CN106353313A - Rapid detection kit for melamine residue in milk product and use method thereof - Google Patents
Rapid detection kit for melamine residue in milk product and use method thereof Download PDFInfo
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- CN106353313A CN106353313A CN201610983887.XA CN201610983887A CN106353313A CN 106353313 A CN106353313 A CN 106353313A CN 201610983887 A CN201610983887 A CN 201610983887A CN 106353313 A CN106353313 A CN 106353313A
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- agent
- detection kit
- melamine
- buffer
- ultrafiltration
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
Abstract
The invention discloses a rapid detection kit for melamine residues in a milk product and a use method thereof. The rapid detection kit comprises treatment agents, a developing agent, an extraction agent, a pipette, a buffer agent, reducing agents and an enzyme agent, wherein the treatment agents are trichloroacetic acid and phenol; the developing agent is a mixture of potassium mercuric iodide, phosphomolybdic acid and thiourea; the extraction agent is a sodium acetate solution; the buffer agent is a Gly-HCl buffer solution; the reducing agents are dithiothreitol and beta-mercaptoethanol; the enzyme agent is phytoesterase. The rapid detection kit is low in cost, simple to prepare, high in detection sensitivity, that is, the sensitivity of the rapid detection kit is tens and thousands of times that in the prior art, remarkably widened in linear detection range, convenient to use, simple, rapid and convenient to operate, and free of complex reagent preparation process in detection, and detection can be completed easily by nonprofessional operators according to the instruction book of the kit.
Description
Technical field
The present invention relates to a kind of test kit, specifically a kind of quick detection kit of melamine in dairy products residual and
Using method.
Background technology
Tripolycyanamide is commonly called as melamine etc., and it is a kind of synthesis organic nitrogen-containing heterocyclic compound, belongs to a kind of industrial chemicals,
Extensively apply in the fields such as textile industry, paper-making industry and packaging material.Tripolycyanamide can be from the link such as ring mirror and packaging material for food
In enter in food, but in general, its content is relatively low.A kind of white crystalline powder of tripolycyanamide, nitrogen content is very high
(66%) protein content can, be significantly improved, people are referred to as " extract of protein ".In addition its simple production process, cost are very low.Cause
This partly illegal manufacturer driven by interests, in the food such as milk add tripolycyanamide, to significantly improve the egg detecting
White matter content.But the artificial tripolycyanamide adding is far beyond the dosis tolerata of people, easily formed water insoluble after entering body
Macromolecular complex deposition and form calculus, damage kidney, severe patient can cause renal failure, research also demonstrates that, melamine
A large amount of absorptions of amine also can cause the infringement of reproductive system, also result in the serious diseases such as bladder cancer simultaneously and occur, serious threat
The healthy and life security of people.
The detection method of the tripolycyanamide in currently available technology is more, exempts from including near-infrared, mid-infrared technology, enzyme connection
Epidemic disease, but there is, testing cost high, false positive rate high shortcoming high to instrument requirements in these methods.Therefore, in order to accurately and effectively
Tripolycyanamide in detection food, it is ensured that the health of food safety and people, is badly in need of seeking a kind of detection rapidly, sensitivity is high
Tripolycyanamide detection method.
Content of the invention
It is an object of the invention to provide a kind of structure melamine in dairy products residual simple, easy to use is quick
Detection kit and using method, to solve the problems, such as to propose in above-mentioned background technology.
For achieving the above object, the following technical scheme of present invention offer:
A kind of quick detection kit of melamine in dairy products residual, including inorganic agent, developer, extractant, dropper, delays
Electuary, reducing agent and enzyme agent, described inorganic agent is trichloroacetic acid and phenol, and described developer is potassium mercuric iodide, phosphomolybdic acid and sulfur
The mixture of urea, described extractant is sodium acetate solution, and described buffer agent is gly-hcl buffer, and described reducing agent is two sulfur
Threitol and beta -mercaptoethanol, described enzyme agent is plant esterase.
