CN104048947B - Method for detecting dioxin content in food - Google Patents
Method for detecting dioxin content in food Download PDFInfo
- Publication number
- CN104048947B CN104048947B CN201410219084.8A CN201410219084A CN104048947B CN 104048947 B CN104048947 B CN 104048947B CN 201410219084 A CN201410219084 A CN 201410219084A CN 104048947 B CN104048947 B CN 104048947B
- Authority
- CN
- China
- Prior art keywords
- english
- pentane
- parts
- silica gel
- drip washing
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Abstract
The invention relates to a method for detecting dioxin content in food. The method is characterized by comprising the following steps: taking an oil sample of a livestock and poultry product to be detected; dissolving the sample in pentane to obtain a mixed solution; heating the mixed solution to volatilize pentane and most oil in the mixed solution in turn, so as to obtain high content dioxin grease; purifying the high content dioxin grease by a leaching method; and performing fluorescence analysis on the by the dioxin obtained by purification to obtain the result of dioxin content in the livestock and poultry product. The invention has the advantage that the detection method provided by the invention does not require sending the sample to the laboratory for testing, and the detection is time-saving, fast, convenient, cheap and easy to operate, so as to reduce a lot of manpower, material and financial resources, for the detection mechanism, save the cost of detection and improve the detection efficiency.
Description
Technical field
The present invention relates to a kind of food quality detection method, particularly to a kind of detection of two English contents in meat product
Method.
Background technology
With the continuous improvement of social development and living standards of the people, food security is increasingly subject to many national government
Widely pay close attention to the common people, the theory of " bread is the staff of life, and food is with An Weixian " is increasingly stronger in consumer, particularly in recent years
People are made more to pay close attention to food " bird flu ", " clenbuterol hydrochloride ", " artificial honey " to occur, the food safety affair such as " two English "
Safety problem.With the epoch and expanding economy, people no longer worry the problem of food shortages, food quality is pacified gradually
Full requirement more and more higher, the especially food security of poultry meat, people have started to from number to the demand of poultry meat products
Amount type changes to quality safety-type.
Two English classes extensively and are permanently present in environment, and are enriched in the pollutant of food chain, and its main source is
Volcano eruption, forest fire, Chemical Manufacture, trash burning, metal smelt and papermaking etc..Two English class materials are not only present in soil
Earth, deposit, in water, and it is present in wild animal, domestic animal, tissue.In food two English class materials be difficult biological
And chemical degradation, therefore there is biological concentration and biomagnification, long-term with environment in vivo can retain two English class things
Matter also has lipophilicity, after entering body, is mainly accumulated in humans and animals in fat and mainly contacts two English classes by food
Material, wherein significant portion come from animal foodstuff!Two English in poultry meat can quickly be detected by this device
Content, improve food safe mass, allow people be able to eat green non-pollution food.
China only a few laboratory possesses the ability of two English contents in meat product to be detected at this stage, and examines
Survey is expensive, method of operating is loaded down with trivial details, time-consuming, and this far can not meet the detection needs to two English.Therefore it is badly in need of a kind of
Can in quick detection meat product two English contents method, to realize being used for quickly detecting the purpose of sample, warp at the scene
Cross and extensively retrieve, not yet find ideal technical scheme.
Content of the invention
The invention aims to solve at this stage in meat product two English content detection exist expensive,
The shortcomings of method of operating is loaded down with trivial details, time-consuming, and propose a kind of food in two English contents detection method.
To achieve these goals, present invention employs following technical scheme：
In a kind of food, the detection method of two English contents is it is characterised in that comprise the following steps：
A, take poultry to be checked, the solid fat of bird product, heating melts into liquid fat, sample from liquid fat；
B, the liquid fat being sampled is completely dissolved in pentane, obtains mixed solution；
C, mixed solution is heated, the temperature of heating is maintained between the boiling point of pentane and grease boiling point, and keeps one section
Time, the pentane in mixed liquor is made all to volatilize；
D, again by without pentane solution continue heating, the preferred 300oC of temperature of heating, the preferred 5-10min of temperature retention time,
Make most grease volatilization in solution, obtain two English greases of high-load；
E, purification：Using elution method by two English greases of high-load, pass sequentially through acidic silica gel post and activated carbon
Post carries out filtering purification；
F, two English obtaining by purification are carried out fluorescence analysis, raiseeed, two English contents in bird product
Result.
