CN106124465A - Polyphosphate new detecting method based on gold nano cluster fluorescent characteristic - Google Patents
Polyphosphate new detecting method based on gold nano cluster fluorescent characteristic Download PDFInfo
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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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- 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
Abstract
Present invention polyphosphate based on gold nano cluster fluorescent characteristic new detecting method, there is provided with bovine serum albumin (BSA) as template, the bovine serum albumin gold nano cluster (BSA AuNCs) using chemical reduction method to synthesize a diameter of 2nm, can send the fluorescence of redness under the scanning that wave-length coverage is 200nm 800nm ultraviolet light.Work as Cu2+BSA AuNCs Cu is generated with the glycine chelate in BSA chain2+Time, fluorescence can quencher；When adding polyphosphate radical (PPy), PPy will be with Cu2+Chelating makes Cu2+From BSA AuNCs Cu2+Surface spins off, and forms BSA AuNCs, and fluorescence recovers.Based on the principle that, the present invention designs BSA AuNCs Cu2+Fluorescent optical sensor, detects content of polyphosphate.This fluorescent optical sensor has selective feature pollution-free, simple, quick, high, and the range of linearity is 10 100nM, and detection is limited to 10nM.Result shows, PPy concentration is the biggest, and accordingly, the fluorescence of system recovers more, and the fluorescence recovery rate of the concentration of PPy and system presents good linear relationship.
Description
Technical field
The present invention relates to one with bovine serum albumin (BSA) as template, use chemical reduction method to synthesize a diameter of 2nm's
Bovine serum albumin-gold nano cluster (BSA-AuNCs), and obtained BSA-AuNCs-Cu by BSA-AuNCs synthesis2+Fluorescence sense
Device, and utilize this BSA-AuNCs-Cu2+The method that content of polyphosphate is detected by fluorescent optical sensor.
Background technology
Polyphosphate, as water-retaining agent and quality improver, is widely used in the storage and transport process of aquatic products
In, play and keep moisture, reduce drop and run off, improve the effect of mouthfeel, pH value, emulsifying can also be regulated simultaneously, buffer, chelate
Metal ion etc..But, during processing of aquatic products, use polyphosphate too much, not only compromise the power of consumer
Benefit, impacts the health of consumers in general especially.
The polyphosphate of excess intake, can promote blood clotting, and its catabolite phosphate is likely to increase absorption person
The probability that cardiovascular and cerebrovascular disease occurs；The excess residual of polyphosphate, also can affect the required unit such as calcium in human body, ferrum, copper, zinc
The absorption equilibrium of element, the constantly accumulation of internal polyphosphate can cause the unbalance of body calcium phosphorus, affect the absorption of calcium, easily lead
Cause osteoporosis.
Along with China's accession to the WTO, the demand for exports amount of aquatic products is increasing, a lot of domestic export enterprises
For improving yield, keeping aquatic products quality, increase economic efficiency and occur in that excess adds polyphosphate in aquatic products
Situation, specify maximum limitation beyond European Union and international food code (CODEX), exceeded standard by again and again circulating a notice of polyphosphate,
Thus formation trade barrier, have a strong impact on the economic interests of Enterprises for Export in China.CODEX (CODEX) allows polyphosphate
In freezing aquatic product (fillet, shrimp etc.) and finished product, Cmax is 1%, and EC regulations allows research on maximum utilized quantity to be 5g/
Kg, food office of the state regulations such as Brazil and Canada Cmax in different final products is 0.1% or 0.5%.Therefore, detection
Content of polyphosphate in aquatic products, it appears particularly significant.
At present the detection method for polyphosphate mainly have thin layer chromatography chromatography, high performance capillary electrophoresis, from
Sub-chromatography and nuclear magnetic resonance method etc., wherein thin layer chromatography chromatography is easy to operate, equipment simple, colour developing is easy, launches simultaneously
Speed is fast, but sensitivity is low, it is impossible to accurate quantitative analysis and loaded down with trivial details time-consumingly, it is impossible to meet modern measure demand；High performance capillary electrophoresis
Method and nuclear magnetic resonance method are all because of reasons such as operating process are loaded down with trivial details, testing staff requires higher and food substrate more complicated, no
Can be widely applied in the middle of food analysis；The chromatography of ions is the side of content of polyphosphate in common detection aquatic products
Method, has analysis efficiency height, speed soon, the advantage that detection sensitivity is high, but qualitative ability is poor.
