CN107892914A - A kind of quick identification and the fluorescent microsphere preparation method for quantitatively detecting mercury ion - Google Patents
A kind of quick identification and the fluorescent microsphere preparation method for quantitatively detecting mercury ion Download PDFInfo
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- CN107892914A CN107892914A CN201711292361.8A CN201711292361A CN107892914A CN 107892914 A CN107892914 A CN 107892914A CN 201711292361 A CN201711292361 A CN 201711292361A CN 107892914 A CN107892914 A CN 107892914A
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- fluorescent microsphere
- microsphere
- mercury ion
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- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
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- C—CHEMISTRY; METALLURGY
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- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/02—Use of particular materials as binders, particle coatings or suspension media therefor
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
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Abstract
Detect the invention discloses a kind of quick identification and quantitatively the fluorescent microsphere preparation method of mercury ion, it is characterised in that comprise the following steps：（1）Fluorescent small molecule is prepared as reactant using citric acid and 3 aminopropyl trimethoxy siloxanes；（2）Using tetraethyl orthosilicate as raw material, ethanol is medium, and preparing monodisperse silica microspheres by ammonia-catalyzed hydrolysis and cohesion is used as molecular vehicle；（3）Using tetrahydrofuran as solvent, the silicone hydroxyl of microsphere surface is substituted by fluorescent small molecule, prepares fluorescent microsphere.（4）Fluorescent microsphere solution is configured by solvent of deionized water.It is its uniform particle diameter, controllable using fluorescent microsphere particle diameter made from method provided by the present invention in the range of 100 ~ 500 nm, to Hg2+With efficient identification and sensitivity, detection is limited to 8.7 nM, and Quantitative detection can be achieved in 0 ~ 25 μM of concentration range.This method has the characteristics of easy to operate, quick, accuracy rate is high, test limit is low.
Description
Technical field
The fluorescent microsphere preparation method of mercury ion is detected the present invention relates to a kind of quick identification and quantitatively, belongs to fluorescence probe
Technical field.
Background technology
Mercury ion（Hg2+）It is a kind of heavy metal that environment and biosystem can be also damaged even in very low concentration
Ion.It can be easily by biomembrane, and Central nervous and internal system cause serious harm therefore, U.S. environment
Protection administration（USEPA）Setting highest in drinking water allows Hg2+Concentration is 2 μ g L−1（About 0.01 μm of ol L 1）.So
The accurately Hg in measure biological tissue and environment water system2+As an important field of research.
Conventional Hg2+Detection technique has atomic absorption spectrum（AAS）, atomic emission spectrum（AES）, atomic fluorescence spectrophotometry
（AFS）And inductivity coupled plasma mass spectrometry（ICP-MS）.However, these technologies need to use the instrument of costliness and complicated sample
Product pre-treatment flow.In recent years, due to easy to operate, the inexpensive and efficiency high of fluorescence spectrum the advantages that, design and synthesize new
Fluorescent molecular probe in Hg2+Detection field has obtained fast development.This patent overcomes the above insufficient, and having prepared one kind makes
With convenience, the Hg that test limit is low, strong antijamming capability can carry out Quantitative detection simultaneously2+Probe, there is very important reality
Border meaning.
The content of the invention
In view of the shortcomings of the prior art, one of present disclosure is to provide a kind of is prepared not by chemical bonding
The preparation method of the fluorescent microsphere of the group containing Representative fluorescence.
The fluorescent microsphere that the two of present disclosure are to provide can be to Hg2+Carry out high efficiency identification and fast quantification inspection
Survey, detection is limited to 8.7 nM, and Quantitative detection can be carried out in 0 ~ 25 μM of concentration range.With existing Hg2+Concentration detection method
Compare, the fluorescent microsphere is to Hg2+Test limit is far below other patents, and it is easy to operate, quick, accuracy rate is high, anti-interference energy
Power is strong.
The preparation method of fluorescent microsphere provided by the present invention, comprises the following steps：
（1）Prepare fluorescent small molecule
Anhydrous citric acid and thionyl chloride are added in tetrahydrofuran, lucifuge stirring a period of time, after revolving purification three times, so
Obtained lemon acyl chlorides and 3- aminopropyl trimethoxy siloxanes are reacted completely in tetrahydrofuran afterwards, after revolving purification, most
Fluorescent small molecule is made afterwards.
