A method for extracting abalone hepatopancreas metabolites

A technology of hepatopancreas and metabolites, which is applied in the field of molecular biology, can solve the problems that are not suitable for high-throughput, large-scale use, cannot effectively remove polysaccharides and pigments, and affect the quantification of important metabolites, so as to achieve the elimination of polysaccharides and pigments Interference, high recovery rate, low cost effect

Active Publication Date: 2022-02-18
SOUTH CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method cannot effectively remove polysaccharides and pigments in abalone hepatopancreas
Polysaccharides and pigments will cover up the signals of some important metabolites in the NMR spectrum, which will seriously affect the quantification of these important metabolites, making subsequent analysis impossible
Although polysaccharides and pigments can be effectively removed using ultrafiltration centrifuge tubes, ultrafiltration centrifuge tubes are expensive and not suitable for high-throughput, large-scale use

Method used

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  • A method for extracting abalone hepatopancreas metabolites
  • A method for extracting abalone hepatopancreas metabolites

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Using the method of the present invention to extract abalone hepatopancreas metabolites

[0034] The specific method includes the following steps:

[0035] S1. Weigh 300 mg abalone hepatopancreas and place in a 5 mL centrifuge tube;

[0036] S2. Add 1.2 mL of methanol and 0.6 mL of water to the centrifuge tube in step S1, and add steel balls, and then place it in a low-temperature tissue grinder and vibrate at 4 °C to break;

[0037] S3. Add 1.2 mL of chloroform and 1.2 mL of water to the centrifuge tube in step S2, vortex at 2500 rpm for 1 min, and place it on ice for 10 min;

[0038] S4. Centrifuge at 10000 g for 10 min at 4 °C after standing still, and transfer the upper layer solution to another centrifuge tube;

[0039] S5. Put the upper layer solution obtained in step S4 into a freeze-dried powder by vacuum freeze-drying in a vacuum freeze dryer at -80°C after quick-freezing with liquid nitrogen;

[0040] S6. Add 0.9 mL methanol and 0.1 mL water (90% ...

Embodiment 2

[0047] Example 2 Using the method of the present invention to extract hepatopancreatic metabolites from abalone

[0048] The specific method includes the following steps:

[0049] S1. Weigh 300 mg abalone hepatopancreas and place in a 5 mL centrifuge tube;

[0050] S2. Add 1.2 mL of methanol and 0.6 mL of water to the centrifuge tube in step S1, and add steel balls, and then place it in a low-temperature tissue grinder and vibrate at 4 °C to break;

[0051] S3. Add 1.2 mL of chloroform and 1.2 mL of water to the centrifuge tube in step S2, vortex at 2500 rpm for 1 min, and place it on ice for 10 min;

[0052] S4. Centrifuge at 10000 g for 10 min at 4 °C after standing still, and transfer the upper layer solution to another centrifuge tube;

[0053] S5. Put the upper layer solution obtained in step S4 into a freeze-dried powder by vacuum freeze-drying in a vacuum freeze dryer at -80°C after quick-freezing with liquid nitrogen;

[0054] S6. Add 0.9 mL methanol and 0.16 mL wat...

Embodiment 3

[0061] Example 3 Using the method of the present invention to extract abalone hepatopancreas metabolites

[0062] The specific method includes the following steps:

[0063] S1. Weigh 300 mg abalone hepatopancreas and place in a 5 mL centrifuge tube;

[0064] S2. Add 1.2 mL of methanol and 0.6 mL of water to the centrifuge tube in step S1, and add steel balls, and then place it in a low-temperature tissue grinder and vibrate at 4 °C to break;

[0065] S3. Add 1.2 mL of chloroform and 1.2 mL of water to the centrifuge tube in step S2, vortex at 2500 rpm for 1 min, and place it on ice for 10 min;

[0066] S4. Centrifuge at 10000 g for 10 min at 4 °C after standing still, and transfer the upper layer solution to another centrifuge tube;

[0067] S5. Put the upper layer solution obtained in step S4 into a freeze-dried powder by vacuum freeze-drying in a vacuum freeze dryer at -80°C after quick-freezing with liquid nitrogen;

[0068] S6. Add 0.9 mL methanol and 0.23 mL water (80%...

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Abstract

The invention relates to the technical field of molecular biology, in particular to a method for extracting hepatopancreas metabolites from abalone. place, centrifuge, and collect the first supernatant; freeze-dry the first supernatant, add methanol aqueous solution, ultrasonic, stand still, centrifuge, collect the second supernatant; collect the remaining precipitate when the second supernatant is added Methanol aqueous solution, ultrasonication, standing still, centrifugation, and collecting the third supernatant; the second supernatant and the third supernatant were mixed and freeze-dried, added phosphate buffer, reconstituted by shaking, and centrifuged at high speed, the obtained fourth The supernatant is water-soluble abalone hepatopancreas metabolites. The invention completely eliminates the interference of polysaccharides and pigments in the abalone hepatopancreas, and has simple operation, low cost, strong stability, and high recovery rate. High animal and plant tissue metabolites can be used for reference.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a method for extracting abalone hepatopancreas metabolites. Background technique [0002] Abalone is known as the "crown of the eight treasures" of marine products. It has high nutritional and medicinal value. It is an important economically cultured shellfish in the coastal areas of my country, and it is also an ideal marine environment monitoring indicator organism. The hepatopancreas is an important digestive tissue of abalone, and it is also an important immune tissue against external environmental stress. The extraction of metabolites in the hepatopancreas is an important link in the study of metabolomics. In nuclear magnetic resonance (NMR)-based metabolomics studies, liquid-liquid extraction using methanol / chloroform / water as the extraction solvent is most widely used in the extraction of polar metabolites. However, this method cannot effectively remove polysac...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/28G01N24/08
CPCG01N1/28G01N24/08
Inventor 卢洁姚托叶灵通王江勇
Owner SOUTH CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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