CN106596617B - One kind is based on the melamine detection method of magnetic resonance imaging (MRI) - Google Patents
One kind is based on the melamine detection method of magnetic resonance imaging (MRI) Download PDFInfo
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Abstract
The invention provides a kind of melamine detection method for being based on magnetic resonance imaging (MRI).The agglomeration of magnetic resonance T1 contrast agent MnO magnetic nano-particles, the relation established between the change in mri contrast agent relaxation time and melamine concentration, to realize the detection to melamine are induced by building melamine.This detection method is not only easy to operate, it is not necessary to which complicated sample pre-treatments, and the cost of material is low, machine is simple to operate efficiently, the shortcomings that compensate for conventional melamine detection method well, and can realize the detection of melamine in actual milk sample.
Description
Technical field
The present invention relates to field of food detection.
Background technology
Food security is always emphasis of concern, and early in September, 2008, the Sanlu milk powder of shock compatriots occurs for China
Event, this is that illegal retailer determines the detection leak of protein content using " triumphant formula nitriding ", by artificially adding three
Poly cyanamid, reach the purpose for improving protein content.So that eater occur disease in the urological system even result in it is dead (Zhou Li,
Xu Xin clouds melamines toxic action and its healthhazard [J] occupations and health, 2010 (23):2857-2859.).
Melamine (Malemine, chemical formula：C3N3(NH2)3), melamine extract of protein is commonly called as, is a kind of three class nitrogen heterocyclic rings
Organic compound, containing three amido functional groups, aqueous solution state carries electropositive.Melamine as a kind of raw material of industry,
The industrial productions such as coating, plastics, binder, weaving, papermaking are widely used in, are not useable for food processing or food addition.At present,
The conventional melamine detection method in dairy products that contains mainly includes：High performance liquid chromatography (HPLC), liquid phase chromatogram-mass spectrometry combination
Usage (HPLC-MS), gas chromatography-mass spectrography (GC-MS) etc..These method instruments all costly, and are located before sample
Reason is complicated, detects time-consuming serious, it is more difficult to realize economic simple efficient application.(the detection side of Wang Ming brightness melamine in dairy products
Method [J] modern farming enterprise's science and technology, 2016 (5):179-179.).
Constantly improve and development with nanometer technology, based on the unique magnetic property of magnetic Nano material, and its is good
Good surface modificability and biocompatibility, magnetic nano-particle is more and more extensive in the application of each research field, for example,
Tumour magnetic heat cure technology, Magnetic cell sorting technology, the drug targeting fortune developed using the magnetic performance of magnetic Nano material
Transferring technology etc., there is good development prospect.Meanwhile the extensive use of nanometer technology has also promoted the new inspection of multi-crossed disciplines
The development of survey technology.At present, based on mri contrast agent image-forming principle, the magnetic nano-particle radiography of research complex functionality
Agent, the method that the influence using its special magnetic property and to the relaxation time detects target analytes come qualitative and quantitative, no
It is only less demanding to sample pre-treatments, and method is simply efficient, has become new breakthrough and research that analysis detection instantly is learned
Focus.
There are 5 azygous electronics outside paramagnetic metal manganese atom core, there is higher magnetic susceptibility, and the poison of manganese ion
Property is low, therefore manganese oxide (MnO) magnetic nano-particle is more and more extensive as the application of nuclear magnetic resonance T1 contrast agent in recent years.
CN103920168 also reports nmr sensor and its preparation based on iron oxide magnetic nano detection of particles melamine
Methods and applications, but its used nmr sensor is T2 contrast agent Fe/Fe3O4Magnetic nano-particle, it was prepared
Need to use harsh synthesis and separation condition in journey：Reaction must be carried out under an inert gas, and reaction product must pass through post
Chromatographic purification.
The content of the invention
Present invention aims at provide a kind of new melamine detection method based on magnetic resonance imaging.Pass through structure three
Poly cyanamid induces the agglomeration of MnO magnetic nano-particles, establishes change and the melamine in mri contrast agent relaxation time
Relation between amine concentration, to realize the detection to melamine.
