CN108760700A - A kind of preparation of fluorescence gold nanoclusters and it is used for tetracycline and copper fluorescence probe - Google Patents
A kind of preparation of fluorescence gold nanoclusters and it is used for tetracycline and copper fluorescence probe Download PDFInfo
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- CN108760700A CN108760700A CN201810531924.2A CN201810531924A CN108760700A CN 108760700 A CN108760700 A CN 108760700A CN 201810531924 A CN201810531924 A CN 201810531924A CN 108760700 A CN108760700 A CN 108760700A
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- fluorescence
- tetracycline
- gold nanoclusters
- quantum dot
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- 239000010931 gold Substances 0.000 title claims abstract description 61
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 title claims abstract description 56
- 229910052737 gold Inorganic materials 0.000 title claims abstract description 56
- 239000004098 Tetracycline Substances 0.000 title claims abstract description 45
- 229960002180 tetracycline Drugs 0.000 title claims abstract description 45
- 229930101283 tetracycline Natural products 0.000 title claims abstract description 45
- 235000019364 tetracycline Nutrition 0.000 title claims abstract description 45
- 150000003522 tetracyclines Chemical class 0.000 title claims abstract description 44
- 239000010949 copper Substances 0.000 title claims abstract description 26
- 239000000523 sample Substances 0.000 title claims abstract description 23
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 title claims abstract description 13
- 229910052802 copper Inorganic materials 0.000 title claims abstract description 13
- 238000002360 preparation method Methods 0.000 title claims abstract description 5
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 claims abstract description 23
- 235000013336 milk Nutrition 0.000 claims abstract description 21
- 210000004080 milk Anatomy 0.000 claims abstract description 21
- 239000005018 casein Substances 0.000 claims abstract description 20
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 claims abstract description 20
- 235000021240 caseins Nutrition 0.000 claims abstract description 20
- 238000001514 detection method Methods 0.000 claims abstract description 19
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- RTZKZFJDLAIYFH-UHFFFAOYSA-N ether Substances CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 12
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- HIXDQWDOVZUNNA-UHFFFAOYSA-N 2-(3,4-dimethoxyphenyl)-5-hydroxy-7-methoxychromen-4-one Chemical compound C=1C(OC)=CC(O)=C(C(C=2)=O)C=1OC=2C1=CC=C(OC)C(OC)=C1 HIXDQWDOVZUNNA-UHFFFAOYSA-N 0.000 claims description 3
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- FDWREHZXQUYJFJ-UHFFFAOYSA-M gold monochloride Chemical compound [Cl-].[Au+] FDWREHZXQUYJFJ-UHFFFAOYSA-M 0.000 claims description 2
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- VLTRZXGMWDSKGL-UHFFFAOYSA-N perchloric acid Chemical compound OCl(=O)(=O)=O VLTRZXGMWDSKGL-UHFFFAOYSA-N 0.000 description 1
- 229920002647 polyamide Polymers 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
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- OFVLGDICTFRJMM-WESIUVDSSA-N tetracycline Chemical compound C1=CC=C2[C@](O)(C)[C@H]3C[C@H]4[C@H](N(C)C)C(O)=C(C(N)=O)C(=O)[C@@]4(O)C(O)=C3C(=O)C2=C1O OFVLGDICTFRJMM-WESIUVDSSA-N 0.000 description 1
- 238000005979 thermal decomposition reaction Methods 0.000 description 1
- 229960002663 thioctic acid Drugs 0.000 description 1
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- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 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"
- G01N21/643—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" non-biological material
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B22—CASTING; POWDER METALLURGY
- B22F—WORKING METALLIC POWDER; MANUFACTURE OF ARTICLES FROM METALLIC POWDER; MAKING METALLIC POWDER; APPARATUS OR DEVICES SPECIALLY ADAPTED FOR METALLIC POWDER
- B22F1/00—Metallic powder; Treatment of metallic powder, e.g. to facilitate working or to improve properties
- B22F1/07—Metallic powder characterised by particles having a nanoscale microstructure
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B22—CASTING; POWDER METALLURGY