A kind of using method of the quick detection kit of melamine in dairy products residual, specifically comprises the following steps that
(1) first, milk product is sealed off on aseptic operating platform, place after drawing 10-20ml in mortar, add extractant
Quickly ground with liquid nitrogen, subsequently draw the solution after 50-100 μ l grinds using dropper and be added drop-wise in 96 hole elisa Plates, so
After be placed on that to carry out cold preservation in -80 DEG C of refrigerators of ultralow temperature stand-by；
(2) then, sodium tartrate, bovine serum albumin, lanthanum chloride, oxalic acid are added in Deoxydization nucleotide sodium solution, fully stir
Mix uniformly, place cryopreservation at lucifuge；
(3) then, 96 stand-by for cold preservation in above-mentioned steps (1) hole elisa Plates are taken out, after defrosting, add buffer agent and reducing agent,
Wherein buffer 4000-6000 times of dilution agent, wherein reduce 600-800 times of dilution agent, at 4 DEG C, stand 15min；
(4) then, the Deoxydization nucleotide sodium solution after will be uniform in above-mentioned steps (2) carries out ultrafiltration, set aside for use after ultrafiltration；
(5) then, inorganic agent, enzyme agent and developer will be added in 96 hole elisa Plates after above-mentioned steps (3) standing, incubate at 37 DEG C
Educate 20min, be subsequently adding the Deoxydization nucleotide sodium solution after ultrafiltration, after fully mixing, observe color change；
(6) last, using standard color comparison card, the color after change is compared.
As the further scheme of the present invention: in step (2), the mass percent concentration of Deoxydization nucleotide sodium solution is
4.6%.
As the further scheme of the present invention: in step (2), bovine serum albumin derives from Zhejiang sky Hangzhoupro biotechnology share
Company limited.
As the further scheme of the present invention: in step (4), ultrafiltration is specially and uses interception to be 200-300kda
Ultrafilter membrane carries out ultrafiltration.
As the further scheme of the present invention: in step (5), enzyme agent needs to be preheating to 37 DEG C before using.
Compared with prior art, the invention has the beneficial effects as follows:
Low cost of the present invention, making are simple, and detection sensitivity is high, and its remolding sensitivity prior art exceeds decades of times and thousands of times,
And detect that the range of linearity is significantly broadening, and easy to use, simple and quick, do not need the reagent preparation mistake of complexity in testing
Journey, layman just can be readily accomplished detection according to kit specification.
Specific embodiment
With reference to specific embodiment, the technical scheme of this patent is described in more detail.
Embodiment 1
A kind of quick detection kit of melamine in dairy products residual, including inorganic agent, developer, extractant, dropper, delays
Electuary, reducing agent and enzyme agent, described inorganic agent is trichloroacetic acid and phenol, and described developer is potassium mercuric iodide, phosphomolybdic acid and sulfur
The mixture of urea, described extractant is sodium acetate solution, and described buffer agent is gly-hcl buffer, and described reducing agent is two sulfur
Threitol and beta -mercaptoethanol, described enzyme agent is plant esterase.
A kind of using method of the quick detection kit of melamine in dairy products residual, specifically comprises the following steps that
(1) first, milk product is sealed off on aseptic operating platform, draw 10ml after place mortar in, add extractant and
Liquid nitrogen is quickly ground, and subsequently draws the solution after 50 μ l grind using dropper and is added drop-wise in 96 hole elisa Plates, then places
Carry out cold preservation in -80 DEG C of refrigerators of ultralow temperature stand-by；
(2) then, sodium tartrate, bovine serum albumin, lanthanum chloride, oxalic acid are added in Deoxydization nucleotide sodium solution, fully stir
Mix uniformly, place cryopreservation at lucifuge；
(3) then, 96 stand-by for cold preservation in above-mentioned steps (1) hole elisa Plates are taken out, after defrosting, add buffer agent and reducing agent,
Wherein buffer 4000 times of dilution agent, wherein reduce 600 times of dilution agent, at 4 DEG C, stand 15min；
(4) then, the Deoxydization nucleotide sodium solution after will be uniform in above-mentioned steps (2) carries out ultrafiltration, set aside for use after ultrafiltration；
(5) then, inorganic agent, enzyme agent and developer will be added in 96 hole elisa Plates after above-mentioned steps (3) standing, incubate at 37 DEG C
Educate 20min, be subsequently adding the Deoxydization nucleotide sodium solution after ultrafiltration, after fully mixing, observe color change；
(6) last, using standard color comparison card, the color after change is compared.
