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Size-exclusion chromatography (HPLC-SEC) technique optimization by simplex method to estimate molecular weight distribution of agave fructans.

Food chemistry (2017-08-03)
Lorena Moreno-Vilet, Stéphane Bostyn, Jose-Luis Flores-Montaño, Rosa-María Camacho-Ruiz
ABSTRACT

Agave fructans are increasingly important in food industry and nutrition sciences as a potential ingredient of functional food, thus practical analysis tools to characterize them are needed. In view of the importance of the molecular weight on the functional properties of agave fructans, this study has the purpose to optimize a method to determine their molecular weight distribution by HPLC-SEC for industrial application. The optimization was carried out using a simplex method. The optimum conditions obtained were at column temperature of 61.7°C using tri-distilled water without salt, adjusted pH of 5.4 and a flow rate of 0.36mL/min. The exclusion range is from 1 to 49 of polymerization degree (180-7966Da). This proposed method represents an accurate and fast alternative to standard methods involving multiple-detection or hydrolysis of fructans. The industrial applications of this technique might be for quality control, study of fractionation processes and determination of purity.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Maltoheptaose, ≥60% (HPLC)
Supelco
Dextran, analytical standard, for GPC, 5,000
Supelco
Nystose, analytical standard
Sigma-Aldrich
Blue Dextran, average mol wt 20,000
Supelco
1-Kestose, analytical standard
Sigma-Aldrich
Maltohexaose, ≥65% (HPLC)