The Raffinose/Sucrose/D-Glucose test kit is for the measurement and analysis of D-glucose, sucrose and raffinose, stachyose and verbascose in seeds and seed meals. Based on the measurement of D-glucose on enzymic hydrolysis of raffinose, stachyose and verbascose to D-glucose, D-fructose and D-galactose.
Colourimetric method for the determination of Raffinose (also
stachyose and verbascose), Sucrose and D-Glucose in
legume seeds, plant materials, foodstuffs and feed
Principle:
(α-galactosidase)
(1) Raffinose + stachyose + verbascose + H2O → D-galactose +
sucrose
(invertase)
(2) Sucrose + H2O → D-glucose + D-fructose
(glucose oxidase)
(3) D-Glucose + H2O + O2 → D-gluconate + H2O2
(peroxidase)
(4) 2H2O2 + p-hydroxybenzoic acid + 4-aminoantipyrine →
quinoneimine + 4H2O
Kit size: 120 assays
Method: Spectrophotometric at 510 nm
Reaction time: ~ 20 min
Detection limit: 100 mg/L
Application examples:
Analysis of grain legumes and other materials containing raffinose,
stachyose and verbascose
Method recognition: Used and accepted in food analysis
Advantages
Q1. Should the pH of the sample be adjusted even for samples in acidic media?
The pH of the assay solution after the sample is added should be the same as that of the assay buffer that is supplied with the kit.
Low sample volumes (e.g. 0.1 mL) are not likely to affect the pH of the assay solution and therefore may not require pH adjustment.
Samples above 0.1 mL are more likely to affect the pH of the assay solution and therefore the pH of these samples should be adjusted as described in the data booklet, prior to addition to the assay.
If you suspect that the Megazyme test kit is not performing as expected such that expected results are not obtained please do the following:
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