Wherein:
In step (2), the mass percent concentration of Deoxydization nucleotide sodium solution is 4.6%.
In step (2), bovine serum albumin derives from Zhejiang Tian Hang biotech inc.
In step (4), ultrafiltration is specially and carries out ultrafiltration using interception for 200-300kda ultrafilter membrane.
In step (5), enzyme agent is using front needing to be preheating to 37 DEG C.
Embodiment 2
A kind of quick detection kit of melamine in dairy products residual, including inorganic agent, developer, extractant, dropper, delays
Electuary, reducing agent and enzyme agent, described inorganic agent is trichloroacetic acid and phenol, and described developer is potassium mercuric iodide, phosphomolybdic acid and sulfur
The mixture of urea, described extractant is sodium acetate solution, and described buffer agent is gly-hcl buffer, and described reducing agent is two sulfur
Threitol and beta -mercaptoethanol, described enzyme agent is plant esterase.
A kind of using method of the quick detection kit of melamine in dairy products residual, specifically comprises the following steps that
(1) first, milk product is sealed off on aseptic operating platform, draw 15ml after place mortar in, add extractant and
Liquid nitrogen is quickly ground, and subsequently draws the solution after 80 μ l grind using dropper and is added drop-wise in 96 hole elisa Plates, then places
Carry out cold preservation in -80 DEG C of refrigerators of ultralow temperature stand-by；
(2) then, sodium tartrate, bovine serum albumin, lanthanum chloride, oxalic acid are added in Deoxydization nucleotide sodium solution, fully stir
Mix uniformly, place cryopreservation at lucifuge；
(3) then, 96 stand-by for cold preservation in above-mentioned steps (1) hole elisa Plates are taken out, after defrosting, add buffer agent and reducing agent,
Wherein buffer 5000 times of dilution agent, wherein reduce 700 times of dilution agent, at 4 DEG C, stand 15min；
(4) then, the Deoxydization nucleotide sodium solution after will be uniform in above-mentioned steps (2) carries out ultrafiltration, set aside for use after ultrafiltration；
(5) then, inorganic agent, enzyme agent and developer will be added in 96 hole elisa Plates after above-mentioned steps (3) standing, incubate at 37 DEG C
Educate 20min, be subsequently adding the Deoxydization nucleotide sodium solution after ultrafiltration, after fully mixing, observe color change；
(6) last, using standard color comparison card, the color after change is compared.
Wherein:
In step (2), the mass percent concentration of Deoxydization nucleotide sodium solution is 4.6%.
In step (2), bovine serum albumin derives from Zhejiang Tian Hang biotech inc.
In step (4), ultrafiltration is specially and carries out ultrafiltration using interception for 200-300kda ultrafilter membrane.
In step (5), enzyme agent is using front needing to be preheating to 37 DEG C.
Embodiment 3
A kind of quick detection kit of melamine in dairy products residual, including inorganic agent, developer, extractant, dropper, delays
Electuary, reducing agent and enzyme agent, described inorganic agent is trichloroacetic acid and phenol, and described developer is potassium mercuric iodide, phosphomolybdic acid and sulfur
The mixture of urea, described extractant is sodium acetate solution, and described buffer agent is gly-hcl buffer, and described reducing agent is two sulfur
Threitol and beta -mercaptoethanol, described enzyme agent is plant esterase.
A kind of using method of the quick detection kit of melamine in dairy products residual, specifically comprises the following steps that
(1) first, milk product is sealed off on aseptic operating platform, draw 20ml after place mortar in, add extractant and
Liquid nitrogen is quickly ground, and subsequently draws the solution after 100 μ l grind using dropper and is added drop-wise in 96 hole elisa Plates, then places
Carry out cold preservation in -80 DEG C of refrigerators of ultralow temperature stand-by；
(2) then, sodium tartrate, bovine serum albumin, lanthanum chloride, oxalic acid are added in Deoxydization nucleotide sodium solution, fully stir
Mix uniformly, place cryopreservation at lucifuge；
(3) then, 96 stand-by for cold preservation in above-mentioned steps (1) hole elisa Plates are taken out, after defrosting, add buffer agent and reducing agent,
Wherein buffer 6000 times of dilution agent, wherein reduce 800 times of dilution agent, at 4 DEG C, stand 15min；
(4) then, the Deoxydization nucleotide sodium solution after will be uniform in above-mentioned steps (2) carries out ultrafiltration, set aside for use after ultrafiltration；
(5) then, inorganic agent, enzyme agent and developer will be added in 96 hole elisa Plates after above-mentioned steps (3) standing, incubate at 37 DEG C
Educate 20min, be subsequently adding the Deoxydization nucleotide sodium solution after ultrafiltration, after fully mixing, observe color change；
(6) last, using standard color comparison card, the color after change is compared.