- B22F—WORKING METALLIC POWDER; MANUFACTURE OF ARTICLES FROM METALLIC POWDER; MAKING METALLIC POWDER; APPARATUS OR DEVICES SPECIALLY ADAPTED FOR METALLIC POWDER
- B22F9/00—Making metallic powder or suspensions thereof
- B22F9/16—Making metallic powder or suspensions thereof using chemical processes
- B22F9/18—Making metallic powder or suspensions thereof using chemical processes with reduction of metal compounds
- B22F9/24—Making metallic powder or suspensions thereof using chemical processes with reduction of metal compounds starting from liquid metal compounds, e.g. solutions
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y30/00—Nanotechnology for materials or surface science, e.g. nanocomposites
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y40/00—Manufacture or treatment of nanostructures
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- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/08—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials
- C09K11/58—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials containing copper, silver or gold
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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"
- G01N2021/6432—Quenching
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Abstract
The invention discloses a kind of preparation of fluorescence gold nanoclusters and it is used for tetracycline and copper fluorescence probe.The present invention is the stabilizer and reducing agent for synthesizing gold nanoclusters with casein and carbon quantum dot isolated in milk, passes through one-step synthesis method fluorescence gold nanoclusters（Au NCs）.Au NCs generate three transmittings under 350nm excitation wavelengths at 433nm, 702nm and 1052nm, and tetracycline has linear quenching effect at 702nm, Cu is added in the Au NCs systems to after being quenched2+, fluorescence restores, and whole process has little influence on the fluorescence intensity at 1052nm；Tetracycline is added in Au NCs, is observed under 365nm ultraviolet lamps, the red fluorescence of Au NCs becomes yellow fluorescence, adds Cu2+Afterwards, yellow fluorescence fades away, and red fluorescence gradually restores.Thereby establish tetracycline and Cu2+Near-infrared ratio fluorescent switch and ultraviolet lower visualization dual signal detect.This method fluorescence spectrum interferes small, high specificity, detection sensitivity high, easy to operate, can be carried out at the same time tetracycline and Cu in nearly red sector2+Detection.
Description
Technical field
The present invention relates to chemical analysis detection technique field, the preparation of specially a kind of fluorescence gold nanoclusters and be used for Fourth Ring
Element and copper fluorescence probe.
Background technology
Noble metal fluorescence nano cluster is a kind of novel fluorescence nano material, is made of several to hundreds of atoms, grain size is about
The characteristics such as unique optics, electricity, chemistry are presented close to Fermi's wavelength of electronics for 0.2~3.0 nm, bioanalysis at
The fields such as picture, environmental monitoring have broad application prospects.Fluorescence gold nanoclusters (AuNCs) have stronger luminescence generated by light, good
Good photostability, big Stokes shift and high fluorescence quantum yield, is to study widest noble metal fluorescence at present
Nano-cluster has been used for the detection of hydrogen peroxide, metal ion, tea polyphenols etc..With bovine serum albumin(BSA) (BSA) for ligand,
The protein hybridization fluorescence gold nanoclusters (AuNCs BSA) with red fluorescence characteristic are prepared under conditions of mild.Currently,
Has the synthesis that gas chromatography is used for AuNCs, such as dendrimer or high polymer (polyamide, polyethyleneimine), ammonia
Base acid (proline, tyrosine, cysteine etc.), transferrins, enzyme (horseradish peroxidase, trypsase, lysozyme,
Pepsin etc.) and some small-molecule substances (lipoic acid, dithiothreitol (DTT)) etc..Because BSA raw materials are easy to get, and are with BSA
Process that ligand synthesizes AuNCs is simple, mild condition, so AuNCs@BSA are widely used in many fields.Casein is ox
Main protein in milk, content are about 2.6 g/100 mL, account for 80% of total protein in milk.Casein contains human body
Required 8 kinds of amino acid is a kind of binding protein of phosphorous calcium.