On the basis of technique scheme, can there is technical scheme further below：
Described elution method comprises the following steps, and acidic silica gel post and activated-charcoal column are placed in a pipeline, forms string
Connection post, first uses pentane drip washing columns in series, then acidic silica gel post is taken out pipeline, using toluene independent drip washing activated-charcoal column,
Afterwards acidic silica gel post is put back to pipeline, the two English greases being passed through high-load carry out drip washing purification.
Described acidic silica gel post is sequentially filled the sulfuric acid silica gel of glass fibre, 17 parts of anhydrous sodium sulfate and 33% from bottom to up
30 parts, populated after pass through pentane drip washing.
Described activated-charcoal column is sequentially filled mineral wool, 5 parts of anhydrous sodium sulfate, 1% 3 parts of X-CARB/ diatomite from bottom to up
With 10 parts of anhydrous sodium sulfate, populated after pass sequentially through 1 part of acetone, the pentane drip washing of 4 parts of toluene and 2 parts.
Described fluorescence analysis comprises the following steps：First 400 μ LPMI1640 are added to concentrate to two purified English greases
Volatilize completely to pentane；Mix after adding the nutrient solution containing two English luciferase cells；3rd mixed liquor uniformly adds
It is added on the orifice plate in 96 holes being placed with cell H1L6.1c2, cultivate 20～24h；Finally nutrient solution is shone by infrared ray
Penetrate, project to fluorescence identifying analysis plates, data extraction is carried out by sepectrophotofluorometer, and connect and carry out data to computer
Process, obtain two English content results, and be shown on screen.
In the meat products such as poultry, fowl, due in its fat two English contents to compare its hetero-organization higher, so in detection
In meat product during two English contents, its grease is taken to do quality testing.
The present invention uses two English pollutants in CALUX method detection food, and the operating procedure of this detection method mainly has：
Sample collection, two English class material extractions, purifying, drying, cell process, cell cracking, sepectrophotofluorometer analysis and number
According to process, concrete operation method is different because of the difference of material, the main distinction between different samples to its contained two
The behaviour of the extraction of English pollutant and purifying does, and method is not mainly the inspection of two English contents in the meat product such as poultry, fowl
Survey.
It is an advantage of the current invention that：Using detection method provided by the present invention, enter without again sample being sent into laboratory
Row detection, and detect save time, quick, convenient, cheap, processing ease, to testing agency decrease substantial amounts of human and material resources,
Financial resources, have saved testing cost, improve detection efficiency.
Specific embodiment
In a kind of food that the present invention provides, the detection method of two English contents is it is characterised in that comprise the following steps：
First, heating obtains two higher English greases of content, and its step is as follows：
A, take the solid fat of the meat products such as poultry to be checked, fowl, heating melts into liquid fat, from the fluid oil obtaining
50g is taken out as sample in fat.
B, first loading pentane in heating dress concentration is put, during the liquid fat of the 50g of acquirement is added heating dress concentration to put,
Wherein pentane and liquid fat press 10:1 ratio mixing, after mixing shakes up mixed liquor so that liquid fat is completely dissolved in
In pentane, obtain mixed solution.
C, heating dress concentration are put and are heated mixed solution, the temperature of heating be maintained at the boiling point of pentane and grease boiling point it
Between, and keep a period of time, the wherein temperature of heating preferably 40oC, the preferred 5-10min of temperature retention time, make the pentane in mixed liquor
All volatilize.
D, again by without pentane mixed solution continue heating, the wherein temperature of heating preferably 300oC, temperature retention time is preferred
5-10min, makes most grease volatilization in solution, obtains two English greases of high-load.
2nd, the high-load obtaining in the first step two English grease is carried out purification using elution method, its step is as follows：
E, the 10mL acidic silica gel post high-load two English grease obtaining in the first step being passed through series connection and 5mL activity
Charcoal post is purified.
The preparation method of described 10mL acidic silica gel post is to be sequentially filled glass fibre, anhydrous sodium sulfate from bottom to up
107g and 33% sulfuric acid silica gel 3.0g, populated after use 30ml pentane drip washing once.
The preparation method of described 5ml activated-charcoal column is, be sequentially filled from bottom to up mineral wool, anhydrous sodium sulfate 0.5g, 1%
X-CARB/ diatomite 0.3g and anhydrous sodium sulfate 1.0g, populated after pass sequentially through the acetone of 5ml, the toluene of 20mL and
The pentane drip washing of 10mL.
