CN106290665A - A kind of detection method of Trans-fatty Acids in Foods - Google Patents
A kind of detection method of Trans-fatty Acids in Foods Download PDFInfo
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- CN106290665A CN106290665A CN201610942770.7A CN201610942770A CN106290665A CN 106290665 A CN106290665 A CN 106290665A CN 201610942770 A CN201610942770 A CN 201610942770A CN 106290665 A CN106290665 A CN 106290665A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/60—Construction of the column
- G01N30/6052—Construction of the column body
- G01N30/6073—Construction of the column body in open tubular form
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Abstract
The invention discloses the detection method of a kind of Trans-fatty Acids in Foods, including: utilize ethanol petroleum ether mixed-liquor return to extract oils and fats；After oil at low temperature cold preservation, it is centrifuged to obtain liquid fat and solid grease；The silver nitrate solution that mass concentration is 2.2% is mixed the 2/3 of filling chromatography column volume with activation 300 mesh silica gel, adds the liquid fat of a column volume, chromatograph, collect and first flow out oils and fats, be combined into trans fats acid crude with solid grease；Take trans fatty acid crude product, add methanolic potassium hydroxide mixed liquor, be heated to reflux obtaining mixed liquor, cooling, add saturated nacl aqueous solution, centrifugal, stay supernatant；In supernatant, add normal hexane, stand, take supernatant dehydration, carry out gas chromatographic analysis.The present invention can completely separate and measure simultaneously the multiple trans fatty acid from low carbon chain to high carbon chain, it is adaptable to carries out qualitative and quantitative detection to common edible oil and containing trans fatty acid kind contained in edible oil converted products, content.
Description
Technical field
The present invention relates to technical field of food detection, the detection method of a kind of Trans-fatty Acids in Foods.
Background technology
The standard method of the mensuration of Trans-fatty Acids in Foods mainly includes gas chromatography and infrared spectrometry.Gas phase color
Spectrometry is the content by first measuring each trans fats acid constituents, and the read group total that then adds up goes out containing of total trans fatty acid
Amount, needs through sample handling processes such as loaded down with trivial details fatty acid saponification, esterifications, in addition it is also necessary to use the fatty acid (methyl ester) of costliness
Standard substance and special fatty acid analysis capillary gas chromatographic column, measure and calculating process is complicated.And infrared spectrum rule is direct
Measure the total content of Trans-fatty Acids in Foods, and the kind of Trans-fatty Acids in Foods and content can not be carried out qualitative, quantitative
Analyze.
Recently, the whole society pays high attention to trans fatty acid, but China does not the most formulate corresponding limit standard,
This just proposes new problem to food production processing enterprise of China, sale intermediate links and government regulation decision-making section, sets up food
In product, the assay method of trans fatty acid becomes urgent needs.
Summary of the invention
It is an object of the invention to provide the detection of the Trans-fatty Acids in Foods that a kind of range is wide and accuracy is high
Method.
For achieving the above object, the present invention provides following technical scheme:
The detection method of a kind of Trans-fatty Acids in Foods, comprises the following steps:
(1) extract oils and fats: take food samples 5～8g, pulverize, add ethanol-petroleum ether mixed liquor 30～35ml, mixing, 56～
58 DEG C of reflux, extract, rotary evaporation obtains oils and fats；
(2) separate trans fatty acid: at a temperature of oils and fats is placed in 2 DEG C after cold preservation 24h, temperature be 4 DEG C, speed be 800～
Low temperature low-speed centrifugal cold preservation oils and fats under conditions of 850r/min, collects liquid fat and solid grease respectively；Then according to nitric acid
Silver is 1:3 with the mass ratio of activation 300 mesh silica gel, is mixed with activation 300 mesh silica gel by the silver nitrate solution that mass concentration is 2.2%
Merge and fill the 2/3 of chromatography column volume, add the liquid fat of a column volume, chromatograph, collect and first flow out oils and fats, treat
When first flowing out reception amount is liquid fat the 12～15% of oils and fats, terminate to collect；This is first flowed out oils and fats and solid grease closes
And, obtain trans fats acid crude；
(3) the trans fats acid crude of certain volume is taken, according to trans fats acid crude: the body of potassium hydroxide-methyl alcohol mixed liquor
Long-pending ratio is 1:15～18, adds potassium hydroxide-methyl alcohol mixed liquor, and mixing, 68～74 DEG C are heated to reflux, and obtain mixed liquor；
(4) mixed liquor is cooled down, according to mixed liquor: the volume ratio of saturated sodium-chloride is 1:4～5, adds saturated nacl aqueous solution,
Centrifugal, abandon lower floor, stay supernatant, standby；
(5) in supernatant, add normal hexane, stand, take its supernatant, after anhydrous cupric sulfate is dehydrated, carry out gas chromatogram
Analyze.
