SPSFAM ERP

10. Pipet 1 mL of this filtrate into a 4 dram vial and evaporate the solution to complete dryness under a flow of nitrogen at 55 ± 3°C. This may be achieved using an N-Evap 112 Nitrogen Evaporator or a heating block. 11. Once vials are completely dry, add 10 mL water to each. Cap and vortex for at least 15 seconds. Allow vials to sit for at least 15 minutes and repeat vortexing procedure. 12. Dilute samples as necessary so that all analytes are within the calibration curve. It may be necessary to perform multiple dilutions on a given sample to accomplish this. 13. Filter the diluted sample using a disposable syringe and 0.45 μm GHP filter into an injection vial.

H. (a)

HPAEC-PAD Analysis IC Operating Conditions.

1. 2. 3. 4. 5. 6. 7. 8. 9. 9.

Column:

Thermo Carbo Pac PA20, 150 × 3.0 mm Thermo Carbo Pac PA20, 30 × 3.0 mm Thermo BorateTrap, 50 × 4.0 mm Thermo IonPac ATC-500, 24 × 9.0 mm

Guard column: Borate Trap:

Anion Trap:

Column temperature: Detector temperature: Working electrode: Reference electrode: Detector waveform:

30 ± 2 °C 30 ± 2 °C

Conventional Au

Ag/AgCl

see Table 3

Mobile phase A: Mobile phase B: Mobile phase C: Mobile phase D:

water

10. 11. 12. 13. 14. 15. 16. 17. 18. 19.

100 mM sodium hydroxide 200 mM sodium hydroxide 600 mM sodium hydroxide

Post-column mobile phase: 200 mM sodium hydroxide

Column flow rate:

0.5 mL/min 1.0 mL/min

Post-column flow rate:

Elution gradient: Injection volume:

see Tables 4, 5, 6, and 7

10 µL

Run time:

30.0 min

System configuration:

see Figure 2

Table 3. PAD waveform, carbohydrate quad potential Time (sec) Potential (V) Integration 0.00 +0.1 0.20 +0.1 Begin 0.40 +0.1 End 0.41 -2.0