CN106085410B - PEI modifies carbon dots-FITC compounds as ratio fluorescent pH probes and Cu2+The application of fluorescence probe - Google Patents
PEI modifies carbon dots-FITC compounds as ratio fluorescent pH probes and Cu2+The application of fluorescence probe Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
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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
- G01N21/62—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
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/14—Macromolecular compounds
- C09K2211/1441—Heterocyclic
- C09K2211/1466—Heterocyclic containing nitrogen as the only heteroatom
Abstract
The invention discloses PEI to modify carbon dots FITC compounds as ratio fluorescent pH probes and Cu2+The application of fluorescence probe.The present invention is raw material using cherry tomato, and fluorescence is made by Hydrothermal Synthesiss changes insensitive carbon dots to pH, and then the amino on PEI modifications CDs, PEI is reacted with isosulfocyanate radical on FITC molecules, forms compound.The response that the fluorescence of FITC changes pH in compound is high, and the ratio probes based on CDs and FITC fluorescence are detected for pH；Amino and Cu on PEI2+Coordination, which occurs, causes the fluorescence intensity of CDs and FITC with Cu in solution2+Concentration is raised and declined, and realizes detection Cu2+.Fluorescence probe selection that the present invention is built is novel, have high sensitivity, selectivity is good, easy to operate, the range of linearity is wide, test limit is low, available for pH and Cu in aqueous solution and biological sample2+Efficient detection.
Description
Technical field
The invention belongs to the crossing domain of chemistry, biology and materialogy, is related to material preparation, biology and environment ion inspection
Survey and pH detection, and in particular to PEI modification carbon dots-FITC compounds preparation as well as ratio fluorescent pH probes and
Cu2+The application of fluorescence probe.
Background technology
Quantum dot typically refers to size and is less than or the semiconductor nano close to its Exciton Bohr Radius, be it is a kind of have it is only
The nano-particle of special light, electrology characteristic.Compared with one-dimensional or two-dimentional carbon nanomaterial, using the carbon dots of nontoxic biological raw material preparation
(CDs) specific surface area with higher and more edge active sites, this makes CDs compared to large-size carbon material in bio-sensing
Possesses more extensive and efficient application prospect with catalytic field.In addition, the photoluminescent property of CDs is repaiied better than no group after amination
The CDs of decorations, and polyethyleneimine (PEI) can regulate and control its optical property as the nitrogen source of CDs.In addition, PEI is as a kind of
Polyamine family macromolecule has good coordination to metal ion, it is had in ion detection, biomedicine and imaging field
There is important application, correlative study becomes one of current research hotspot.
H in solution+Concentration or pH value are to characterize the important parameter of SOLUTION PROPERTIES, it not only influences other materializations of solution
Matter, while also influence reaction speed and direction, therefore, the measurement of pH value industrial and agricultural production, medicine, bioengineering, environment and
The fields such as scientific research are of great significance.Detected compared to traditional pH test paper and pH glass electrodes, photochemistry pH sensors are then
Have the advantages that to may be reused, the time is fast, small, electromagnetism interference performance is strong, pollution-free, all kinds of chemical reactions,
Telemetry and telecommand of dangerous situation such as environmental monitoring, biomedical sector and underground mine, weapon test etc. has wide
Application prospect.Fluorescein isothiocynate (FITC) is widely used in biomedical sector as fluorescence probe, and FITC is as glimmering
Light pH indicator its fluorescence intensity in the relatively low solution of pH is small, its fluorescence intensity increases in the solution of pH higher, detection spirit
Sensitivity is high.