As the further scheme of the present invention: in described step (1), the ethanol in ethanol-petroleum ether mixed liquor and stone
The volume ratio of oil ether is 1:0.42～0.45.
As the further scheme of the present invention: in described step (1), evaporating temperature is 62～65 DEG C, and vacuum is
0.05～0.06MPa.
As the further scheme of the present invention: described step (2) also includes, when after chromatographic column 5 column volumes of chromatography, use
50 DEG C of edible wine fine purifiation posts, use eluate recovered alcohol again.
As the further scheme of the present invention: in described step (3), the hydroxide in potassium hydroxide-methyl alcohol mixed liquor
Potassium: methanol=1g:7～10ml.
As the further scheme of the present invention: in described step (5), the working condition of gas chromatogram is: DB-5 quartz
Capillary column；Carrier gas is the helium of flow velocity 2.5～3.5ml/min, and split ratio is 80～150:1；Injector temperature 280 DEG C；Sample introduction
Measure 0.5 μ L；Heating schedule is: initial temperature is 50～70 DEG C, and keeps 3～4min, then rises to 200 DEG C with 8 DEG C/min, connects
And rise to 230 DEG C with 6 DEG C/min, finally rise to 280 DEG C with 4 DEG C/min, keep 2～3min.
Compared with prior art, the invention has the beneficial effects as follows: the inventive method can completely separate and measure from low simultaneously
Carbochain is to the multiple trans fatty acid of high carbon chain, it is adaptable to common edible oil and containing contained trans in edible oil converted products
Fatty acid species, content carry out qualitative and quantitative detection, and range is wide and accuracy is high.
Detailed description of the invention
Below in conjunction with the embodiment of the present invention, the technical scheme in the embodiment of the present invention is clearly and completely described,
Obviously, described embodiment is only a part of embodiment of the present invention rather than whole embodiments.Based in the present invention
Embodiment, the every other embodiment that those of ordinary skill in the art are obtained under not making creative work premise, all
Belong to the scope of protection of the invention.
Embodiment 1
In the embodiment of the present invention, the detection method of a kind of Trans-fatty Acids in Foods, comprise the following steps:
(1) extract oils and fats: take food samples 5g, pulverize, add ethanol-petroleum ether mixed liquor (wherein ethanol and body of petroleum ether
Long-pending ratio is 1:0.45) 30ml, mixing, 58 DEG C of reflux, extract, rotary evaporation obtains oils and fats；Evaporating temperature is 62 DEG C, and vacuum is
0.06MPa；
(2) separate trans fatty acid: at a temperature of oils and fats is placed in 2 DEG C after cold preservation 24h, temperature be 4 DEG C, speed be 800r/min
Under conditions of low temperature low-speed centrifugal cold preservation oils and fats, collect liquid fat and solid grease respectively；Then according to silver nitrate and activation
The mass ratio of 300 mesh silica gel is 1:3, and the silver nitrate solution that mass concentration is 2.2% is mixed with activation 300 mesh silica gel and filled
The 2/3 of chromatographic column volume, adds the liquid fat of a column volume, chromatographs, and collects and first flows out oils and fats, treats first effluent oil
Fat reception amount is liquid fat 15% time, terminate collect；This is first flowed out oils and fats and solid grease merges, obtain trans fat
Fat acid crude；
(3) the trans fats acid crude of certain volume is taken, according to trans fats acid crude: the body of potassium hydroxide-methyl alcohol mixed liquor
Long-pending ratio is 1:15, adds potassium hydroxide-methyl alcohol mixed liquor (potassium hydroxide: methanol=1g:10ml), mixing, and 68 DEG C are heated to reflux,
Obtain mixed liquor；
(4) mixed liquor is cooled down, according to mixed liquor: the volume ratio of saturated sodium-chloride is 1:5, add saturated nacl aqueous solution, from
The heart, abandons lower floor, stays supernatant, standby；
(5) in supernatant, add normal hexane, stand, take its supernatant, after anhydrous cupric sulfate is dehydrated, carry out gas chromatogram
Analyze；The working condition of gas chromatogram is: DB-5 quartz wool buttress shaft；Carrier gas is the helium of flow velocity 2.5ml/min, and split ratio is
150:1；Injector temperature 280 DEG C；Sample size 0.5 μ L；Heating schedule is: initial temperature is 50 DEG C, and keeps 4min, then with
8 DEG C/min rises to 200 DEG C, then rises to 230 DEG C with 6 DEG C/min, finally rises to 280 DEG C with 4 DEG C/min, keeps 2min.
