Method for preparing reduced type glucoraphanin by enteric bacterium metabolism and detection method

A glucoraphanin and detection method technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of complex purification process, low purity, low conversion efficiency of reduced glucoraphanin, and achieve high degradation rate and conversion High efficiency and ease of purification difficulty

Active Publication Date: 2019-03-22
SHANDONG UNIV OF TRADITIONAL CHINESE MEDICINE
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention aims at the shortcomings of low conversion efficiency of reduced glucoraphanin, complex purification process and low purity, and provides a method for preparing high-purity reduced glucoraphanin. The method has simple purification process, high purity, high conversion efficiency, Low energy consumption, no pollution

Method used

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  • Method for preparing reduced type glucoraphanin by enteric bacterium metabolism and detection method
  • Method for preparing reduced type glucoraphanin by enteric bacterium metabolism and detection method
  • Method for preparing reduced type glucoraphanin by enteric bacterium metabolism and detection method

Examples

Experimental program
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Embodiment 1

[0036]Embodiment 1: A method for preparing reduced radishin by metabolism of intestinal bacteria, comprising the following steps:

[0037] (1) Preparation of anaerobic culture medium

[0038] Solution A: Accurately weigh 0.2925g of dipotassium hydrogen phosphate, add distilled water to 37.5ml, and dissolve.

[0039] Solution B: Accurately weigh 0.1763g of dipotassium hydrogen phosphate, 0.4425g of sodium chloride, 0.4500g of ammonium sulfate, 0.0450g of calcium chloride, MgSO 4 ·H 2 O 0.0938g, add distilled water to 37.5ml, dissolve.

[0040] Solution C: Accurately weigh 4g of sodium carbonate, add distilled water to 50ml, and dissolve.

[0041] Then add 0.5g of L-cysteine, 2ml of L-ascorbic acid with a volume fraction of 25%, 1g of beef extract, 1g of tryptone, 1g of nutrient agar, add distilled water to 1L, adjust the pH to 7.0-7.5 with hydrochloric acid, and obtain anaerobic culture Store the solution in a refrigerator at 4°C for later use.

[0042] (2) Preparation of ...

Embodiment 2

[0058] Embodiment 2: A method for preparing reduced radishin by metabolism of intestinal bacteria, comprising the following steps:

[0059] (1) Preparation of Enterobacteriaceae Culture Solution

[0060] Please refer to Example 1 for the experimental steps of preparing enterobacteria culture solution;

[0061] (2) Preparation of sample solution

[0062] 2.1 Preparation of test solution samples

[0063] a. Crush the decoction pieces of fried radish seeds into coarse powder, add 20 times the amount, extract for 15-30 minutes, extract 3 times, combine the extracts, and concentrate to 0.5g·mL -1 , put the concentrated fried radish seed extract on AB-8 macroporous adsorption resin, the sample volume is 0.5-0.75BV, and the sample flow rate is 0.5-1BV·h -1 , eluted with water, the elution flow rate is 1~2BV·h -1 , the elution volume is 3-5BV, the water eluate is collected, concentrated, and dried under reduced pressure to obtain the effective fraction of glucosinolate.

[0064] ...

Embodiment 3

[0069] Embodiment 3: A method for preparing reduced radishin by metabolism of intestinal bacteria, comprising the following steps:

[0070] (1) Preparation of Enterobacteriaceae Culture Solution

[0071] Please refer to Example 1 for the experimental steps of preparing enterobacteria culture solution;

[0072] (2) Sample preparation

[0073] 2.1 Preparation of test solution samples

[0074]a. Crush the decoction pieces of fried radish seeds into coarse powder, add 20 times the amount, extract for 15-30 minutes, extract 3 times, combine the extracts, and concentrate to 0.5g·mL -1 To obtain the fried radish seed extract, the fried radish seed extract is concentrated under reduced pressure to a mass concentration of 0.1-0.2 g·mL -1 , through D301 chlorine-type anion exchange resin, the sample volume is 7BV, and the adsorption flow rate of the sample solution is 1-3BV h -1 , the amount of water elution is 3BV, the concentration is 5% ~ 10% NaCl solution 3 ~ 5BV elution, the el...

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Abstract

The invention provides a method for preparing reduced type glucoraphanin by enteric bacterium metabolism and a detection method. The method comprises the following steps: preparing an anaerobic culture solution and an intestinal bacteria solution, mixing and incubating the anaerobic culture solution and the intestinal bacteria solution to obtain an intestinal bacteria culture solution, then addinga glucoraphanin extract into the intestinal bacteria solution, thus obtaining a crude product solution containing metabolite under the action of intestinal flora and utilizing sephadex to separate and purify the metabolite to obtain the highly purified reduced type glucoraphanin. The reduced type glucoraphanin obtained by the invention has high degree of purity and a good purification effect.

Description

technical field [0001] The invention relates to a method for preparing reduced glucoraphanin by metabolism of intestinal bacteria and a detection method, belonging to the field of traditional Chinese medicine. Background technique [0002] Cruciferous vegetables are rich in reduced glucoraphanin and myrosinase. Studies have found that reduced glucoraphanin can be hydrolyzed by myrosinase and converted into reduced sulforaphane through non-enzymatic intramolecular rearrangement reactions. Reduced sulforaphane (4-methylthio-3-butenyl isothiocyanate), an isothiocyanate lipid compound that induces phase II detoxification enzyme activity, will not only increase the body's resistance to carcinogens It can also inhibit the killing of cancer cells. Reduced sulforaphane is the phytochemical component with the strongest anticancer activity found in cruciferous vegetables, and it has strong inhibitory effects on breast cancer, lung cancer, prostate cancer and various cancers. [0003...

Claims

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Application Information

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IPC IPC(8): C12P19/64G01N30/60
CPCC12P19/64G01N30/6052
Inventor 朱立俏盛华刚张茜周洪雷
Owner SHANDONG UNIV OF TRADITIONAL CHINESE MEDICINE
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