CN103175818B - Method for detecting melamine - Google Patents
Method for detecting melamine Download PDFInfo
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- CN103175818B CN103175818B CN201310087869.XA CN201310087869A CN103175818B CN 103175818 B CN103175818 B CN 103175818B CN 201310087869 A CN201310087869 A CN 201310087869A CN 103175818 B CN103175818 B CN 103175818B
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- solution
- melamine
- value
- milk
- fluorescent value
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- 229920000877 Melamine resin Polymers 0.000 title claims abstract description 34
- JDSHMPZPIAZGSV-UHFFFAOYSA-N melamine Chemical compound NC1=NC(N)=NC(N)=N1 JDSHMPZPIAZGSV-UHFFFAOYSA-N 0.000 title claims abstract description 31
- 238000000034 method Methods 0.000 title claims abstract description 16
- 239000000243 solution Substances 0.000 claims abstract description 33
- 235000013336 milk Nutrition 0.000 claims abstract description 12
- 239000008267 milk Substances 0.000 claims abstract description 12
- 210000004080 milk Anatomy 0.000 claims abstract description 12
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 9
- YQGOJNYOYNNSMM-UHFFFAOYSA-N eosin Chemical compound [Na+].OC(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C(O)=C(Br)C=C21 YQGOJNYOYNNSMM-UHFFFAOYSA-N 0.000 claims abstract description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 5
- 230000005284 excitation Effects 0.000 claims abstract description 4
- 229940046892 lead acetate Drugs 0.000 claims abstract description 4
- 239000010453 quartz Substances 0.000 claims abstract description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 4
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 claims abstract description 4
- 239000007788 liquid Substances 0.000 claims description 7
- KVYRCBOUKXJXDK-UHFFFAOYSA-N 3,4-dimethylphenazine-1,2-diamine hydrochloride Chemical compound Cl.C1=CC=CC2=NC3=C(C)C(C)=C(N)C(N)=C3N=C21 KVYRCBOUKXJXDK-UHFFFAOYSA-N 0.000 claims description 3
- 239000007853 buffer solution Substances 0.000 claims description 3
- 238000012417 linear regression Methods 0.000 claims description 3
- 239000006228 supernatant Substances 0.000 claims description 3
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 3
- 239000012498 ultrapure water Substances 0.000 claims description 3
- 239000006171 Britton–Robinson buffer Substances 0.000 abstract 1
- PGSADBUBUOPOJS-UHFFFAOYSA-N neutral red Chemical compound Cl.C1=C(C)C(N)=CC2=NC3=CC(N(C)C)=CC=C3N=C21 PGSADBUBUOPOJS-UHFFFAOYSA-N 0.000 abstract 1
- 238000009210 therapy by ultrasound Methods 0.000 abstract 1
- 238000004458 analytical method Methods 0.000 description 3
- 235000019219 chocolate Nutrition 0.000 description 3
- 238000002189 fluorescence spectrum Methods 0.000 description 3
- 150000007974 melamines Chemical class 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 238000004611 spectroscopical analysis Methods 0.000 description 2
- 238000003556 assay Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 239000003317 industrial substance Substances 0.000 description 1
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 1
- IJGRMHOSHXDMSA-UHFFFAOYSA-N nitrogen Substances N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 235000020185 raw untreated milk Nutrition 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- -1 triazines nitrogen heterocyclic ring organic compound Chemical class 0.000 description 1
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- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The invention discloses a method for detecting melamine. The method comprises the following steps of: sequentially adding 0.05-5.0mL of 0.1-0.5g/L melamine solution, 1.0-2.5ml of 4.0*10<-6>mol/L eosin Y-solution, 0.5-1.0ml of 2.0*10<-4>mol/L neutral red solution and 0.3-1.0ml of Britton-Robinson buffer solution of which the pH value is 3.5 into a 10ml colorimetric tube, using the solution without melamine as the reagent blank, fixing the volume to a specific scale by using super-pure water, reacting for 30min in a water bath of 30 DEG C, subsequently respectively measuring the fluorescence value F538nm of the melamine-containing solution and the fluorescence value F0 of the reagent blank and calculating the difference value that deltaF538nm is equal to F538nm subtracted by the F0 value at 538nm on a fluorophotometer by using a 1cm quartz cuvette in a mode that 520nm is used as the excitation wavelength; preparing 30ml of milk, adding 20.0ml of 1% trichloroacetic acid solution and 8.0ml of 2% lead acetate solution, and carrying out ultrasound treatment for 20min; centrifuging a part of the solution for 10min, taking 1ml of the supernate, and measuring the fluorescence value and calculating the content of the melamine in the milk according to the same way. The method is simple, convenient and rapid.