Cu2+Played a crucial role in environmental area and biology or chemical system, effectively detect Cu2+
It is necessary.Literature research shows that the CDs of PEI modifications can specifically detect Cu2+, Yongqiang Dong etc. devise one
Fluorescence probe of the kind based on PEI modifications CDs, with Cu2+Concentration increases, the increase of CDs fluorescence intensities, and then can be to Cu2+Determined
Measure fixed (Yongqiang Dong, Ruixue Wang, Geli Li, Congqiang Chen, Yuwu Chi, * and Guonan
Chen., Anal.Chem., 2012,84,6220-6224).The Fe that Goon, I.Y. etc. are modified using PEI3O4NPs quantifies detection
Hg2+(Goon, I.Y., Zhang, C.C., Lim, M.；, Gooding, J.J., Amal, R.Langmuir 2010,26,12247-
12252).Fangkai Du etc. construct fluorescence resonance energy transfer pH sensors, and CDs is supplied as fluorescence resonance energy transfer
Body, FITC are energy acceptor, realize to the Sensitive Detection of pH (Fangkai Du., Yunhao Ming., Fang Zeng.,
Changmin Yu and Shuizhu Wu.,Nanotechnology.,2013,24,365101)。
But above-mentioned detection largely pertains only to the detection of one pack system, and the more of detection object are often related in practical application
Component detects, such as the heavy metal and pH value of sewage, generally requires multiple detection probes or detection device, so that detection
It is cumbersome, so, explore a kind of multi-functional detection means and have a good application prospect.
The content of the invention
For problem of the prior art, it is an object of the invention to provide a kind of multi-functional detection means come detect pH and
Cu2+。
To achieve the above object, the present invention relates to following technical scheme：
First, the present invention provides a kind of PEI and modifies carbon dots-FITC compounds as ratio fluorescent pH probes and Cu2+Fluorescence
The application of probe.
The PEI modifies carbon dots-FITC compounds：On the basis of PEI modifies carbon dots, further it is coupled FITC and is prepared into
Arrive；The preparation of PEI modification carbon dotss belongs to the state of the art, such as Yongqiang Dong (Yongqiang Dong,
Ruixue Wang,Geli Li,Congqiang Chen,Yuwu Chi,and Guonan Chen.,Anal.Chem.2012,
84,6220-6224) disclosure, introduces the application in this together.
In preferable technical solution, PEI modification carbon dots-FITC compounds are prepared via a method which：It is molten to carbon dots (CDs)
Polyethyleneimine (PEI) is added in liquid, the PEI modifications for carrying out carbon dots is uniformly mixed, then adds fluorescein isothiocynate
(FITC) it is coupled, PEI modification carbon dots-FITC compounds is made.
In preferred technical solution, the carbon dots (CDs) is prepared via a method which：Cherry tomato is squeezed the juice, weigh β-
Cyclodextrin powder is added in holy girl's fruit juice, is stirred evenly, autoclave hydro-thermal reaction, and reactant centrifuges, and is collected upper strata and is disperseed
Liquid, with membrane filtration, removes bulky grain；The filtered fluid of upper strata dispersion liquid is poured into rotary evaporation in Rotary Evaporators, adds third
Ketone and ethanol extraction, liquid separation, centrifuges, lower sediment thing is carbon dots (CDs).
Although in relation to pH and Cu2+The preparation method of fluorescence probe it has been reported that but so far, there has been no based on
PEI modifications carbon dots-FITC composites are at the same time for detecting pH and Cu2+Domestic and foreign literature and patent report.The present invention
On the basis of early-stage study, it is found that PEI modification carbon dots-FITC compounds serve not only as ratio fluorescent pH probes and carry out pH
Detection, and double fluorescent quenchings based on CDs and FITC can effectively detect Cu2+Concentration.Specifically, the present invention utilizes
Amino on PEI, can not only be used for amination modification CDs, and can specificity and isosulfocyanate radical reaction on FITC.Work as pH of buffer
When different, CDs fluorescence intensities are almost unchanged, therefore can be used as reference, and FITC fluorescence intensities gradually rise and increase with pH, so
CDs can be used for qualitative detection pH value of solution with FITC ratio fluorescents；The coupling of FITC and PEI modification carbon dotss, does not interfere with PEI not only
With Cu2+Coordination combine, and be based on PEI and Cu2+Combination can cause respectively at PEI combine CDs and FITC double fluorescence
Quenching effect, lifting detection signal, works as Cu2+In the presence of, Cu2+It can be specifically bound with PEI, with Cu2+Concentration raises so that
CDs and FITC fluorescence intensities decrease at the same time, cause fluorescent quenching, fitting system CDs and FITC fluorescence intensities and Cu2+Concentration
Between relation, can build and pH and Cu are detected based on CDs-FITC compounds at the same time2+Fluoroscopic examination means.