Carbon quantum dot is the new carbon of a kind of carbon such as graphene quantum dot and carbon nano dot.It has excellent
Optical property, adjustable excitation and transmitting behavior, higher fluorescent stability, lower toxicity and good biocompatibility,
It is widely used in more and more fields.The abundant hydroxyl in carbon quantum dot surface becomes gold nanoclusters conjunction
At the good reducing agent of middle surface modification and stabilizer.Currently, casein and carbon quantum dot is used to be closed as reducing agent and stabilizer
It at the method for gold nanoclusters, has not been reported, more has no report of the synthetic material for two kinds of ratio fluorescent probes.
The present invention synthesizes fluorescent carbon quantum dot using the stem of noble dendrobium rich in polysaccharide as carbon quantum raw material, by a step thermal decomposition method,
Reducing agent and protective agent using carbon quantum dot and casein as synthesis fluorescence gold nanoclusters synthesize fluorescence gold nanoclusters.?
Under the excitation wavelength of 350nm, gold nanoclusters are simultaneously in blue light region（443nm）With nearly red light district（714nm,1052nm）Generate transmitting
Spectrum constructs nearly red light district ratio fluorescent sensor using fluorescence gold nanoclusters as tetracycline and the fluorescence probe of copper.Method
Novelty, high specificity, high sensitivity.
Invention content
The purpose of the present invention is to provide one kind synthesizing fluorescence using carbon quantum dot and casein as reducing agent and protective agent
Gold nanoclusters method, and for the fluorescence probe of highly sensitive, highly selective tetracycline and copper.
The purpose of the present invention is be achieved through the following technical solutions：
The synthesis of fluorescence gold nanoclusters includes the following steps：By the isolated casein solution of fresh milk and carbon quantum dot and chlorine
1 mol/L NaOH solution tune is added dropwise after magnetic agitation reacts 1-2 min in 37 DEG C of isometric mixing in auric acid solution
PH value is saved to 12-13, is stirred to react 12 h under the conditions of 37 DEG C, solution colour becomes dark-brown from glassy yellow.
The isolated casein method of the fresh milk includes：50mL Fresh Milks are taken, in water bath with thermostatic control
40 DEG C are heated to, is slowly added into 10% acetum while stirring, makes milk pH=4.6-4.8, after placing cooling, clarification,
The Buchner funnel of the nylon cloth of 200 mesh filters, and is washed successively with each 30-50ml of isometric mixed liquor of ethyl alcohol, ethyl alcohol and ether
Twice, 30-50mL ether washs at twice, last vacuum filter, the casein for spontaneously drying white.
The synthetic method of the carbon quantum dot includes：The dendrobium candidum powder for weighing 2-5g dryings, is scattered in 90-
In the hot water of 150mL 60-70 DEG C and it is stirred continuously, it is to be mixed uniformly to add 10-20mL absolute ethyl alcohols, 1-3mL afterwards in the pasty state
Ethylenediamine after stirring evenly, is transferred to polytetrafluoroethyllining lining reaction kettle after 180 DEG C of heating 9-12h, natural cooling, centrifugation,
Large granular impurity is removed, the carbon quantum dot of N doping is obtained after crossing 0.22 μm of filter membrane.Gained quantum dot is placed in 60 DEG C of vacuum to do
Drying for 24 hours, obtains carbon quantum dot in dry case.
The centrifugation is that 15-20min is carried out at rotating speed 8000-10000rpm.
The casein, carbon quantum dot, gold chloride weight ratio be 15-20：5-10：1.