In recent years, Nano-technology Development is rapid.Fluorogold nanocluster (gold nanocluster, Au NCs) is represented
The size gone out is little, good water solubility, hypotoxicity, surface are prone to advantages such as modifying, fluorescent stability is strong so that it is for bioanalysis and doctor
Learn diagnosis research and provide new indicia means, open new application.Utilize the Fluorescence Quenching Principle of gold nano cluster
The fluorescent probe of design can specifically detect the chemistry in environment and biological preparation etc..2009, Xie etc. utilized Ox blood serum egg
White BSA has the BSA-AuNCs of red fluorescence as templated synthesis, adds Cu2+The fluorescent quenching of BSA-AuNCs can be made,
Adding metal-chelator then can make fluorescence recover.Thus speculate, add metal-chelator sodium polyphosphate PPy and can make BSA-
AuNCs-Cu2+Fluorescence recover.The present invention, with BSA as template, uses the biological that the method for electronation prepares diameter 2nm
Sensor BSA-AuNCs, is determined by optimal detection, explores BSA-AuNCs-Cu2+Testing mechanism to PPy.
Summary of the invention
The present invention as template, uses the Ox blood serum egg that chemical reduction method synthesizes a diameter of 2nm with bovine serum albumin (BSA)
In vain-gold nano cluster (BSA-AuNCs), can send the glimmering of redness under the scanning that wave-length coverage is 200nm-800nm ultraviolet light
Light.Work as Cu2+BSA-AuNCs-Cu is generated with the glycine chelate in BSA chain2+Time, fluorescence can quencher；When adding polyphosphate radical
(PPy), time, PPy will be with Cu2+Chelating makes Cu2+From BSA-AuNCs-Cu2+Surface spins off, and forms BSA-AuNCs, and fluorescence is extensive
Multiple.Based on the principle that, inventor designs BSA-AuNCs-Cu2+Fluorescent optical sensor, examines content of polyphosphate
Survey.This fluorescent optical sensor has selective feature pollution-free, simple, quick, high, and the range of linearity is 10-100nM, detection
It is limited to 10nM.Test result indicate that, PPy concentration is the biggest, and accordingly, the fluorescence of system recovers more, and the concentration of PPy be
The fluorescence recovery rate of system presents good linear relationship.
On the one hand, the present invention relates to one with bovine serum albumin (BSA) as template, use chemical reduction method synthesis diameter
Bovine serum albumin-gold nano cluster (BSA-AuNCs) for 2nm.
On the other hand, the present invention relates to the preparation method of this bovine serum albumin-gold nano cluster (BSA-AuNCs), described
Method is included in HAuCl at 37 DEG C4Solution joins in BSA solution, after being stirred vigorously, adds NaOH solution regulation pH value extremely
12, by this mixed liquor water-bath 12h at 37 DEG C.More specifically, HAuCl4The concentration of solution is that the concentration of 1.0M, BSA solution is
50mg/mL, the concentration of NaOH solution is 1.0M.
On the other hand, the present invention relates to BSA-AuNCs-Cu2+Fluorescent optical sensor, it comprises BSA-AuNCs solution, pH is 6
NaAc-HAc buffer, Cu2+Solution.Specifically, described Cu2+The concentration of solution is 10 μMs.
On the other hand, the present invention relates to BSA-AuNCs-Cu2+Fluorescent optical sensor use in measuring content of polyphosphate
On the way.
