CN101458240A - Thin-layer chromatography developing agent for detecting melamine residual and analytical method - Google Patents
Thin-layer chromatography developing agent for detecting melamine residual and analytical method Download PDFInfo
- Publication number
- CN101458240A CN101458240A CNA2008102367804A CN200810236780A CN101458240A CN 101458240 A CN101458240 A CN 101458240A CN A2008102367804 A CNA2008102367804 A CN A2008102367804A CN 200810236780 A CN200810236780 A CN 200810236780A CN 101458240 A CN101458240 A CN 101458240A
- Authority
- CN
- China
- Prior art keywords
- parts
- thin
- sample
- weight
- melamine
- 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.)
- Pending
Links
Landscapes
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
Abstract
The invention discloses a thin-layer chromatographic developing solvent for detecting melamine residue and an analytic method thereof, and relates to a developing agent for detecting the melamine residue in food and the analytic method thereof. The developing agent consists of n-butyl alcohol, ethyl acetate and the like. The analytic method comprises the following steps: pretreating a silica gel plate, preparing the developing agent, preparing a color-developing agent, dialyzing and separating a test sample, analyzing the sample through thin layer chromatography, developing color by spraying the color-developing agent and finishing the chromatographic analysis. The method is a qualitative identification method of melamine which is added and remained in the food such as milk products, feeding stuff and the like, and is repeatedly tested, and the method has the advantages of simple and convenient operation, high sensitivity, good reproducibility, clear spots and good separation effect. Compared with negative samples, the negative samples have no interference, which proves that the method has exclusiveness and feasibility. The developing solvent and the method can be employed to accurately and conveniently identify whether the test sample contains the melamine without experienced experts and expensive detection equipment.
Description
Technical field
The wood invention relates to a kind of developping agent and analytical approach that detects melamine residual amount in the food.
Background technology
Melamine (Melanmine) is called for short triamine, formal name used at school three ammonia triazines, and another name melamine, cyanuramide, three polyamide are a kind of important azacyclo-Organic Chemicals, white crystalline powder is tasteless.This chemical reagent often is used to produce plastics, glue and fire retardant, and in part Asian countries, it also is used to make chemical fertilizer.It is a kind of chemical substance of forbidding being used for food, feed, can kidney failure take place and cause death after humans and animals is edible.
Melamine detection adopts the described high-efficient liquid phase technique of " State Standard of the People's Republic of China " (GB/T 22400-2008) in the raw milk, concrete grammar is: take by weighing the 15g raw milk sample (accurately to 0.01g) that mixes, place 50mL tool plug scale test tube, add the 30mL acetonitrile, thermal agitation 6min adds water and is settled to full scale, fully leaves standstill 3min behind the mixing, after the filtration of disposable syringe absorption supernatant usefulness pin type filtrator, as the efficient liquid phase chromatographic analysis sample.Sample carries out liquid phase analysis, carries out the method for qualitative identification according to the retention time consistance.According to the retention time of melamine standard substance, determine whether contain melamine in the sample.
Other dairy produces are not then had unified standard, use for reference the detection method of U.S. FDA, concrete grammar is: a. takes by weighing powdered milk sample 5g (or milk 10mL), add 50mL 1% trichloroacetic acid extract, fully mixing adds 2mL2% lead acetate solution, ultrasonic 20min; B. get part solution and be transferred in the 10mL centrifuge tube, the centrifugal 10min of 8000rpm/min.Activation of solid-phase extraction column PCX pillar and balance: 3ml methyl alcohol, 3ml water; Last sample: add extract 3ml; Drench Xian: 3ml water, 3ml methyl alcohol discards and drenches Xian's liquid and will lean on for a short time and drain; Wash-out: 5ml 5% ammonification methyl alcohol (v/v) wash-out.(the preparation of 5% ammonification methyl alcohol: 5ml ammoniacal liquor+95ml methyl alcohol); Concentrate: 50 ℃, nitrogen dries up, and 20% methanol is settled to 2ml.Sample carries out liquid phase analysis, carries out the method for qualitative identification according to the retention time consistance.According to the retention time of melamine standard substance, determine whether contain melamine in the sample.
Existing GB melamine detection method is a chromatography, but this method needs perfect laboratory, experienced technician, and complicated and expensive instrument, and also dairy produce pre-treatment process is very loaded down with trivial details, is not suitable for the examination of a large amount of samples.
Summary of the invention
The objective of the invention is at above-mentioned present situation, aim to provide a kind of easy and simple to handle, cheaply, a kind of plate chromatography developing solvent and analytical approach that is used to detect melamine residual accurately and fast.
