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A method for detecting and determining covalently linked sulfate groups of Cordyceps polysaccharides

A Cordyceps polysaccharide and sulfate technology, applied in the field of medicine, can solve the problems of large sample consumption, high detection limit, and the existence form of sulfur element cannot be explained, and achieves the effect of high detection sensitivity and less sample consumption.

Active Publication Date: 2015-08-19
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods have many shortcomings in the detection of sulfate radicals. For example, the elemental analysis method has high requirements on the purity of the sample, and has no ability to distinguish impurities. The form (organic sulfur, inorganic sulfur) cannot be specified
The barium sulfate gelatin turbidimetric method consumes a lot of samples, the detection limit is high, and the reproducibility of the results is not good
Neither of these two methods can rule out the interference of impurities, and it is impossible to determine whether the sulfur element comes from polysaccharides, medium or protein, and the sample consumption is large
At present, there is no report about Cordyceps polysaccharides containing covalently linked sulfate groups, which is due to the limitation of detection methods.

Method used

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  • A method for detecting and determining covalently linked sulfate groups of Cordyceps polysaccharides
  • A method for detecting and determining covalently linked sulfate groups of Cordyceps polysaccharides
  • A method for detecting and determining covalently linked sulfate groups of Cordyceps polysaccharides

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Experimental program
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Effect test

Embodiment 1

[0026] Strain amplification: PDA solid medium was used to amplify the strain of Paecilomyces batata, and cultured at 27°C for 7 days;

[0027] Seed liquid culture: pick mycelium from the solid medium and put it in 150mL potato liquid medium, put it on a shaker at 180r / min, and cultivate it at 27°C for 10 days;

[0028] Take 6mL seed solution and add 1ug / mL Na 2 34 SO 4 In 200mL potato liquid medium, cultured at 27°C for 10 days.

Embodiment 2

[0030] Strain amplification and seed liquid culture steps are the same as in Example 1;

[0031] Take 6mL seed solution and add 1000ug / mLNa 2 34 SO 4 In 200mL potato liquid medium, cultured at 27°C for 10 days.

Embodiment 3

[0033] Strain amplification and seed liquid culture steps are the same as in Example 1;

[0034] Take 6mL seed solution and add Na-free 2 34 SO 4 In 200mL potato liquid medium, cultured at 27°C for 10 days.

[0035] The mycelium that embodiment 1-3 cultivates extracts Cordyceps polysaccharide according to the following method:

[0036] Concentrate the culture solution to 1 / 4 volume, add absolute ethanol until the final concentration of ethanol is 80%, reflux at 80° C. for 3 h to degrease. After cooling, it was centrifuged for 10 min, and the precipitate was collected. This was repeated three times to obtain a degreased product. Take 2 g of the defatted product, add 40 mL of water, stir magnetically for 3 h, centrifuge for 10 min, and repeat three times. Dialyze the supernatant, then concentrate the dialysate, add 4 times of absolute ethanol for ethanol precipitation, and obtain the Cordyceps polysaccharide.

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Abstract

The invention belongs to the medicine field, and relates to a method for detecting and determining a covalently linked sulfate radical in cordyceps polysaccharide. The detection steps include: (1) adding Na2<34>SO4 in a liquid culture medium; (2) adding a cordyceps seed liquid into the culture medium of the step (1) for culture; (3) extracting the cordyceps polysaccharide from cordyceps militaris obtained in the step (2); (4) carrying out medium acid degradation of the cordyceps polysaccharide extracted in the step (3), and labeling a degradation product by aniline; and (5) adopting liquid chromatography-mass spectrometry to detect the sulfate radical covalently linked with saccharide in the product obtained in the step (4). The concentration of Na2<34>SO4 added in the culture medium is 1-1000 [mu]g / mL. The method has less using amount of samples, has high sensitivity and safe use, and provides a new index for characterization of biological activity of the cordyceps polysaccharide.

Description

technical field [0001] The invention belongs to the field of medicine, and relates to a method for detecting and determining covalently linked sulfate radicals of Cordyceps polysaccharides. Background technique [0002] Cordyceps (Cordyceps), also known as Cordyceps, Cordyceps, Summer Grass Cordyceps. Belongs to the Kingdom Fungi, Phylum Fungi, Class Ascomycetes, Hypocreales, Ergotaceae, Cordyceps. It is a kind of insect-bacteria complex formed by parasitizing on the larvae of bat moths. It is mainly distributed in high-altitude areas of about 4,000 meters above sea level in Qinghai, Tibet, Sichuan, Yunnan, Guizhou, and Gansu. The natural Cordyceps growth environment is special and the price is relatively high. People have used biological fermentation technology to cultivate Cordyceps mycelium to replace Cordyceps sinensis. Its composition and medicinal value are similar to those of Cordyceps sinensis. Cordyceps polysaccharide is the main active ingredient of Cordyceps sin...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02
Inventor 张丽娟曾洋洋韩章润邱培菊兰莹
Owner OCEAN UNIV OF CHINA