After 10mL acidic silica gel post and 5mL activated-charcoal column prepare, they are connected in a pipeline, form series connection
Post, first uses pentane drip washing columns in series, by the Impurity removal in two posts；Again acidic silica gel post is taken out pipeline, using toluene list
Solely drip washing activated-charcoal column, isolates PCDD/Fs, makes to pass through filtering two inch and can increase it with toluene drip washing activated-charcoal column
Rate；Finally acidic silica gel post is put back to pipeline, be passed through two English greases of high-load, leacheate rotary evaporation is concentrated, then uses
Pentane is settled to the sample of 4mL concentrate.
3rd, two English that the purification in second step is obtained carry out fluorescence analysis
F, from the sample being settled to 4mL take out 1mL be placed in test tube, then add 400 μ LPMI1640 dense in test tube
Contracting, adds the nutrient solution of 400 μ L bis- English luciferase cell to be uniformly mixed so as to obtain mixing after the pentane in sample volatilizees completely
Liquid, described English luciferase cell adopts the FBS of 1% Penicilllin-streptomycin+8% for prior art.
G, on the orifice plate in 96 holes being placed with cell H1L6.1c2, every hole all add 190 μ L mixed liquors culture 20
The temperature setting of～24h, wherein culture is 37oC, the CO being 5% containing volume fraction in nutrient solution2.
After i, culture, the nutrient solution of every in the hole is collected in test tube, test tube is irradiated by infrared ray, by light after irradiation
Line projection, to fluorescence identifying analysis plates, carries out data extraction by sepectrophotofluorometer, and connects and carry out data to computer
Process, obtain two English content results, and be shown on screen.Wherein adopt chemiluminescence ELIASA to measure luminosity, pass through
Luminosity calculates PCDD concentration.
Cleaning Principle
The principle using two English class materials in CALUX method detection food of the present invention is built upon two English class materials and enters
On the basis of entering the principle of produced biological agent after living organism.Two English enter cells after, first with cytoplasm in
Aryl hydrocarbon receptor AhR combine, form dioxin-AhR conjugate, this conjugate proceeds to nucleus, in core, aromatic hydrocarbon again
Acceptor 2 heat shock protein subunit Hsp90 successively in connection, 1 p23 and 1 XAP2 protein ligands departs from, and thin
Assemble in karyon, then Dioxin-AhR mixture is formed with the aryl hydrocarbon receptor consideration convey seat protein combination in nucleus again
There is the dioxin-AhR-AR NT albumen dimer of height adhesion with DNA, this dimer specifically combines in two English again
On two English response element DREs in the enhancers upstream of response gene, thus starting the transcriptional expression of these target genes, and
Lead to a series of biochemical changes relevant with these gene expressions and toxic action.
CALUX method is a kind of to detect in food two English class materials lifes with the recombinant cell containing luciferase reporter gene
Thing analysis method.It is intended to first when making in this way build the recombinant plasmid containing luciferase reporter gene, then will recombinate
Plasmid transfection makes the recombinant cell lines of stable transfection in zooblast, is finally detected in food extracts with recombinant cell
Two English class materials.In theory, the available much different clones of CALUX method detection, as long as these intracellular contain
Functional sound aryl hydrocarbon receptor signal transducting system, but the reaction to various two English class materials for the actually distinct clone
Can be different.At present, conventional cell line has rat hepatoma cell strain, murine hepatocarcinoma cell strain and human hepatocarcinoma cells' strain.
When building plasmid, first by different number of two English response element DREs insertions containing luciferase reporter gene
Made recombinant plasmid by the region of MMTV promoter regulation in plasmid, then recombinant plasmid is proceeded to zooblast and make stable transfection
Clone.During detection, process recombinant cell with the food extracts containing two English class materials, when cells contacting is to two English class things
After matter, the aryl hydrocarbon receptor in cell can be activated, and can lead to the expression of luciferase reporter gene further.Luciferase base
Fluorescence produced by the expression of cause then can be detected with sepectrophotofluorometer.Because recombinant cell touches after two English class materials
The fluorescence volume of expressed luciferase is directly proportional to the content of two English class materials within the specific limits, therefore passes through to pass through
Amount of fluorescence detected is calculating the amount of dioxins.
