CN106966380B - The preparation method of nano cages fluorescence probe and its in Fe3+Application in detection - Google Patents
The preparation method of nano cages fluorescence probe and its in Fe3+Application in detection Download PDFInfo
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- 239000000523 sample Substances 0.000 title claims abstract description 71
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- 238000002360 preparation method Methods 0.000 title claims abstract description 23
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- 235000019441 ethanol Nutrition 0.000 claims abstract description 20
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- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims abstract description 16
- 230000005284 excitation Effects 0.000 claims abstract description 16
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- 229960000935 dehydrated alcohol Drugs 0.000 claims abstract description 8
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- 238000010791 quenching Methods 0.000 claims description 10
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- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 22
- 239000001576 FEMA 2977 Substances 0.000 description 20
- AKYHKWQPZHDOBW-UHFFFAOYSA-N (5-ethenyl-1-azabicyclo[2.2.2]octan-7-yl)-(6-methoxyquinolin-4-yl)methanol Chemical compound OS(O)(=O)=O.C1C(C(C2)C=C)CCN2C1C(O)C1=CC=NC2=CC=C(OC)C=C21 AKYHKWQPZHDOBW-UHFFFAOYSA-N 0.000 description 15
- LOUPRKONTZGTKE-UHFFFAOYSA-N cinchonine Natural products C1C(C(C2)C=C)CCN2C1C(O)C1=CC=NC2=CC=C(OC)C=C21 LOUPRKONTZGTKE-UHFFFAOYSA-N 0.000 description 15
- 229960003110 quinine sulfate Drugs 0.000 description 15
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 14
- 229910052799 carbon Inorganic materials 0.000 description 13
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- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 description 2
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- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
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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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- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/08—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials
- C09K11/65—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials containing carbon
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- C01—INORGANIC CHEMISTRY
- C01P—INDEXING SCHEME RELATING TO STRUCTURAL AND PHYSICAL ASPECTS OF SOLID INORGANIC COMPOUNDS
- C01P2004/00—Particle morphology
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- C01P—INDEXING SCHEME RELATING TO STRUCTURAL AND PHYSICAL ASPECTS OF SOLID INORGANIC COMPOUNDS
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Abstract
The present invention provides a kind of preparation methods of nano cages fluorescence probe, and the specific steps are be baked by raw material of fish, will extract in dehydrated alcohol by roasted fish soaking；After extraction processing, liquid phase is collected, rotary evaporation removes ethyl alcohol；It is redissolved using chloroform, extraction degreasing, extraction is until aqueous phase solution clear repeatedly, collect water phase, cross G25 sephadex column chromatography, collect the part that fluorescence intensity is more than or equal to 80, excitation wavelength 330nm, launch wavelength 390nm, freeze-drying, obtained powdered substance, as nano cages fluorescence probe；The present invention also provides above-mentioned nano cages fluorescence probes in Fe3+Application in detection.Nano cages fluorescence probe made from this is hollow structure and has a multilayer ring-type Shells, and detection sensitivity and stability with higher, fluorescence lifetime is long, quantum yield is high；Fe provided by the invention3+The minimum detection line of concentration detection method is up to 3.96 μM.
Description
Technical field
The present invention relates to the preparation of fluorescence probe and applied technical fields, the in particular to preparation of nano cages fluorescence probe
Method and its in Fe3+Application in detection.
Background technique
Nano cages (Carbon nanonages, CNCs) are that a kind of inside made of being coated as graphite linings is hollow carbon
Nano material.Most of CNC surface texture is similar to porous carbon, it has unique hollow pore structure, thus physical chemistry
Performance is more unique, and such as specific surface area height, corrosion resistance is strong, and conductivity is good, and bio-compatibility is good etc., can be applied to much lead
Domain, including the storage of pharmaceutical carrier, data, the depollution of environment, capacitance material, lithium battery, hydrogen storage material, protected protein matter and catalyzing enzyme
Deng.Up to the present, the preparation method of nano cages mainly has chemical vapour deposition technique, template and Plasma Polymerization
It is complicated Deng, preparation method, it is not easy to operate.Therefore, it is imperative for developing a kind of preparation method of nano cages being simple and efficient
's.