Embodiment 2
In the embodiment of the present invention, the detection method of a kind of Trans-fatty Acids in Foods, comprise the following steps:
(1) extract oils and fats: take food samples 8g, pulverize, add ethanol-petroleum ether mixed liquor (wherein ethanol and body of petroleum ether
Long-pending ratio is 1:0.42) 35ml, mixing, 56 DEG C of reflux, extract, rotary evaporation obtains oils and fats；Evaporating temperature is 65 DEG C, and vacuum is
0.05MPa；
(2) separate trans fatty acid: at a temperature of oils and fats is placed in 2 DEG C after cold preservation 24h, temperature be 4 DEG C, speed be 850r/min
Under conditions of low temperature low-speed centrifugal cold preservation oils and fats, collect liquid fat and solid grease respectively；Then according to silver nitrate and activation
The mass ratio of 300 mesh silica gel is 1:3, and the silver nitrate solution that mass concentration is 2.2% is mixed with activation 300 mesh silica gel and filled
The 2/3 of chromatographic column volume, adds the liquid fat of a column volume, chromatographs, and collects and first flows out oils and fats, treats first effluent oil
Fat reception amount is liquid fat 12% time, terminate collect；This is first flowed out oils and fats and solid grease merges, obtain trans fat
Fat acid crude；
(3) the trans fats acid crude of certain volume is taken, according to trans fats acid crude: the body of potassium hydroxide-methyl alcohol mixed liquor
Long-pending ratio is 1:18, adds potassium hydroxide-methyl alcohol mixed liquor (potassium hydroxide: methanol=1g:7ml), mixing, and 74 DEG C are heated to reflux,
Mixed liquor；
(4) mixed liquor is cooled down, according to mixed liquor: the volume ratio of saturated sodium-chloride is 1:4, add saturated nacl aqueous solution, from
The heart, abandons lower floor, stays supernatant, standby；
(5) in supernatant, add normal hexane, stand, take its supernatant, after anhydrous cupric sulfate is dehydrated, carry out gas chromatogram
Analyze；The working condition of gas chromatogram is: DB-5 quartz wool buttress shaft；Carrier gas is the helium of flow velocity 3.5ml/min, and split ratio is
80:1；Injector temperature 280 DEG C；Sample size 0.5 μ L；Heating schedule is: initial temperature is 70 DEG C, and keeps 3min, then with 8
DEG C/min rises to 200 DEG C, then rises to 230 DEG C with 6 DEG C/min, finally rises to 280 DEG C with 4 DEG C/min, keeps 3min.
