Dietary Fiber Assay Kit （食物纖維測(cè)定試劑盒）
Grade: for Food Analysis
Storage condition: Keep at 2-10 degrees C.
Manufacturer: Wako Pure Chemical Industries, Ltd.
Cat. No. Pkg. Size
291-59701 100Tests
Summary
This product is for research use only. Do not administer it to human.
A simple and modified version of Prosky's method. Compared to conventional Prosky's method, it assures high measurement accuracy. Furthermore, enzyme treatment time is shortened.
Kit Contents
Thermostable alpha-Amylase Solution 1 vial x 20 mL
Protease Solution 1 vial x 20 mL
Amyloglucosidase Solution 1 vial x 20 mL
Diatomaceous Earth 1 vial x 100 g
Measurement Outline
Sample dissolved in MES/Tris buffer
Digestion with thermostable alpha-amylase
Digestion with protease and amyloglucosidase
for 30 minutes at pH 6.3 and 60 degrees C
Ethanol Precipitation
Leave 1 hour
Filtration
Wash with ethanol and acetone
Dry overnight
Kjeldahl Protein Determination and Ash Determination
Total Dietary Fiber Determination
Principle
1、Determination of total Dietary Fiber Contents
Digestible components in samples, such as starch and protein, are hydrolyzed
with alpha-amylase, protease, and amyloglucosidase, precipitate undecomposed residual macromolecule by adding quadruple volume 95% Ethanol, and then trapped with a glass filter crucible containing diatomaceous earth.
Total Dietary Fiber Contents are determined by deducting weight of protein and ash content from the dry weight of trapped residue on a glass filter crucible.
2、Fractional Analysis of Soluble Dietary Fiber Contents and Insoluble Dietary Fiber Contents
Digestible components in samples, such as starch and protein, are hydrolyzed with alpha-amylase, protease, and amyloglucosidase, then filtrated with a glass filter crucible containing diatomaceous earth.
Insoluble Dietary Fiber Contents are determined by deducting weight of protein and ash content from the dry weight of trapped insoluble residue on a glass filter crucible.
Meanwhile, add a quadruple volume 95% Ethanol into the filtrate, precipitate undecomposed residual soluble macromolecule, and trap with a glass filter crucible containing diatomaceous earth. Soluble Dietary Fiber Contents are determined by deducting weight of protein and ash content from the dry weight of the trapped precipitated residue on a glass filter crucible.
Total Dietary Fiber Contents are defined as the sum of Insoluble Dietary Fiber Contents and Soluble Dietary Fiber Contents.
3、Determination of Low-molecular Soluble Dietary Fiber
Specific Low-molecular Soluble Dietary Fibers, which are not precipitated even by addition of a quadruple volume of 95% Ethanol, are determined by high-performance liquid chromatography.
After digestible components such as starch and protein in samples are hydrolyzed with alpha-amylase, protease, and amyloglucosidase, the undecomposed residual macromolecule are precipitated by adding a quadruple volume of 95% Ethanol. Then filter it with a glass filter crucible containing diatomaceous earth.
Proteins, organic acids and inorganic salts in the filtrate are removed with ion-exchange resin, and then the solution is applied to high-performance liquid chromatography. Low-molecular Soluble Dietary Fiber Content is determined by the ratio of the peak area of the sample with that of glucose (internal standard) on the chromatogram.
Meanwhile, Insoluble and Soluble Macromolecule Dietary Fiber Content is determined by deducting weight of protein and ash content from the dry weight of the trapped precipitated residue on a glass filter crucible.
Total Dietary Fiber Contents is defined as sum of Low-molecular Soluble Dietary Fiber Contents and Insoluble and Soluble Macromolecular Dietary Fiber Contents.
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