The implementation of the object of the invention is that a kind of plate chromatography developing solvent that is used to detect melamine residual is made up of following compositions in portion by weight: 4-8 parts of normal butyl alcohols, 1-4 parts in ethyl acetate, water 0.1-1 part.
A kind of thin-layer chromatographic analysis method that is used to detect melamine residual, concrete steps are as follows:
1) pre-service of silica gel plate: handle silica gel thin-layer plate with chloroform, washed with methanol liquid
2) preparation developping agent: getting parts by weight is 4-8 parts of normal butyl alcohols, 1-4 parts in ethyl acetate, and water 0.1-1 part or concentration are that KCl aqueous solution 0.05-1 part of 0.2-0.5% is mixed with developping agent,
3) preparation of developer: get parts by weight and be 1-5 part of 0.01-2% HgCl
2The 0.1-2% diphenylcarbazone ethanolic solution of ethanolic solution and 1-5 parts is hybridly prepared into compound developer.
4) tested sample pre-service: the sample separation of dialysing, get the dislysate evaporate to dryness, add water-soluble the separating that its weight 50-500 doubly measures, as sample to be tested.
5) thin-layer chromatographic analysis: at first with the sample to be tested of step 4), point sample is put into thin layer plate the expansion cylinder of pouring developping agent into again to the silica gel plate of handling through step 1), and ascending development takes out thin layer plate to the upper edge, dries up solvent.
6) colour developing: the thin layer plate that launches in the step 5) is sprayed the developer of step 3) preparation, under 105 ℃, heating 10min, stratographic analysis is finished in colour developing.
The present invention is as food, qualitative identification method as interpolation and residual melamine in dairy produce, the feed etc., through repetition test, this method is easy and simple to handle, highly sensitive, favorable reproducibility, clear spot, good separating effect, through the negative sample contrast, negative sample is noiseless, proves that the method has specificity and feasibility.Use the present invention not need veteran reviewer and the high checkout equipment of price, can differentiate whether contain melamine in the sample accurately and easily.
Description of drawings
Fig. 1 is that the thin layer of melamine in the milk powder is differentiated figure,
Feminine gender and the positive thin layer contrast discriminating figure of Fig. 2 for not containing melamine in the milk powder.
Embodiment
Developping agent of the present invention is made up of following compositions in portion by weight: 4-8 parts of normal butyl alcohols, 1-4 parts in ethyl acetate, water 0.1-1 part.
Also can be made up of following compositions in portion by weight: 4-8 parts of normal butyl alcohols, 1-4 parts in ethyl acetate, concentration are KCl aqueous solution 0.05-1 part of 0.2-0.5%.
Enumerate specific embodiments of the invention below:
Example 1, get a silica gel thin-layer plate, put into and be added with 1ml chloroform and methyl alcohol (1:1 volume ratio), be expanded to the upper edge, taking-up is dried, and at 110 ℃ of heating baking 15min, cools off standby.
The 2.5g powdered milk sample is separated with 10ml is water-soluble, or got 10ml liquid milk sample, add in the dialysis treatment device, the 10-20min that dialyses under ultrasonic and heating condition gets the dislysate evaporate to dryness, adds water-soluble the separating of 200 times of amounts of its weight, as sample to be tested.
Get 2mg melamine sample and be dissolved in the 1L water, make reference substance solution.
Checkout procedure, test according to thin-layered chromatography (2005 editions appendix VIB of Chinese Pharmacopoeia), get need testing solution 25u l silica gel plate point sample, 6 parts of normal butyl alcohols, 2 parts in ethyl acetate and water are the developping agent expansion for 0.5 part, and taking-up is dried, and spray is with compound chromogenic reagent, 100 ℃ of heating colour developing 5min are to clear spot.Compound developer is 2 part of 0.1% HgCl
2The mixed liquor of 0.5% diphenylcarbazone ethanolic solution of ethanolic solution and 4 parts.
The results are shown in Figure 1, Fig. 2: in the test sample chromatogram, on the corresponding position of melamine reference substance, show the spot of same color; In the negative milk sample chromatogram, with melamine chromatogram relevant position on, no same color spot.Illustrate in the milk sample and whether contain melamine.
1,3 is powdered milk sample among Fig. 1, and 2 is the melamine reference substance.1,3 positive samples among Fig. 2,2 negative samples.
Example 2, with example 1, different is, adds water-soluble the separating of 50 times of amounts of its weight, 8 parts of normal butyl alcohols, 2 parts in ethyl acetate and water are developping agent for 0.1 part, compound developer is 1 part of 0.01% HgCl
2The mixed liquor of 0.1% diphenylcarbazone ethanolic solution of ethanolic solution and 1 part.
