The presence of galactosyl-sucrose oligosaccharides (e.g. raffinose, stachyose and verbascose) in a sample will lead to overestimation of fructan results, as exo-inulinase will hydrolyse the linkage between glucose and fructose of galactosyl-sucrose oligosaccharides. To remove galactosyl-sucrose oligosaccharides add 50 μL of α-galactosidase (Megazyme cat. no. E-AGLANP) at 200 U/mL in 50 mM sodium acetate buffer (pH 4.5) to 0.2 mL of solution to be analysed and incubate for 30 min at 40°C before addition of the Sucrase/Amylase working mixture (Enzymes Solution 1). This enzyme gives complete hydrolysis of D-galactose from galactosyl-sucrose oligosaccharides and sucrose is then removed by an incubation with sucrase followed by borohydride reduction. Please adjust the calculations accordingly (change of factor 1.1/0.2 to 1.15/0.2).
Modified on: Thu, 10 May, 2018 at 3:37 PM
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