SPSFAM ERP

Method Sugar Profile by High Performance Anion Exchange Chromatography with Pulsed Amperometric Detection [ Applicable to determination of galactose, glucose, sucrose, fructose, lactose, isomaltulose, and maltose in food products, pet food and animal feed, infant formula, and dietary supplements. ] Principle Sugars in the sample are extracted with a mixture of equal parts of water and ethanol, with the addition of an internal standard. Sample extracts may be treated with Carrez reagents to precipitate proteins. Sample extracts are filtered through a molecular weight cut-off filter, dried, reconstituted in water, diluted within the calibration range and filtered prior to the analysis using high- performance anion exchange chromatography with pulsed amperometric detection (HPAEC- PAD). (a) HPAEC-PAD .—ICS-5000 (or an equivalent system) with electrochemical (PAD) detection (Thermo; CA, USA) equipped with gold working electrode and silver/silver chloride reference electrode. (b) Analytical balances.— accurate to at least 0.00001 g. (c) Volumetric flasks.— Class A, glass, assorted sizes. (d) Erlenmeyer flasks with ground glass stoppers . — 500 mL. (e) Volumetric pipets.— Class A, glass, assorted sizes. (f) Filter paper.— 15cm, fluted, qualitative P8, coarse. (g) Molecular weight cut-off filter.— Amicon Ultra-4, Regenerated Cellulose, 10kDa (or equivalent). (h) Centrifuge.— capable of at least 1,500 g (RCF) with 20 and 30 mm diameter tube adapters. (i) Magnetic stir plates. (j) Bottle top dispenser.— 100 ± 0.6 mL, adjustable with graduations. (k) Desiccator.— containing activated silica gel desiccant for storage of reference standards. (l) Nitrogen evaporation system with heating capabilities.— N-Evap 112 (or equivalent system) (Organimation Associates, Inc.). (m) Vortex mixer. (n) Plastic centrifuge tubes.— assorted sizes . (o) Serological pipets.— 25 mL, glass or disposable. (p) pH meter. (q) Various sized adjustable volume positive displacement pipets with disposable tips. (r) Electronic repeater pipet (Eppendorf) or equivalent. (s) LC autosampler vials and caps. (t) 4-dram vials and caps, Borosilicate glass, Teflon lined caps (VWR) or equivalent. A. Apparatus