Oligofructose

Native INULIN (INULIN ST)

High Molecular Weight INULIN (INULIN HP)

DP Degree of polymerization

FIGURE 3.7 Schematic representation of the DP distribution of the fructan chains in oligofructose, native inulin, high molecular weight inulin, and oligofructose-enriched inulin as analyzed by HPAEC-PAD.

Oligofructose-enriched INULIN (SYNERGY 1)

FIGURE 3.7 Schematic representation of the DP distribution of the fructan chains in oligofructose, native inulin, high molecular weight inulin, and oligofructose-enriched inulin as analyzed by HPAEC-PAD.

• CGC quantification (see above) of sucrose glucose and fructose before (to quantify native mono- and disaccharides) and after (to quantify inulin) enzymatic (Novozym 230) hydrolysis of all oligomers and polymers53

• Oxidation of native glucose by glucose oxidase (EC 1.1.3.4) enzymatic determination of native fructose, followed by hydrolysis of inulin by inulinase (EC 3.2.1.7), and quantification of inulin-derived fructose by using a series of biochemical reactions that include its phosphorylation to fructose-6-phosphate by hexokinase (EC 2.7.1.1) and ATP, followed by isomerization to glucose-6-phosphate by phosphoglucose isomerase (EC 5.3.1.9), oxidation to gluconate-6-phosphate by glucose-6-phos-phate-NADP+ oxidoreductase (EC 1.1.1.49), and finally the spectrophotometry (340 nm) determination of NADPH53

• Fractionation of the hot water extract of the plant or food product into three extracts used, respectively, to measure free glucose, fructose, and sucrose (after yeast D-glucosidase hydrolysis), glucose from starches and other D-gluco-oligo/polysaccharrides (after Aspergillus niger D-amyloglu-cosidase hydrolysis), and total fructose and glucose from inulin (after fructanase hydrolysis) and other oligo/polysaccharides (after D-amyloglu-cosidase hydrolysis), respectively, followed by spectrophotometric measurement of NADPH, as described above, and calculation of glucose and fructose released from fructans by the differences54

Hydrolysis of sucrose (by sucrase) to fructose and glucose, and starch to glucose (by a mixture of D-amylase, pullulanase and maltase), and reduction of the hexoses (using alkaline borohydride solution) to sugar alcohols, followed by hydrolysis of inulin (by a mixture of purified exo- and endo-inulinases) to fructose and glucose that are then measured with a spectrophotometer after reaction with para-hydroxybenzoic acid hydrazide.55, 56

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