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Method for extracting residual protein in edible vegetable oil

A technology for edible vegetable oil and edible oil, applied in the biological field, can solve problems such as unreported, and achieve the effects of wide applicability, low cost and high yield

Inactive Publication Date: 2014-08-20
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, methods for extracting sufficient residual protein from edible oils for subsequent qualitative and quantitative studies have not been reported

Method used

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  • Method for extracting residual protein in edible vegetable oil

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] (1) Add 36-44ml edible oil and 9-11ml extract to the centrifuge tube and mix well; the extract consists of 0.2% SDS, pH 7.5, 0.1 M Tris-HCl, 0.5 M NaCl, 10 mM DTT;

[0035] (2) Centrifuge the mixture of edible oil and extract obtained in step (1) with a centrifugal force of 8000-12000g for 4-6 min; (3) Take the mixture of edible oil and extract obtained in step (2) after centrifugation Add 2.5-3.5 ml 50% TCA to the lower aqueous phase, mix well, and place in ice bath for 15-25 min;

[0036] (4) Centrifuge the mixture of the lower aqueous phase and TCA obtained in step (3) with a centrifugal force of 8000-12000g for 8-12 min;

[0037] (5) Discard the supernatant of the mixture of the lower aqueous phase and TCA after centrifugation in step (4), add 1.5-2.5 ml of 80% cold acetone to wash the precipitate, and recover the precipitate;

[0038] (6) Centrifuge the precipitate washed with cold acetone obtained in step (5) with a centrifugal force of 10000-14000g for 4-6 min, ...

Embodiment 2

[0041] (1) Add 38-42ml edible oil and 9.5-10.5ml extract to the centrifuge tube and mix well; the extract consists of 0.2% SDS, pH 7.5, 0.1 M Tris-HCl, 0.5 M NaCl, 10 mM DTT;

[0042] (2) Centrifuge the mixture of edible oil and extract obtained in step (1) with a centrifugal force of 9000-11000g for 4.5-5.5min;

[0043] (3) Take the lower aqueous phase of the mixture of edible oil and extract after centrifugation in step (2), add 2.75-3.25 ml 50% TCA, mix well, and place in ice bath for 17.5-22.5 min;

[0044] (4) Centrifuge the mixture of the lower aqueous phase and TCA obtained in step (3) with a centrifugal force of 9000-11000g for 9-11 min;

[0045] (5) Discard the supernatant of the mixture of the lower aqueous phase and TCA after centrifugation in step (4), add 1.75-2.25 ml of 80% cold acetone to wash the precipitate, and recover the precipitate;

[0046] (6) Centrifuge the precipitate washed with cold acetone obtained in step (5) with a centrifugal force of 11000-1300...

Embodiment 3

[0049] (1) Add 36ml of a certain brand of pressed peanut oil and 9ml of extract to the centrifuge tube and mix well; the extract consists of 0.2% SDS, pH 7.5, 0.1 M Tris-HCl, 0.5 M NaCl, 10 mM DTT;

[0050] (2) Centrifuge the mixture of edible oil and extract obtained in step (1) with a centrifugal force of 8000g for 4 minutes;

[0051] (3) Take the lower aqueous phase of the mixture of edible oil and extract after centrifugation in step (2), add 2.5 ml 50% TCA, mix well, and place in ice bath for 15 minutes;

[0052] (4) Centrifuge the mixture of lower aqueous phase and TCA obtained in step (3) with a centrifugal force of 8000g for 8 min;

[0053] (5) Discard the supernatant of the mixture of the lower aqueous phase and TCA after centrifugation in step (4), add 1.5 ml of 80% cold acetone to wash the precipitate, and recover the precipitate;

[0054] (6) Centrifuge the precipitate washed with cold acetone obtained in step (5) with a centrifugal force of 10,000g for 4 minutes,...

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Abstract

The present invention provides a method for extracting residual protein in edible vegetable oil. According to the method, a SDS extraction solution is adopted to separate residual protein in edible oil, concentration and purification are performed, an electrophoresis technology is adopted to analyze the protein so as to obtain the electrophoregram of the residual protein, and a mass spectrometry technology can further be applied to identify the type of the main residual protein.

Description

technical field [0001] The invention relates to a method for extracting residual protein in edible vegetable oil, which belongs to the field of biotechnology. Background technique [0002] Residual trace protein in edible oil is a potential allergen, and it can also be used as a 'molecular imprint' to distinguish different types, sources and even brands of edible oil, and can also be used as an auxiliary detection indicator for the quality and production process of edible oil. Due to the physical and chemical properties of edible oil and the characteristics of its production process, it is very difficult to extract and separate residual protein from non-polar edible oil. [0003] For a long time, the quality of various edible oils in the market has been uneven, and the situation of substituting inferior or fake ones for real is more serious. For example, low-priced or inferior oils are mixed with high-quality oils for sale, which damages the interests of consumers , endange...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/30C07K1/14
Inventor 王伟吴晓林熊二辉
Owner HENAN AGRICULTURAL UNIVERSITY
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