Example 3, with example 1, different is, adds water-soluble the separating of 500 times of amounts of its weight, 8 parts of normal butyl alcohols, 4 parts in ethyl acetate and water are developping agent for 1 part, compound developer is 5 part of 2% HgCl
2The mixed liquor of 0.5% diphenylcarbazone ethanolic solution of ethanolic solution and 2 parts.
Example 4, with example 1, different is, adds water-soluble the separating of 100 times of amounts of its weight, 4 parts of normal butyl alcohols, 1 part in ethyl acetate and 0.5% KCl aqueous solution are developping agent for 0.05 part, compound developer is 1 part of 0.02% HgCl
2The mixed liquor of 0.2% diphenylcarbazone ethanolic solution of ethanolic solution and 1 part.
Example 5, with example 1, different is, adds water-soluble the separating of 200 times of amounts of its weight, 6 parts of normal butyl alcohols, 2 parts in ethyl acetate and 0.2% KCl aqueous solution are developping agent for 0.4 part, compound developer is 3 part of 0.1% HgCl
2The mixed liquor of 1% diphenylcarbazone ethanolic solution of ethanolic solution and 5 parts.
Example 6, with example 1, different is, adds water-soluble the separating of 300 times of amounts of its weight, 6 parts of normal butyl alcohols, 3 parts in ethyl acetate and 0.2% KCl aqueous solution are developping agent for 1 part, compound developer is 4 part of 0.3% HgCl
2The mixed liquor of 0.3% diphenylcarbazone ethanolic solution of ethanolic solution and 3 parts.
Claims (4)
1, a kind of plate chromatography developing solvent that is used to detect melamine residual is characterized in that being made up of following compositions in portion by weight: 4-8 parts of normal butyl alcohols, 1-4 parts in ethyl acetate.
2, a kind of plate chromatography developing solvent that is used to detect melamine residual according to claim 1, it is characterized in that following compositions in portion by weight composition: 4-8 parts of normal butyl alcohols, 1-4 parts in ethyl acetate, water 0.1-1 part or concentration are KCl aqueous solution 0.05-1 part of 0.2-0.5%.
3, a kind of thin-layer chromatography developer that is used to detect melamine residual is characterized in that following compositions in portion by weight composition: get parts by weight and be 1-5 part of 0.01-2% HgCl
2The 0.1-2% diphenylcarbazone ethanolic solution of ethanolic solution and 1-5 parts is hybridly prepared into compound developer.
4, carry out the method for thin-layer chromatographic analysis with the described a kind of plate chromatography developing solvent that is used to detect melamine residual of claim 1, it is characterized in that concrete steps are as follows:
1) pre-service of silica gel plate: handle silica gel thin-layer plate with chloroform, washed with methanol liquid
2) preparation developping agent: getting parts by weight is 4-8 parts of normal butyl alcohols, 1-4 parts in ethyl acetate, and water 0.1-1 part or concentration are that KCl aqueous solution 0.05-1 part of 0.2-0.5% is mixed with developping agent.
3) preparation of developer: get parts by weight and be 1-5 part of 0.01-2% HgCl
2The 0.1-2% diphenylcarbazone ethanolic solution of ethanolic solution and 1-5 parts is hybridly prepared into compound developer.
4) tested sample pre-service: the sample separation of dialysing, get the dislysate evaporate to dryness, add water-soluble the separating that its weight 50-500 doubly measures, as sample to be tested.
5) thin-layer chromatographic analysis: at first with the sample to be tested of step 4), point sample is put into thin layer plate the expansion cylinder of pouring developping agent into again to the silica gel plate of handling through step 1), and ascending development takes out thin layer plate to the upper edge, dries up solvent.
