Research method of antioxidant activity of hemp seed protein hydrolysate
A technology of anti-oxidative activity and hemp seed protein, which is applied in the research field of hemp seed protein, can solve the problems of difficult access to its enzyme cleavage site, high equipment requirements, and difficult operating conditions, so as to increase protease cleavage sites, The effect of improving enzymatic hydrolysis efficiency and reducing enzymatic hydrolysis time
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Embodiment 1
[0045] Example 1: The ability to scavenge DPPH free radicals includes the following steps. Take 0.3 mL of sample solutions of different concentrations (concentrations 10 mg / mL, 15 mg / mL, 20 mg / mL, 25 mg / mL, 30 mg / mL) and add them to 2.7 mL of 0.0001 mol / LDPPH freshly prepared ethanol solution, mix vigorously, place the reaction mixture in the dark at room temperature for 30min, measure the absorbance at 517nm with a UV-visible spectrophotometer, use DPPH ethanol solution without sample as a blank control, and the inhibition rate can be used The following formula expresses:
[0046] Inhibition rate%=
[0047] In the formula: A t=0 represents the absorbance of the blank;
[0048] A t=30min It represents the absorbance after adding the hydrolyzate and keeping it warm for 30 minutes.
[0049] according to figure 1 As shown, with the increase of the concentration of each sample, the ability of each sample to scavenge DPPH free radicals increases, and each sample shows a cer...
Embodiment 2
[0051] Embodiment two: total reducing power measurement comprises the following steps, get the 1mL sample solution of concentration 10mg / mL, 15mg / mL, 20mg / mL, 25mg / mL, 30mg / mL respectively with 2.5mL phosphate buffer (0.2M, pH6. 6) Mix with 2.5mL potassium ferricyanide (1%) solution, keep the mixture at 50°C for 20min, then add 2.5mL10% trichloroacetic acid, centrifuge at 3000r / min for 10min with a centrifuge, absorb 2mL supernatant after centrifugation Add 2.5mL distilled water and 0.5mL (0.1%) ferric chloride to the test tube, mix well, use distilled water as a reference, and measure the spectrophotometric value at 700nm with a UV spectrophotometer. High absorbance indicates strong reducing power.
[0052] according to figure 2 As shown, the hemp seed protein and the hemp seed polypeptide under different treatment conditions have a certain reducing ability, and the total reducing ability is in the order of HMR
Embodiment 3
[0054] Example 3: The ability to scavenge hydroxyl radicals HO includes the following steps. After adding 1 mL of 6 mmol / L ferrous sulfate and 1 mL of 6 mmol / L salicylic acid-ethanol in a 10 mL test tube, sample solutions of different concentrations (concentration 2 mg / mL, 4mg / mL, 6mg / mL, 8mg / mL, 10mg / mL) 1mL was added to the test tube respectively, then 1mL of 0.1% hydrogen peroxide was added, and the total volume was made up to 5mL with distilled water, and no sample was added to the control tube solution, the bottom tube does not add , after shaking well, keep it in a water bath at 37°C for 30 minutes, measure the absorbance value at 510nm with a UV spectrophotometer, and the formula for calculating the clearance rate is:
[0055] Clearance %=
[0056] In the formula: : Absorbance value of hydroxyl radical system without adding hydrolyzate;
[0057] : the absorbance value of the hydroxyl radical system added to the hydrolyzate;
[0058] : No addition in the hyd...
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