Since metal ion plays an important role in environment and the ecosystem, the design of metal ion inducing probes with
Synthesis has attracted a large amount of concerns of researcher.Wherein iron is the metallic element of content second in nature to the entire ecosystem
There is highly important effect, it plays key player in many biochemical processes, as cell metabolism, enzymatic, electronics turn
Shifting, the synthesis of oxidation reaction, O_2 transport, DNA and RNA.Excessive or insufficient Fe3+It will lead to the unbalance and disease of body, example
Such as decline of anaemia, intelligence, arthritis, heart failure, diabetes and cancer.Therefore, it is necessary to develop one kind easily and effectively
Method detect the Fe in food3+.Compared to needing complex instrument or the conventional method of cumbersome sample preparation procedure such as
Electrochemical process and mass spectrography, fluorescence detection method show uniqueness in highly selective, high sensitivity and aspect easy to operate
Advantage.
Summary of the invention
The purpose of the present invention is to provide a kind of preparation methods of nano cages fluorescence probe, break through traditional preparation method,
Reduction production cost is low, it is simple, controllable to realize process；Meanwhile providing a kind of utilization nano cages fluorescence probe quantitative detection Fe3+
The method of concentration.
In order to achieve the above objectives, the present invention provides a kind of preparation method of nano cages fluorescence probe, specific steps are as follows:
Using fish as raw material, 150~300 DEG C of 10~120min of baking；
2~48h will be extracted in dehydrated alcohol by roasted fish soaking, it is described by roasted fish and added ethyl alcohol
Final concentration of every gram of fish 1ml ethyl alcohol；After extraction processing, liquid phase is collected, rotary evaporation removes ethyl alcohol；
It is redissolved using chloroform, extraction degreasing, extraction is until aqueous phase solution clear, collects water phase, as repeatedly
Slightly mention solution；
The solution that slightly mentions is crossed into G25 sephadex column chromatography, fluorescence intensity is collected and is more than or equal to 80, excitation wavelength
The part of 330nm, launch wavelength 390nm, freeze-drying, obtained powdered substance, as nano cages fluorescence probe.
The nano cages fluorescence probe that the present invention collects is slightly to mention the biggish part of fluorescence intensity in solution.
The preferred turbot of the fish, conger pile.
The fish that the present invention uses are first passed through baking processing and are extracted again using ethyl alcohol, and nano cages effectively can be slightly mentioned.
Product made from the above method is hollow and with multilayer ring-type Shells, diameter by projecting Electronic Speculum to can detect it
Nanometer basket structure less than 25nm.
A large amount of hydroxyl and amino are contained in the nano cages surface, and luminous efficiency is high, good water solubility.
The present invention also provides above-mentioned nano cages fluorescence probes in Fe3+Application in detection, includes the following steps:
(1) by the nano cages fluorescence probe be dissolved in water obtained concentration be 2~50mg/ml nano cages fluorescence visit
Needle aqueous solution；Configure a series of Fe of known concentrations3+Solution, to the Fe3+The nano cages fluorescence probe is added in solution
Aqueous solution obtains Fe3+Standard sample solution；
The Fe3+Fe in solution3+Concentration range be 20~200 μM；
The nano cages fluorescence probe aqueous solution and Fe3+The volume ratio of solution is 1:0.5~5；
The Fe3+The preferred FeCl of solution3Solution；
The concentration of the nano cages fluorescence probe aqueous solution is preferably 10mg/ml；
(2) the nano cages fluorescence probe aqueous solution made from step (1) is added into water, as blank control group；
The Fe made from the adding proportion of the nano cages fluorescence probe aqueous solution and step (1)3+The addition of standard sample solution
Ratio is identical；
Fluorescence intensity in examination criteria solution is compared with control group, according to Fe3+Solution concentration and corresponding fluorescent quenching
Intensity modeling, obtains Fe3+Standard curve between concentration and fluorescence intensity；
The actual conditions of the fluorescence intensity detection are as follows:
Cover the sepectrophotofluorometer of 280~700nm using Detection wavelength range；
Excitation wavelength is 280~500nm；It is preferred that excitation wavelength 330nm；
The wavelength of fluorescence emission peak is 350~690nm；It is preferred that launch wavelength 390nm；
Fluorescent quenching intensity is calculated as ((F0-F1)/F0), the wherein fluorescence intensity F in standard solution1, blank control group
Fluorescence intensity be F0。
Under preferred embodiment, the modeling process, with Fe3+Solution concentration is abscissa, corresponding Fe3+Standard sample solution obtains
The fluorescent quenching intensity arrived is ordinate, obtains Fe3+Concentration and Standardization curve for fluorescence intensity.
