Gel permeation chromatographic method for simplified determination of relative molecular weight of amylose and amylopectin of paddy rice
A technology of gel permeation chromatography and amylose, which is applied in the field of starch determination, can solve the problems of starch loss, etc., and achieve the effects of good data repeatability, good application prospects, and simplified starch purification steps
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Embodiment 1
[0024] A gel permeation chromatography for simplifying the determination of the relative molecular weight of rice amylose and amylopectin, the steps are as follows:
[0025] Step 1: the pretreatment of rice flour: adopt polished rice flour to cross the rice flour of 0.42mm fineness sieve;
[0026] Step 2: Polish rice flour to remove fat and soluble sugar
[0027] Weigh 50mg of polished rice flour into a 10ml centrifuge tube, add 30ml of 100% chromatographically pure methanol, place in a boiling water bath for 10 minutes, centrifuge at 10,000g for 5 minutes, and pour off the suspension. Repeat step 2, 2-3 times for the precipitate, and combine the precipitate.
[0028] Step 3: Weigh 10mg of the precipitate into a 2ml centrifuge tube, add 0.2ml of 0.25mol / L NaOH, and bake in an oven at 50°C for 2 hours. After mixing, add 80 μl of 0.5 mol / L sodium acetate (pH=4.0), then add 20 μl of isoamylase (enzyme activity greater than 250 U) and react in a constant temperature water bath a...
Embodiment 2
[0033] A gel permeation chromatography for simplifying the determination of the relative molecular weight of rice amylose and amylopectin, the steps are as follows:
[0034] Step 1: the pretreatment of rice flour: adopt polished rice flour to cross the rice flour of 0.42mm fineness sieve;
[0035] Step 2: Polish rice flour to remove fat and soluble sugar
[0036] Weigh 100mg of polished rice flour into a 10ml centrifuge tube, add 60ml of 100% chromatographically pure methanol, place in a boiling water bath for 10 minutes, centrifuge at 10,000g for 5 minutes, and pour off the suspension. Repeat step 2 for the precipitate, 2 times, and combine the precipitate.
[0037] Step 3: Weigh 20mg of the precipitate into a 2ml centrifuge tube, add 0.4ml of 0.25mol / L NaOH, and bake in an oven at 50°C for 2 hours. After mixing, add 80 μl of 0.5 mol / L sodium acetate (pH=4.0), then add 20 μl of isoamylase (enzyme activity greater than 250 U) and react in a constant temperature water bath at...
Embodiment 3
[0041] A gel permeation chromatography for simplifying the determination of the relative molecular weight of rice amylose and amylopectin, the steps are as follows:
[0042] Step 1: the pretreatment of rice flour: adopt polished rice flour to cross the rice flour of 0.42mm fineness sieve;
[0043] Step 2: Polish rice flour to remove fat and soluble sugar
[0044] Weigh 100mg of polished rice flour into a 10ml centrifuge tube, add 60ml of 100% chromatographically pure methanol, place in a boiling water bath for 10 minutes, centrifuge at 10,000g for 5 minutes, and pour off the suspension. Repeat steps 2 and 3 times for the precipitate, and combine the precipitate.
[0045] Step 3: Weigh 20mg of the precipitate into a 2ml centrifuge tube, add 0.4ml of 0.25mol / L NaOH, and bake in an oven at 50°C for 2 hours. After mixing, add 80 μl of 0.5 mol / L sodium acetate (PH=4.0), then add 20 μl of isoamylase (enzyme activity greater than 250 U) and react in a constant temperature water bat...
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