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Sea pumpkin and sea pumpkin product sea pumpkin polysaccharide content determination method

A technology of sea cucumber polysaccharide and determination method, applied in the directions of measuring device, biological test, material inspection product, etc., can solve the problems of complex variety and difficult to satisfy, and achieve the effects of good stability, short detection time and high sensitivity

Inactive Publication Date: 2008-10-15
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the variety of sea cucumber products currently on the market is complex, so it is difficult to use simple spectrophotometry or weighing methods to meet the requirements of establishing quality standards for sea cucumber products with sea cucumber polysaccharides as evaluation indicators.

Method used

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  • Sea pumpkin and sea pumpkin product sea pumpkin polysaccharide content determination method
  • Sea pumpkin and sea pumpkin product sea pumpkin polysaccharide content determination method
  • Sea pumpkin and sea pumpkin product sea pumpkin polysaccharide content determination method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0010] The mensuration of polysaccharide content in the sea cucumber peptide of embodiment 1

[0011] (1) Accurately weigh 6g of sample A sea cucumber peptide, grind it finely, add 30ml of sodium acetate buffer solution with a concentration of 0.05M / L, pH 6 and 100mg of papain, react at 37°C for 24h, centrifuge, and take Add 2 times the volume of ethanol to the clear liquid to precipitate the sea cucumber polysaccharide produced after enzymatic hydrolysis, dissolve the precipitate obtained by centrifugation with water, and dilute to 3ml;

[0012] (2) Take 0.5ml of the obtained liquid and add an equal volume of 2mol / L trifluoroacetic acid (TFA) aqueous solution to hydrolyze at 120°C for 2 hours, dry the obtained hydrolyzate with nitrogen, neutralize with 0.3M aqueous sodium hydroxide solution, and After distilling to 2ml, it was used as the sample test solution.

[0013] (3) Accurately weigh the fucose standard substance with water to prepare a 1 mmol / L aqueous solution as a s...

Embodiment 2

[0017] The mensuration of sea cucumber polysaccharide content in dry sea cucumber of embodiment 2

[0018] (1) Accurately weigh 10 g of commercially available dry ginseng (Apostichopus japonicus) sample B, add 60 ml of sodium acetate buffer solution with a concentration of 0.05 M / L and a pH of 7.5 and 1 g of trypsin for enzymolysis at 60°C for 24 hours, and digest the obtained enzyme After the solution was centrifuged, 3 times the volume of ethanol was used to precipitate the sea cucumber polysaccharide in the obtained supernatant, and the obtained precipitate was adjusted to 5 ml.

[0019] (2) Take 0.5ml of the above liquid, add an equal volume of 6mol / L TFA, seal the tube with nitrogen, and hydrolyze at 110°C for 8h; Sample for test solution.

[0020] (3) Get 250 μ L of the test solution, carry out derivation and liquid chromatography separation according to (3)-(5) steps in Example 1, and obtain the relevant working curve y=9675x+9.0095 (R 2 = 0.9986). The measured peak ...

Embodiment 3

[0021] The mensuration of sea cucumber polysaccharide content in the sea cucumber granule of embodiment 3

[0022] (1) Take 1 g each of sea cucumber granule sample C and sea cucumber granule sample D, add 60 ml of potassium acetate buffer solution with a concentration of 0.05M / L and pH of 6.5 and add 1 mg of papain for enzymolysis, centrifuge the obtained enzymatic hydrolyzate, and use 2 times the volume of ethanol to precipitate the sea cucumber polysaccharide in the obtained supernatant, centrifuge, and settle the precipitate to 100ml.

[0023] (2) Take 0.5ml of the above liquid, add an equal volume of 1mol / L trifluoroacetic acid, seal the tube with nitrogen, and hydrolyze at 110°C for 1 hour; dry the hydrolyzate with nitrogen, neutralize it with 0.3M sodium hydroxide, and set the volume to 2ml Provide the test solution for the sample.

[0024] Get 250 μ L of test solution, carry out derivation and liquid chromatography separation according to (3)-(5) step in embodiment 1, ...

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Abstract

The invention discloses a method for determining sea cucumber polysaccharide content in a sea cucumber or a sea cucumber product. The method is characterized in that the sea cucumber or the sea cucumber product is enzymolyzed by protease; the sea cucumber polysaccharide produced by the enzymolysis is separated out by adopting alcohol precipitation; then tested solution is prepared by hydrolysis of trifluoroacetate; the tested solution is prepared into fucose standard water solution which is derived by 1-phenyl-3-methyl-5-pyrazolone; the peak area of fucose derivative in the obtained derivatives is measured by high performance liquid chromatography; and the sea cucumber polysaccharide content in the sea cucumber product is obtained by calculating with the fucose as a reference. The invention provides a simple, accurate, reliable and standardized quantitative analysis method for determining the sea cucumber polysaccharide in various sea cucumbers and sea cucumber products.

Description

technical field [0001] The invention relates to a method for determining macromolecular compounds, in particular to a method for determining the content of sea cucumber polysaccharides in sea cucumbers and sea cucumber products. Background technique [0002] Sea cucumber polysaccharide is an important active ingredient in sea cucumber. The structure of polysaccharides in sea cucumbers is complex and varies with the species of sea cucumbers and the growth environment. The polysaccharide structures in different species of sea cucumbers are different, and the contents of sea cucumber fucoidan sulfate (SC-FUC) and sea cucumber chondroitin sulfate (SC-CHS) in the same species are also different. In addition, sea cucumbers also contain a large amount of complex components such as protein, polysaccharides, fats, and trace elements, which will inevitably bring many difficulties to the determination of total polysaccharides in sea cucumbers. [0003] Existing at present about the r...

Claims

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Application Information

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IPC IPC(8): G01N33/48G01N30/02
Inventor 薛长湖陈士国尹利昂李兆杰薛勇徐杰董平常耀光
Owner OCEAN UNIV OF CHINA
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