Description
Technical field
The present invention relates to a kind of method of measuring melamine, particularly energy shifts the method for fluorescence spectrum method for measuring melamine.
Background technology
Melamine is a kind of important triazines nitrogen heterocyclic ring organic compound, is commonly used for industrial chemicals, is not useable for food processing or food additives.China is after Sanlu milk powder case in 2008 occurs, and national regulation: in liquid milk (comprising raw milk), milk powder, other prescription emulsifiable powders, the Limited Doses of melamine is 2.5mg/kg, must not sell without exception higher than the product of 2.5mg/kg.Therefore the new method that content of melamine in a kind of mensuration milk quick, accurate, with low cost is set up in research has great importance.At present, the assay method of melamine is mainly high performance liquid chromatography, GC-MS(gas chromatography-mass spectrography), Liquid Chromatography-Mass Spectrometry etc.But these method instruments are expensive, be difficult for promoting.Fluorescent spectrometry has that analysis speed is fast, easy and simple to handle, analysis precision advantages of higher, in actual analysis, is widely used.Eosin W or W S-dimethyl diaminophenazine chloride, as energy transfer system, has been applied to protein measuring, but utilizes this energy transfer system to adopt fluorescence spectrum method for measuring melamine, there is not yet report.
Summary of the invention
The fluorescent spectrometry that the object of this invention is to provide a kind of mensuration melamine shifting based on eosin W or W S-dimethyl diaminophenazine chloride energy.
Concrete steps are:
(1) in 6 10mL color comparison tubes, add successively the melamine solution of 0.0,0.07,1.0,2.0,3.0,5.0 mL 0.1～0.5g/L, then in every color comparison tube, add respectively 1.0～2.5 mL 4.0 * 10
-6the eosin W or W S of mol/L (EY) solution, 0.5～1.0mL 2.0 * 10
-4the Bloomsbury of the dimethyl diaminophenazine chloride of mol/L (NR) solution and 0.3～1.0mL pH3.5 is smooth-Robison (Britton-Robinson, B-R) buffer solution, the solution that does not add melamine of take is reagent blank, then with ultrapure water, 6 color comparison tubes are settled to scale, shake up, in the water bath with thermostatic control of 30 ℃, react after 30 min, on fluorophotometer, use 1cm quartz colorimetric utensil, take 520nm as excitation wavelength, at 538nm place, measure respectively the fluorescent value F containing melamine solution
538nmfluorescent value F with reagent blank
0, calculate fluorescent value difference DELTA F
538nm=F
538nm– F
0value, its fluorescent value difference DELTA F
538nmρ is linear within the scope of 1.4～100mg/L with melamine mass concentration, and equation of linear regression is: Δ F
538nm=
0.7736ρ
+ 17.33, correlation coefficient r=0.9992, detects and is limited to 1.1mg/L.
(2) get milk 30 mL, add the lead acetate solution that trichloroacetic acid solution that 20.0 mL mass percent concentrations are 1% and 8.0 mL mass percent concentrations are 2%, ultrasonic 20 min.Then get part solution in centrifuge tube, centrifugal 10 min, get supernatant as liquid to be measured.