（2）Prepare silicon dioxide microsphere
Tetraethyl orthosilicate, deionized water, ammoniacal liquor are added to the in the mixed solvent of alcohols and water, stirring reaction for a period of time, passes through
Centrifugation, washing, pulverized after drying, silicon dioxide microsphere is made.
（3）Prepare fluorescent microsphere
Silicon dioxide microsphere obtained above and the fluorescent small molecule are added in solvent, after reaction completely, centrifuges, wash
Wash, dry, fluorescent microsphere is made.
（4）Configure fluorescent microsphere solution
Take appropriate fluorescent microsphere dissolving in deionized water, heat fully dissolving.
As optimal technical scheme, during synthetic silica microballoon, described absolute ethyl alcohol and positive silicic acid second
The volume ratio of ester is（50~200）：1, preferably 100:1.
As optimal technical scheme, during synthetic silica microballoon, the volume of described absolute ethyl alcohol and water
Than（8~12）:1, preferably 10：1.
As optimal technical scheme, during fluorescent microsphere is synthesized, the particle diameter of the silicon dioxide microsphere for 100 ~
500 nm, preferably 150 nm.
As optimal technical scheme, during fluorescent microsphere is synthesized, the fluorescent microsphere preparation method has copolymerization method,
Investment and chemical bonding etc., preferably chemical bonding.
As optimal technical scheme, during fluorescent microsphere is synthesized, the silicon dioxide microsphere and fluorescent small molecule
With mass ratio 1:（0.06~0.12）With reference to preferably 1:0.10.
As optimal technical scheme, during fluorescent microsphere is synthesized, described solvent is water, toluene, tetrahydrofuran,
Absolute ethyl alcohol etc., preferably tetrahydrofuran.
As optimal technical scheme, during fluorescent microsphere is synthesized, configuration fluorescent microsphere solution detection Hg2+When, institute
State and heated 5 ~ 15 minutes at 60 ~ 100 DEG C, concentration is 1 ~ 10 μ g/ml, preferably 100 DEG C, is heated 10 minutes, concentration is 2 μ
g/ml。
Compared with prior art, the advantage of the invention is that compared with single probe, the good dispersion of the fluorescent microsphere, surely
Qualitative height, luminous efficiency is high, can directly be entrained in silicon rubber and fluorescent device is made.Meanwhile because silica itself is excellent
Good nontoxicity and biocompatibility, it is set to can be used for the direct detection of organism, in addition, the spherical junctions that the fluorescent microsphere is unique
Structure, it will have potential application value in following other field.
Brief description of the drawings
Fig. 1 is fluorescent microsphere scanning electron microscopic picture prepared by embodiment one.
The fluorescent microsphere that Fig. 2 is prepared for embodiment one prepares schematic diagram.
Fig. 3 is block diagram of the fluorescent microsphere to each ion identification of the preparation of embodiment one.
Fig. 4 is the fluorescent microsphere of the preparation of embodiment one to Fe3+The block diagram shielded.
Fig. 5 is the fluorescent microsphere of the preparation of embodiment one to Hg2+The fluorescence spectra of identification.
Fig. 6 is the fluorescent microsphere of the preparation of embodiment one to Hg2+The linear relationship chart of identification.
Such scheme is described further below in conjunction with specific embodiment.It should be understood that these embodiments are to be used to illustrate
The present invention and be not limited to limit the scope of the present invention.The implementation condition used in embodiment can be done according to actual conditions into
One step
Adjustment, unreceipted implementation condition is usually the condition in routine test.By the following examples come illustrate the present invention
Specifically
Implementation process.
Embodiment one
（1）Prepare fluorescent small molecule
1 g anhydrous citric acids and 1.2 ml thionyl chlorides are placed in the flask full of nitrogen, stirred 12 hours at -5 DEG C, will
The purification of gained lemon acyl chlorides revolving three times, obtains pure lemon acyl chlorides.Then lemon acyl chlorides is dissolved in 40 ml tetrahydrofurans, stirred
3.8 ml 3- aminopropyl trimethoxy siloxanes are added dropwise under the conditions of mixing and continue reaction 3 hours, small point of neat fluorescent is obtained after purification
Son.
