v2

Page 2 of 4

AsureQuality Laboratory Services

4. REAGENTS

1.

2 M Sodium Hydroxide

: Weigh 80g of sodium hydroxide into a 1L volumetric flask, dissolve

in water and make up to mark.

2.

0.15 M Sodium Phosphate Buffer

: Weigh 9.15g of

sodium dihydrogen phosphate

dihydrate and 16.31g of disodium hydrogen phosphate dihydrate into a 1L volumetric flask.

Dissolve in water and make up to mark. Adjust pH to 7 with 2M sodium hydroxide.

3.

0.1% Phosphoric acid:

In a 1L volumetric flask, add 500 mL water. Add 1.2 mL of ortho-

phosphoric acid. Mix and make up to mark with water.

5. APPARATUS

1.Whatman Glass Microfiber Filters, CAT No. 1820-125

2.Biopharm Immunoaffinity column Pack P82 / P82B or equivalent

3.SPE Manifold with accessories

4.Autoclave set at 121

0

C

5.Centrifuge (variable speed)

6.Analytical Balance 4 dp

7.Amber glass screw cap bottle, 100mL

8.Horizontal shaker

9.Volumetric flasks, 1L, 250mL, 100mL, 10mL

10. Pipettors, calibrated, 10.0mL, 5.0mL, 1.0mL, 200μL, 100μL and 50μL.

11. Measuring cylinder, 100mL, 50mL

12. Reacti-vials

13. Reacti-therm heating block with nitrogen blow down

14. Ultrasonic bath set at 50

0

C

15. Centrifuge tubes, 50mL

16. Vortex mixer

17. Syringe filter, PTFE 0.45μm (HP045AN)

18. Disposable syringes, 10mL and 1mL

19. HPLC vials, 2mL with glass inserts (200μL)

6. SAMPLE PREPARATION

Note: Please see Table 1 for weight and loading volume for different product range.

Slurry may be used wherever product heterogeneity is expected.

Slurry: Reconstitute the 25g powder with warm water (~50ºC) to a total weight of 200g. Mix

thoroughly on a horizontal shaker for 15 minutes and then sonicate at 50

0

C for 10 minutes.

Cool to room temperature.

1. Weigh sample / slurry into a 100mL amber glass screw cap bottle. Refer to Table 1.

2. Add 0.15 M sodium phosphate buffer to a volume of 50mL.

3. Swirl gently to mix.

4. Autoclave the sample preparation at 121

0

C for 25 minutes.

Bio-02 (February 2016)

FOR ERP USE ONLY

DO NOT DISTRIBUTE