(3) get the milk liquid 1mL to be measured after step (2) is processed, according to the method for step (1), measure fluorescent value, calculate the content of melamine in milk.
The inventive method is easy, quick.
Accompanying drawing explanation
Fig. 1 is the fluorescence spectrum figure of embodiment of the present invention blank and 20 mg/L melamines (MEL).
Mark a in figure: reagent blank; B:4.0 * 10
-6mol/L EY-2.0 * 10
-4mol/L NR-pH3.5 B-R-20 mg/L MEL.
Embodiment
Embodiment:
(1) in 6 10mL color comparison tubes, add successively the melamine solution of 0.0,0.07,1.0,2.0,3.0,5.0 mL 0.2g/L, then in every color comparison tube, add respectively 2.0 mL 4.0 * 10
-6the eosin W or W S of mol/L (EY) solution, 0.5mL 2.0 * 10
-4the Bloomsbury of the dimethyl diaminophenazine chloride of mol/L (NR) solution and 0.5mL pH3.5 is smooth-Robison (Britton-Robinson, B-R) buffer solution, the solution that does not add melamine of take is reagent blank, then with ultrapure water, 6 color comparison tubes are settled to scale, shake up, in the water bath with thermostatic control of 30 ℃, react after 30 min, on fluorophotometer, use 1cm quartz colorimetric utensil, take 520nm as excitation wavelength, at wavelength 538nm place, measure respectively the fluorescent value F containing melamine solution
538nmfluorescent value F with reagent blank
0, calculate fluorescent value difference DELTA F
538nm=F
538nm– F
0value, its fluorescent value difference DELTA F
538nmρ is linear within the scope of 1.4～100mg/L with melamine mass concentration, and equation of linear regression is: Δ F
538nm=
0.7736ρ
+ 17.33, correlation coefficient r=0.9992, detects and is limited to 1.1mg/L.
(2) measure respectively each 30 mL of Mongolia Ox and Erie's plain chocolate, add respectively the lead acetate solution that trichloroacetic acid solution that 20.0 mL mass percent concentrations are 1% and 8.0 mL mass percent concentrations are 2%, ultrasonic 20 min; Then get part solution in centrifuge tube, centrifugal 10 min, get supernatant as liquid to be measured.
(3) get Mongolia Ox and each 1mL of Erie's plain chocolate liquid to be measured after step (2) is processed, according to step (1) method, measure fluorescent value, calculate the content of melamine in Mongolia Ox, Erie's plain chocolate; Do recovery testu simultaneously, the results are shown in Table 1:
Claims (1)
1. measure a method for melamine, it is characterized in that concrete steps are:
(1) in 6 10mL color comparison tubes, add successively the melamine solution of 0.0,0.07,1.0,2.0,3.0,5.0 mL 0.1～0.5g/L, then in every color comparison tube, add respectively 1.0～2.5 mL 4.0 * 10
-6the eosin W or W S of mol/L is EY solution, 0.5～1.0mL 2.0 * 10
-4the dimethyl diaminophenazine chloride of mol/L be the Bloomsbury of NR solution and 0.3～1.0mL pH3.5 smooth-Robison is B-R buffer solution, the solution that does not add melamine of take is reagent blank, then with ultrapure water, 6 color comparison tubes are settled to scale, shake up, in the water bath with thermostatic control of 30 ℃, react after 30 min, on fluorophotometer, use 1cm quartz colorimetric utensil, take 520nm as excitation wavelength, at 538nm place, measure respectively the fluorescent value F containing melamine solution
538nmfluorescent value F with reagent blank
0, calculate fluorescent value difference DELTA F
538nm=F
538nm– F
0value, its fluorescent value difference DELTA F
538nmρ is linear within the scope of 1.4～100mg/L with melamine mass concentration, and equation of linear regression is: Δ F
538nm=
0.7736ρ
+ 17.33, correlation coefficient r=0.9992, detects and is limited to 1.1mg/L;
(2) get milk 30 mL, add the trichloroacetic acid solution that 20.0 mL mass percent concentrations are 1%, and 8.0 mL mass percent concentrations be 2% lead acetate solution, ultrasonic 20 min; Then get part solution in centrifuge tube, centrifugal 10 min, get supernatant as liquid to be measured;
(3) get the milk liquid 1mL to be measured after step (2) is processed, according to the method for step (1), measure fluorescent value, calculate the content of melamine in milk.