（2）Prepare silicon dioxide microsphere
500 ml absolute ethyl alcohols, 50 ml water and 15 ml ammoniacal liquor are sequentially added in beaker, use magnetic stirrer at room temperature
Equal 30 min.Then 5 ml tetraethyl orthosilicates (TEOS) are slowly dropped in well mixed above-mentioned solution, use is added dropwise
Polyethylene film sealed beaker mouth.6 h are stirred, make reaction complete, then are obtained through a series of post processing (centrifugation, washing, drying)
To silicon dioxide microsphere.
（3）Prepare fluorescent microsphere
The g of silicon dioxide microsphere 1 obtained above is taken to be added in 40 ml tetrahydrofurans, stirring makes its dispersed, adds
The lower normal-temperature reaction of 0.10 g fluorescent small molecules stirring 4 hours, centrifugation, washing, dries, fluorescent microsphere is made.
（4）Fluorescent microsphere solution is prepared by solvent of deionized water, concentration is 2 μ g/ml, is heated 10 minutes at 100 DEG C,
The Hg detected2+0 ~ 80 μM of concentration range.A length of 360 nm of optimum excitation wave is set, the fluorescence intensity of recording solution is with Hg2+
Change in concentration collection of illustrative plates.Its detection is limited to 8.7 nM, 0 ~ 25 μM of the range of linearity.
Embodiment two
（1）Prepare fluorescent small molecule
1 g anhydrous citric acids and 1 ml thionyl chlorides are placed in the flask full of nitrogen, stirred 8 hours at -5 DEG C, revolving carries
Pure lemon acyl chlorides is obtained after pure.Then lemon acyl chlorides is dissolved in 40 ml tetrahydrofurans, 3.5 ml 3- is added dropwise under stirring condition
Aminopropyl trimethoxy siloxane continues reaction 4 hours, and neat fluorescent small molecule is obtained after purification.
（2）Prepare silicon dioxide microsphere
500 ml absolute ethyl alcohols, 50 ml water and 15 ml ammoniacal liquor are sequentially added in beaker, use magnetic stirrer at room temperature
Equal 1 h.Then 6 ml tetraethyl orthosilicates (TEOS) are slowly dropped in well mixed above-mentioned solution, be added dropwise with poly- second
Alkene diaphragm seal beaker mouth.8 h are stirred, make reaction complete, then two are obtained through a series of post processing (centrifugation, washing, drying)
Silicon oxide microsphere.
（3）Prepare fluorescent microsphere
The g of silicon dioxide microsphere 1 obtained above is taken to be added in 40 ml toluene, stirring makes its dispersed, adds 0.08 g
The lower normal-temperature reaction of fluorescent small molecule stirring 6 hours, centrifugation, washing, dries, fluorescent microsphere is made.
（4）Fluorescent microsphere solution is prepared by solvent of deionized water, concentration is 3 μ g/ml, is heated 10 minutes at 90 DEG C,
The Hg detected2+0 ~ 80 μM of concentration range.A length of 360 nm of optimum excitation wave is set, the fluorescence intensity of recording solution is with Hg2+
Change in concentration collection of illustrative plates.Its detection is limited to 20.5 nM, 0 ~ 20 μM of the range of linearity.
Embodiment three
（1）Prepare fluorescent small molecule
1 g anhydrous citric acids and 1.8 ml thionyl chlorides are placed in the flask full of nitrogen, stirred 12 hours at -5 DEG C.Rotation
Pure lemon acyl chlorides is obtained after steaming purification.Then lemon acyl chlorides is dissolved in 40 ml tetrahydrofurans, 4 ml is added dropwise under stirring condition
3- aminopropyl trimethoxy siloxanes continue reaction 5 hours, and neat fluorescent small molecule is obtained after purification.
（2）Prepare silicon dioxide microsphere
500 ml absolute ethyl alcohols, 50 ml water and 20 ml ammoniacal liquor are sequentially added in beaker, use magnetic stirrer at room temperature
Equal 1 h.Then 8 ml tetraethyl orthosilicates (TEOS) are slowly dropped in well mixed above-mentioned solution, be added dropwise with poly- second
Alkene diaphragm seal beaker mouth.10 h are stirred, make reaction complete, then two are obtained through a series of post processing (centrifugation, washing, drying)
Silicon oxide microsphere.
