Broccoli polypeptide component microwave-assisted extracting and activity measuring method

A microwave-assisted, extraction method technology, applied in the field of extraction, can solve the problems of long extraction time of protein and polypeptide, reduced activity of polypeptide products, large amount of organic solvent, etc., and achieve the effect of reducing operating costs, high recovery rate, and complete separation

Inactive Publication Date: 2016-10-26
JINLIN MEDICAL COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Using organic solvent precipitation method to extract proteins and polypeptides takes a long time, the amount of organic solvents is large, an

Method used

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  • Broccoli polypeptide component microwave-assisted extracting and activity measuring method
  • Broccoli polypeptide component microwave-assisted extracting and activity measuring method
  • Broccoli polypeptide component microwave-assisted extracting and activity measuring method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] see figure 1 , The method for microwave-assisted extraction of broccoli polypeptide components of the present invention mainly includes the following steps.

[0015] S100: Preparation by liquid nitrogen grinding. Wash broccoli with distilled water, take flower buds, dry in the shade, and grind with liquid nitrogen to prepare.

[0016] S200: Microwave radiation. Get 100 parts (grams) of broccoli after grinding, add 10% of fresh weight of PVP, 300% of fresh weight of Tris-HCL solution, solution concentration 55mmol / L-65mmol / L, pH value 6-7, mix , put it into a microwave oven, the ratio of broccoli weight (grams) to microwave power (watts) is 1:5-9, preferably 1:8, the power of microwaves is crucial to the extraction of broccoli polypeptides, and the power is too large It will destroy the polypeptide structure, and if the power is too small, the active ingredient can not flow out freely. For this reason, the inventor of the application has found that the ratio of brocco...

Embodiment 2

[0021] Further, the present application also discloses a method for determining the activity of broccoli polypeptide components, comprising the following steps:

[0022] S600: Take 1-3ml of broccoli polypeptide crude extract and apply it to Sephadex G-25 gel filtration chromatography (16mm×1000mm), elute with distilled water, flow rate 1mL min -1 , monitored with a protein and nucleic acid detector, with a wavelength of 280nm, and collected each elution peak.

[0023] S700: Take 100 microliters of each component of the broccoli polypeptide, put it in a test tube, add 5ml of Coomassie brilliant blue solution, shake it well, let it stand for 10 minutes, and measure the absorbance value of each component of the broccoli polypeptide at 595nm.

[0024] S800: Take the sample solution of each separated component of broccoli polypeptide or distilled water (control) and 0.05mol·L -1 pH=8.2 Tris-HCl buffer solution was mixed in a test tube, and placed in a constant temperature water ba...

experiment example

[0026] The advantages of the extraction method in Example 1 of the present invention can be further illustrated through the comparison of this experimental example.

[0027] Experimental process conditions:

[0028]Traditional organic solvent extraction method: wash the broccoli, cut off the flower buds, and dry in the shade for later use. Weigh the wet weight buds, add PVP according to 10% of the fresh weight, grind with liquid nitrogen, add Tris-HCl extract with a concentration of 62.5mmol / L, place at 4°C for 1h, remove precipitation, take the supernatant, add a certain volume of -20 Pre-cool the acetone at -20°C, remove the supernatant, and collect the precipitate. The precipitation was placed at -20°C for 20 minutes. Distilled water was added to dissolve the precipitate to obtain the crude extract of broccoli polypeptide.

[0029] Microwave-assisted extraction method: Wash the broccoli, take the flower buds, and dry them in the shade for later use. Weigh the wet buds, ...

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Abstract

The invention discloses a broccoli polypeptide component microwave-assisted extracting and activity measuring method. The method comprises the following steps: step 100, liquid nitrogen grinding and preparation: drying broccolis in the shade, and grinding broccolis in liquid nitrogen; step 200, radiating broccolis by microwaves; step 300, separating the supernate; step 400, adding an organic solvent; and step 500, separating the broccoli polypeptide component. The provided method has the advantages that the extraction time is effectively shortened, the operation cost is reduced, the using amount of organic solvent is reduced, the method is environment-friendly, the extraction efficiency is prominently improved, and more active components are obtained.

Description

technical field [0001] The invention relates to an extraction method, in particular to a microwave-assisted extraction and activity determination method of broccoli polypeptide components. Background technique [0002] Broccoli is a very popular vegetable. It is delicious, nutritious, and has high medicinal value. Its vitamin C content is very rich, and it also has anti-cancer effects. The average nutritional value and disease prevention effect of broccoli far exceed other vegetables. Broccoli polypeptide is an effective anticancer ingredient in broccoli. Polypeptide compounds widely exist in nature. Among them, the research on polypeptides with certain biological activities has always been a main direction of drug development. The active components of broccoli peptides are applied in anti-tumor cell experiments, and have the functions of inhibiting tumor cell proliferation and promoting tumor cell apoptosis. At present, organic solvent precipitation method is generally us...

Claims

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Application Information

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IPC IPC(8): G01N1/34G01N30/14G01N30/00G01N30/02
CPCG01N1/34G01N30/00G01N30/02G01N30/14
Inventor 徐俊杰吕士杰李妍朱文赫张巍芦晓晶刘洋
Owner JINLIN MEDICAL COLLEGE
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