The fluorescence gold nanoclusters of synthesis are used for the fluorescence probe of tetracycline and copper, and specific method includes：
（1）The excitation wavelength and launch wavelength of fluorescence gold nanoclusters determine：The fluorescence gold nanoclusters of synthesis are dissolved in aqueous solution
In, it is scanned to 800nm 300, fluorescence gold nanoclusters are under 350nm excitation wavelengths, in 433nm, 702nm and 1052nm
Place generates three transmittings；
（2）Near-infrared ratio fluorescent switch detects tetracycline and Cu respectively2+：It is molten in the tetracycline containing 0-20 μM of concentration range
In liquid, tetracycline has linear quenching effect, Cu to fluorogold nanocluster fluorescence intensity at 702nm2+To glimmering after quenching
Light gold nanoclusters system has fluorescence restitution, and fluorescence sensitivity is linear in 0-30 μM of range, and whole process is hardly
The fluorescence intensity at 1052nm is influenced, near-infrared ratio fluorescent probe is thus established and carries out tetracycline and Cu2+Detection；
（3）Cu2+Visual detection：Tetracycline is added in fluorescence gold nanoclusters, is observed under 365nm ultraviolet lamps, fluorescence
The red fluorescence of gold nanoclusters becomes yellow fluorescence, adds Cu2+Afterwards, yellow fluorescence fades away, and red fluorescence gradually restores,
Cu is carried out according to the depth of fluorescent red2+Assay；
Step（2）The fluorescence gold nanoclusters dosage is 100-300 μ L；
Step（3）The fluorescence gold nanoclusters dosage 100-200 μ L.
The advantage of the invention is that：
1. the present invention uses casein and is to synthesize the reduction of fluorescence gold nanoclusters with the carbon quantum of the stem of noble dendrobium synthesis rich in polysaccharide
The fluorogold nano-cluster of agent and protective agent, synthesis has blue light region（443nm）With nearly red light district（702nm,1052nm）Three
Launch wavelength；
2, tetracycline has the fluorogold nanocluster fluorescence intensity of synthesis linear quenching effect, Cu at 702nm2+To quenching
Fluorescence gold nanoclusters system later has fluorescence restitution, and fluorescence sensitivity is linear, and whole process has little influence on
Thus fluorescence intensity at 1052nm establishes near-infrared ratio fluorescent probe and carries out tetracycline and Cu2+Detection；
3, using tetracycline to the quenching effect to fluorescence gold nanoclusters, under 365nm ultraviolet lights, the red of fluorescence gold nanoclusters
Fluorescence becomes yellow fluorescence, then due to Cu2+To the restitution of fluorescence, red fluorescence gradually restores, according to the depth of fluorescent red
Shallow carry out Cu2+Visual measurement；
4, near-infrared ratio fluorescent probe carries out tetracycline and Cu2+Detection be happened near infrared region, interference is small, high specificity,
High sensitivity, method are unique.
Description of the drawings
Fig. 1 is the scanning electron microscope (SEM) photograph that fluorescence gold nanoclusters are synthesized in embodiment 1.
Fig. 2 is the excitation-emission spectral schematic spectrum of synthesis fluorescence gold nanoclusters in embodiment 1.
Fig. 3 is the ratio fluorescent probe of tetracycline with relative intensity of fluorescence ratio I702/I1052Collection of illustrative plates is illustrated to tetracycline.
Fig. 4 is Cu2+Ratio fluorescent probe with relative intensity of fluorescence ratio I702/I1052To Cu2+Concentration illustrates collection of illustrative plates.
Specific implementation mode
Explanation, but this hair are described in further detail to technical scheme of the present invention below in conjunction with specific embodiments
Bright protection domain is not limited to that.