Secondly, the present invention provides a kind of detection pH and Cu2+Detection method, this method includes：
PEI is modified carbon dots-FITC compounds to detect for pH：PEI modification carbon dots-FITC compounds are slow from different pH's
Fliud flushing mixes, the ratio fluorescent intensity I of measurement systemFITC/ICDs(IFITCFor fluorescence intensities of the FITC in the buffer solution of different pH
Value, ICDsFor fluorescence intensity levels of the CDs in the buffer solution of different pH), system ratio fluorescent intensity and the relation of pH are fitted, into
The detection of row pH value of solution to be measured；
PEI modification carbon dots-FITC compounds are used for Cu2+Detection：By the identical pH of PEI modification carbon dots-FITC compounds
Buffer solution dilution, measurement system fluorescence intensity, then adds Cu2+Hatching, then the fluorescence intensity of measurement system, are fitted system
CDs and FITC fluorescence intensities and Cu2+Relation between concentration, carries out solution C u to be measured2+The detection of concentration.
The PEI modifies carbon dots-FITC compounds：On the basis of PEI modifies carbon dots, further it is coupled FITC and is prepared into
Arrive, it is preferred that PEI modification carbon dots-FITC compounds are prepared via a method which：Polyethylene is added into carbon dots (CDs) solution
Imines (PEI), is uniformly mixed the PEI modifications for carrying out carbon dots, then adds fluorescein isothiocynate (FITC) and is coupled, is made
PEI modifies carbon dots-FITC compounds.
In preferable technical solution, the carbon dots (CDs) is prepared via a method which：Cherry tomato is squeezed the juice, weighs β-ring
Dextrin powder is added in holy girl's fruit juice, is stirred evenly, autoclave hydro-thermal reaction, and reactant centrifuges, and collects upper strata dispersion liquid,
With membrane filtration, bulky grain is removed；The filtered fluid of upper strata dispersion liquid is poured into rotary evaporation in Rotary Evaporators, add acetone and
Ethanol extracts, liquid separation, centrifuges, lower sediment thing is carbon dots (CDs).
It is placed on specifically, cherry tomato is squeezed the juice in clean beaker, weighs a certain amount of beta-cyclodextrin powder and add holy girl
In fruit juice, by magnetic agitation, autoclave hydro-thermal reaction, centrifuges, and collects upper strata dispersion liquid, with membrane filtration, removes big
Particle, the filtered fluid of upper strata dispersion liquid is poured into Rotary Evaporators, at a certain temperature rotary evaporation into flask without bright
Aobvious liquid, adds acetone and ethanol extraction, and liquid separation, centrifuges, repeat the step three times and more than, lower sediment thing is carbon
Point (CDs).
The present invention prepares carbon dots using cherry tomato and beta-cyclodextrin hydro-thermal method, explores new carbon dots and prepares material, made
Standby carbon dots has excellent optical property, and quantum yield is high, its fluorescence intensity is suitable for the need of relative broad range pH detection stability
Will, and the preparation method technique is simple, efficient, and raw material are nontoxic, are easy to get.