In addition it is also possible to recombinant plasmid carries out transient transfection to detect two English class things in food extracts to cell
Matter, so can save the time making needed for stable transfected cells.But the test repeatability of transient transfection is not so good as stable turning
Contaminate the good of clone detection, and instantaneously proceed to the gene of cell and would generally lose after 72h, therefore be only suitable for short-term research
With.
Claims (5)
1. in a kind of food the detection method of two English contents it is characterised in that comprising the following steps：
A, take poultry to be checked, the solid fat of bird product, heating melts into liquid fat, sample from liquid fat；
B, the liquid fat being sampled is completely dissolved in pentane, obtains mixed solution；
C, mixed solution is heated, the temperature of heating is maintained between the boiling point of pentane and grease boiling point, and keeps a period of time,
The pentane in mixed liquor is made all to volatilize；
D, again by without pentane solution continue heating, the temperature 300 of heatingoC, temperature retention time 5-10min, make big portion in solution
Divide grease volatilization, obtain two English greases of high-load；
E, purification：Using elution method by two English greases of high-load, pass sequentially through acidic silica gel post and activated-charcoal column enters
Row filters purification；
F, two English obtaining by purification are carried out fluorescence analysis, raiseeed, in bird product two English contents knot
Really.
2. detection method according to claim 1 it is characterised in that：Described elution method comprises the following steps, acid silicon
Glue post and activated-charcoal column are placed in a pipeline, form columns in series, first use pentane drip washing columns in series, then by acidic silica gel post
Take out pipeline, using toluene independent drip washing activated-charcoal column, finally acidic silica gel post is put back to pipeline, be passed through two English of high-load
Grease carries out drip washing purification.
3. detection method according to claim 1 and 2 it is characterised in that：Described acidic silica gel post is filled out from bottom to up successively
Fill 30 parts of the sulfuric acid silica gel of glass fibre, 17 parts of anhydrous sodium sulfate and 33%, populated after pass through pentane drip washing.
4. detection method according to claim 1 and 2 it is characterised in that：Described activated-charcoal column is sequentially filled from bottom to up
Mineral wool, 5 parts of anhydrous sodium sulfate, 1% 3 parts of X-CARB/ diatomite and 10 parts of anhydrous sodium sulfate, populated after pass sequentially through 1 part
Acetone, the pentane drip washing of 4 parts of toluene and 2 parts.
5. detection method according to claim 1 it is characterised in that：Described fluorescence analysis comprises the following steps：First to
Two purified English greases addition 400 μ LPMI1640 are concentrated into pentane and volatilize completely；Add containing two English luciferases
Mix after the nutrient solution of cell；3rd mixed liquor is uniformly added on the orifice plate in 96 holes being placed with cell H1L6.1c2, training
Support 20～24h；Finally nutrient solution is irradiated by infrared ray, project to fluorescence identifying analysis plates, by fluorescence spectrophotometry
Meter carries out data extraction, and connects and carry out data processing to computer, obtains two English content results, and is shown on screen.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410219084.8A CN104048947B (en) | 2014-05-23 | 2014-05-23 | Method for detecting dioxin content in food |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410219084.8A CN104048947B (en) | 2014-05-23 | 2014-05-23 | Method for detecting dioxin content in food |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104048947A CN104048947A (en) | 2014-09-17 |
| CN104048947B true CN104048947B (en) | 2017-02-22 |
Family
ID=51502066
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410219084.8A Expired - Fee Related CN104048947B (en) | 2014-05-23 | 2014-05-23 | Method for detecting dioxin content in food |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104048947B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109253908A (en) * | 2017-07-12 | 2019-01-22 | 上海宝钢工业技术服务有限公司 | The fast purification method of dioxin analytic process in soil |