（3）Prepare fluorescent microsphere
The g of silicon dioxide microsphere 1 obtained above is taken to be added in 40ml absolute ethyl alcohols, stirring makes its dispersed, adds 0.12
The lower normal-temperature reaction of g fluorescent small molecules stirring 8 hours, centrifugation, washing, dries, fluorescent microsphere is made.
（4）Fluorescent microsphere solution is prepared by solvent of deionized water, concentration is 4 μ g/ml, is heated 5 minutes at 100 DEG C,
The Hg detected2+0 ~ 80 μM of concentration range.A length of 360 nm of optimum excitation wave is set, the fluorescence intensity of recording solution is with Hg2+
Change in concentration collection of illustrative plates.Its detection is limited to 20.2 nM, 0 ~ 30 μM of the range of linearity.
Example IV
（1）Prepare fluorescent small molecule
1g anhydrous citric acids and 2ml thionyl chlorides are placed in the flask full of nitrogen, stirred 12 hours at -5 DEG C.Revolving carries
Pure lemon acyl chlorides is obtained after pure.Then lemon acyl chlorides is dissolved in 40ml tetrahydrofurans, 4.5 ml 3- ammonia is added dropwise under stirring condition
Propyl trimethoxy siloxanes continues reaction 6 hours, and neat fluorescent small molecule is obtained after purification.
（2）Prepare silicon dioxide microsphere
500 ml absolute ethyl alcohols, 80 ml water and 15 ml ammoniacal liquor are sequentially added in beaker, use magnetic stirrer at room temperature
Equal 2 h.Then 10 ml tetraethyl orthosilicates (TEOS) are slowly dropped in well mixed above-mentioned solution, be added dropwise with poly-
Vinyl film sealed beaker mouth.12 h are stirred, make reaction complete, then are obtained through a series of post processing (centrifugation, washing, drying)
Silicon dioxide microsphere.
（3）Prepare fluorescent microsphere
The g of silicon dioxide microsphere 1 obtained above is taken to be added in 40 ml tetrahydrofurans, stirring makes its dispersed, adds
The lower normal-temperature reaction of 0.12g fluorescent small molecules stirring 8 hours, centrifugation, washing, dries, fluorescent microsphere is made.
（4）Fluorescent microsphere solution is prepared by solvent of deionized water, concentration is 10 μ g/ml, heats 8 minutes, enters at 80 DEG C
The Hg of row detection2+0 ~ 100 μM of concentration range.A length of 360 nm of optimum excitation wave is set, the fluorescence intensity of recording solution is with Hg2+
Change in concentration collection of illustrative plates.Its detection is limited to 36.8 nM, 0 ~ 40 μM of the range of linearity.
Applicant states that the present invention illustrates the detailed process equipment of the present invention and technological process by above-described embodiment,
But the invention is not limited in above-mentioned detailed process equipment and technological process, that is, it is above-mentioned detailed not mean that the present invention has to rely on
Process equipment and technological process could be implemented.Person of ordinary skill in the field it will be clearly understood that any improvement in the present invention,
The addition of equivalence replacement and auxiliary element to each raw material of product of the present invention, selection of concrete mode etc., all fall within the present invention's
Within the scope of protection domain and disclosure.
Claims (8)
1. a kind of quick identification and the fluorescent microsphere preparation method for quantitatively detecting mercury ion, it is characterised in that comprise the following steps：
（1）Prepare fluorescent small molecule
Anhydrous citric acid and thionyl chloride are added in tetrahydrofuran, lucifuge stirring a period of time, after revolving purification three times, so
Obtained lemon acyl chlorides and 3- aminopropyl trimethoxy siloxanes are reacted completely in tetrahydrofuran afterwards, after revolving purification, most
Fluorescent small molecule is made afterwards；
（2）Prepare silicon dioxide microsphere
Tetraethyl orthosilicate, deionized water, ammoniacal liquor is added to the in the mixed solvent of absolute ethyl alcohol and water, during one section of stirring reaction
Between, pulverized through centrifuging, wash, after drying, silicon dioxide microsphere is made；
（3）Prepare fluorescent microsphere
Silicon dioxide microsphere obtained above and the fluorescent small molecule are added in solvent, after reaction completely, centrifuges, wash
Wash, dry, fluorescent microsphere is made；
（4）Configure fluorescent microsphere solution
Take appropriate fluorescent microsphere dissolving in deionized water, heat fully dissolving.