Technical scheme is as follows：
A kind of melamine detection method based on magnetic resonance imaging, comprises the following steps：
1) the MnO nano materials being modified by Succimer, i.e. MnO-DMSA are prepared, preparation method is：
Manganese acetate (Mn (Ac)2) with trioctylamine (TOA) and oleic acid (OA) according to mol ratio it is (1.5-10):17:(1.5-
6), in 290-320 DEG C of isothermal reaction 10-30min.Reaction product MnO nanocrystals are cleaned 2-3 times with ethanol.
Dimercaptosuccinic acid (DMSA) is weighed again, while measures dimethyl sulfoxide (DMSO) solution, is configured to 1-10mg/mL's
DMSA/DMSO solution As.Then MnO nanocrystal is taken, is scattered in again in ethanol solution, the material for obtaining 1-10mg/ml is molten
Liquid B.Solution A and solution B will be appealed by volume (1-5)：After (1-5) fully shaking mixes, vortex oscillation is overnight (to be preferably
800-1000r/min).Product is collected by centrifugation, is cleaned each 2~3 times with absolute ethyl alcohol and deionized water, finally by the MnO- of collection
DMSA is scattered in deionized water.
2) MnO-DMSA nano materials are configured to standard liquid, it is standard liquid one；Prepare melamine mark simultaneously
Quasi- solution, it is standard liquid two；The concentration of standard liquid one is 0.1-10mg/mL, and the concentration of standard liquid two is 10-4-mM-10- 2mM；
3) standard liquid one of different series volume and the standard liquid of different series volume are added in different test tubes
Two；Then add appropriate pure water so that the often liquor capacity polishing in pipe to same volume, this is volume one；To each it try
Pipe, which is acutely vortexed, makes material fully mix；
4) agar solution of volume two is added to above-mentioned all test tubes again, be acutely vortexed 2-10min, until material fully shakes
Even fixation, agar solution quality volume fraction are 0.5-5%；Volume one：Volume two is 1:5-5:1；
5) sample at ambient temperature, is placed in magnetic resonance imager and carries out magnetic resonance imaging experiment；Obtain melamine
Corresponding MnO-DMSA relaxation time 1/T1 and concentration curve variation relation figure；
6) by testing sample wiring solution-forming, mixed with standard liquid one, detect the relaxation of MnO-DMSA corresponding to melamine
Henan time 1/T1, the curve that control step 5) obtains, obtains melamine concentration.
In preferred embodiment, the concentration of standard liquid one is 0.5-5mg/mL, and the concentration of standard liquid two is 10-4mM-
10-2mM。
In preferred embodiment, volume one is 100-1000 μ L.
In preferred embodiment, in step 5), the volume one of volume two and step 3) is equal.
In preferred embodiment, step 2)：MnO-DMSA is configured to the solution that concentration is 1mg/mL respectively, matched somebody with somebody simultaneously
Concentration processed is 10-3MM melamine solution；
Step 3) takes 1.5mL 40, test tube, is divided into 5 groups；Then pipette successively respectively 1mg/mL MnO-DMSA 50,
35th, 25,16,12.5,8,6.25,4 μ L are added in 1 group of above-mentioned 8 test tube, other 4 groups of repetition aforesaid operations, and each volume
Corresponding label 1-8；
Component under each concentration of step 4) be sequentially added into the 10-3mM of above-mentioned preparation melamine solution 0,5,
7.5th, for 10,15,20,25 μ L in 7 test tubes, test tube corresponds to label 1-1,1-2,1-3,1-4,1-5,1-6,1-7 etc..Then add
Appropriate pure water so that the often liquor capacity polishing in pipe to 500 μ L；
6) the μ L of agar solution 500 that quality volume fraction is 2% are added to above-mentioned all test tubes again, be acutely vortexed 2min,
Until material fully shakes up fixation, it is easy to the detection in MRI relaxation time.
In preferred embodiment, step 7) using Varian 7.0T/160mm magnetic resonance imagers carry out magnetic resonance into
As experiment.
In preferred embodiment, test parameter setting：TR=3000ms, TE=15ms, the visual field are 45 × 45mm.
