A chive seed antioxidant hexapeptide and its preparation method and application
A leek seed, anti-oxidation technology, applied in the food field, can solve the problem of no reports of active peptides
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Embodiment 1
[0018] The separation and purification of the antioxidant hexapeptide in the present invention includes two steps of Sephadex G-25 gel filtration chromatography and reversed-phase high-performance liquid chromatography (RP-HPLC).
[0019] Preparation of antioxidative hexapeptide hydrolyzate of leek seed: using acid protease under the optimal hydrolysis conditions (pH3.02, enzyme addition 1345.7U, substrate concentration 2% (w / v)) for 6.45h , inactivated in a boiling water bath for 10 minutes, centrifuged at 12000 rpm for 20 minutes, and the supernatant was freeze-dried, which was the leek seed protein enzymatic hydrolysis product;
[0020] Sephadex G-25 Gel Filtration Chromatography: Dissolve the lyophilized powder of leek seed enzymatic hydrolyzate in deionized water, and centrifuge at 12000 rpm for 15 min. The supernatant was filtered through a 0.22 μm pore size microfiltration membrane. Sephadex G-50 gel column (1.6cm×100cm) was equilibrated with deionized water, and the f...
Embodiment 2
[0025] The natural antioxidant hexapeptide activity obtained in Example 1 is studied:
[0026] DPPH free radical scavenging effect: 0.4 mL samples of different concentrations and an equal volume of 4% ( W / V ) DPPH absolute ethanol solution was mixed thoroughly, and kept at room temperature in the dark for 30 min, and the absorbance value was measured at 517 nm. Deionized water and reduced glutathione were used instead of samples as blank control and positive control, respectively. The DPPH free radical scavenging activity of the sample is calculated according to the following formula (1):
[0027]
[0028] In the formula, A 0 : Absorbance value of blank control group; A s : Absorbance value of sample group
[0029] ABTS free radical scavenging effect: prepare 7 mmol / L ABTS with distilled water + solution and 2.45 mmol / L potassium persulfate solution (must be placed at room temperature for 16 h before use), ABTS + Mix with potassium persulfate solution at a volume rat...
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