Storage Conditions: Reagent 1 at -20℃, Reagent 2 at -20℃, Reagent 3 at 4℃, Reagent 4 at -20℃, Standard at 4℃

Detection Method: Visible Spectrophotometry

Unit: Box

Specification: 50T

 

 

Resistant starch (RS) refers to starch that cannot be enzymatically hydrolyzed in the human small intestine and is partially or completely fermented in the large intestine. RS is a component of total dietary fiber. The method for determining resistant starch involves incubating a sample with α-amylase and amyloglucosidase (AMG) in an oscillating water bath at 37℃ for 16 hours. During this period, the combined action of the two enzymes solubilizes and hydrolyzes the non-resistant starch into D-glucose. After incubation, an equal volume of ethanol or industrial methylated spirits (IMS, denatured ethanol) is added to stop the reaction. The solution is then centrifuged, and the supernatant is collected while the flocculent mass remaining at the bottom represents the RS in the sample. The flocculent mass is washed with aqueous IMS or ethanol (50% v/v), followed by centrifugation. The washing and centrifugation steps are repeated, and the supernatants obtained after each centrifugation are combined with the previously collected supernatant. The residual liquid in the test tube is carefully poured out, and the flocculent mass is placed in an ice-water bath. 2M KOH is added to dissolve the mass, and the mixture is vigorously stirred using a magnetic stirrer. The solution is neutralized with acetate buffer, and AMG is used to quantitatively hydrolyze the starch into glucose. The D-glucose is measured using the glucose oxidase/peroxidase reagent (GOPOD), which also determines the RS content in the sample. The determination of non-resistant starch (soluble starch) can be achieved by pooling the supernatants, diluting them to a final volume of 100ml, and then measuring the D-glucose using GOPOD.

Application:

For the detection of resistant starch.

Notes:

Different reagents should be stored at specified temperatures. This method requires a sample with an RS content greater than 2% w/w. If the RS content is greater than 2% w/w, the typical standard deviation is ±5%. For samples with less than 2% w/w RS, the error is higher.

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