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Method for determining glycogen content in oyster tissues

A technology of glycogen and content, applied in the direction of measuring device, color/spectral characteristic measurement, material analysis by observing the influence of chemical indicators, etc., can solve the problems of difficult operation process and low operation efficiency, and achieve reasonable operation process , accurate measurement results, stable and accurate glycogen content

Pending Publication Date: 2020-06-09
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is true that this sampling method with a fixed amount will facilitate the subsequent calculation of glycogen content, but when large quantities of samples need to be weighed, the operating efficiency of this sampling method is too low. Analytical balance (100,000th place balance, precision 0.01mg) is the most ideal instrument for weighing, while laboratory routine analytical balance is one ten thousandth balance (precision 0.1mg), it is difficult to use conventional analytical balance in the actual operation process

Method used

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  • Method for determining glycogen content in oyster tissues
  • Method for determining glycogen content in oyster tissues
  • Method for determining glycogen content in oyster tissues

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] 1. Reagents and instruments

[0072] (1) Reagents:

[0073] Anthrone (Anthrone>98.0% (GC), Japan, TOKYO CHEMICAL INDUSTRY CO.,LTD);

[0074] Glucose standard (D(+)-anhydrous glucose standard, China, Beijing Suolaibao Technology Co., Ltd.);

[0075] Potassium hydroxide (excellent grade, China, Kemiou Chemical Reagent Co., Ltd.);

[0076] Concentrated sulfuric acid (excellent grade, China, Sinopharm Chemical Reagent Co., Ltd.).

[0077] (2) Instrument:

[0078] Microplate reader (Multiskan Sky Microplate Spectrophotometer, USA, ThermoScientific TM );

[0079] Microplate ( 96-well Clear Flat Bottom Polystyrene NBSMicroplate, USA, Corning); choosing an ordinary flat-bottom 96-well plate can meet the needs of routine homogeneous non-binding absorption light analysis experiments, without using expensive special surface-treated microplates.

[0080] Freeze-dryer (Operating Manual Freeze-dryerALPHA 1-4LDplus, Germany, CHRIST);

[0081] Pipette gun (Eppendorf plus, th...

Embodiment 2

[0128] Determination of glycogen content (dry weight) in the same tissue of the same oyster individual with different sampling amounts

[0129] 1. Preparation of glycogen test solution

[0130] (1) Take an appropriate amount of fresh oyster adductor muscle, gills, mantle, hepatopancreas, gonads, and lips, and put them in a 1.5mL EP tube (the tissue in the EP tube should not be too much, preferably within 1g, to prevent freeze-drying Incomplete), freeze-dried for 48 hours in the main drying process of a freeze-dryer;

[0131] (2) Grinding the freeze-dried sample into powder, wherein the adductor muscle and the mantle are more difficult to grind into powder, and only need to be flocculent;

[0132] (3) Analytical balance Weigh 18.0-22.0 mg of the freeze-dried tissue of two oysters, and the weighed amount is shown in Table 1 below.

[0133] Table 1. Tissue weight after freeze-drying of two oysters in mg

[0134]

[0135] (2) Add the above sample into a 10mL graduated test t...

Embodiment 3

[0172] Effects of the ratio of different color reagents and glycogen detection solution on the determination results

[0173] Among them, the 0.2% anthrone sulfuric acid reagent is the chromogenic agent, and the glycogen detection solution is the glycogen test solution diluted in the measurement process.

[0174] (1) Preparation of glycogen test solution

[0175] Six kinds of oyster tissues were taken to prepare the glycogen test solution, and the specific steps were the same as the preparation of the glycogen test solution in the above-mentioned Example 2.

[0176] (2) Set the ratio of different chromogens to the glycogen detection solution, and the addition amount of each component in the reaction system is shown in Table 5.

[0177] Table 5. The ratio of different color reagents to glycogen detection solution

[0178]

[0179] (3) Standard curve creation

[0180] Different reaction systems should prepare their own standard curves, and the specific steps are the same a...

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Abstract

The invention provides a method for determining glycogen content in oyster tissues, and belongs to the field of determination of glycogen content in animal tissues, and the method comprises the following steps: preparing a to-be-determined solution, making a standard curve, and determining glycogen content of a sample. In the sample weighing step, a dry weight sample of any mass in an interval range of 18.0-22.0 mg is weighed, for example, a high-precision analytical balance is used to make the sample accurate to 2 bits after a decimal point; meanwhile, a whole set of operation process for determining the glycogen content by an anthrone colorimetric method under a small reaction system (300mu L system) is constructed. The method has the advantages of simple sampling, less used chemical reagents, small reaction system, detection cost saving, and experiment risk reduction.

Description

technical field [0001] This research field relates to the determination method of glycogen content in animal tissue, especially the operation method of anthrone colorimetric method to determine the glycogen content of oyster tissue. Background technique [0002] Glycogen, also known as animal starch, is the main form of sugar stored in humans and other animals. It is a kind of polysaccharide, which is formed by the dehydration condensation of glucose. The main biological function is as an energy storage material for animals. Studies have shown that glycogen has biological activities such as hangover protection, anti-fatigue, and anti-tumor. Glycogen is also the most direct and effective energy storage substance in most aquatic shellfish, and plays a vital role in its physiological metabolism. [0003] The glycogen content of oysters usually accounts for 20%-40% of its dry weight, which can be directly absorbed and utilized by the human body. A large number of studies have ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31G01N21/78
CPCG01N21/31G01N21/78G01N2021/775
Inventor 陈夕刘志鸿吴彪周丽青孙秀俊杨爱国
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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