The present invention with BSA as template, uses the method for electronation to be successfully prepared the fluorescin of diameter 2nm by research
Thing sensor BSA-AuNCs, in order to detect the content of PPy.Result shows, gold nano cluster forms BSA-under the effect of BSA
AuNCs, has the fluorescence of redness.Add Cu2+Time, Cu2+BSA-AuNCs-Cu is generated with the glycine chelate in BSA chain2+, cause
Make fluorescent quenching.Being subsequently added into metal-chelator PPy, PPy has higher and Cu2+The ability of chelating, with Cu2+Chelate and make
Cu2+Spin off from BSA-AuNCs surface, make system fluorescence recover, and along with the increasing of the polyphosphoric acids ion concentration added
Greatly, accordingly, the fluorescence of system recovers the most.Based on the principle that, the present invention devises a kind of BSA-AuNCs fluorescence
Biosensor, for detecting the content of PPy in aquatic products.The method range of linearity is 10nM-100nM, and detection is limited to 10nM.
Therefore, it is simple that detection method has technological process, easy to operate, and PPy is highly sensitive, efficient, quick in detection, has reality
By advantages such as property.
Accompanying drawing explanation
Fig. 1 (a) shows the ultraviolet spectrogram of BSA-AuNCs solution.
Fig. 1 (b) shows the Electronic Speculum figure (low multiple) of the BSA-AuNCs that mean size is 2nm.
Fig. 1 (c) shows Cu2+There is chemical reaction with BSA-AuNCs, form BSA-AuNCs-Cu2+Electronic Speculum figure (high
Multiple).
Fig. 2 shows that the design program of biological sensor BSA-AuNCs illustrates with testing mechanism.
Fig. 3 (a) shows the fluorescence spectrum figure of BSA-AuNCs.
Fig. 3 (b) shows the fluorescence spectrum figure of EDTA.
Fig. 4 shows variable concentrations Cu2+Fluorescence spectrum figure.
Fig. 5 shows the relation in response time and system fluorescent value.
Fig. 6 (a) and (b) show the content of BSA-AuNCs biological sensor detection PPy.
Detailed description of the invention
Below, the accompanying drawing in the embodiment of the present invention will be combined, the technical scheme in the embodiment of the present invention will be carried out detailed
Describe, it is clear that described embodiment is only a part of embodiment of the present invention rather than whole embodiments.Based on this
Embodiment in bright, the every other reality that those of ordinary skill in the art are obtained on the premise of not making creative work
Execute example, broadly fall into the scope of protection of the invention.
It should be noted that in this article, term " includes ", " comprising " or its any other variant are intended to non-row
Comprising of his property, so that include that the process of a series of key element, method, article or equipment not only include those key elements, and
And also include other key elements being not expressly set out, or also include intrinsic for this process, method, article or equipment
Key element.
The present invention as template, uses the Ox blood serum egg that chemical reduction method synthesizes a diameter of 2nm with bovine serum albumin (BSA)
In vain-gold nano cluster (BSA-AuNCs), can send the glimmering of redness under the scanning that wave-length coverage is 200nm-800nm ultraviolet light
Light.Work as Cu2+BSA-AuNCs-Cu is generated with the glycine chelate in BSA chain2+Time, fluorescence can quencher；When adding polyphosphate radical
(PPy), time, PPy will be with Cu2+Chelating makes Cu2+From BSA-AuNCs-Cu2+Surface spins off, and forms BSA-AuNCs, and fluorescence is extensive
Multiple.Based on the principle that, inventor designs BSA-AuNCs-Cu2+Fluorescent optical sensor, examines content of polyphosphate
Survey.This fluorescent optical sensor has selective feature pollution-free, simple, quick, high, and the range of linearity is 10-100nM, detection
It is limited to 10nM.Result shows, PPy concentration is the biggest, and accordingly, the fluorescence of system recovers more, and the concentration of PPy and system
Fluorescence recovery rate presents good linear relationship.
1. reagent and instrument
Table 1: experiment reagent
Table 2: experimental apparatus
Note: all of chemical substance purity more than 99%, and should purify the most further.Used by each step
Water should be ultra-pure water.
The preparation of 2.BSA-AuNCs
At 37 DEG C, by the HAuCl of 15mL 10.0mM4Solution adds in 15mL, 50mg/mL BSA solution, acutely stirs
Mix.After 2min, the NaOH regulation pH value of addition 1.3mL, 1.0M is to 12, and this mixed liquor is water-bath 12h at 37 DEG C.The color of solution
Light brown will be become from light yellow, then become dark-brown.Finally, BSA-AuNCs solution is made, and is diluted to ultra-pure water
200mL, and preserve with standby at 4 DEG C.