The present invention provides a kind of detection pH and Cu2+The construction method of the detection architecture of concentration, includes the following steps：
1) cherry tomato is squeezed the juice and be placed in beaker, weighed beta-cyclodextrin powder and add in holy girl's fruit juice, stirred by magnetic force
Mix, autoclave hydro-thermal reaction, centrifuge, collect upper strata dispersion liquid, with membrane filtration, bulky grain is removed, by upper strata dispersion liquid
Filtered fluid pour into Rotary Evaporators, rotary evaporation without obvious liquid, adds acetone and ethanol extraction into flask, liquid separation,
Centrifuge, repeat the step three times and more than, lower sediment thing is carbon dots (CDs)；
2) polyethyleneimine (PEI) is added into the CDs solution of preparation, magnetic agitation under room temperature, it is glimmering to add isothiocyanic acid
Light element (FITC), continues magnetic agitation, and PEI modification carbon dots-FITC compounds are made；
3) that the PEI of preparation modification carbon dots-FITC compounds are divided into two parts is spare,
Part I PEI modifies carbon dots-FITC compounds and is built for pH detection architectures：By CDs-FITC compounds and not
Phosphate buffer with pH mixes, the ratio fluorescent intensity (I of measurement systemFITC/ICDs), it is fitted system ratio fluorescent intensity
With the relation of pH, the pH ratio fluorescent probe in detecting systems based on PEI modification carbon dots-FITC compounds are built；
Part II PEI modification carbon dots-FITC compounds are used for Cu2+Detection architecture is built：CDs-FITC compounds are used
The phosphate buffer dilution of identical pH, measurement system fluorescence intensity, then adds various concentrations Cu2+Hatching, then measurement system
Fluorescence intensity, be fitted system fluorescence intensity and Cu2+Relation between concentration, builds based on PEI modification carbon dots-FITC compounds
Cu2+Fluorescence probe detection architecture.
In preferable technical solution, the concrete technology condition of step (1) is：The volume of holy girl's fruit juice is 10~100mL, β-
The quality of cyclodextrin powder is 0.1~1.0g, and the time of magnetic agitation is 1~10min, and hydrothermal temperature is 100~200
DEG C, the hydro-thermal reaction time is 2~12h, the rotating speed of the centrifugation after hydro-thermal reaction and time be respectively 5000~16000rpm,
3~15min, for the temperature set in Rotary Evaporators as 25~80 DEG C, the volume for adding acetone is 5~50mL, adds ethanol
Volume is 5~50mL, and the rotating speed of the centrifugation after liquid separation and time are respectively 5000~16000rpm, 3~15min.
In preferable technical solution, the concrete technology condition of step (2) is：CDs concentration and volume be respectively 0.1~
1.0mgmL-1, 1~10mL, PEI mass fractions and volume are respectively 0.01~0.1%, 0.1~1.0mL, and the magnetic agitation time is
30min~6h, FITC concentration and volume are respectively 0.01~0.1mM, 5~50mL, and the magnetic agitation time is 30min~3h.
In preferable technical solution, the concrete technology condition of step (3) is：Part I PEI modification carbon dotss-FITC is compound
The volume of thing is that the volume of 20~200 μ L, 10mM phosphate buffer solutions is 1~10mL, the pH of phosphate buffer for 3.2~
9.0；The volume of Part II PEI modification carbon dots-FITC composites is 20~200 μ L, phosphate buffer solution volume
For 1~10mL, the pH of phosphate buffer is 7.4, Cu2+Concentration be 0.01~10.0 μM, 1~10min of brooding time.
The present invention achieves following beneficial effect：
(1) present invention discover that PEI modification carbon dots-FITC compounds serve not only as ratio fluorescent pH probes and carry out pH's
Detection, and double fluorescent quenchings based on CDs and FITC, can effectively detect Cu2+Concentration, provides a kind of multi-functional for this
Detection means, and in order to expand the H of FITC+Sensor application scope, increase sensor service life and raising are sensitive
Degree, the present invention devise the ratio fluorescent sensor based on CDs-FITC and are used for while detect pH and Cu2+。
(2) present invention prepares material using new carbon dots, and prepared carbon dots has excellent optical property, it is wider
Scope pH stablizes, and passes through IFITC/ICDsThe detection of ratio fluorescent intensity, can improve Detection accuracy and detection efficiency.
(3) the method for the present invention mild condition, equipment is simple, easily operated, CDs-FITC compounds can pass through Hydrothermal Synthesiss
Method, rotary evaporation purification are directly made, and raw material availability is high, and cost is low.
