CN103992370A - Method for extracting residual protein in edible vegetable oil - Google Patents
Method for extracting residual protein in edible vegetable oil Download PDFInfo
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- CN103992370A CN103992370A CN201410124871.4A CN201410124871A CN103992370A CN 103992370 A CN103992370 A CN 103992370A CN 201410124871 A CN201410124871 A CN 201410124871A CN 103992370 A CN103992370 A CN 103992370A
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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
The present invention relates to a kind of method of extracting residual protein in edible vegetable oil, belong to biological technical field.
Background technology
In edible oil, residual trace protein is potential anaphylactogen, also can be used as ' molecular imprinting '---distinguish different sorts, the even edible oil of brand of originating, also can be used as the auxiliary detection index of edible oil quality and production technique.Due to physico-chemical property and the production technology characteristic thereof of edible oil, from nonpolar edible oil, the protein of extraction, separating residual is very difficult.
For a long time, on market, the quality of various edible oils ginseng is time uneven, and the situation of adulterating, passing a fake product off as a genuine one is comparatively serious, as low price or poor quality oil mixed in oil of high quality and sold, infringement consumer's interests, harm consumer health.At present, from edible oil, extract enough residual proteins and have not yet to see report for the method for follow-up quantitative and qualitative analysis research.
Summary of the invention
The technical problem to be solved in the present invention is to extract the residual protein of edible oil, for addressing the above problem, the present invention a kind of fast and convenient method of extracting residual protein from edible oil is provided.
Technical scheme of the present invention realizes in the following manner:
A method of extracting residual protein in edible vegetable oil, it is comprised of following steps:
(1) in centrifuge tube, add 36～44ml edible oil and 9 ~ 11ml extracting solution to mix;
(2) edible oil step (1) being obtained and the mixed solution of extracting solution are with centrifugal 4～6 min of centrifugal force of 8000～12000g;
(3) get lower floor's water of the mixed solution of the edible oil of step (2) after centrifugal and extracting solution, add 2.5～3.5 ml 50%TCA, after fully mixing, ice bath 15～25 min;
(4) lower floor's water step (3) being obtained and the mixture of TCA are with centrifugal 8～12 min of centrifugal force of 8000～12000g;
(5) the lower floor's water after step (4) is centrifugal and the mixture of TCA are abandoned supernatant, add 1.5～2.5 ml 80% cold acetones to rinse precipitation, reclaim precipitation;
(6) precipitation that the cold acetone that step (5) obtained rinses is with centrifugal 4～6 min of centrifugal force of 10000～14000g, abandons supernatant after centrifugal, obtains protein precipitation;
(7) room temperature is dried step (6) and is obtained protein precipitation.
A method of extracting residual protein in edible vegetable oil, it is comprised of following steps:
(1) in centrifuge tube, add 38～42ml edible oil and 9.5 ~ 10.5ml extracting solution to mix;
(2) edible oil step (1) being obtained and the mixed solution of extracting solution are with the centrifugal 4.5～5.5min of centrifugal force of 9000～11000g;
(3) get lower floor's water of the mixed solution of the edible oil of step (2) after centrifugal and extracting solution, add 2.75～3.25 ml 50%TCA, after fully mixing, ice bath 17.5～22.5 min;
(4) lower floor's water step (3) being obtained and the mixture of TCA are with centrifugal 9～11 min of centrifugal force of 9000～11000g;
(5) the lower floor's water after step (4) is centrifugal and the mixture of TCA are abandoned supernatant, add 1.75～2.25 ml 80% cold acetones to rinse precipitation, reclaim precipitation;
(6) precipitation that the cold acetone that step (5) obtained rinses is with centrifugal 4.5～5.5 min of centrifugal force of 11000～13000g, abandons supernatant after centrifugal, obtains protein precipitation;
(7) room temperature is dried step (6) and is obtained protein precipitation.
A method of extracting residual protein in edible vegetable oil, it is comprised of following steps:
(1) in centrifuge tube, add 40ml edible oil and 10ml extracting solution to mix;
(2) edible oil step (1) being obtained and the mixed solution of extracting solution are with the centrifugal 5min of centrifugal force of 10000g;
(3) get lower floor's water of the mixed solution of the edible oil of step (2) after centrifugal and extracting solution, add 3 ml 50%TCA, after fully mixing, ice bath 20 min;
(4) lower floor's water step (3) being obtained and the mixture of TCA are with centrifugal 10 min of centrifugal force of 10000g;
(5) the lower floor's water after step (4) is centrifugal and the mixture of TCA are abandoned supernatant, add 2 ml 80% cold acetones to rinse precipitation, reclaim precipitation;
(6) precipitation that the cold acetone that step (5) obtained rinses is with the centrifugal 5min of centrifugal force of 12000 g, abandons supernatant after centrifugal, obtains protein precipitation;
(7) room temperature is dried step (6) and is obtained protein precipitation.