Embodiment 1：The assay operating procedure of tetracycline and copper is as follows in egg：
（1）The separation method of fresh milk casein：50mL Fresh Milks are taken, 40 DEG C are heated in water bath with thermostatic control, while stirring
It mixes side and is slowly added into 10% acetum, make milk pH=4.6-4.8, after placing cooling, clarification, the cloth of the nylon cloth of 200 mesh
Family name's funnel filters, and is washed twice successively with each 30-50ml of isometric mixed liquor of ethyl alcohol, ethyl alcohol and ether, 30-50 milliliters of ether
It washs at twice, last vacuum filter, the casein for spontaneously drying white；
（2）Carbon quantum dot（CDs）Synthesis：The dendrobium candidum powder for weighing 2-5g dryings, is scattered in 90-150mL 60-70 DEG C
Hot water in and be stirred continuously, to be mixed uniformly to add 10-20mL absolute ethyl alcohols afterwards in the pasty state, 1-3mL ethylenediamines, stirring is equal
After even, polytetrafluoroethyllining lining reaction kettle is transferred to after 180 DEG C of heating 9-12h, natural cooling, 10000rpm centrifuges 15min,
Large granular impurity is removed, the carbon quantum dot of N doping is obtained after crossing 0.22 μm of filter membrane, using quinine sulfate as reference substance, fluorescence volume
Sub- yield 25%.It is for 24 hours, spare that gained quantum dot is placed in drying in 60 DEG C of vacuum drying chambers；
（3）Fluorescence gold nanoclusters（AuNCs）Synthesis：By step（1）Obtained 20mg/mL casein solutions 3mL and step（2）
For obtained 3mg/mL carbon quantum dots 10mL and 3.4 mg/1 mL of mL chlorauric acid solutions in 37 DEG C of isometric mixing, magnetic agitation is anti-
After answering 1-2 min, 1 mol/L NaOH solution is added dropwise and adjusts pH value to 12-13, is stirred to react under the conditions of 37 DEG C
12 h, solution colour become dark-brown from glassy yellow；
（4）The excitation wavelength and launch wavelength of fluorescence gold nanoclusters determine：The fluorescence gold nanoclusters of synthesis are dissolved in aqueous solution
In, it is scanned to 1200nm 300, in 350nm, maximum emission wavelength exists the maximum excitation wavelength of fluorescence gold nanoclusters respectively
At 433nm, 702nm and 1052nm；
（5）Fluorescence response of the fluorescence gold nanoclusters to tetracycline：In tetracycline (TeT) solution containing 0-20 μM of concentration range
In, the 100 μ L of fluorescence gold nanoclusters of synthesis are added, fluorescence spectrum is under the illumination excitation of 350nm wavelength, and probe is with relative fluorescence
Intensity rate I702/I1052It maps to tetracycline concentration, obtains linear relationship I702/I1052= 0.03210+0.0191C（μM）
(r=0.9968), detection are limited to 0.015 μM (S/N=3), the concentration for quantitatively detecting tetracycline；
（6）Fluorescence gold nanoclusters are to Cu2+Fluorescence response：In the fluorescent quenching with 100 μ L AuNCs containing 15 μM of concentration
In TeT- AuNCs systems, Cu is added2+, fluorescence spectrum is under the illumination excitation of 350nm wavelength, and fluorescence sensitivity is in 0-30 μM of model
It encloses, probe is with relative intensity of fluorescence ratio I702/I1052To Cu2+Concentration is mapped, and linear relationship I is obtained702/I1052= 0.2142+
0.0067C（μM）(r=0.9970), detection is limited to 0.05 μM (S/N=3), for quantitatively detecting Cu2+；
（7）Cu2+Visual detection：In the fluorescent quenching TeT- AuNCs systems with 100 μ L AuNCs containing 15 μM of concentration
In, 10-100 μM of Cu is added2+, fluorescence spectrum is under the illumination excitation of 365nm wavelength, and yellow fluorescence fades away, red fluorescence
Gradually restore, the strong and weak progress Cu restored according to red fluorescence2+Visual detection；
（8）Egg sample tetracycline measures：Take egg white 50mL that 100mL methanol-acetones (1 are added：1, V/V) extracts reagent, it is homogeneous
3h is stood in refrigerator afterwards, after being filtered to remove precipitation, is evaporated under reduced pressure in draught cupboard and is concentrated into 5mL, by step（5）Carry out Fourth Ring
The ratio fluorescent of element measures, and substitutes into linear relationship, obtains 0.33 μM；
（9）Egg sample copper measures：0. 4g of egg white is weighed in polytetrafluoroethylene (PTFE) digester, adds the dense HNO of people respectively3 2.0mL
And H2O21.0mL is put into micro-wave oven, is placed in 5 grades (100 % power, 650 W), and 15min is digested, and extraction sample is disappeared
Change liquid in 50mL volumetric flasks, is diluted to scale.Take 5mL by step（6）Carry out Cu2+Ratio fluorescent measure, substitute into linear close
System, obtains 3.75 μM；Step is pressed simultaneously（7）Carry out Cu2+Visual detection, as a result unanimously.
