SPSFAM ERP

11. After 15 minutes, centrifuge samples that do not have a clear zone present for five minutes at 3000 RPM 8 . 12. Transfer a sample aliquot, avoiding precipitate and fat, into a 3 cc disposable syringe equipped with a 0.20 µm 25 mm syringe filter. 13. Dispense filtered sample into a 2 mL auto sampler vial, and enclose with a Teflon coated crimp seal 9 . Occasionally, samples may need to be prefiltered through a 0.45 um filter first, and subsequently through a 0.2 um filter. Salt Removal For soy sauce and fermented condiments, meat snacks, premixes and fortified grains an additional cleanup step is required to remove the cations. 14. At step 6, insert a 10 ml disposable syringe into the aqueous central layer and collect 6 mL of sample 15. Attach a 25 mm 0.45 mm syringe filter, and filter the sample into a clean vessel for each sample (e.g test tube). 16. Dispense 4 mL of the filtered sample from step 15 onto a dry strata ABW SPE cartridge to elute a minimum of 3 mL into a clean test tube by gravity (or light pipet bulb encouragement) 17. Continue with step 8 as outlined above 18. Analyze by RI-HPLC according to the following parameters on the chromatography system that is equipped with a 0.5 um column prefilter, then a guard column, followed by the analytical amino column. Injection volume: 10 µL Flow Rate: 0.5 mL min, isocratic Column temperature: ambient Sample temperature: ambient Mobile Phase startup~75 % acetonitrile/25 % water, up to 82 % acetonitrile/18 % water over time. Typical Chromatogram of the 0.5 mg/ml Sugar Standard

8 Centrifuge maltodextrin or soluble corn fiber samples to precipitate these out of solution. Milk protein isolate concentrates should be centrifuged at 6000 RPM for 20 minutes. 9 Alternatively, a 0.1 um 25 mm syringe filter membrane may be used at the discretion of the chemist