Advantage of the present invention is as follows：
There are 5 azygous electronics outside paramagnetic metal manganese atom core, there is higher magnetic susceptibility, and the poison of manganese ion
Property is low, therefore manganese oxide (MnO) magnetic nano-particle is more and more extensive as the application of nuclear magnetic resonance T1 contrast agent in recent years.This
In invention, we induce the agglomeration of MnO magnetic nano-particles by building melamine, establish mri contrast agent relaxation
Relation between the change of Henan time and melamine concentration, to realize the detection to melamine.Compared with background technology, this hair
Nuclear magnetic resonance detector is nuclear magnetic resonance T1 contrast agent MnO magnetic nano-particles used by bright, and it is synthesized in gentle experiment
Under the conditions of can complete；Product separation process also only needs to be cleaned with ethanol and pure water.Meanwhile prepared by the present invention
MnO magnetic nano-particles have very strong antijamming capability, in the actual food product detection of complexity, can be used directly to detect city
The melamine in milk is sold, the sample pre-treatments that detection process need not be complicated, the cost of material is low, machine height simple to operate
Effect, the shortcomings that compensate for conventional melamine detection method well.
Brief description of the drawings
Fig. 1 is the transmission electron microscope photo of the MnO nano-particles prepared in embodiment 1, from figure 1 it appears that prepared
MnO nano particle diameters than more uniform.
Fig. 2 is transmission electron microscope (TEM) photo of the MnO-DMSA nano-particles prepared in embodiment 1.
Fig. 3 is MnO (A) and modified (B) before modified infrared fourier conversion infrared spectrogram (Fourier
Transform infrared spectroscopy, FTIR), 2920cm-1 and 2849cm-1 absorption intensity disappear substantially,
I.e. modified MnO-DMSA has not had-CH2-, illustrates that oleic acid molecular is replaced by DMSA substantially, that is, has obtained MnO-
DMSA compound particles.
Fig. 4 is that MnO-DMSA is used for melamine detection mechanism schematic diagram.
Fig. 5 is to add after melamine MnO transmission electron microscopes (TEM) picture reunited, it can be seen that a large amount of particles
Reunite.
Fig. 6 is MnO-DMSA 1/T1 corresponding to 0~315ppb melamines and concentration curve variation diagram.
Fig. 7 is the variation diagram for the r1 that 0~315ppb melamines correspond to MnO-DMSA.
Fig. 8 is the variation diagram of the relaxation rate r1 of the MnO obtained by detection melamine in certain commercially available brand milk powder.
Embodiment
With reference to embodiment and accompanying drawing, the present invention is described further.
Embodiment 1：
A kind of new melamine detection method based on magnetic resonance imaging.
1) the MnO nano-particles of DMSA modifications are prepared：Weigh 0.2862g manganese acetate (Mn (Ac)2), the three of 5.8mL are pungent
Amine (TOA), 1.55mL oleic acid (OA), as in reaction heating mantle, are warming up to 310 DEG C of isothermal reactions in round-bottomed flask
30min.Reaction terminates rear product MnO nanocrystals and cleaned 2-3 times with ethanol.
15mg dimercaptosuccinic acid (DMSA) is weighed again, while measures 3mL dimethyl sulfoxide (DMSO) solution, is configured to
5mg/mL DMSA/DMSO solution As.Then 15mgMnO nanocrystal is taken, is scattered in again in 5mL ethanol solution, is obtained
Material solution B.After solution A and the mixing of solution B fully shaking will be appealed, vortex oscillation is overnight (900r/min).Production is collected by centrifugation
Thing, cleaned each 2~3 times with absolute ethyl alcohol and deionized water, finally by the scattered (1mg/ in deionized water of the MnO-DMSA of collection
ml)。
2) will be configured to concentration respectively by the modified MnO nano materials (MnO-DMSA) of Succimer (DMSA) is
1mg/mL solution, while compound concentration is 10-3MM melamine solution；
1.5mL 40, test tube is taken, is divided into 5 groups.Then pipette successively respectively 1mg/mL MnO-DMSA 50,35,25,
16th, 12.5,8,6.25,4L is added in 1 group of above-mentioned 8 test tube, other 4 groups of repetition aforesaid operations, and each volume is corresponding marks
Number 1~8；
3) component under each concentration is sequentially added into the 10 of above-mentioned preparation-3MM melamine solution 0,5,7.5,
10th, 15,20, for 25L in 7 test tubes, test tube corresponds to label 1-1,1-2,1-3,1-4,1-5,1-6,1-7 etc..Then add appropriate
Pure water so that the often liquor capacity polishing in pipe to 500 μ L；Each test tube is acutely vortexed 2min so that material fully mixes；
4) the μ L of agar solution 500 that quality volume fraction is 2% are added to above-mentioned all test tubes again, be acutely vortexed 2min,
Until material fully shakes up fixation, it is easy to the detection in MRI relaxation time.
