Gas chromatographic method for simultaneously detecting contents of fatty acids in blood and liver tissue of mouse

A gas chromatography and liver tissue technology, applied in the field of gas chromatography, can solve the problems of high boiling point, strong polarity, instability, etc., and achieve the effects of wide application range, good separation, and good detection effect.

Inactive Publication Date: 2015-08-19
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Fatty acids extracted from tissues (especially long-chain fatty acids with more than 12 carbons) are prone to loss during analysis due to their high boiling point, strong polarity, and instability at high temperatures.

Method used

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  • Gas chromatographic method for simultaneously detecting contents of fatty acids in blood and liver tissue of mouse
  • Gas chromatographic method for simultaneously detecting contents of fatty acids in blood and liver tissue of mouse
  • Gas chromatographic method for simultaneously detecting contents of fatty acids in blood and liver tissue of mouse

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The gas chromatography method detection of fatty acid content in embodiment 1 mouse blood and liver tissue

[0030] 1. Experimental method

[0031] 1.1 Preparation of reagents

[0032] Preparation of 0.5mol / L KOH-methanol solution: Accurately weigh 2.8g of KOH, dissolve in about 80mL of anhydrous methanol, heat to dissolve, cool to room temperature, dilute to 100mL, and store in a refrigerator at 4°C.

[0033] Physiological saline: Accurately weigh 9g of NaCl, dissolve it in water to a volume of 1000mL, and sterilize the obtained solution by moist heat at 120°C for 15min.

[0034]Preparation of mixed standards: Accurately weigh C12:0, C14:0, C15:0, C16:0, C17:0, C18:0, C18:1, C18:2, C18:3, C20:0, C22: 0, C23:0, C20:1, C20:2, C20:3, C20:4, C22:6 each 1mg of fatty acid standard substance, mix and make the mixture of fatty acid standard substance.

[0035] Preparation of fatty acid standard solution: Precisely weigh C12:0, C14:0, C15:0, C16:0, C17:0, C18:0, C18:1, C18:2...

experiment example 1

[0089] Experimental Example 1 Detection of fatty acid content in serum and liver of NAFLD mice

experiment example 2

[0097] Experimental example 2 Chromatographic condition optimization experiment

[0098] 1. Experimental method

[0099] The fatty acid standard mixture was directly saponified and methylated according to Example 1.

[0100] The chromatographic conditions are the same as in Example 1, and 7 processes are established for the heating program:

[0101]1: The initial column temperature is 50°C, keep for 1min, the heating rate is 5°C / min, rise to 70°C and keep for 5min, 15°C / min rise to 170°C, keep for 2min, 10°C / min rise to 200°C, keep for 10min, 2°C / min Min rises to 230°C and keeps for 10 minutes, 5°C / min rises to 240°C and keeps for 1min.

[0102] 2: The initial column temperature is 50°C, keep for 2min, the heating rate is 15°C / min, rise to 170°C and keep for 2min, 10°C / min rise to 200°C and keep for 10min, 2°C / min rise to 240°C and keep for 5min.

[0103] 3: The initial column temperature is 50°C, keep it for 2min, the heating rate is 20°C / min, raise it to 170°C and keep it...

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Abstract

The invention discloses a gas chromatographic method for simultaneously detecting contents of fatty acids in the blood and the liver tissue of a mouse, and belongs to the field of gas chromatographic detection on the contents of the fatty acids. The gas chromatographic method comprises the following steps: (1) collecting the blood of the mouse to be detected, and separating the serum, or extracting the fatty acid in the liver tissue of the mouse to be detected; (2) performing saponification methyl esterification on the separated serum or the extracted fatty acid by the steps (1); and (3) performing qualitative and quantitative measurement by gas chromatography. By optimization of the temperature and the time of saponification methyl esterification and the heating program of a chromatographic condition, the fatty acids in the serum and the liver of the mouse are well separated. The method is suitable for detecting fatty acids of various types and large in boiling point range; by the programmed heating and a high-polarity column, the fatty acids in the tissue can be well separated; the gas chromatographic method has the advantages of high degree of accuracy, stable reproducibility, high sensitivity and the like.

Description

technical field [0001] The invention relates to a gas chromatographic method, in particular to a gas chromatographic method for simultaneously detecting fatty acid content in mouse blood and liver tissue, and belongs to the field of gas chromatographic detection of fatty acid content. Background technique [0002] Fatty acid is an aliphatic carboxylic acid compound produced by the decomposition reaction of natural oil and water. It is an important component of lipids, so the properties of fatty acids will greatly affect the characteristics of triglycerides, phospholipids and glycolipids. There are more than 200 kinds of natural fatty acids found so far, which are widely found in animal and vegetable oils. According to the number of carbon atoms in the molecular structure, it can be divided into short-chain fatty acids (2-6), medium-chain fatty acids (8-12), and long-chain fatty acids (14-26); according to the number of double bonds of carbon atoms (saturation) It can be div...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88
Inventor 张秀英李睿李春艳
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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