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Ergothioneine detection method

A technology for ergothioneine and its detection method, which is applied in the technical fields of analytical chemistry and biochemistry, can solve problems such as the lack of standards for ergothioneine detection methods, achieve ideal retention and elution effects, strong versatility, and stable detection conditions Effect

Active Publication Date: 2015-07-01
TIANJIN SINONOCY BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] So far there is no standard method for the detection of ergothioneine

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Preparation of Pleurotus otreatus CGMCC No.6232 mycelium fermentation broth

[0026] Slant medium: PDA medium (Becton, Dickinson and Company).

[0027] Liquid seed medium (g / L): corn flour 30g / L, soybean meal 15g / L, α-amylase 54U / L, KH 2 PO 4 3g / L, MgSO 4 ·7H 2 O1.5g / L, the rest is water, sterilized at 121°C for 20 minutes, and the liquid volume in a 500mL Erlenmeyer flask is 150mL.

[0028] Liquid fermentation medium (g / L): glycerol 50g / L, casein peptone 35g / L, KH 2 PO 4 3g / L, MgSO 4 ·7H 2 O1.5g / L, the rest is water, the rest is water, sterilized at 121°C for 20min, and the liquid volume in a 500mL Erlenmeyer flask is 150mL.

[0029] The bacterial lawn of strain CGMCC No.6232 from the slant medium was picked and inserted into the seed medium, and cultured on a shaker at 25° C. at 150 rpm for 4 days to obtain a seed solution. The seed solution was inserted into the fermentation medium with an inoculation amount of 5% by volume, and cultured on a shak...

Embodiment 2

[0030] Embodiment 2: the pretreatment of ergothioneine fermentation liquid sample

[0031] After the submerged fermentation of Pleurotus ostreatus CGMCC No.6232 mycelia in Example 1, the mycelia fermentation liquid was placed in a 90°C water bath and leached for 15 minutes under stirring at 200 rpm, and the ergothioneine was leached to Extracellular. Filtrate, collect the filtrate, and use an ultrafiltration membrane with a molecular weight cut-off of 4kDa for ultrafiltration, and the obtained ultrafiltration permeate is the sample to be tested for high-pressure liquid phase detection.

Embodiment 3

[0032] Embodiment 3: the pretreatment of ergothioneine aqueous suspension sample

[0033] After the submerged fermentation of Pleurotus ostreatus CGMCC No.6232 mycelium in Example 1, the mycelium was collected by filtration, and added according to the weight of wet mycelium: water (g wet mycelium: mL water) 1:15 water to make an aqueous suspension of mycelium. The mycelia aqueous suspension is placed in a water bath at 85° C., stirred and leached at 300 rpm for 10 minutes, and the ergothioneine is leached into the aqueous solution. Filtrate, collect the filtrate, and use an ultrafiltration membrane with a molecular weight cut-off of 4kDa for ultrafiltration, and the obtained ultrafiltration permeate is the sample to be tested for high-pressure liquid phase detection.

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Abstract

The invention relates to a high pressure liquid chromatography detection method for ergothioneine. According to the invention, an hydrophilic interaction chromatography (HILIC) analysis chromatographic column is employed for performing a high pressure liquid chromatography analysis on a sample containing ergothioneine, the chromatographic column employs a HILIC filler as a fixed phase, a solution of acetonitrile and water (v / v) with ratio of (80-95): (20-5) is taken as a mobile phase, the pH value of the solution is 5.6-8.0, and the column temperature is 25-45 DEG C. the invention also relates to a quantitative determination method of ergothioneine.

Description

technical field [0001] The invention belongs to the fields of biochemical technology and analytical chemistry, and relates to a method for determining ergothioneine and its content. Background technique [0002] Ergothioneine (L-Ergothioneine, EGT) is a rare natural chiral amino acid strong antioxidant, a unique cell physiological protection agent, with anti-oxidation, chelating metal ions, preventing ultraviolet radiation and regulating biological energy , gene expression, immune system and many other functions. Due to the special biological functions and pharmacological activities of ergothioneine, such as: anti-inflammation, defense against nerve tissue degeneration, prevention of copper-induced oxidative damage to DNA and proteins, inhibition of developmental defects, protection of the liver, etc., and humans and animals will not It is generally recognized as safe (GRAS) products without any toxic side effects. Therefore, ergothioneine has broad application prospects in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
Inventor 姜文侠刘琦王洪宇刘伟李运华
Owner TIANJIN SINONOCY BIOLOGICAL TECH CO LTD
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