Embodiment 3
In the embodiment of the present invention, the detection method of a kind of Trans-fatty Acids in Foods, comprise the following steps:
(1) extract oils and fats: take food samples 6g, pulverize, add ethanol-petroleum ether mixed liquor (wherein ethanol and body of petroleum ether
Long-pending ratio is 1:0.44) 32ml, mixing, 57 DEG C of reflux, extract, rotary evaporation obtains oils and fats；Evaporating temperature is 63 DEG C, and vacuum is
0.055MPa；
(2) separate trans fatty acid: at a temperature of oils and fats is placed in 2 DEG C after cold preservation 24h, temperature be 4 DEG C, speed be 820r/min
Under conditions of low temperature low-speed centrifugal cold preservation oils and fats, collect liquid fat and solid grease respectively；Then according to silver nitrate and activation
The mass ratio of 300 mesh silica gel is 1:3, and the silver nitrate solution that mass concentration is 2.2% is mixed with activation 300 mesh silica gel and filled
The 2/3 of chromatographic column volume, adds the liquid fat of a column volume, chromatographs, and collects and first flows out oils and fats, treats first effluent oil
Fat reception amount is liquid fat 14% time, terminate collect；This is first flowed out oils and fats and solid grease merges, obtain trans fat
Fat acid crude；
(3) the trans fats acid crude of certain volume is taken, according to trans fats acid crude: the body of potassium hydroxide-methyl alcohol mixed liquor
Long-pending ratio is 1:16, adds potassium hydroxide-methyl alcohol mixed liquor (potassium hydroxide: methanol=1g:8ml), mixing, and 70 DEG C are heated to reflux,
Mixed liquor；
(4) mixed liquor is cooled down, according to mixed liquor: the volume ratio of saturated sodium-chloride is 1:4.5, adds saturated nacl aqueous solution,
Centrifugal, abandon lower floor, stay supernatant, standby；
(5) in supernatant, add normal hexane, stand, take its supernatant, after anhydrous cupric sulfate is dehydrated, carry out gas chromatogram
Analyze；The working condition of gas chromatogram is: DB-5 quartz wool buttress shaft；Carrier gas is the helium of flow velocity 3.0ml/min, and split ratio is
120:1；Injector temperature 280 DEG C；Sample size 0.5 μ L；Heating schedule is: initial temperature is 60 DEG C, and keeps 3.5min, then
Rise to 200 DEG C with 8 DEG C/min, then rise to 230 DEG C with 6 DEG C/min, finally rise to 280 DEG C with 4 DEG C/min, keep 2.5min.
In the food that the various embodiments described above are surveyed, the content of variety classes trans fatty acid is as shown in table 1, table 1 can see
Go out: the inventive method can completely separate and measure simultaneously the multiple trans fatty acid from low carbon chain to high carbon chain, it is adaptable to often
See edible oil and carry out qualitative and quantitative detection containing trans fatty acid kind contained in edible oil converted products, content, using model
Enclose wide and accuracy is high.
The each embodiment of table 1 records result (content of fatty acid: g/100g)
| Fatty acid | Embodiment 1 | Embodiment 2 | Embodiment 3 |
| C16:0 | 12.63 | 13.08 | 13.42 |
| C18:0 | 4.06 | 4.32 | 5.75 |
| C18:1 | 29.84 | 29.13 | 28.52 |
| C18:2 | 51.06 | 49.61 | 48.84 |
| C18:3 | 8.23 | 8.86 | 9.43 |
| C20:0 | 0.39 | 0.44 | 0.46 |
| C20:1 | 0.45 | 0.41 | 0.36 |
It is obvious to a person skilled in the art that the invention is not restricted to the details of above-mentioned one exemplary embodiment, and do not carrying on the back
In the case of the spirit or essential attributes of the present invention, it is possible to realize the present invention in other specific forms.Therefore, no matter from which
From the point of view of a bit, all should regard embodiment as exemplary, and be nonrestrictive, the scope of the present invention is wanted by appended right
Ask rather than described above limits, it is intended that all changes that will fall in the implication of equivalency and scope of claim
Include in the present invention.
Although moreover, it will be appreciated that this specification is been described by according to embodiment, but the most each embodiment only wraps
Containing an independent technical scheme, this narrating mode of description is only that for clarity sake those skilled in the art should
Description can also be formed those skilled in the art through appropriately combined as an entirety, the technical scheme in each embodiment
May be appreciated other embodiments.