5) sample at ambient temperature, is placed in Varian 7.0T/160mm magnetic resonance imagers and carries out magnetic resonance imaging
Experiment.Test parameter is set：TR=3000ms, TE=15ms, the visual field are 45 × 45mm.
Fig. 1 is the TEM photos of the MnO nano-particles prepared in embodiment 1, from figure 1 it appears that prepared MnO
Nano particle diameter is than more uniform.
Fig. 2 is the TEM photos of the MnO-DMSA nano-particles prepared in embodiment 1.
The FTIR that Fig. 3 is MnO (A) and modified (B) before modified schemes, 2920cm-1 and 2849cm-1 absorption intensity base
This disappearance, i.e., modified MnO-DMSA and do not have-CH2-, illustrates that oleic acid molecular is replaced by DMSA substantially, it was demonstrated that
MnO-DMSA compound particles are arrived.
Fig. 5 is the TEM pictures for adding after melamine the MnO to reunite, it can be seen that group occurs for a large amount of particles
It is poly-.
Fig. 6 is that the relaxation time (1/T1) of MnO-DMSA corresponding to 0~315ppb melamines and concentration curve change are closed
System's figure, it can be seen that as concentration is higher, corresponding r1 is lower.
Embodiment 2：
A kind of new melamine detection method based on magnetic resonance imaging, with embodiment 1, difference is to walk step
It is rapid 1) in, the isothermal reaction time for preparing MnO nano-particles is 20min.
Embodiment 3：
A kind of new melamine detection method based on magnetic resonance imaging, with embodiment 1, difference is to walk step
It is rapid 1) in, add 1.7g manganese acetate (Mn (Ac)2)。
Embodiment 4：
A kind of new melamine detection method based on magnetic resonance imaging, with embodiment 1, difference is to walk step
It is rapid 2) in, prepare melamine solution concentration as 2 × 10-3MM, in step 3), then add appropriate pure water so that every
Liquor capacity polishing in pipe is to 1000 μ L.
Embodiment 5：
A kind of new melamine detection method based on magnetic resonance imaging, with embodiment 1, difference is to walk step
It is rapid 1) in, prepare melamine solution concentration as 0.5 × 10-3MM, in step 3), then add appropriate pure water so that
Often the liquor capacity polishing in pipe is to 250 μ l.
Embodiment 6：
A kind of new melamine detection method based on magnetic resonance imaging, with embodiment 1, difference is to walk step
It is rapid 4) in all test tubes add the μ L of agar solution 500 that quality volume fractions are 1%, be acutely vortexed 2min.
Embodiment 7：
A kind of new melamine detection method based on magnetic resonance imaging, with embodiment 1, difference is to walk step
It is rapid 3) in, be sequentially added into the component under each concentration the 10-3mM of above-mentioned preparation μ L of melamine solution 0,50 μ L in
In 8 test tubes, certain commercially available brand milk power solution polishing that concentration is 100mg/mL is then added into each test tube to 500 μ L.Fig. 8
To detect the relaxation rate r1 of the MnO obtained by melamine change in certain commercially available brand milk powder.It can be seen that work as
When detecting the melamine in milk powder, r1 values are reduced to 6.1mM/s there occurs obvious change from 8.4.
