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Method for determining compositions of fatty acids and acylglycerols at alpha and beta positions of structured lipid by combining solid-phase extraction and gas chromatography

A gas chromatography and solid phase extraction small column technology is applied in the field of detecting structural lipids, which can solve the problems of large consumption of organic solvents, time-consuming, and great influence on operation level, and achieve the effect of less consumption of organic solvents.

Inactive Publication Date: 2014-09-17
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

The plate preparation, balance, separation, extraction and other operations of the TLC method are relatively complicated. The tested samples are exposed to the air after spotting, which increases the oxidation probability of fatty acids, especially for some long-chain polyunsaturated fatty acids (LCPUFAs). larger
In addition, because the TLC method is difficult to quantify, and has disadvantages such as a long time and being greatly affected by the operator's operation level, it can only be used as an auxiliary method for qualitative analysis, thus limiting its further application
However, the NP-HPLC instrument is expensive, and the retention time of the corresponding standard needs to be recorded first. When performing sample analysis, each component needs to be collected separately according to the retention time of each component, and the fatty acid is detected by GC, so there is a large consumption of organic solvents. , time-consuming and other disadvantages

Method used

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  • Method for determining compositions of fatty acids and acylglycerols at alpha and beta positions of structured lipid by combining solid-phase extraction and gas chromatography
  • Method for determining compositions of fatty acids and acylglycerols at alpha and beta positions of structured lipid by combining solid-phase extraction and gas chromatography
  • Method for determining compositions of fatty acids and acylglycerols at alpha and beta positions of structured lipid by combining solid-phase extraction and gas chromatography

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Embodiment 1: a kind of method for rapidly detecting α, β position fatty acid in lard, it comprises the steps:

[0056] 1) Sample preparation: Weigh 300mg lard in a 30mL glass test tube, add 20mg porcine pancreatic lipase, 2mL Tris-HCl buffer (pH=8, 2mol / L), shake well, add 0.5mL sodium cholate solution ( 1g / L), 0.2mL CaCl 2 Solution (220g / L), after vortex mixing, shake and react in a water bath at 40°C for 5min, add 1mL anhydrous ether, 1mL HCl solution (6mol / L), shake evenly, centrifuge at 5000r / min for 2min, take the upper layer , blown dry with nitrogen to obtain the sample;

[0057] 2) Column activation: Take a small solid phase extraction column (the filler in the column is 2g florisil), place it on the solid phase extraction device, add 20mL of n-hexane to equilibrate the activation of the column;

[0058] 3) Sample loading: Add 8 mL of n-hexane to dissolve the sample obtained in step 1), and vortex to mix. Take 2mL sample solution and transfer it to the solid...

Embodiment 2

[0063] Embodiment 2: A kind of method utilizing the glyceride composition in the 1,3-arachidonic acid-diacylglycerol (1,3-ARA-DAG) structure lipid that utilizes solid-phase extraction small column rapid separation enzymatic method to synthesize, it Including the following steps:

[0064] 1) Sample preparation: Weigh 1.00 g of arachidonic acid (ARA) and 0.15 g of glycerol, add 0.12 g of immobilized lipase LipozymRMIM (accounting for 10% of the mass of the substrate), and react for 3 hours under magnetic stirring in a constant temperature water bath at 50°C to obtain SLs reaction mixture;

[0065] 2) Column activation: Take a small solid phase extraction column (the filler in the column is 2g florisil), place it on the solid phase extraction device, add 10mL of n-hexane to balance and activate the column;

[0066] 3) Sample loading: Weigh 50 mg of the SLs reaction product synthesized in step 1) into a test tube, add 5 mL of n-hexane, and vortex to mix. Transfer the mixed sampl...

Embodiment 3

[0069] Embodiment 3: a method for rapidly detecting α, β fatty acid in high erucic acid rapeseed oil, it comprises the steps:

[0070] 1) Sample preparation: Accurately weigh 50 mg of rapeseed oil in a 30 mL glass test tube, add 20 mg of porcine pancreatic lipase, 2 mL of Tris-HCl buffer solution (pH=8, 2 mol / L), shake well, add 0.5 mL of sodium cholate Solution (1g / L), 0.2mL CaCl2 solution (220g / L), after vortex mixing, shake and react in a water bath at 40°C for 5min, add 1mL anhydrous ether, 1mLHCl solution (6mol / L), shake evenly , centrifuge at 5000r / min for 2min, take the upper layer, blow dry with nitrogen to obtain a sample;

[0071] 2) Column activation: Take a small solid phase extraction column (the filler in the column is 2g florisil), place it on the solid phase extraction device, add 20mL of n-hexane to equilibrate the activation of the column;

[0072] 3) Sample loading: Add 4 mL of n-hexane to dissolve the sample obtained in step 1), and vortex to mix. Take 2m...

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Abstract

The invention discloses a method for determining compositions of fatty acids and acylglycerols at alpha and beta positions of structured lipid by combining solid-phase extraction and gas chromatography. The method comprises utilizing pure arachidonic acid-triacylglycerol (ARA-TAG), docosahexaenoic acid-diacylglycerol (DAH-DAG), linolenic acid- monoacylglycerol (Ln-MAG) and an oleic acid-free fatty acid (O-FFA) standard sample, and optimizing eluent elution conditions with different polarity ratios, so as to effectively separate TAG, DAG, MAG and FFA in mixtures in a same SPE (Solid Phase Extraction) microcolumn; and the method is characterized by also comprising the following steps: 1) separation of a mixed standard sample and analysis on compositions of fatty acids; and 2) analysis on compositions of fatty acids and acylglycerols at alpha and beta positions of the structured lipid. The method has the advantages of simple operation, low sample demanded amount, less consumption amount of a needed solvent, accurate qualitative quantitative analysis results, low detection limit and short analysis time, and is capable of realizing high-flux detection on TAG, DAG, MAG and FFA compositions and content in structured lipid and compositions of fatty acids at alpha and beta positions of structured lipid.

Description

technical field [0001] The invention relates to a method for detecting structural lipids, in particular to a method for combining solid-phase extraction and gas chromatography to determine fatty acids and glycerides at α and β positions of structural lipids. Background technique [0002] Lipid is the main component of human diet and occupies an important position in the three major energy nutrients of human body. With the continuous improvement of people's living standards, human beings have put forward new requirements for the nutrition of lipids in the diet. Eating good oil and eating nutritious and healthy oil has become a new demand for Chinese consumers to pay attention to their own health and ensure normal nutritional intake. [0003] Studies have shown that the content of TAGs in oil is as high as 95%. The composition of fatty acids in TAGs, the length of carbon chain, the number and position of double bonds are important factors that affect the physical and chemica...

Claims

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Application Information

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IPC IPC(8): G01N30/90G01N30/88
Inventor 董绪燕刘四磊吴琳陈洪魏芳吕昕钟娟
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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