Wherein:
In step (2), the mass percent concentration of Deoxydization nucleotide sodium solution is 4.6%.
In step (2), bovine serum albumin derives from Zhejiang Tian Hang biotech inc.
In step (4), ultrafiltration is specially and carries out ultrafiltration using interception for 200-300kda ultrafilter membrane.
In step (5), enzyme agent is using front needing to be preheating to 37 DEG C.
Low cost of the present invention, make simple, detection sensitivity is high, its remolding sensitivity prior art exceeds decades of times and thousands of
Times, and detect that the range of linearity is significantly broadening, and easy to use, simple and quick, do not need the reagent preparation of complexity in testing
Process, layman just can be readily accomplished detection according to kit specification.
Above the better embodiment of this patent is explained in detail, but this patent is not limited to above-mentioned embodiment party
Formula, in the ken that one skilled in the relevant art possesses, can also be on the premise of without departing from this patent objective
Various changes can be made.
Claims (6)
1. a kind of melamine in dairy products residual quick detection kit it is characterised in that include inorganic agent, developer,
Extractant, dropper, buffer agent, reducing agent and enzyme agent, described inorganic agent is trichloroacetic acid and phenol, and described developer is mercuric iodixde
The mixture of potassium, phosphomolybdic acid and thiourea, described extractant is sodium acetate solution, and described buffer agent is gly-hcl buffer, described
Reducing agent is dithiothreitol, DTT and beta -mercaptoethanol, and described enzyme agent is plant esterase.
2. a kind of using method of the quick detection kit of melamine in dairy products residual as claimed in claim 1, its
It is characterised by, specifically comprise the following steps that
(1) first, milk product is sealed off on aseptic operating platform, place after drawing 10-20ml in mortar, add extractant
Quickly ground with liquid nitrogen, subsequently draw the solution after 50-100 μ l grinds using dropper and be added drop-wise in 96 hole elisa Plates, so
After be placed on that to carry out cold preservation in -80 DEG C of refrigerators of ultralow temperature stand-by；
(2) then, sodium tartrate, bovine serum albumin, lanthanum chloride, oxalic acid are added in Deoxydization nucleotide sodium solution, fully stir
Mix uniformly, place cryopreservation at lucifuge；
(3) then, 96 stand-by for cold preservation in above-mentioned steps (1) hole elisa Plates are taken out, after defrosting, add buffer agent and reducing agent,
Wherein buffer 4000-6000 times of dilution agent, wherein reduce 600-800 times of dilution agent, at 4 DEG C, stand 15min；
(4) then, the Deoxydization nucleotide sodium solution after will be uniform in above-mentioned steps (2) carries out ultrafiltration, set aside for use after ultrafiltration；
(5) then, inorganic agent, enzyme agent and developer will be added in 96 hole elisa Plates after above-mentioned steps (3) standing, incubate at 37 DEG C
Educate 20min, be subsequently adding the Deoxydization nucleotide sodium solution after ultrafiltration, after fully mixing, observe color change；
(6) last, using standard color comparison card, the color after change is compared.
3. the using method of the quick detection kit of melamine in dairy products residual according to claim 2, it is special
Levy and be, in step (2), the mass percent concentration of Deoxydization nucleotide sodium solution is 4.6%.
4. the using method of the quick detection kit of melamine in dairy products residual according to claim 2, it is special
Levy and be, in step (2), bovine serum albumin derives from Zhejiang Tian Hang biotech inc.
5. the using method of the quick detection kit of melamine in dairy products residual according to claim 2, it is special
Levy and be, in step (4), ultrafiltration is specially and carries out ultrafiltration using interception for 200-300kda ultrafilter membrane.