(4) the present invention provides a kind of technique is simple, yield is high, production cost is low, it is environmentally protective, be easy to operate and control
A kind of ratio fluorescent pH probes and Cu based on carbon dots/fluorescein isothiocynate compound2+The system of fluorescence probe detection architecture
Preparation Method, compared with prior art, probe in detecting system of the present invention have high sensitivity, and detection range is wide, and it is excellent that test limit is low etc.
Point, in having a extensive future for the fields such as biochemical analysis, medical diagnosis, environmental monitoring.
Brief description of the drawings
Ratio fluorescent pH probe and Cu of the Fig. 1 based on carbon dots/fluorescein isothiocynate compound2+The preparation side of fluorescence probe
Method and its principle schematic；
Fig. 2：PEI modifies fluorescence emission spectrum of the carbon dots-FITC compounds under condition of different pH；
Fig. 3：The ratio fluorescent intensity and the Linear Fit Chart of corresponding pH of PEI modification carbon dots-FITC compounds；
Fig. 4：PEI modifies carbon dots-FITC compounds in various concentrations Cu2+In the presence of fluorescence emission spectrogram of compound；
Fig. 5：FITC and CDs relative intensity of fluorescence and corresponding Cu in PEI modification carbon dots-FITC compounds2+Line under concentration
Property fitted figure.
Embodiment
Mode by the following examples further illustrates the present invention, but does not therefore limit the present invention to the reality
Apply among a scope.The experimental method of actual conditions is not specified in the following example, is conventionally selected with condition
Select.
Embodiment 1
Fresh cherry tomato juice 30mL is squeezed using juice extractor, is placed in clean beaker；Weigh 0.5g beta-cyclodextrin powder
Add in holy girl's fruit juice, magnetic agitation 5min, transposition autoclave hydro-thermal reaction 5h, 120 DEG C of reaction temperature；Centrifuge after reaction
Separate (10000rpm, 15min), collect upper strata dispersion liquid, filter membrane is filtered, and removes bulky grain；Upper strata dispersion liquid is poured into
In Rotary Evaporators, rotary evaporation is separately added into the acetone and second of 20mL into flask without obvious liquid at a temperature of 40 DEG C
Alcohol, centrifuges (12000rpm, 5min)；Repeat the centrifugation step three times, lower sediment thing is CDs；Prepare 1.0mg mL- 1CDs aqueous solution 10mL, add 0.1% PEI 0.2mL, magnetic agitation 0.5h under room temperature；Add the FITC 10mL of 0.05mM
Continue magnetic agitation 1h and PEI modification carbon dots-FITC compounds are made.
The compound of above-mentioned preparation is divided into two parts：A part is used for pH and detects, the PEI modification carbon dotss-FITC of 200 μ L
Compound is mixed with the 10mM phosphate buffers of 3mL differences pH (3.2~9.0)；The fluorescence intensity of measurement system, is fitted system
The relation of ratio fluorescent intensity and pH, builds the pH ratio fluorescent probe in detecting bodies based on PEI modification carbon dots-FITC compounds
System.It is additionally useful for Cu2+Detection, by 200 μ L PEI modification carbon dots-FITC compounds with 3mL phosphate buffers (10mM,
PH7.4) dilute, the fluorescence intensity of measurement system, then add 0.01~10.0 μM of Cu2+Hatching 10min, then measurement system
Fluorescence intensity, is fitted system fluorescence intensity and Cu2+Relation between concentration, builds based on PEI modification carbon dots-FITC compounds
Cu2+Fluorescence probe detection architecture.