2. a kind of quick identification as claimed in claim 1 and the fluorescent microsphere preparation method for quantitatively detecting mercury ion, its feature
It is step（2）Described absolute ethyl alcohol and the volume ratio of tetraethyl orthosilicate are（50~200）：1.
3. a kind of quick identification as claimed in claim 1 and the fluorescent microsphere preparation method for quantitatively detecting mercury ion, its feature
It is step（2）Described absolute ethyl alcohol and the volume ratio of water are（8~12）:1.
4. a kind of quick identification as claimed in claim 1 and the fluorescent microsphere preparation method for quantitatively detecting mercury ion, its feature
It is step（2）The particle diameter of the silicon dioxide microsphere is 100 ~ 500nm.
5. a kind of quick identification as claimed in claim 1 and the fluorescent microsphere preparation method for quantitatively detecting mercury ion, its feature
It is step（3）It is described that fluorescent small molecule is grafted on silicon dioxide microsphere by chemical bonding.
6. a kind of quick identification as claimed in claim 1 and the fluorescent microsphere preparation method for quantitatively detecting mercury ion, its feature
It is step（3）The silicon dioxide microsphere and the mass ratio of fluorescent small molecule are 1:（0.06~0.12）.
7. a kind of quick identification as claimed in claim 1 and the fluorescent microsphere preparation method for quantitatively detecting mercury ion, its feature
It is step（3）Described solvent is water, toluene, tetrahydrofuran, absolute ethyl alcohol etc..
8. a kind of quick identification as claimed in claim 1 and the fluorescent microsphere preparation method for quantitatively detecting mercury ion, its feature
It is step（4）Described configuration fluorescent microsphere solution detection Hg2+When, using deionized water as solvent, at 60 ~ 100 DEG C heat 5 ~
15 minutes, concentration was 1 ~ 10 μ g/ml.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111665224A (en) * | 2019-11-26 | 2020-09-15 | 山东立菲生物产业有限公司 | Method for preparing and detecting mercury ions based on fluorescent microspheres |
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| CN104987346A (en) * | 2015-04-30 | 2015-10-21 | 济南大学 | Method for preparing fluorescence polyamino compound |
| CN105018070A (en) * | 2014-04-18 | 2015-11-04 | 天津工业大学 | Preparation method of mercury ion rate probe based on rhodamine derivative and quantum dot fluorescent microsphere |
| CN105693532A (en) * | 2016-01-21 | 2016-06-22 | 济南大学 | Preparation method of fluorescent ammonia carboxylate |
| CN105968003A (en) * | 2016-06-07 | 2016-09-28 | 济南大学 | Citrate fluorescent compound and application thereof to mercury ion detection |
| CN107245332A (en) * | 2017-07-13 | 2017-10-13 | 济南大学 | A kind of organo-mineral complexing fluorescent microsphere preparation method |
-
2017
- 2017-12-08 CN CN201711292361.8A patent/CN107892914A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105018070A (en) * | 2014-04-18 | 2015-11-04 | 天津工业大学 | Preparation method of mercury ion rate probe based on rhodamine derivative and quantum dot fluorescent microsphere |
| CN104987346A (en) * | 2015-04-30 | 2015-10-21 | 济南大学 | Method for preparing fluorescence polyamino compound |
| CN105693532A (en) * | 2016-01-21 | 2016-06-22 | 济南大学 | Preparation method of fluorescent ammonia carboxylate |
| CN105968003A (en) * | 2016-06-07 | 2016-09-28 | 济南大学 | Citrate fluorescent compound and application thereof to mercury ion detection |
| CN107245332A (en) * | 2017-07-13 | 2017-10-13 | 济南大学 | A kind of organo-mineral complexing fluorescent microsphere preparation method |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111665224A (en) * | 2019-11-26 | 2020-09-15 | 山东立菲生物产业有限公司 | Method for preparing and detecting mercury ions based on fluorescent microspheres |
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