(3) Fe to be measured is taken3+Solution, to the Fe to be measured3+It is glimmering that the nano cages made from step (1) are added in solution
Light probe aqueous solution, as Fe to be measured3+Solution example；The adding proportion and step of the nano cages fluorescence probe aqueous solution
(1) Fe made from3+The adding proportion of standard sample solution is identical；
Using detection method identical with step (2), Fe to be measured is obtained3+The fluorescence intensity of solution example, in conjunction with step (2)
Obtained Fe3+Concentration and Standardization curve for fluorescence intensity obtain Fe to be measured3+The Fe of solution example3+Concentration.
The method of the present invention detects Fe3+The minimum detection line of concentration is 3.96 μM, be can be used for containing low concentration Fe3+Sample
Detection.
Beneficial effects of the present invention are as follows:
1, the fish that the present invention uses are first passed through baking processing and are extracted again using ethyl alcohol, effectively can slightly mention nano cages.
2, the preparation method of nano cages fluorescence probe of the present invention is compared with prior synthesizing method, and raw material is easy to get, cost
Low, preparation process is simply controllable, breaks through traditional preparation method；Nano cages fluorescence probe obtained is hollow structure and has more
The cyclic annular Shells of layer, the nano cages fluorescence probe detection sensitivity with higher and stability, fluorescence lifetime length, quantum
Yield is high.
3, Fe provided by the invention3+Photoluminescent property of the concentration detection method based on nano cages fluorescence probe, utilizes Fe3+
Nano cages fluorescent quenching is acted on, can get Fe3+The standard curve of concentration, and Fe3+Concentration Testing high sensitivity is minimum
Line is detected up to 3.96 μM；Fe provided by the invention3+Concentration detection method simple to operate, high sensitivity with detection process
And selectivity is good, testing result is intuitive, can quantitative detection the characteristics of.
4, traditional fluorescence detection will use organic dyestuff or semiconductor-quantum-point, in contrast, the present invention uses
Nano cages fluorescence probe raw material it is natural, have many advantages, such as good water-soluble, biocompatibility and hypotoxicity, there is application
The potentiality of concentration of metal ions detection in biological sample.
Detailed description of the invention
Specific embodiments of the present invention will be described in further detail with reference to the accompanying drawing.
Fig. 1 is the TEM figure of nano cages fluorescence probe prepared by the embodiment 1 of the method for the present invention.
Fig. 2 is the TEM figure of nano cages fluorescence probe prepared by the embodiment 2 of the method for the present invention.
The TEM figure of nano cages fluorescence probe prepared by the embodiment 3 of Fig. 3 the method for the present invention.
The TEM figure of nano cages fluorescence probe prepared by the embodiment 4 of Fig. 4 the method for the present invention.
The TEM figure of nano cages fluorescence probe prepared by the embodiment 5 of Fig. 5 the method for the present invention.
Fig. 6 is the nano cages fluorescence probe 3 in embodiment 3 to Fe3+Sensitivity analysis figure.
Specific embodiment
With reference to embodiments, a specific embodiment of the invention is described in more details, so as to more preferably
Ground understands the advantages of the solution of the present invention and its various aspects.However, specific embodiments described below and embodiment are only
It is for illustrative purposes, rather than limiting the invention.
Embodiment 1: the preparation of nano cages fluorescence probe
By by the conger pile of 180 DEG C of baking 30min, roasted whole fish soaking will be passed through in dehydrated alcohol, every gram of fish 1ml
Ethyl alcohol extracts 15h, slightly mentions nano cages；Rotary evaporation is redissolved using chloroform, is extracted three times, taken off to ethyl alcohol is completely removed
Rouge, until water phase, is slightly mentioned solution and cross G25 sephadex column chromatography, collected fluorescence intensity and be higher than 80 by aqueous phase solution clear
(excitation wavelength=330nm, launch wavelength=390 nm) part, freeze-drying, obtained powdered substance.