Claims (6)
1. the detection method of a Trans-fatty Acids in Foods, it is characterised in that comprise the following steps:
(1) extract oils and fats: take food samples 5～8g, pulverize, add ethanol-petroleum ether mixed liquor 30～35ml, mixing, 56～
58 DEG C of reflux, extract, rotary evaporation obtains oils and fats；
(2) separate trans fatty acid: at a temperature of oils and fats is placed in 2 DEG C after cold preservation 24h, temperature be 4 DEG C, speed be 800～
Low temperature low-speed centrifugal cold preservation oils and fats under conditions of 850r/min, collects liquid fat and solid grease respectively；Then according to nitric acid
Silver is 1:3 with the mass ratio of activation 300 mesh silica gel, is mixed with activation 300 mesh silica gel by the silver nitrate solution that mass concentration is 2.2%
Merge and fill the 2/3 of chromatography column volume, add the liquid fat of a column volume, chromatograph, collect and first flow out oils and fats, treat
When first flowing out reception amount is liquid fat the 12～15% of oils and fats, terminate to collect；This is first flowed out oils and fats and solid grease closes
And, obtain trans fats acid crude；
(3) the trans fats acid crude of certain volume is taken, according to trans fats acid crude: the body of potassium hydroxide-methyl alcohol mixed liquor
Long-pending ratio is 1:15～18, adds potassium hydroxide-methyl alcohol mixed liquor, and mixing, 68～74 DEG C are heated to reflux, and obtain mixed liquor；
(4) mixed liquor is cooled down, according to mixed liquor: the volume ratio of saturated sodium-chloride is 1:4～5, adds saturated nacl aqueous solution,
Centrifugal, abandon lower floor, stay supernatant, standby；
(5) in supernatant, add normal hexane, stand, take its supernatant, after anhydrous cupric sulfate is dehydrated, carry out gas chromatogram
Analyze.
The detection method of Trans-fatty Acids in Foods the most according to claim 1, it is characterised in that described step (1)
In, the ethanol in ethanol-petroleum ether mixed liquor is 1:0.42～0.45 with the volume ratio of petroleum ether.
The detection method of Trans-fatty Acids in Foods the most according to claim 1, it is characterised in that described step (1)
In, evaporating temperature is 62～65 DEG C, and vacuum is 0.05～0.06MPa.
The detection method of Trans-fatty Acids in Foods the most according to claim 1, it is characterised in that described step (2)
Also include, when after chromatographic column 5 column volumes of chromatography, with 50 DEG C of edible wine fine purifiation posts, eluate recovered alcohol is used again.
The detection method of Trans-fatty Acids in Foods the most according to claim 1, it is characterised in that described step (3)
In, the potassium hydroxide in potassium hydroxide-methyl alcohol mixed liquor: methanol=1g:7～10ml.
The detection method of Trans-fatty Acids in Foods the most according to claim 1, it is characterised in that described step (5)
In, the working condition of gas chromatogram is: DB-5 quartz wool buttress shaft；Carrier gas is the helium of flow velocity 2.5～3.5ml/min, split ratio
It is 80～150:1；Injector temperature 280 DEG C；Sample size 0.5 μ L；Heating schedule is: initial temperature is 50～70 DEG C, and keeps 3
～4min, then rise to 200 DEG C with 8 DEG C/min, then rise to 230 DEG C with 6 DEG C/min, finally rise to 280 DEG C with 4 DEG C/min,
Keep 2～3min.
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| CN107132296A (en) * | 2017-06-30 | 2017-09-05 | 华仁药业股份有限公司 | It is a kind of at the same determine special medicine purposes formula food in content of fatty acid method |
| CN110470755A (en) * | 2019-08-09 | 2019-11-19 | 凭祥出入境检验检疫局综合技术服务中心 | A kind of method of quick detection Trans-fatty Acids in Foods |
| CN112326821A (en) * | 2020-10-27 | 2021-02-05 | 杨振全 | Method for detecting lipid content of biological liver tissue |
| CN114740128A (en) * | 2022-05-17 | 2022-07-12 | 湖南东方天润农业科技有限公司 | Method for detecting content of trans-oleic acid for edible oil production |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107132296A (en) * | 2017-06-30 | 2017-09-05 | 华仁药业股份有限公司 | It is a kind of at the same determine special medicine purposes formula food in content of fatty acid method |
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| CN112326821A (en) * | 2020-10-27 | 2021-02-05 | 杨振全 | Method for detecting lipid content of biological liver tissue |
| CN114740128A (en) * | 2022-05-17 | 2022-07-12 | 湖南东方天润农业科技有限公司 | Method for detecting content of trans-oleic acid for edible oil production |
| CN114740128B (en) * | 2022-05-17 | 2024-03-15 | 湖南东方天润农业科技有限公司 | Method for detecting trans-oleic acid content for edible oil production |
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| Publication number | Publication date |
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| CN106290665B (en) | 2018-08-31 |
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