3. the detection method of polyphosphoric acids salt ion
It is the NaAc-HAc buffer of 6,10 μMs of Cu by 1.00mL BSA-AuNCs solution, 1.00mL pH2+Solution and
The polyphosphate (PPy) of 1.00mL variable concentrations adds in the color comparison tube of 10mL, is diluted to 10mL with ultra-pure water and quickly mixes
Even, stand 20min, use fluorescence indexing to measure its fluorescent value.Do reagent blank test simultaneously.
△ F=F1-F0……………………………………………(1)
In formula:
The relative intensity of fluorescence of △ F system；
The fluorescence intensity of F1 solution；
The fluorescence intensity of F0 blank solution
Fluorescent quenching rate=△ F/F0……………………………………(2)
In formula:
The relative intensity of fluorescence of △ F system；
The fluorescence intensity of F0 blank solution
4. result and discussion
The ultraviolet spectral analysis of 4.1BSA-AuNCs
Taking the above-mentioned BSA-AuNCs solution prepared, use ultraviolet spectrophotometer to be scanned, scanning wavelength scope is
200nm-800nm, show that shown in ultraviolet spectrogram such as Fig. 1 (a), wherein, abscissa represents that scanning wavelength, vertical coordinate represent corresponding
Absorbance.It is seen that in the wave-length coverage of 200nm-800nm, BSA-AuNCs does not has an absworption peak, but one
The smooth curve downwardly extended.Being analyzed this, occur that the reason of this situation is, synthesized gold nano cluster size is very
Little, the characteristic absorption peak of large scale gold nano grain will not be produced, thus be not inhale in the wave-length coverage of 200nm-800nm
Receive peak.
It is the BSA-AuNCs of 2nm that Fig. 1 (b) presents mean size under Electronic Speculum, and they are spherical in shape, has preferably dispersion
Property.Work as Cu2+When joining BSA-AuNCs system, Cu2+There is chemical reaction with BSA-AuNCs, form BSA-AuNCs-Cu2+
Shown in (such as Fig. 1 (c)), and volume the most correspondingly becomes greatly, fluorescence generation quencher.As past BSA-AuNCs-Cu2+During middle addition PPy,
The size of BSA-AuNCs will return to about 2nm, this is because PPy and Cu2+Chelate, Cu2+From BSA-
AuNCs-Cu2+In remove and make the volume of BSA-AuNCs be reduced to original size, fluorescent characteristic is recovered simultaneously.Above-mentioned detection
Mechanism illustrates well, from BSA-AuNCs, BSA-AuNCs-Cu2+To BSA-AuNCs-Cu2+-PPy, their granular size
Changing, correspondingly, its fluorescent characteristic also changes.
The mechanism of 4.2BSA AuNCs biological sensor analyte detection polyphosphoric acids radical ion
Utilize protein-bovine serum albumin (bovine serumalbumin, BSA) as template fabricated in situ BSA-
Au NCs, this cluster is made up of 25 gold atoms, has red fluorescence (λ em=640nm), and fluorescence quantum yield (QY) is
6%.Free state Cu2+Can be combined with the sulfydryl in protein, the activity of interference sulfydryl.Cu2+Can be with the paddy on gold nano cluster surface
Guang sweet peptide (glutathine, GSH) protective layer generation coordination (GSH:Cu2+=2:1), this effect cause Au NCs reunite from
And cause fluorescence to reduce.After it should be noted that addition metal ion chelation agent[13], by Cu2+The fluorescence of suppression can get again extensive
Multiple.
The design program of above-mentioned fluorescence induction apparatus is with the diagram of testing mechanism as shown in Figure 2.BSA-AuNCs and Cu2+In conjunction with
Become BSA-AuNCs-Cu2+, cause BSA-AuNCs fluorescence to reduce；In said system add metal-chelator PPy, PPy with
Cu2+Chelating, makes Cu2+Discharging from BSA-AuNCs surface, BSA-AuNCs fluorescent value is recovered.