It is hollow and is less than 100nm with multilayer ring-type Shells, diameter by projects Electronic Speculum to can detect it to have
Structure nano cages.
Nano cages fluorescent yield is measured by known formula, formula are as follows: Φ1=Φ2I1A2η21/I2A1η22It is required that inhaling
Luminosity A1, A2Less than 0.05, to reduce error.It is measured respectively under identical shooting condition using quinine sulfate as reference substance
Integrated fluorescence intensities (the I of sample and reference substance1/I2) and Same Wavelength under the conditions of incident light absorbing state (A2/A1)。I2-
The integrated area value of quinine sulfate fluorescence intensity；The integrated area value of the fluorescence intensity of I1- coffee；A2Quinine sulfate it is ultraviolet
Absorption value；A1The ultraviolet absorption value of coffee；η2-The refractive index of quinine sulfate solvent；η1The refractive index of the solvent of coffee.Through counting
Calculate prepared nano cages quantum yield is 14.66%.
Fluorescence lifetime passes through formula R (τ)=B1e(-τ/τ1) 6.75ns is calculated to obtain, wherein AiIt is time resolution decay time
(τi) percentage.
Tem analysis (Fig. 1) is carried out to nano cages prepared by this method, it is known that this method can successfully prepare hollow carbon and receive
Rice cage material.The nano cages cavity is about 10-20nm, and the carbon layers having thicknesses of shell are 3-5 nm, and have good knot
Brilliant degree, spacing of lattice about 0.34nm.
Embodiment 2: the preparation of nano cages fluorescence probe
By by the conger pile of 280 DEG C of baking 15min, roasted whole fish soaking will be passed through in dehydrated alcohol, every gram of fish 1ml
Ethyl alcohol extracts 12h, slightly mentions nano cages；Rotary evaporation is redissolved using chloroform, is extracted three times, taken off to ethyl alcohol is completely removed
Rouge, until water phase, is slightly mentioned solution and cross G25 sephadex column chromatography, collected fluorescence intensity and be higher than 80 by aqueous phase solution clear
(excitation wavelength=330nm, launch wavelength=390 nm) part, freeze-drying, obtained powdered substance.
It is hollow and is less than 100nm with multilayer ring-type Shells, diameter by projects Electronic Speculum to can detect it to have
Structure nano cages.
Nano cages fluorescent yield is measured by known formula, formula are as follows: Φ1=Φ2I1A2η21/I2A1η22It is required that inhaling
Luminosity A1, A2Less than 0.05, to reduce error.It is measured respectively under identical shooting condition using quinine sulfate as reference substance
Integrated fluorescence intensities (the I of sample and reference substance1/I2) and Same Wavelength under the conditions of incident light absorbing state (A2/A1)。I2-
The integrated area value of quinine sulfate fluorescence intensity；The integrated area value of the fluorescence intensity of I1- coffee；A2Quinine sulfate it is ultraviolet
Absorption value；A1The ultraviolet absorption value of coffee；η2-The refractive index of quinine sulfate solvent；η1The refractive index of the solvent of coffee.Through counting
Calculate prepared nano cages quantum yield is 15.61%.
Fluorescence lifetime passes through formula R (τ)=B1e(-τ/τ1) 6.54ns is calculated to obtain, wherein AiIt is time resolution decay time
(τi) percentage.
Tem analysis (Fig. 2) is carried out to nano cages prepared by the present embodiment, it is known that hollow carbon can successfully be prepared by this method
Nanocages material.The nano cages cavity is about 10-20nm, and the carbon layers having thicknesses of shell are 3-5 nm, and are had good
Crystallinity, spacing of lattice about 0.33nm.