Embodiment 2
Fresh cherry tomato juice 50mL is squeezed using juice extractor, is placed in clean beaker；Weigh 0.7g beta-cyclodextrin powder
Add in holy girl's fruit juice, magnetic agitation 7min, transposition autoclave hydro-thermal reaction 3h, 140 DEG C of reaction temperature；Centrifuge after reaction
Separate (12000rpm, 10min), collect upper strata dispersion liquid, filter membrane is filtered, and removes bulky grain；Upper strata dispersion liquid is poured into
In Rotary Evaporators, rotary evaporation is separately added into the acetone and second of 30mL into flask without obvious liquid at a temperature of 60 DEG C
Alcohol, centrifuges (12000rpm, 10min)；Repeat the centrifugation step three times, lower sediment thing is CDs；Prepare 0.5mg
mL-1CDs aqueous solution 10mL, add 0.1% PEI 0.1mL, magnetic agitation 1h under room temperature；Add 0.05mM FITC 5mL after
PEI modification carbon dots-FITC compounds are made in continuous magnetic agitation 2h.
The compound of above-mentioned preparation is divided into two parts：A part is used for pH and detects, the PEI modification carbon dotss-FITC of 100 μ L
Compound is mixed with the 10mM phosphate buffers of 2mL differences pH (3.2~9.0)；The fluorescence intensity of measurement system, is fitted system
The relation of ratio fluorescent intensity and pH, builds the pH ratio fluorescent probe in detecting bodies based on PEI modification carbon dots-FITC compounds
System.It is additionally useful for Cu2+Detection, 100 μ L CDs-FITC compounds 5mL phosphate buffers (10mM, pH7.4) are diluted, are surveyed
The fluorescence intensity of amount system, then adds 0.01~10.0 μM of Cu2+Hatch 10min, then the fluorescence intensity of measurement system, fitting
System fluorescence intensity and Cu2+Relation between concentration, builds the Cu based on PEI modification carbon dots-FITC compounds2+Fluorescence probe is examined
Survey system.
Embodiment 3
Fresh cherry tomato juice 80mL is squeezed using juice extractor, is placed in clean beaker；Weigh 0.8g beta-cyclodextrin powder
Add in holy girl's fruit juice, magnetic agitation 10min, transposition autoclave hydro-thermal reaction 6h, 140 DEG C of reaction temperature；After reaction from
The heart separates (12000rpm, 10min), collects upper strata dispersion liquid, and filter membrane is filtered, and removes bulky grain；Upper strata dispersion liquid is fallen
Enter in Rotary Evaporators, at a temperature of 60 DEG C rotary evaporation into flask without obvious liquid, be separately added into 20mL acetone and
Ethanol, centrifuges (12000rpm, 10min)；Repeat the centrifugation step three times, lower sediment thing is CDs；Prepare 0.5mg
mL-1CDs aqueous solution 10mL, add 0.1% PEI 0.2mL, magnetic agitation 1h under room temperature；Add 0.05mM FITC 5mL after
PEI modification carbon dots-FITC compounds are made in continuous magnetic agitation 2h.
The compound of above-mentioned preparation is divided into two parts：A part is used for pH and detects, the PEI modification carbon dotss-FITC of 150 μ L
Compound is mixed with the 10mM phosphate buffers of 5mL differences pH (3.2~9.0)；The fluorescence intensity of measurement system, is fitted system
The relation of ratio fluorescent intensity and pH, builds the pH ratio fluorescent probe in detecting bodies based on PEI modification carbon dots-FITC compounds
System.It is additionally useful for Cu2+Detection, by 150 μ L PEI modification carbon dots-FITC compounds with 5mL phosphate buffers (10mM,
PH7.4) dilute, the fluorescence intensity of measurement system, then add 0.01~10.0 μM of Cu2+Hatching 10min, then measurement system
Fluorescence intensity, is fitted system fluorescence intensity and Cu2+Relation between concentration, builds based on PEI modification carbon dots-FITC compounds
Cu2+Fluorescence probe detection architecture.
It is 4.6~7.7 that above-mentioned detection probe, which is used for the pH detection ranges of linearity,；Cu2+The scope of detection is 0.01~12.45 μM
It is 7nM with Monitoring lower-cut.
The above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
Member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications also should be regarded as
Protection scope of the present invention.