Embodiment 2：The assay step of tetracycline and copper is in environmental water sample：
（1）The separation method of fresh milk casein：：With 1 step of embodiment (1)；
（2）Carbon quantum dot（CDs）Synthesis：With 1 step of embodiment (2)；
（3）Fluorescence gold nanoclusters（AuNCs）Synthesis：With 1 step of embodiment (3)；
（4）The excitation wavelength and launch wavelength of fluorescence gold nanoclusters determine：With 1 step of embodiment (4)；
（5）Fluorescence response of the fluorescence gold nanoclusters to tetracycline：With 1 step of embodiment (5)；
（6）Fluorescence gold nanoclusters are to Cu2+Fluorescence response：With 1 step of embodiment (6)；
（7）Cu2+Visual detection：With 1 step of embodiment (7)；
（8）Tetracycline measures in environmental water sample：Environmental water sample 5mL is taken, by step（5）The ratio fluorescent for carrying out tetracycline measures,
Linear relationship is substituted into, is not detected；
（9）Cu in environmental water sample2+It measures：Environmental water sample 5mL is taken, by step（6）Carry out Cu2+Ratio fluorescent measure, substitute into line
Sexual intercourse obtains 9.01 μM；Step is pressed simultaneously（7）Carry out Cu2+Visual detection, as a result unanimously.
Embodiment 3：Tetracycline and copper content testing step are in milk sample：
（1）The separation method of fresh milk casein：：With 1 step of embodiment (1)；
（2) carbon quantum dot（CDs）Synthesis：With 1 step of embodiment (2)；
（3）Fluorescence gold nanoclusters（AuNCs）Synthesis：With 1 step of embodiment (3)；
（4）The excitation wavelength and launch wavelength of fluorescence gold nanoclusters determine：With 1 step of embodiment (4)；
（5）Fluorescence response of the fluorescence gold nanoclusters to tetracycline：With 1 step of embodiment (5)；
（6）Fluorescence gold nanoclusters are to Cu2+Fluorescence response：With 1 step of embodiment (6)；
（7）Cu2+Visual detection：With 1 step of embodiment (7)；
（8）Milk sample tetracycline measures：The accurate defatted milk 0.1g that weighs is added 4.00 in having in 10 mL tool plug test tubes of scale
ML papain solutions, oscillation are set and are digested in 65 DEG C of insulating boxs overnight, and next day takes out enzymolysis liquid cooling, with 3000r/min
Centrifuge 15 min, 1 mL of Aspirate supernatant, by step（5）The ratio fluorescent for carrying out tetracycline measures, and substitutes into linear relationship, obtains
0.12μM；
（9）Milk sample Cu2+It measures：5.00 mL HNO are added in thermophilic digestion pipe in the accurate 5.00 mL milk samples that pipette3
With 2.00 mL HClO4, a bead is added, nozzle puts Bent nose funnel, stands 24 h, sets and be heated to red gas on electric furnace
Body disperses, and removes cooling, then proceedes to be heated to emitting dense white cigarette, be cooled to room temperature, and a small amount of double distilled water, digestion to sample is added
Cooling is taken out after becoming supernatant liquid, adds 10% HNO3Solution is settled to 50 mL.Take 5mL by step（6）Carry out Cu2+Ratio
Fluoremetry substitutes into linear relationship, obtains 1.05 μM；Step is pressed simultaneously（7）Carry out Cu2+Visual detection, as a result unanimously.
As shown in Figure 1：The scanning electron microscope (SEM) photograph of fluorescence gold nanoclusters is synthesized, nanometer particle size is about 7nm.
As shown in Figure 2：Fluorescence gold nanoclusters prepared by the present invention, when excitation wavelength is 433nm, in 702nm and
There is maximum emission wavelength at 1052nm.