Described extracting solution is by 0.2% SDS, pH 7.5,0.1 M Tris-HCl, and 0.5 M NaCl, 10 mM DTT form.
With respect to prior art, the present invention is easy fast, and 1 h can complete; Suitability is wide, and the extraction that can be used for residual protein in various edible oils is separated; Cost is low, does not need expensive test kit; Productive rate is high.
Accompanying drawing explanation
Fig. 1 is the SDS-PAGE electrophoretic analysis collection of illustrative plates of residual protein in certain brand squeezing peanut oil.
Embodiment
Embodiment 1
(1) in centrifuge tube, add 36～44ml edible oil and 9 ~ 11ml extracting solution to mix; Extracting solution is by 0.2% SDS, pH 7.5,0.1 M Tris-HCl, and 0.5 M NaCl, 10 mM DTT form;
(2) edible oil step (1) being obtained and the mixed solution of extracting solution are with centrifugal 4～6 min of centrifugal force of 8000～12000g; (3) get lower floor's water of the mixed solution of the edible oil of step (2) after centrifugal and extracting solution, add 2.5～3.5 ml 50%TCA, after fully mixing, ice bath 15～25 min;
(4) lower floor's water step (3) being obtained and the mixture of TCA are with centrifugal 8～12 min of centrifugal force of 8000～12000g;
(5) the lower floor's water after step (4) is centrifugal and the mixture of TCA are abandoned supernatant, add 1.5～2.5 ml 80% cold acetones to rinse precipitation, reclaim precipitation;
(6) precipitation that the cold acetone that step (5) obtained rinses is with centrifugal 4～6 min of centrifugal force of 10000～14000g, abandons supernatant after centrifugal, obtains protein precipitation;
(7) room temperature is dried step (6) and is obtained protein precipitation.
Embodiment 2
(1) in centrifuge tube, add 38～42ml edible oil and 9.5 ~ 10.5ml extracting solution to mix; Extracting solution is by 0.2% SDS, pH 7.5,0.1 M Tris-HCl, and 0.5 M NaCl, 10 mM DTT form;
(2) edible oil step (1) being obtained and the mixed solution of extracting solution are with the centrifugal 4.5～5.5min of centrifugal force of 9000～11000g;
(3) get lower floor's water of the mixed solution of the edible oil of step (2) after centrifugal and extracting solution, add 2.75～3.25 ml 50%TCA, after fully mixing, ice bath 17.5～22.5 min;
(4) lower floor's water step (3) being obtained and the mixture of TCA are with centrifugal 9～11 min of centrifugal force of 9000～11000g;
(5) the lower floor's water after step (4) is centrifugal and the mixture of TCA are abandoned supernatant, add 1.75～2.25 ml 80% cold acetones to rinse precipitation, reclaim precipitation;
(6) precipitation that the cold acetone that step (5) obtained rinses is with centrifugal 4.5～5.5 min of centrifugal force of 11000～13000g, abandons supernatant after centrifugal, obtains protein precipitation;
(7) room temperature is dried step (6) and is obtained protein precipitation.
Embodiment 3
(1) in centrifuge tube, add 36ml brand squeezing peanut oil and 9ml extracting solution to mix; Extracting solution is by 0.2% SDS, pH 7.5,0.1 M Tris-HCl, and 0.5 M NaCl, 10 mM DTT form;
(2) edible oil step (1) being obtained and the mixed solution of extracting solution are with the centrifugal 4min of centrifugal force of 8000g;
(3) get lower floor's water of the mixed solution of the edible oil of step (2) after centrifugal and extracting solution, add 2.5 ml 50%TCA, after fully mixing, ice bath 15min;
(4) lower floor's water step (3) being obtained and the mixture of TCA are with centrifugal 8 min of centrifugal force of 8000g;
(5) the lower floor's water after step (4) is centrifugal and the mixture of TCA are abandoned supernatant, add 1.5 ml 80% cold acetones to rinse precipitation, reclaim precipitation;
(6) precipitation that the cold acetone that step (5) obtained rinses is with the centrifugal 4min of centrifugal force of 10000g, abandons supernatant after centrifugal, obtains protein precipitation;
(7) room temperature is dried step (6) and is obtained protein precipitation.Can extract 28.5 μ g residual proteins.