Embodiment 3: the preparation of nano cages fluorescence probe
By by the turbot of 230 DEG C of baking 30min, roasted whole fish soaking will be passed through in dehydrated alcohol, every gram of fish
1ml ethyl alcohol extracts 12h, slightly mentions nano cages；Rotary evaporation is redissolved, extraction three to ethyl alcohol is completely removed using chloroform
It is secondary, degreasing, until water phase, is slightly mentioned solution and cross G25 sephadex column chromatography, it is high to collect fluorescence intensity by aqueous phase solution clear
In 80 (excitation wavelength=330nm, launch wavelength=390nm) parts, freeze-drying, obtained powdered substance.
It is hollow and is less than 100nm with multilayer ring-type Shells, diameter by projects Electronic Speculum to can detect it to have
Structure nano cages.
Nano cages fluorescent yield is measured by known formula, formula are as follows: Φ1=Φ2I1A2η21/I2A1η22It is required that inhaling
Luminosity A1, A2Less than 0.05, to reduce error.It is measured respectively under identical shooting condition using quinine sulfate as reference substance
Integrated fluorescence intensities (the I of sample and reference substance1/I2) and Same Wavelength under the conditions of incident light absorbing state (A2/A1)。I2-
The integrated area value of quinine sulfate fluorescence intensity；The integrated area value of the fluorescence intensity of I1- coffee；A2Quinine sulfate it is ultraviolet
Absorption value；A1The ultraviolet absorption value of coffee；η2-The refractive index of quinine sulfate solvent；η1The refractive index of the solvent of coffee.Through counting
Calculate prepared nano cages quantum yield is 16.37%.
Fluorescence lifetime passes through formula R (τ)=B1e(-τ/τ1) 6.82ns is calculated to obtain, wherein AiIt is time resolution decay time
(τi) percentage.
Tem analysis (Fig. 3) is carried out to nano cages prepared by the present embodiment, it is known that hollow carbon can successfully be prepared by this method
Nanocages material.The nano cages cavity is about 10-20nm, and the carbon layers having thicknesses of shell are 3-6 nm, and are had good
Crystallinity, spacing of lattice about 0.33nm.
Embodiment 4: the preparation of nano cages fluorescence probe
By by the turbot of 150 DEG C of baking 90min, roasted whole fish soaking will be passed through in dehydrated alcohol, every gram of fish
1ml alcohol steep 12h, slightly mentions nano cages；Rotary evaporation is redissolved, extraction degreasing to ethyl alcohol is completely removed using chloroform,
Degreasing three times is extracted, until water phase, is slightly mentioned solution and cross G25 sephadex column chromatography, collect fluorescence by aqueous phase solution clear
Intensity is higher than 80 (excitation wavelength=330nm, launch wavelength=390nm) parts, freeze-drying, obtained powdered substance.
It is hollow and is less than 100nm with multilayer ring-type Shells, diameter by projects Electronic Speculum to can detect it to have
Structure nano cages.
Nano cages fluorescent yield is measured by known formula, formula are as follows: Φ1=Φ2I1A2η21/I2A1η22It is required that inhaling
Luminosity A1, A2Less than 0.05, to reduce error.It is measured respectively under identical shooting condition using quinine sulfate as reference substance
Integrated fluorescence intensities (the I of sample and reference substance1/I2) and Same Wavelength under the conditions of incident light absorbing state (A2/A1)。I2-
The integrated area value of quinine sulfate fluorescence intensity；The integrated area value of the fluorescence intensity of I1- coffee；A2Quinine sulfate it is ultraviolet
Absorption value；A1The ultraviolet absorption value of coffee；η2-The refractive index of quinine sulfate solvent；η1The refractive index of the solvent of coffee.Through counting
Calculate prepared nano cages quantum yield is 18.14%.
Fluorescence lifetime passes through formula R (τ)=B1e(-τ/τ1) 7.92ns is calculated to obtain, wherein AiIt is time resolution decay time
(τi) percentage.
Tem analysis (Fig. 4) is carried out to nano cages prepared by the present embodiment, it is known that hollow carbon can successfully be prepared by this method
Nanocages material.The nano cages cavity is about 10-25nm, and the carbon layers having thicknesses of shell are 3-5 nm, and are had good
Crystallinity, spacing of lattice about 0.32nm.