As shown in Fig. 3 (a), a curve represents that by 1.00mL BSA-AuNCs solution, 1.00mL pH be the NaAc-HAc of 6
Buffer, 10 μMs of Cu2+Solution mixes and is diluted to 10mL, stands 20min, uses the fluorescence spectrum figure that fluorescence protractor records,
Wherein excitation wavelength is 400nm, and sweep limits is 560-660nm, and maximum absorption band is at 618nm；Another curve represent by
1.00mL BSA-AuNCs solution, 1.00mL pH are the NaAc-HAc buffer of 6,10 μMs of Cu2+And the mixing of 100nM PPy solution
And it is diluted to 10mL, and stand 20min, use the fluorescence spectrum figure that fluorescence protractor records, wherein excitation wavelength is 400nm, sweeps
The scope of retouching is 560-660nm, and maximum absorption band is at 618nm.By Fig. 3 (a) it is apparent that at BSA-AuNCs-Cu2+In solution
After adding PPy, fluorescent spectrum curve up moves, i.e. fluorescent value increases.This is because PPy has higher and Cu2+Chelating
Ability, when system add PPy time, Cu2+To spin off from BSA-AuNCs surface and make fluorescence recover.According to BSA-
AuNCs-Cu2+These characteristics, the fluorescence induction apparatus of a novel detection PPy just designs.
Based on above-mentioned BSA-AuNCs-Cu2+The Cleaning Principle of biological sensor thing, it has been found that: the metal chelating of interpolation
Mixture concentration is the biggest, accordingly, and BSA-AuNCs-Cu2+The fluorescence of solution system recovers the biggest, and presents certain linear pass
System.According to above-mentioned discovery, now carry out checking test: the metal-chelator EDTA of preparation variable concentrations, concentration is respectively 1 μM, 2 μ
M, 3 μMs, 4 μMs, 5 μMs, 6 μMs, 7 μMs, 8 μMs, 9 μMs, 10 μMs, be separately added into 1.00mL BSA-AuNCs solution, 1.00mL10 μM
Cu2+Solution and the NaAc-HAc buffer that 1.00mL pH is 6, miscible merging is diluted to 10mL, stands 20min, uses fluorescence to divide
Its fluorescent value is not surveyed in degree score, and the data obtained processes, and draws shown in curve chart such as Fig. 3 (b).It can be seen that with
The increase of EDTA concentration, the value of (F1-F0)/F0 increases the most therewith.
The optimal external condition of 4.3 detection polyphosphoric acids radical ions
In order to avoid being disturbed by other extraneous factors during detection, make the testing result of polyphosphoric acids ion concentration
It is the most accurate, it is therefore necessary to ensure the detection of polyphosphoric acids ion concentration is carried out under optimal external condition.For making reality
Testing and the most successfully carry out, during the optimal external condition exploring detection polyphosphoric acids ion concentration, unified setting is anti-
Answering temperature is room temperature (25 DEG C), uses unitary variant method, mainly have studied amount of reagent and response time in susceptiveness, selection
The impact of the aspects such as property, accuracy and repeatability.
And Cu 4.3.1BSA-AuNCs2+The impact of concentration
For research Cu2+The concentration impact on system fluorescent value, now uses control variate method, by changing Cu2+Concentration,
The fluorescent value situation of change of viewing system, to seek Cu optimal in detection system fluorescent value2+Concentration.
Take 1 μM, 2 μMs, 3 μMs, 4 μMs, 5 μMs, 6 μMs, 7 μMs, 8 μMs, 9 μMs, the Cu of 10 μMs2+Solution, is separately added into 1.00mL
BSA-AuNCs solution and the NaAc-HAc buffer that 1.00mL pH is 6, solution mixes and is diluted to 10mL, stands 20min, makes
Surveying its fluorescent value respectively with fluorescence protractor, wherein excitation wavelength is 400nm, and scanning wavelength is 500nm-700nm, absorption maximum
Peak is at 618nm.The data obtained processes, and show that curve chart is as shown in Figure 4.As shown in Figure 4, along with Cu2+The increase of concentration,
Correspondingly, the value of vertical coordinate also increases, and presents good linear relationship.I.e. along with Cu2+The increase of concentration, correspondingly, system
Relative fluorescence decline, work as Cu2+When concentration is 10 μMs, the relative fluorescence of system reaches minimum.Therefore, we are selected examines
The Cu that examining system fluorescent value is optimal2+Concentration is 10 μMs.