Embodiment 4
(1) in centrifuge tube, add 38ml brand squeezing peanut oil and 9.5ml extracting solution to mix; Extracting solution is by 0.2% SDS, pH 7.5,0.1 M Tris-HCl, and 0.5 M NaCl, 10 mM DTT form;
(2) edible oil step (1) being obtained and the mixed solution of extracting solution are with the centrifugal 4.5min of centrifugal force of 9000g;
(3) get lower floor's water of the mixed solution of the edible oil of step (2) after centrifugal and extracting solution, add 2.75 ml 50%TCA, after fully mixing, ice bath 17.5min;
(4) lower floor's water step (3) being obtained and the mixture of TCA are with centrifugal 9 min of centrifugal force of 9000g;
(5) the lower floor's water after step (4) is centrifugal and the mixture of TCA are abandoned supernatant, add 1.75 ml 80% cold acetones to rinse precipitation, reclaim precipitation;
(6) precipitation that the cold acetone that step (5) obtained rinses is with the centrifugal 4.5min of centrifugal force of 11000g, abandons supernatant after centrifugal, obtains protein precipitation;
(7) room temperature is dried step (6) and is obtained protein precipitation.Can extract 29.25 μ g residual proteins.
Embodiment 5
(1) in centrifuge tube, add 40ml brand squeezing peanut oil and 10ml extracting solution to mix; Extracting solution is by 0.2% SDS, pH 7.5,0.1 M Tris-HCl, and 0.5 M NaCl, 10 mM DTT form;
(2) edible oil step (1) being obtained and the mixed solution of extracting solution are with the centrifugal 5min of centrifugal force of 10000g;
(3) get lower floor's water of the mixed solution of the edible oil of step (2) after centrifugal and extracting solution, add 3ml 50%TCA, after fully mixing, ice bath 20min;
(4) lower floor's water step (3) being obtained and the mixture of TCA are with centrifugal 10 min of centrifugal force of 10000g;
(5) the lower floor's water after step (4) is centrifugal and the mixture of TCA are abandoned supernatant, add 2 ml 80% cold acetones to rinse precipitation, reclaim precipitation;
(6) precipitation that the cold acetone that step (5) obtained rinses is with the centrifugal 5min of centrifugal force of 12000g, abandons supernatant after centrifugal, obtains protein precipitation;
(7) room temperature is dried step (6) and is obtained protein precipitation.Can extract 30 μ g residual proteins.
Embodiment 6
(1) in centrifuge tube, add 42ml brand squeezing peanut oil and 10.5ml extracting solution to mix; Extracting solution is by 0.2% SDS, pH 7.5,0.1 M Tris-HCl, and 0.5 M NaCl, 10 mM DTT form;
(2) edible oil step (1) being obtained and the mixed solution of extracting solution are with the centrifugal 5.5min of centrifugal force of 11000g;
(3) get lower floor's water of the mixed solution of the edible oil of step (2) after centrifugal and extracting solution, add 3.25ml 50%TCA, after fully mixing, ice bath 22.5min;
(4) lower floor's water step (3) being obtained and the mixture of TCA are with centrifugal 11 min of centrifugal force of 11000g;
(5) the lower floor's water after step (4) is centrifugal and the mixture of TCA are abandoned supernatant, add 2 .25ml 80% cold acetones to rinse precipitation, reclaim precipitation;
(6) precipitation that the cold acetone that step (5) obtained rinses is with the centrifugal 5.5min of centrifugal force of 13000g, abandons supernatant after centrifugal, obtains protein precipitation;
(7) room temperature is dried step (6) and is obtained protein precipitation.Can extract 30.75 μ g residual proteins.
Embodiment 7
(1) in centrifuge tube, add 44ml brand squeezing peanut oil and 11ml extracting solution to mix; Extracting solution is by 0.2% SDS, pH 7.5,0.1 M Tris-HCl, and 0.5 M NaCl, 10 mM DTT form;
(2) edible oil step (1) being obtained and the mixed solution of extracting solution are with the centrifugal 6min of centrifugal force of 12000g;
(3) get lower floor's water of the mixed solution of the edible oil of step (2) after centrifugal and extracting solution, add 3.5ml 50%TCA, after fully mixing, ice bath 25min;
(4) lower floor's water step (3) being obtained and the mixture of TCA are with centrifugal 12 min of centrifugal force of 12000g;
(5) the lower floor's water after step (4) is centrifugal and the mixture of TCA are abandoned supernatant, add 2 .5ml 80% cold acetones to rinse precipitation, reclaim precipitation;
(6) precipitation that the cold acetone that step (5) obtained rinses is with the centrifugal 6min of centrifugal force of 14000g, abandons supernatant after centrifugal, obtains protein precipitation;
(7) room temperature is dried step (6) and is obtained protein precipitation.Can extract 31.5 μ g residual proteins.
Above-described is only the preferred embodiment of the present invention; should be understood that; for a person skilled in the art; do not departing under general idea prerequisite of the present invention; can also make some changes and improvements; these also should be considered as protection scope of the present invention, and these can not affect effect of the invention process and practical applicability.