Embodiment 5: the preparation of nano cages fluorescence probe
By by the turbot of 250 DEG C of baking 10min, roasted whole fish soaking will be passed through in dehydrated alcohol, every gram of fish
1ml alcohol steep 12h, slightly mentions nano cages；Rotary evaporation is redissolved, extraction degreasing to ethyl alcohol is completely removed using chloroform,
Degreasing three times is extracted, until water phase, is slightly mentioned solution and cross G25 sephadex column chromatography, collect fluorescence by aqueous phase solution clear
Intensity is higher than 80 (excitation wavelength=330nm, launch wavelength=390nm) parts, freeze-drying, obtained powdered substance.
It is hollow and is less than 100nm with multilayer ring-type Shells, diameter by projects Electronic Speculum to can detect it to have
Structure nano cages.
Nano cages fluorescent yield is measured by known formula, formula are as follows: Φ1=Φ2I1A2η21/I2A1η22It is required that inhaling
Luminosity A1, A2Less than 0.05, to reduce error.It is measured respectively under identical shooting condition using quinine sulfate as reference substance
Integrated fluorescence intensities (the I of sample and reference substance1/I2) and Same Wavelength under the conditions of incident light absorbing state (A2/A1)。I2-
The integrated area value of quinine sulfate fluorescence intensity；The integrated area value of the fluorescence intensity of I1- coffee；A2Quinine sulfate it is ultraviolet
Absorption value；A1The ultraviolet absorption value of coffee；η2-The refractive index of quinine sulfate solvent；η1The refractive index of the solvent of coffee.Through counting
Calculate prepared nano cages quantum yield is 13.64%.
Fluorescence lifetime passes through formula R (τ)=B1e(-τ/τ1) 5.94ns is calculated to obtain, wherein AiIt is time resolution decay time
(τi) percentage.
Tem analysis (Fig. 5) is carried out to nano cages prepared by the present embodiment, it is known that hollow carbon can successfully be prepared by this method
Nanocages material.The nano cages cavity is about 12-25nm, and the carbon layers having thicknesses of shell are 2-6 nm, and are had good
Crystallinity, spacing of lattice about 0.33nm.
Embodiment 6:Fe3+The foundation of standard curve between concentration and fluorescence intensity
Nano cages fluorescence probe 3 in Example 3 is configured to the nano cages solution that concentration is 10mg/ml, takes
The nano cages solution that 1ml is prepared is added in 5ml water, as blank control group；In addition it takes in 15 centrifuge tubes and is separately added into again
Then the nano cages solution that 1ml is prepared sequentially adds 5ml concentration difference 10,20,30,40,50,60 in 15 centrifuge tubes,
70,80,90,100,110,120,150,200 μM of FeCl3Aqueous solution, after being sufficiently mixed, obtained nano cages and Fe3+'s
After standard solution stands 2min, covered in the fluophotometer examination criteria solution of 280~700nm using Detection wavelength range
Fluorescence intensity F1, testing conditions are that excitation wavelength is 330nm, and launch wavelength 390nm, the fluorescence intensity of blank control group is F0(swash
Hair wavelength is 330nm, launch wavelength 390nm), draw out fluorescent quenching intensity ((F0-F1)/F0) and Fe3+Mark between concentration
Directrix curve.
Fig. 6 is the nano cages fluorescence probe 3 in embodiment 3 to Fe3+Sensitivity analysis figure, it is seen that nano cages fluorescence
Probe is with Fe3+The increase of concentration, fluorescence intensity gradually decrease, and illustrate the nano cages fluorescence probe to Fe3+Concentration it is very quick
Sense；But as [Fe3+When] >=120 μM, the fluorescence intensity of nano cages no longer reduces substantially, so nano cages are to [Fe3+] inspection
Surveying the upper limit is 120 μM；With Fe3+Solution concentration range is abscissa at 20-120 μM, with fluorescent quenching intensity ((F0-F1)/F0) be
Equation of linear regression y=0.00868x-0.0981, coefficient R can be made in ordinate2=0.993, detectable limit is 3.96 μ
M。
FeCl is added in Xiang Shuizhong3, Fe known to compound concentration3+Solution ([Fe3+]=100 μM) it is used as sample to be tested, then
The volume ratio of nano cages fluorescence probe 3 in Example 3, nano cages solution and solution to be detected is 1:5, as to be measured
Fe3+Solution example detects its fluorescence intensity, obtains fluorescence intensity change value；It is substituted into the standard curve of the present embodiment, is counted
Calculation obtains Fe3+99.1 μM of concentration, the rate of recovery illustrates that probe in detecting accuracy is high up to 99.1%.