6. the using method of the quick detection kit of melamine in dairy products residual according to claim 2, it is special
Levy and be, in step (5), enzyme agent is using front needing to be preheating to 37 DEG C.
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| CN201610983887.XA CN106353313B (en) | 2016-11-09 | 2016-11-09 | A kind of remaining quick detection kit of melamine in dairy products and application method |
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| CN101881759A (en) * | 2009-05-05 | 2010-11-10 | 天津生机集团股份有限公司 | Tripolycyanamide rapid detection kit and application thereof |
| CN102519951A (en) * | 2011-12-06 | 2012-06-27 | 江南大学 | Color reagent made from colorimetry nanometer material and used for detecting melamine and application of color reagent |
| CN102954959A (en) * | 2011-08-29 | 2013-03-06 | 中国科学院上海有机化学研究所 | Melamine colorimetric detection method based on enzymatic amino hydrolysis reaction |
| CN102998305A (en) * | 2011-09-19 | 2013-03-27 | 青岛大学 | Method for detecting melamine in raw milk by utilizing chromogenic reaction |
| TWI412494B (en) * | 2009-12-31 | 2013-10-21 | Ind Tech Res Inst | Detection kit of melamine and detection method thereof |
| US8637269B2 (en) * | 2009-07-10 | 2014-01-28 | Bioo Scientific Corporation | Method for the detection of melamine |
| CN103645275A (en) * | 2013-12-03 | 2014-03-19 | 中山鼎晟生物科技有限公司 | Melamine detection reagent and detection method |
| CN104459009A (en) * | 2014-12-09 | 2015-03-25 | 苏州东辰林达检测技术有限公司 | Quick detection kit for melamine and using method thereof |
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2016
- 2016-11-09 CN CN201610983887.XA patent/CN106353313B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101881759A (en) * | 2009-05-05 | 2010-11-10 | 天津生机集团股份有限公司 | Tripolycyanamide rapid detection kit and application thereof |
| US8637269B2 (en) * | 2009-07-10 | 2014-01-28 | Bioo Scientific Corporation | Method for the detection of melamine |
| TWI412494B (en) * | 2009-12-31 | 2013-10-21 | Ind Tech Res Inst | Detection kit of melamine and detection method thereof |
| CN102954959A (en) * | 2011-08-29 | 2013-03-06 | 中国科学院上海有机化学研究所 | Melamine colorimetric detection method based on enzymatic amino hydrolysis reaction |
| CN102998305A (en) * | 2011-09-19 | 2013-03-27 | 青岛大学 | Method for detecting melamine in raw milk by utilizing chromogenic reaction |
| CN102519951A (en) * | 2011-12-06 | 2012-06-27 | 江南大学 | Color reagent made from colorimetry nanometer material and used for detecting melamine and application of color reagent |
| CN103645275A (en) * | 2013-12-03 | 2014-03-19 | 中山鼎晟生物科技有限公司 | Melamine detection reagent and detection method |
| CN104459009A (en) * | 2014-12-09 | 2015-03-25 | 苏州东辰林达检测技术有限公司 | Quick detection kit for melamine and using method thereof |
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| CN106353313B (en) | 2019-03-29 |
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Effective date of registration: 20190605 Address after: 530000 High-tech Zone, Nanning City, Guangxi Zhuang Autonomous Region, No. 1 High-tech Second Road, Science Park, Guangxi University Patentee after: Agricultural University Food Factory of Guangxi University Address before: 214070 Creative Park No. 100 Dicui Road, Wuxi City, Jiangsu Province, Phase III A Building 303 Patentee before: BEOSEN (JIANGSU) FOOD SAFETY TECHNOLOGY Co.,Ltd. |
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Effective date of registration: 20220729 Address after: 543003 building a-5, Donghua industrial community, Wuzhou Industrial Park, Guangxi Zhuang Autonomous Region Patentee after: Wuzhou Zhongjin Technology Co.,Ltd. Address before: 530000 High-tech Zone, Nanning City, Guangxi Zhuang Autonomous Region, No. 1 High-tech Second Road, Science Park, Guangxi University Patentee before: Agricultural University Food Factory of Guangxi University |
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