Claims (4)
1. a method of extracting residual protein in edible vegetable oil, is characterized in that: it is comprised of following steps:
(1) in centrifuge tube, add 36～44ml edible oil and 9 ~ 11ml extracting solution to mix;
(2) edible oil step (1) being obtained and the mixed solution of extracting solution are with centrifugal 4～6 min of centrifugal force of 8000～12000g;
(3) get lower floor's water of the mixed solution of the edible oil of step (2) after centrifugal and extracting solution, add 2.5～3.5 ml 50%TCA, after fully mixing, ice bath 15～25 min;
(4) lower floor's water step (3) being obtained and the mixture of TCA are with centrifugal 8～12 min of centrifugal force of 8000～12000g;
(5) the lower floor's water after step (4) is centrifugal and the mixture of TCA are abandoned supernatant, add 1.5～2.5 ml 80% cold acetones to rinse precipitation, reclaim precipitation;
(6) precipitation that the cold acetone that step (5) obtained rinses is with centrifugal 4～6 min of centrifugal force of 10000～14000g, abandons supernatant after centrifugal, obtains protein precipitation;
(7) room temperature is dried step (6) and is obtained protein precipitation.
2. the method for residual protein in extraction edible vegetable oil according to claim 1, is characterized in that: it is comprised of following steps:
(1) in centrifuge tube, add 38～42ml edible oil and 9.5 ~ 10.5ml extracting solution to mix;
(2) edible oil step (1) being obtained and the mixed solution of extracting solution are with the centrifugal 4.5～5.5min of centrifugal force of 9000～11000g;
(3) get lower floor's water of the mixed solution of the edible oil of step (2) after centrifugal and extracting solution, add 2.75～3.25 ml 50%TCA, after fully mixing, ice bath 17.5～22.5 min;
(4) lower floor's water step (3) being obtained and the mixture of TCA are with centrifugal 9～11 min of centrifugal force of 9000～11000g;
(5) the lower floor's water after step (4) is centrifugal and the mixture of TCA are abandoned supernatant, add 1.75～2.25 ml 80% cold acetones to rinse precipitation, reclaim precipitation;
(6) precipitation that the cold acetone that step (5) obtained rinses is with centrifugal 4.5～5.5 min of centrifugal force of 11000～13000g, abandons supernatant after centrifugal, obtains protein precipitation;
(7) room temperature is dried step (6) and is obtained protein precipitation.
3. the method for residual protein in extraction edible vegetable oil according to claim 1 and 2, is characterized in that: it is comprised of following steps:
(1) in centrifuge tube, add 40ml edible oil and 10ml extracting solution to mix;
(2) edible oil step (1) being obtained and the mixed solution of extracting solution are with the centrifugal 5min of centrifugal force of 10000g;
(3) get lower floor's water of the mixed solution of the edible oil of step (2) after centrifugal and extracting solution, add 3 ml 50%TCA, after fully mixing, ice bath 20 min;
(4) lower floor's water step (3) being obtained and the mixture of TCA are with centrifugal 10 min of centrifugal force of 10000g;
(5) the lower floor's water after step (4) is centrifugal and the mixture of TCA are abandoned supernatant, add 2 ml 80% cold acetones to rinse precipitation, reclaim precipitation;
(6) precipitation that the cold acetone that step (5) obtained rinses is with the centrifugal 5min of centrifugal force of 12000 g, abandons supernatant after centrifugal, obtains protein precipitation;
(7) room temperature is dried step (6) and is obtained protein precipitation.
4. according to the method for residual protein in the extraction edible vegetable oil described in claim 1 or 2 or 3, it is characterized in that: described extracting solution is by 0.2% SDS, pH 7.5,0.1 mol/L Tris-HCl, 0.5 mol/L NaCl, 10 mmol/L DTT form.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101171952A (en) * | 2007-03-15 | 2008-05-07 | 云南神宇新能源有限公司 | Method for extracting jatropha curcas seed endosperm protein |
| CN101696238A (en) * | 2009-10-27 | 2010-04-21 | 广东省农业科学院作物研究所 | Plant total protein extracting solution and application thereof |
| CN102288565A (en) * | 2011-07-29 | 2011-12-21 | 河南省农科院农副产品加工研究所 | Method for rapidly detecting adulteration of sesame oil |
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- 2014-03-31 CN CN201410124871.4A patent/CN103992370A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101171952A (en) * | 2007-03-15 | 2008-05-07 | 云南神宇新能源有限公司 | Method for extracting jatropha curcas seed endosperm protein |
| CN101696238A (en) * | 2009-10-27 | 2010-04-21 | 广东省农业科学院作物研究所 | Plant total protein extracting solution and application thereof |
| CN102288565A (en) * | 2011-07-29 | 2011-12-21 | 河南省农科院农副产品加工研究所 | Method for rapidly detecting adulteration of sesame oil |
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