Embodiment 7:Fe3+The foundation of standard curve between concentration and fluorescence intensity
Nano cages fluorescence probe 4 in Example 4 is configured to the nano cages solution that concentration is 30mg/ml, takes
The nano cages solution that 1ml is prepared is added in 4ml water, as blank control group；In addition it takes in 15 centrifuge tubes and is separately added into again
The nano cages solution that 1ml is prepared, it is concentration difference 10,20,30,40 that 4ml concentration is then sequentially added in 15 centrifuge tubes,
50,60,70,80,90,100,110,120,150,200 μM of FeCl3Aqueous solution, after being sufficiently mixed, obtained nano cages
With Fe3+Standard solution stand 2min after, be F with the fluorescence intensity in fluophotometer examination criteria solution1(excitation wavelength is
330nm, launch wavelength 390nm), the fluorescence intensity of blank control group is F0(excitation wavelength 330nm, launch wavelength
390nm), fluorescent quenching intensity ((F is drawn out0-F1)/F0) and Fe3+Standard curve between concentration, equation of linear regression y=
0.00938x-0.0284, coefficient R2=0.995, detection range is 10-110 μM, and detectable limit is 4.01 μM.
FeCl is added in Xiang Shuizhong3, Fe known to compound concentration3+Solution ([Fe3+]=100 μM), then in Example 4
The volume ratio of nano cages fluorescence probe 4, nano cages solution and solution to be detected is 1:4, as Fe to be measured3+Solution example,
Its fluorescence intensity is detected, fluorescence intensity change value is obtained；It is substituted into the standard curve of the present embodiment, Fe is calculated3+'s
98.3 μM of concentration, the rate of recovery illustrates that the probe in detecting accuracy is high up to 98.3%.
Embodiment 8:Fe3+The foundation of standard curve between concentration and fluorescence intensity
Nano cages fluorescence probe 5 in Example 5 is configured to the nano cages solution that concentration is 50mg/ml, takes
The nano cages solution that 1ml is prepared is added in 3ml water, as blank control group；In addition it takes in 15 centrifuge tubes and is separately added into again
The nano cages solution that 1ml is prepared, it is concentration difference 10,20,30,40 that 3ml concentration is then sequentially added in 15 centrifuge tubes,
50,60,70,80,90,100,110,120,150,200 μM of FeCl3Aqueous solution, after being sufficiently mixed, obtained nano cages
With Fe3+Standard solution stand 2min after, with the fluorescence intensity F in fluophotometer examination criteria solution1(excitation wavelength is
330nm, 390 nm of launch wavelength), the fluorescence intensity of blank control group is F0(excitation wavelength 330nm, launch wavelength
390nm), fluorescent quenching intensity ((F is drawn out0-F1)/F0) and Fe3+Standard curve between concentration, equation of linear regression y=
0.00749x+0.0234, coefficient R2=0.996, detection range is 20-100 μM, and detectable limit is 4.12 μM.
FeCl is added in Xiang Shuizhong3, Fe known to compound concentration3+Solution ([Fe3+]=100 μM), then in Example 5
The volume ratio of nano cages fluorescence probe 5, nano cages solution and solution to be detected is 1:3, as Fe to be measured3+Solution example,
Its fluorescence intensity is detected, fluorescence intensity change value is obtained；It is substituted into the standard curve of the present embodiment, Fe is calculated3+'s
101.5 μM of concentration, the rate of recovery illustrates that the probe in detecting accuracy is high up to 101.5%.
The method of the present invention raw material is easy to get, is at low cost, process is simply controllable, i.e., carries out simple ethyl alcohol extraction, rotation to grilled fish
It steams, extraction, gel chromatography, gained nano cages are hollow and are less than the knot of 25nm with multilayer ring-type Shells, diameter
The feature that structure, fluorescence lifetime are long, quantum efficiency is high, result is reproducible.