Fig. 3 is the ratio fluorescent probe of tetracycline with relative intensity of fluorescence ratio I702/I1052Collection of illustrative plates is illustrated to tetracycline,
Illustrate the linear relationship having had；
Fig. 4 is Cu2+Ratio fluorescent probe with relative intensity of fluorescence ratio I702/I1052To Cu2+Concentration illustrates collection of illustrative plates, explanation to have
Good linear relationship.
Claims (6)
1. a kind of preparation of fluorescence gold nanoclusters and be used for tetracycline and copper fluorescence probe, which is characterized in that fluorescence gold nanoclusters
Synthesis includes the following steps：By the isolated casein solution of fresh milk and carbon quantum dot and chlorauric acid solution at 37 DEG C etc.
After magnetic agitation reacts 1-2 min, 1 mol/L NaOH solution adjusting pH value to 12-13 is added dropwise in volume mixture,
12 h are stirred to react under the conditions of 37 DEG C, solution colour becomes dark-brown from glassy yellow.
2. the isolated casein method of fresh milk according to claim 1, includes：50mL Fresh Milks are taken,
It is heated to 40 DEG C in water bath with thermostatic control, is slowly added into 10% acetum while stirring, makes milk pH=4.6-4.8, places cold
But after, clarifying, the Buchner funnel of the nylon cloth of 200 mesh filters, and uses isometric mixed liquor of ethyl alcohol, ethyl alcohol and ether each successively
30-50mL is washed twice, 30-50mL ether washs at twice, last vacuum filter, the casein for spontaneously drying white.
3. the synthetic method of carbon quantum dot according to claim 1, includes：The dendrobium candidum powder of 2-5g dryings is weighed,
It is scattered in the hot water of 90-150mL 60-70 DEG C and is stirred continuously, to be mixed uniformly to add 10-20mL afterwards in the pasty state anhydrous
Ethyl alcohol, 1-3mL ethylenediamines after stirring evenly, are transferred to polytetrafluoroethyllining lining reaction kettle and heat 9-12h in 180 DEG C, naturally cold
But after, centrifugation removes large granular impurity, obtains the carbon quantum dot of N doping after crossing 0.22 μm of filter membrane, gained quantum dot is placed in
Drying for 24 hours, obtains carbon quantum dot in 60 DEG C of vacuum drying chambers.
4. the weight ratio of casein according to claim 1, carbon quantum dot, gold chloride is 15-20：5-10：1.
5. the synthetic method of carbon quantum dot according to claim 3, it is characterised in that：Centrifugation is in rotating speed 8000-
15-20min is carried out under 10000rpm.
6. a kind of fluorescence gold nanoclusters include for tetracycline and copper fluorescence probe, specific method：
（1）The excitation wavelength and launch wavelength of fluorescence gold nanoclusters determine：The fluorescence gold nanoclusters of synthesis are dissolved in aqueous solution
In, it is scanned to 800nm 300, fluorescence gold nanoclusters are under 350nm excitation wavelengths, in 433nm, 702nm and 1052nm
Place generates three transmittings；
（2）Near-infrared ratio fluorescent switch detects tetracycline and Cu respectively2+：It is molten in the tetracycline containing 0-20 μM of concentration range
In liquid, tetracycline has linear quenching effect, Cu to fluorogold nanocluster fluorescence intensity at 702nm2+To glimmering after quenching
Light gold nanoclusters system has fluorescence restitution, and fluorescence sensitivity is linear in 0-30 μM of range, and whole process is hardly
The fluorescence intensity at 1052nm is influenced, near-infrared ratio fluorescent probe is thus established and carries out tetracycline and Cu2+Detection；
（3）Cu2+Visual detection：Tetracycline is added in fluorescence gold nanoclusters, is observed under 365nm ultraviolet lamps, fluorogold
The red fluorescence of nano-cluster becomes yellow fluorescence, adds Cu2+Afterwards, yellow fluorescence fades away, and red fluorescence gradually restores, root
Cu is carried out according to the depth of fluorescent red2+Assay.
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