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| CN201310087869.XA CN103175818B (en) | 2013-03-19 | 2013-03-19 | Method for detecting melamine |
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|---|---|---|---|
| CN201310087869.XA CN103175818B (en) | 2013-03-19 | 2013-03-19 | Method for detecting melamine |
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| CN103175818A CN103175818A (en) | 2013-06-26 |
| CN103175818B true CN103175818B (en) | 2014-12-10 |
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| CN201310087869.XA Active CN103175818B (en) | 2013-03-19 | 2013-03-19 | Method for detecting melamine |
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Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104880444A (en) * | 2015-05-21 | 2015-09-02 | 桂林理工大学 | Method for determining kanamycin sulfate |
| CN105158225B (en) * | 2015-09-16 | 2017-11-17 | 盐城工学院 | A kind of method of two-photon fluorescence excitation detection melamine |
| CN106501243B (en) * | 2016-10-01 | 2019-10-11 | 桂林理工大学 | A method of melamine is quickly detected with molecular engram test strips |
| CN108152233B (en) * | 2018-01-03 | 2020-07-31 | 桂林理工大学 | Method for measuring melamine based on methyl blue |
| CN108254344B (en) * | 2018-01-03 | 2020-07-31 | 桂林理工大学 | Method for measuring melamine by using crystal violet |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5955002A (en) * | 1997-11-12 | 1999-09-21 | Spectra Group Limited, Inc. | Method for determining properties of a polymer coating or film cured by cationic polymerization |
| CN101576543A (en) * | 2009-04-21 | 2009-11-11 | 新疆出入境检验检疫局检验检疫技术中心 | Method for fast detecting melamine in dry milk |
| CN102608086A (en) * | 2012-01-12 | 2012-07-25 | 吉林大学 | Method for detecting melamine in milk on basis of inner-filter effect of fluorescence between CdTe quantum dots and AuNPs |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS649345A (en) * | 1987-06-30 | 1989-01-12 | Shimadzu Corp | Method of analyzing porphyrins |
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2013
- 2013-03-19 CN CN201310087869.XA patent/CN103175818B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5955002A (en) * | 1997-11-12 | 1999-09-21 | Spectra Group Limited, Inc. | Method for determining properties of a polymer coating or film cured by cationic polymerization |
| CN101576543A (en) * | 2009-04-21 | 2009-11-11 | 新疆出入境检验检疫局检验检疫技术中心 | Method for fast detecting melamine in dry milk |
| CN102608086A (en) * | 2012-01-12 | 2012-07-25 | 吉林大学 | Method for detecting melamine in milk on basis of inner-filter effect of fluorescence between CdTe quantum dots and AuNPs |
Non-Patent Citations (5)
| Title |
|---|
| JP昭64-9345A 1989.01.12 * |
| 刘保生等.曙红-中性红荧光能量转移抑制法测定肝素.《分析试验室》.2009,第28卷(第9期), * |
| 刘金彦等.荧光光谱法测定奶制品中的三聚氰胺.《分析试验室》.2010,第29卷(第9期), * |
| 曙红-中性红荧光能量转移抑制法测定肝素;刘保生等;《分析试验室》;20090930;第28卷(第9期);第16-18页 * |
| 荧光光谱法测定奶制品中的三聚氰胺;刘金彦等;《分析试验室》;20100930;第29卷(第9期);第26-27页 * |
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