Claims (4)
1. one kind detection pH and Cu2+The construction method of the detection architecture of concentration, includes the following steps：
1）Cherry tomato is squeezed the juice and is placed in beaker, beta-cyclodextrin powder is weighed and adds in holy girl's fruit juice, it is high by magnetic agitation
Kettle hydro-thermal reaction is pressed, is centrifuged, collects upper strata dispersion liquid, with membrane filtration, bulky grain is removed, by the filtering of upper strata dispersion liquid
Liquid is poured into Rotary Evaporators, and rotary evaporation, without obvious liquid, adds acetone and ethanol extraction, liquid separation, centrifugation point into flask
From, repeat the step three times and more than, lower sediment thing is carbon dots CDs；
2）Polyethyleneimine PEI is added into the CDs solution of preparation, magnetic agitation under room temperature, adds fluorescein isothiocynate
FITC, continues magnetic agitation, and PEI modification carbon dots-FITC compounds are made；
3）It is spare that the PEI of preparation modification carbon dots-FITC compounds are divided into two parts,
Part I PEI modifies carbon dots-FITC compounds and is built for pH detection architectures：By CDs-FITC compounds and different pH
Phosphate buffer mixing, the ratio fluorescent intensity I of measurement systemFITC/ICDs, it is fitted system ratio fluorescent intensity and pH
Relation, builds the pH ratio fluorescent probe in detecting systems based on PEI modification carbon dots-FITC compounds；
Part II PEI modification carbon dots-FITC compounds are used for Cu2+Detection architecture is built：By CDs-FITC compounds with identical
The phosphate buffer dilution of pH, measurement system fluorescence intensity, then adds various concentrations Cu2+Hatching, then measurement system are glimmering
Luminous intensity, is fitted system fluorescence intensity and Cu2+Relation between concentration, builds the Cu based on PEI modification carbon dots-FITC compounds2+
Fluorescence probe detection architecture.
2. according to the method described in claim 1, it is characterized in that, step（1）Concrete technology condition be：The body of holy girl's fruit juice
Product is 10 ~ 100 mL, and the quality of beta-cyclodextrin powder is 0.1 ~ 1.0 g, and time of magnetic agitation is 1 ~ 10 min, hydro-thermal reaction
Temperature is 100 ~ 200 DEG C, and the hydro-thermal reaction time is 2 ~ 12 h, and the rotating speed and time centrifuged after hydro-thermal reaction is respectively 5000
~ 16000 rpm, 3 ~ 15 min, for the temperature set in Rotary Evaporators as 25 ~ 80 DEG C, the volume for adding acetone is 5 ~ 50 mL,
The volume for adding ethanol is 5 ~ 50 mL, and the rotating speed and time centrifuged after liquid separation is respectively 5000 ~ 16000 rpm, 3 ~ 15
min。
3. according to the method described in claim 1, it is characterized in that, step（2）Concrete technology condition be：CDs concentration and body
Product is respectively 0.1 ~ 1.0 mg mL-1, 1 ~ 10 mL, PEI mass fractions and volume are respectively 0.01 ~ 0.1 %, 0.1 ~ 1.0 mL,
The magnetic agitation time is the h of 30 min ~ 6, and FITC concentration and volume are respectively 0.01 ~ 0.1 mM, 5 ~ 50 mL, the magnetic agitation time
For the h of 30 min ~ 3.
4. according to the method described in claim 1, it is characterized in that, step（3）Concrete technology condition be：Part I PEI
The volume of modification carbon dots-FITC compounds is 20 ~ 200 μ L, and the volumes of 10 mM phosphate buffer solutions is 1 ~ 10 mL, phosphoric acid
The pH of salt buffer is 3.2 ~ 9.0；The volume of Part II PEI modification carbon dots-FITC composites is 20 ~ 200 μ L,
Phosphate buffer solution volume is 1 ~ 10 mL, and the pH of phosphate buffer is 7.4, Cu2+Concentration be 0.01 ~ 10.0 μM, incubate
Change 1 ~ 10 min of the time.
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| CN107253961B (en) * | 2017-06-29 | 2019-08-13 | 湖南科技大学 | It is a kind of can ratio test cysteine water soluble fluorescence sensor preparation and application |
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