6) colour developing: the thin layer plate that launches in the step 5) is sprayed the developer of step 3) preparation, under 105 ℃, heating 10min, stratographic analysis is finished in colour developing.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008102367804A CN101458240A (en) | 2008-12-11 | 2008-12-11 | Thin-layer chromatography developing agent for detecting melamine residual and analytical method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008102367804A CN101458240A (en) | 2008-12-11 | 2008-12-11 | Thin-layer chromatography developing agent for detecting melamine residual and analytical method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101458240A true CN101458240A (en) | 2009-06-17 |
Family
ID=40769225
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2008102367804A Pending CN101458240A (en) | 2008-12-11 | 2008-12-11 | Thin-layer chromatography developing agent for detecting melamine residual and analytical method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101458240A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102175812A (en) * | 2011-01-17 | 2011-09-07 | 四川大学 | Thin-layer chromatographic developer for detecting residual amount of tripolycyanamide and application of thin-layer chromatographic developer |
| CN102998305A (en) * | 2011-09-19 | 2013-03-27 | 青岛大学 | Method for detecting melamine in raw milk by utilizing chromogenic reaction |
| CN106918586A (en) * | 2017-02-15 | 2017-07-04 | 中国人民解放军第二军医大学 | A kind of method that various dopants are detected simultaneously in milk powder |
| CN108226369A (en) * | 2016-12-15 | 2018-06-29 | 江苏维赛科技生物发展有限公司 | A kind of remaining thin-layered chromatography detection method of high poison antistaling agent thiabendazole |
-
2008
- 2008-12-11 CN CNA2008102367804A patent/CN101458240A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102175812A (en) * | 2011-01-17 | 2011-09-07 | 四川大学 | Thin-layer chromatographic developer for detecting residual amount of tripolycyanamide and application of thin-layer chromatographic developer |
| CN102175812B (en) * | 2011-01-17 | 2014-10-22 | 四川大学 | Thin-layer chromatographic developer for detecting residual amount of tripolycyanamide and application of thin-layer chromatographic developer |
| CN102998305A (en) * | 2011-09-19 | 2013-03-27 | 青岛大学 | Method for detecting melamine in raw milk by utilizing chromogenic reaction |
| CN108226369A (en) * | 2016-12-15 | 2018-06-29 | 江苏维赛科技生物发展有限公司 | A kind of remaining thin-layered chromatography detection method of high poison antistaling agent thiabendazole |
| CN106918586A (en) * | 2017-02-15 | 2017-07-04 | 中国人民解放军第二军医大学 | A kind of method that various dopants are detected simultaneously in milk powder |
| CN106918586B (en) * | 2017-02-15 | 2019-10-15 | 中国人民解放军第二军医大学 | A kind of method that a variety of dopants detect simultaneously in milk powder |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105974041A (en) | Method for detecting perfluorinated compounds by using solid phase microextraction with liquid phase chromatography-tandem mass spectrum | |
| CN101458240A (en) | Thin-layer chromatography developing agent for detecting melamine residual and analytical method | |
| CN106706834A (en) | Method for rapidly detecting multiple biogenic amines in dairy product by virtue of combination of high performance thin layer chromatography and adjustable surface enhanced raman spectroscopy | |
| CN107462650A (en) | The detection method of environmental hormone in human urine | |
| CN110174470A (en) | The high-flux detection method of marine biotoxins in a kind of aquatic products | |
| CN109632978A (en) | A kind of Poria cocos reference extract and its preparation method and application | |
| CN102062768A (en) | Method for quickly detecting 4-methylimidazole in food | |
| CN104535548B (en) | Method for rapidly detecting sulfonamide antibacterial medicines in milk by using in-tube solid-phase micro-extraction technology | |
| Boyer et al. | Detection of testosterone propionate administration in horse hair samples | |
| CN104297399A (en) | Method for detecting byproducts 4-methylimidazole and 2-acetyl-4-hydroxy-butylimidazole in caramel pigment | |
| CN103823003B (en) | The high performance liquid chromatography of total formaldehyde content in a kind of wood-based plate | |
| CN103983706B (en) | Precolumn derivatization and high-efficiency liquid-phase fluorescence detection method for tetrodotoxin and kit based on the method | |
| CN108088715A (en) | Moutan bark reference extract and its preparation method and application | |
| CN104991031A (en) | Rapid multiple information thin-layer identification method of smilax glabra and water extract thereof | |
| CN105158372B (en) | Method for determining urocanic acid and ethyl ester thereof in cosmetics | |
| CN103278586A (en) | Extracting and detecting method for dicyandiamide component in dairy products | |
| CN106950317A (en) | A kind of method for closing the specific aromatic amine that azo dyes discharges in phase chromatographic tandem mass spectrometric determination Cigarette paper | |
| CN104849383A (en) | Method for determining nitroimidazole drug in bee pollen powder through combination of rapid solvent extraction-gel chromatography purification-LC/MS/MS | |
| CN114216983B (en) | Method for detecting residual amount of prochloraz in animal food by liquid chromatography-tandem mass spectrometry | |
| Korte et al. | Comparison of RapiTest® with Emit® d, au™ and GC-MS for the Analysis of Drugs in Urine | |
| CN108982712A (en) | Salmeterol, datro, the method for Ao Dateluo in a kind of measurement pork | |
| CN102178698B (en) | Method for detecting traditional Chinese medicine | |
| CN104634889A (en) | Method for measuring formaldehyde in cosmetics by using online derivatization | |
| CN104280484A (en) | Pretreatment method for chlorpromazine detection in animal product | |
| Hansen | Sample preparation and separation techniques for bioanalysis of morphine and related substances |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Open date: 20090617 |