The foregoing is only a preferred embodiment of the present invention, but scope of protection of the present invention is not limited thereto,
Anyone skilled in the art within the technical scope of the present disclosure, according to the technique and scheme of the present invention and its
Inventive concept is subject to equivalent substitution or change, should be covered by the protection scope of the present invention.
Claims (3)
1. a kind of preparation method of nano cages fluorescence probe, which is characterized in that specific steps are as follows:
Using fish as raw material, 150~300 DEG C of 10~120min of baking；
The fish are turbot, conger pile；
2~48h will be extracted in dehydrated alcohol by roasted fish soaking, it is described dense by roasted fish and the end of added ethyl alcohol
Degree is every gram of fish 1ml ethyl alcohol；After extraction processing, liquid phase is collected, rotary evaporation removes ethyl alcohol；
It is redissolved using chloroform, extraction degreasing, extraction is until aqueous phase solution clear, collects water phase, as slightly mention repeatedly
Solution；
By it is described slightly mention solution cross G25 sephadex column chromatography, collect fluorescence intensity be more than or equal to 80, excitation wavelength 330nm,
The part of launch wavelength 390nm, freeze-drying, obtained powdered substance, as nano cages fluorescence probe.
2. the nano cages fluorescence probe of claim 1 the method preparation is in Fe3+Application in detection, which is characterized in that packet
Include following steps:
(1) by the nano cages fluorescence probe be dissolved in water obtained concentration be 2~50mg/ml nano cages fluorescence probe water
Solution；Configure a series of Fe of known concentrations3+Solution, to the Fe3+It is water-soluble that the nano cages fluorescence probe is added in solution
Liquid obtains Fe3+Standard sample solution；
The Fe3+Fe in solution3+Concentration range be 20~200 μM；
The nano cages fluorescence probe aqueous solution and Fe3+The volume ratio of solution is 1:0.5~5；
(2) the nano cages fluorescence probe aqueous solution made from step (1) is added into water, as blank control group；It is described
The Fe made from the adding proportion of nano cages fluorescence probe aqueous solution and step (1)3+The adding proportion of standard sample solution
It is identical；
Fluorescence intensity in examination criteria solution is compared with control group, according to Fe3+Solution concentration and corresponding fluorescent quenching intensity
Modeling, obtains Fe3+Standard curve between concentration and fluorescence intensity；
The actual conditions of the fluorescence intensity detection are as follows:
Cover the sepectrophotofluorometer of 280~700nm using Detection wavelength range；
Excitation wavelength is 280~500nm；
The wavelength of fluorescence emission peak is 350~690nm；
(3) Fe to be measured is taken3+Solution, to the Fe to be measured3+The nano cages fluorescence probe made from step (1) is added in solution
Aqueous solution, as Fe to be measured3+Solution example；The adding proportion of the nano cages fluorescence probe aqueous solution and step (1) are made
The Fe3+The adding proportion of standard sample solution is identical；
Using detection method identical with step (2), Fe to be measured is obtained3+The fluorescence intensity of solution example is obtained in conjunction with step (2)
Fe3+Concentration and Standardization curve for fluorescence intensity obtain Fe to be measured3+The Fe of solution example3+Concentration.
3. the nano cages fluorescence probe of method preparation is in Fe according to claim 23+Application in detection, feature exist
In the modeling process, with Fe3+Solution concentration is abscissa, corresponding Fe3+The fluorescent quenching intensity that standard sample solution obtains
For ordinate, Fe is obtained3+Concentration and Standardization curve for fluorescence intensity.
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| Presence of Amorphous Carbon Nanoparticles in Food Caramels;Md Palashuddin Sk et al.;《SCIENTIFIC REPORTS》;20120426;第2卷;第383-387页 |
| The presence of carbon nanostructures in bakery products induces metabolic stress in human mesenchymal stem cells through CYP1A and p53 gene expression;Ahmed M.Al-Hadi et al.;《Environmental Toxicology and Pharmacology》;20151202;第41卷;第103-112页 |
| Theranostic carbon dots derived from garlic with efficient anti-oxidative effects towards macrophages;Chuanxu Yang et al.;《 RSC Adv.》;20151109;第5卷;第97836-97840页 |
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