A chive seed antioxidant tripeptide and its preparation and application
A technology for oxidizing tripeptides and leek seeds, which is applied in the fields of food and biomedicine, and can solve the problems of no reports of active peptides
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Embodiment 1
[0027] The separation and purification of the antioxidant tripeptide in the present invention includes two steps of SephadexG-50 gel filtration chromatography and reversed-phase high-performance liquid chromatography (RP-HPLC).
[0028] Extraction of leek seed extract: The leek seeds are pulverized by a high-speed traditional Chinese medicine grinder, soaked in 10% (V / V) acetic acid, assisted by ultrasonic extraction for 20 minutes, stirred and extracted at room temperature for 6 hours, centrifuged at 12000rpm for 15 minutes, and the supernatant was freeze-dried, - Store at 80°C.
[0029] SephadexG-50 Gel Filtration Chromatography: The leek seed extract freeze-dried powder was obtained according to the above method, dissolved in deionized water, and centrifuged at 12000rpm for 15min. The supernatant was filtered through a 0.22 μm pore size microfiltration membrane. SephadexG-50 gel column (1.6cm×100cm) was equilibrated with deionized water, and the filtered sample was loaded ...
Embodiment 1
[0034] The natural antioxidant tripeptide activity obtained in the implementation example 1 is studied:
[0035] DPPH free radical scavenging effect: 0.4mL samples of different concentrations were fully mixed with an equal volume of 4% (W / V) DPPH anhydrous ethanol solution, kept at room temperature in the dark for 30min, and the absorbance value was measured at 517nm. 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):
[0036] (1)
[0037] In the formula, A 0 : Absorbance value of blank control group; A s : Absorbance value of sample group
[0038] ABTS free radical scavenging effect: prepare 7mmol / L ABTS with distilled water + solution and 2.45mmol / L potassium persulfate solution (must be placed at room temperature for 16h before use), ABTS + Mix with potassium persulfate solution at a volu...
Embodiment 3
[0047] The protective effect of the antioxidant tripeptide of the present invention on hydrogen peroxide-induced normal liver cell damage:
[0048] The revived normal hepatocytes (LO2) were treated with modified RPMI-1640 medium containing 10% fetal bovine serum, 2.0mmol / L glutamate and 100U / mL penicillin-streptomycin in a humidified medium containing 5% carbon dioxide , adherent culture at 37°C for 24-48 hours. After digesting with trypsin at 37°C, add RPMI-1640 medium to stop the digestion, add an appropriate amount of RPMI-1640 medium for blowing to fully disperse the cells, inoculate into a 96-well plate and continue culturing for 24 hours, so that the final concentration of the cells is 1.5 ×10 5 / hole. The cells were treated with different concentrations of tripeptide GSQ for 24 hours, and then treated with 300 μmol / L hydrogen peroxide for 24 hours, and the number of viable hepatocytes in each well was detected by MTT method.
[0049] From the test results, it can be ...
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