4.3.2pH with the impact in response time
Existing related researcher's research shows, when the BR buffer solution that 1.0mL pH is 6, the dissolved liquid of Phosphate,
C7H15NO4S、C8H18N2O4S, NaAc-HAc and Tris-Hcl respectively as buffer solution, the △ F value of system is respectively 7.9,
16.4,6.3,27.3,36.5 and 31.3, accordingly, RSD% (relative standard deviation) is 4.3,3.7,4.1,2.9,2.3 and 3.0.
Result shows, when the NaAc-HAc that pH is 6 is as buffer, the △ F of system reaches maximum.The 1mL NaAc-of different pH
The impact of system △ F value has been made assessment by HAc buffer solution the most.When pH is 5, BSA-AuNCs is precipitated out from solution,
PH now is close to the isoelectric point, IP of BSA.Being in the range of 6-10 at Ph, △ F reduces along with the increase of pH.This is because Cu2+
Can hydrolyze in a large number under the conditions of high ph-values (pH ＞ 6.0), reduce the fluorescence recovery rate of system.The △ F value of system reaches when pH is 6
To maximum.Therefore, the pH of selecting system reaction is 6.
It is the NaAc-HAc buffer of 6,10 μMs of Cu by 1.00mL BSA-AuNCs solution, 1.00mL pH2+Solution mixes,
And it is diluted to 10mL, and stand the different time respectively, use fluorescence protractor detection fluorescent value, wherein excitation wavelength is 400nm,
Sweep limits is 550nm-750nm, and maximum absorption band is at 618nm.Data are carried out Treatment Analysis, obtain " time m-fluorescence
Value " graph of a relation is as shown in Figure 5.As can be seen from Figure 5, during solution left standstill 0-6min, the fluorescent value of solution drastically declines；During standing
Between more than 10min time, the fluorescent value of solution tends to be steady.Therefore, we can unify response time of the system that takes is 20min.
BSA-AuNCs biological sensor analyte detection polyphosphoric acids ion concentration, must enter under optimal external condition
OK, report according to result of study above and relevant document, we can determine whether that optimal detection condition is: reaction temperature is room
Temperature (25 DEG C), reaction system pH is 6, and the response time is 20min, Cu2+Concentration is 10 μMs.
The detection to PPy of the 4.4BSA-AuNCs biological sensor
BSA-AuNCs-Cu2+After middle addition metal-chelator, the fluorescence being quenched can be recovered, and PPy is metal-chelating
Agent, principle can make BSA-AuNCs-Cu2+The fluorescence of quencher is restored.
Take the PPy solution of variable concentrations, be separately added into 1.00mL BSA-AuNCs solution, 1.00mL pH is the NaAc-of 6
HAc buffer and the Cu of 1.00mL10 μM2+Solution, solution mixes and is diluted to 10mL, stands 20min, uses fluorescence protractor
Surveying its fluorescent value respectively, wherein excitation wavelength is 400nm, and sweep limits is 550nm-750nm, and maximum absorption band is at 618nm.
The data obtained processes, and draws shown in curve chart such as Fig. 6 (a).Knowable to Fig. 6 (a), PPy concentration and fluorescent value present well
Linear relationship, along with the increase of polyphosphoric acids ion concentration, its fluorescent value the most correspondingly increases.I.e. add PPy, make BSA-
AuNCs-Cu2+The fluorescence of quencher is increased, and the increase of the concentration along with the PPy added, BSA-AuNCs-Cu2+It is quenched
Fluorescence increase more.Therefore, it is possible to use the content of PPy is detected by BSA-AuNCs biological sensor thing.
This experiment the data obtained is arranged again, draws shown in curve chart such as Fig. 6 (b).As seen from the figure, along with
The increase of PPy concentration, correspondingly, the value of vertical coordinate also increases, and presents good linear relationship, fully demonstrates BSA-
The principle of AuNCs biological sensor analyte detection PPy: Cu2+BSA-AuNCs, this work is generated with the glycine chelate in BSA chain
Fluorescence is caused to reduce with causing AuNCs to reunite；After adding metal ion chelation agent PPy, PPy and Cu2+Chelating makes Cu2+From
BSA-AuNCs surface spins off, and forms BSA-AuNCs, and fluorescence recovers.