Claims (6)
1. a kind of melamine detection method based on magnetic resonance imaging, comprises the following steps：
1) the MnO nano materials being modified by Succimer, i.e. MnO-DMSA are prepared, its preparation method is：
Manganese acetate is (1.5-10) according to mol ratio with oleic acid with trioctylamine:17:(1.5-6), in 290-320 DEG C of isothermal reaction
10-30min, reaction product MnO nanocrystals are cleaned 2-3 times with ethanol；
Dimercaptosuccinic acid is weighed again, while measures dimethyl sulfoxide solution, is configured to 1-10mg/mL DMSA/DMSO solution As；
Then MnO nanocrystal is taken, is scattered in again in ethanol solution, obtains 1-10mg/ml material solution B；By above-mentioned solution
A and solution B are by volume (1-5)：After (1-5) fully shaking mixes, vortex oscillation is overnight；Product is collected by centrifugation, with anhydrous second
Alcohol and deionized water are cleaned each 2-3 times, finally that the MnO-DMSA of collection is scattered in deionized water；
2) MnO-DMSA nano materials are configured to standard liquid, it is standard liquid one；It is molten to prepare melamine standard simultaneously
Liquid, it is standard liquid two；The concentration of standard liquid one is 0.1-10mg/mL, and the concentration of standard liquid two is 10-4mM-10-2mM；
3) standard liquid one of different series volume and the standard liquid two of different series volume are added in different test tubes；With
Appropriate pure water is added afterwards so that the often liquor capacity polishing in pipe to same volume, this is volume one；Each test tube is violent
Vortex makes material fully mix；
4) agar solution of volume two is added to above-mentioned all test tubes again, be acutely vortexed 2-10min, until material fully shakes up admittedly
Fixed, agar solution quality volume fraction is 0.5-5%；Volume one：Volume two is 1:5-5:1；
5) sample at ambient temperature, is placed in magnetic resonance imager and carries out magnetic resonance imaging experiment；It is corresponding to obtain melamine
MnO-DMSA relaxation time 1/T1 and concentration curve variation relation figure；
6) by testing sample wiring solution-forming, mixed with standard liquid one, when detecting MnO-DMSA relaxation corresponding to melamine
Between 1/T1, the obtained curve of control step 5), obtain melamine concentration.
A kind of 2. melamine detection method based on magnetic resonance imaging as claimed in claim 1, it is characterised in that：Standard is molten
The concentration of liquid one is 0.5-5mg/mL, and the concentration of standard liquid two is 10-4mM-10-2mM。
A kind of 3. melamine detection method based on magnetic resonance imaging as claimed in claim 1, it is characterised in that：Volume one
For 100-1000 μ L.
A kind of 4. melamine detection method based on magnetic resonance imaging as claimed in claim 1, it is characterised in that：Step 4)
In, the volume one of volume two and step 3) is equal.
5. a kind of melamine detection method based on magnetic resonance imaging as described in any one of Claims 1-4, its feature exist
In：Step 5) carries out magnetic resonance imaging experiment using the T/160mm magnetic resonance imagers of Varian 7.0.
A kind of 6. melamine detection method based on magnetic resonance imaging as claimed in claim 5, it is characterised in that：Test ginseng
Number setting：TR=3000ms, TE=15ms, the visual field are 45 × 45mm.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103920168A (en) * | 2014-03-17 | 2014-07-16 | 上海师范大学 | Nuclear magnetic resonance sensor for detecting melamine based on magnetic nanoparticles, and preparation method and application thereof |
| CN104436222A (en) * | 2014-12-11 | 2015-03-25 | 厦门大学 | Synthesis method of MnO nanometer material of T1 contrast agent |
| CN104764706A (en) * | 2015-04-03 | 2015-07-08 | 上海师范大学 | Melamine dual-mode sensor based on Au-Fe3O4 composite nanoparticles and preparation method thereof |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007064175A1 (en) * | 2005-12-02 | 2007-06-07 | Industry-Academic Cooperation Foundation, Yonsei University | Magnetic resonance imaging contrast agents containing water-soluble nanoparticles of manganese oxide or manganese metal oxide |
| US20120114564A1 (en) * | 2007-01-30 | 2012-05-10 | Seoul National University Industry Foundation | Mri t1 contrasting agent comprising manganese oxide nanoparticle |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103920168A (en) * | 2014-03-17 | 2014-07-16 | 上海师范大学 | Nuclear magnetic resonance sensor for detecting melamine based on magnetic nanoparticles, and preparation method and application thereof |
| CN104436222A (en) * | 2014-12-11 | 2015-03-25 | 厦门大学 | Synthesis method of MnO nanometer material of T1 contrast agent |
| CN104764706A (en) * | 2015-04-03 | 2015-07-08 | 上海师范大学 | Melamine dual-mode sensor based on Au-Fe3O4 composite nanoparticles and preparation method thereof |
Non-Patent Citations (2)
| Title |
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| MnO-Labeled Cells: Positive Contrast Enhancement in MRI;Mathieu L. et al.;《The Journal of Physical Chemistry》;20121003;第116卷(第44期);13228-13238 * |
| 单分散MnO纳米粒子的可控制备;刘新杰 等;《陕西师范大学学报（自然科学版）》;20130731;第41卷(第4期);49-54 * |
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