| CN114459859B (en) * | 2022-02-16 | 2022-09-09 | 北京卢米斯生物科技有限公司 | High-flux dioxin automatic detection device based on luciferase reporter gene method |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2002310912A (en) * | 2001-04-11 | 2002-10-23 | Toshiba Corp | Device for measuring concentration of trace amount of matter |
| CN1731150A (en) * | 2005-08-23 | 2006-02-08 | 中国科学院武汉病毒研究所 | A kind of biochip method that detects dioxin-type chemical species |
| CN102520154A (en) * | 2011-12-08 | 2012-06-27 | 环境保护部华南环境科学研究所 | Method for detecting dioxins in environment |
| CN102967563A (en) * | 2012-11-26 | 2013-03-13 | 东莞博莱特生物科技有限公司 | Device and method for acquiring dioxin toxoid signals |
| CN102967495A (en) * | 2012-11-21 | 2013-03-13 | 北京普立泰科仪器有限公司 | Sample pretreatment device and sample treatment method |
-
2014
- 2014-05-23 CN CN201410219084.8A patent/CN104048947B/en not_active Expired - Fee Related
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2002310912A (en) * | 2001-04-11 | 2002-10-23 | Toshiba Corp | Device for measuring concentration of trace amount of matter |
| CN1731150A (en) * | 2005-08-23 | 2006-02-08 | 中国科学院武汉病毒研究所 | A kind of biochip method that detects dioxin-type chemical species |
| CN102520154A (en) * | 2011-12-08 | 2012-06-27 | 环境保护部华南环境科学研究所 | Method for detecting dioxins in environment |
| CN102967495A (en) * | 2012-11-21 | 2013-03-13 | 北京普立泰科仪器有限公司 | Sample pretreatment device and sample treatment method |
| CN102967563A (en) * | 2012-11-26 | 2013-03-13 | 东莞博莱特生物科技有限公司 | Device and method for acquiring dioxin toxoid signals |
Non-Patent Citations (2)
| Title |
|---|
| 食品中二噁英类化合物痕量检测研究进展;廖桢葳 等;《食品研究与开发》;20110930;第32卷(第9期);第231-235页 * |
| 食品中二恶英类化合物的污染与分析;廖涛 等;《食品与机械》;20080731;第24卷(第4期);第153-157,165页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104048947A (en) | 2014-09-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104820003B (en) | A kind of papery microfluidic system and method for Detecting Pesticide | |
| CN104048947B (en) | Method for detecting dioxin content in food | |
| CN102140489A (en) | Method for testing cytotoxicity in full smoke contamination of cigarette | |
| Cope et al. | The occurrence of electrocortin in human urine | |
| CN113657799B (en) | Method for evaluating environmental health risk of benzo [ a ] pyrene in soil and animal model | |
| CN103969362A (en) | Method for quantitatively detecting FQs(fluroquinolones) in chicken manure | |
| CN105758948A (en) | Application of triptolide in identification presence of tripterygium glycoside honey in honey as feature identification material | |
| CN107058386A (en) | A kind of preparation method of transgenic zebrafish | |
| Batabyal et al. | Social coping styles of lizards are reactive and not proactive in urban areas | |
| York et al. | Bioenergy Underground: Challenges and opportunities for phenotyping roots and the microbiome for sustainable bioenergy crop production | |
| CN103255196A (en) | Method for detecting effect of Coptis chinensis alkaloid monomer on tumor cells | |
| Yang et al. | Disturbance-level-dependent post-disturbance succession in a Eurasian steppe | |
| CN110954688B (en) | Method for determining bioavailability of nickel in soil and food for mice | |
| CN101907562B (en) | Quick measuring method of pyrethrin | |
| CN204439437U (en) | The cleaning equipment of Detecting Pesticide | |
| CN110346494B (en) | Method for simultaneously measuring contents of 4 effective components in erigeron breviscapus | |
| CN101904949A (en) | Plant extract liquor and preparation method thereof | |
| CN103947606B (en) | Knock in the non-human mammal model of uncoupling proteins 1-luciferase gene and construction method thereof and application | |
| CN110749578A (en) | Method for detecting content of dioxin in meat food | |
| Drake et al. | Relationships between salmon abundance and tree-ring δ15N: three objective tests | |
| CN101249068A (en) | Ivermectin long-acting injection and method of preparing the same | |
| CN107736362B (en) | Method for preparing botanical molluscicide by sodium alginate embedding method | |
| CN102218095B (en) | Processing method of myrobalan-processed kusnezoff monkshood root as mongolian medicine | |
| CN108181372A (en) | The detection method of nitre sulphur ketone residual quantity in corn | |
| CN105021436B (en) | A kind of pesticide extracting solution and its application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20170612 Address after: East Donghua road 233000 Anhui Longzihu district city of Bengbu province No. 9 Patentee after: Anhui Science and Technology University Address before: High tech Zone in Anhui city of Bengbu Province Xinghua Road 233010 No. 168 Patentee before: Meng Lingqi |
|
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20170222 Termination date: 20170523 |