In sum, the present invention, with BSA as template, uses the method for electronation to be successfully prepared the fluorescence of diameter 2nm
Biosensor BSA-AuNCs, in order to detect the content of PPy.Test result indicate that, gold nano cluster is shape under the effect of BSA
Become BSA-AuNCs, there is the fluorescence of redness.Add Cu2+Time, Cu2+BSA-AuNCs-is generated with the glycine chelate in BSA chain
Cu2+, cause fluorescent quenching.Being subsequently added into metal-chelator PPy, PPy has higher and Cu2+The ability of chelating, with Cu2+Chela
Close and make Cu2+Spin off from BSA-AuNCs surface, make system fluorescence recover, and along with the polyphosphoric acids radical ion added is dense
The increase of degree, accordingly, the fluorescence of system recovers the most.Based on the principle that, devise BSA-AuNCs biological
Sensor, for detecting the content of PPy in aquatic products.The method range of linearity is 10nM-100nM, and detection is limited to 10nM, detection
PPy is sensitive, efficient, quick, has practicality.
Claims (9)
1. a new method for polyphosphate based on gold nano cluster fluorescent characteristic detection, with bovine serum albumin (BSA) be
Template, uses bovine serum albumin-gold nano cluster (BSA-AuNCs) solution of a diameter of 2nm of chemical reduction method synthesis.
2. one kind with bovine serum albumin (BSA) as template, use the Ox blood serum egg of a diameter of 2nm of chemical reduction method synthesis
In vain-gold nano cluster (BSA-AuNCs) solution.
The preparation method of bovine serum albumin the most according to claim 2-gold nano cluster solution, described method is included at 37 DEG C
By HAuCl4Solution joins in BSA solution, and after being stirred vigorously, this mixed liquor, to 12, is existed by addition NaOH solution regulation pH value
Water-bath 12h at 37 DEG C.
The preparation method of bovine serum albumin-gold nano cluster the most according to claim 3, wherein HAuCl4The concentration of solution is
The concentration of 1.0M, BSA solution is 50mg/mL, and the concentration of NaOH solution is 1.0M.
5.BSA-AuNCs-Cu2+Fluorescent optical sensor, it comprises the BSA-AuNCs solution of claim 2, pH is the NaAc-HAc of 6
Buffer, Cu2+Solution.
BSA-AuNCs-Cu the most according to claim 52+Fluorescent optical sensor, described Cu2+The concentration of solution is 10 μMs.
7. according to the BSA-AuNCs-Cu of claim 5 or 62+Fluorescent optical sensor purposes in measuring content of polyphosphate.
Purposes the most according to claim 7, wherein, polyphosphate is present in aquatic products.
9., according to the purposes of claim 7 or 8, wherein the detection range of content of polyphosphate is 10nM-100nM.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109400633A (en) * | 2017-08-17 | 2019-03-01 | 清华大学 | Gold nano cluster and its high yield preparation method |
| CN109705841A (en) * | 2018-12-24 | 2019-05-03 | 山西大学 | A kind of transferrins is the gold nano cluster and its preparation method and application of template |
| CN110161002A (en) * | 2019-04-28 | 2019-08-23 | 浙江工业大学 | A kind of ratio fluorescence imaging system and its construction method, application |
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| CN109400633B (en) * | 2017-08-17 | 2020-05-22 | 清华大学 | Gold nanocluster and high-yield preparation method thereof |
| CN109705841A (en) * | 2018-12-24 | 2019-05-03 | 山西大学 | A kind of transferrins is the gold nano cluster and its preparation method and application of template |
| CN109705841B (en) * | 2018-12-24 | 2021-07-02 | 山西大学 | Gold nanocluster with transferrin as template and preparation method and application thereof |
| CN110161002A (en) * | 2019-04-28 | 2019-08-23 | 浙江工业大学 | A kind of ratio fluorescence imaging system and its construction method, application |
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