SPIFAN Nutrients ERP Book_9-29-15

FOS-04 FOR ERP USE ONLY DO NOT DISTRIBUTE

Sample Preparation 1. Powder sample reconstitution (if needed) – Accurately weight 5.0 g ± 10% into a 100 mL beaker (record powder weight) . Tare and deliver 40 g of lab water to beaker (record water weight ). Allow to stir for 30 minutes, or until dissolved. 2. Sample dilution – Samples require differing dilutions according to their individual fructan content as follows. Record all weights to four decimal places. This aliquot can be shared with the quantitative fructan methodology in Part 2.

Table 1-1 Sample Size Guidelines

Sample type

Fructan Level (as-is)

SW 1

(g)

Volumetric Flask Size (mL)

Powder Product Powder Product Powder Product RTF Product * RTF Product * RTF Product *

0.27-1.0%

10 g± 10% 7 g± 10% 4 g± 10% 10 g± 10% 7 g± 10% 4 g± 10% 0.4 g± 10%

50

1.1-10% 11-45%

200 500

0.03-0.20% 0.21-1.0%

50

200 500

1.1-5.0% 45-100%

Commodity 1000 * For concentrated liquid (CL) products dilute 10 mL of product with 10 mL of laboratory water and then treat the resultant material per RTF guidelines.

3. Filtration and analysis – After bringing the samples to volume, filter through a 0.45 µm nylon syringe filter into an autosampler vial (prepared samples can be stored in vials at 2-10 °C for 5 days) .

Instrument Conditions 1. Gradient – Fructans are eluted using a gradient of NaOH and sodium acetate at 1.0 mL/min as per figure 1-1 below. Column and detector compartment are maintained at 20 ° C. a. Please note that a Carbopac PA1 guard is used for this procedure rather than the borate trap used in part 2 for quantitative fructan determination. 2. Injection – A single 4 µL injection of each sample test solution is made. At the start of each sequence, 6 precision injections are made to ensure that system is stable, followed by the 3 retention time standards. Samples are bracketed with standards after every 6 injections. Autosampler is maintained at 10 ° C. 3. Electrochemical detector parameters – This method utilizes the carbohydrate triple waveform referred to in Thermo/Dionex Tech Note 21, Waveform B (4). Note this waveform is not appropriate for disposable gold electrodes! The reference electrode is set to AgCl mode.

Figure 1-1 Qualitative ID Gradient

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