CN106959290A - A kind of Ratio-type rare-earth fluorescent probe and the application for detecting bacillus anthracis biomarker - Google Patents
A kind of Ratio-type rare-earth fluorescent probe and the application for detecting bacillus anthracis biomarker Download PDFInfo
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Abstract
The invention discloses a kind of Ratio-type rare-earth fluorescent probe Tb/DPA@SiO2Eu/GMP and visualization method for preparing test paper and its detection the pyridinedicarboxylic acid of bacillus anthracis biomarker -2,6（DPA）The application of aspect.Synthesized by reverse microemulsion method after the fluorescence probe, the quantitative detection to DPA concentration is realized using Time-resolved fluorescence assay method.Further, by the way that filter paper is immersed in fluorescence probe solution, a kind of portable visualization test paper is prepared for, the test paper can be applied to a kind of cell phone software for recognizing color to detect DPA concentration.The present invention changes traditional detection pattern conventional at present, cost-effective without complicated operating process.The characteristics of present invention has accurate, sensitive, high selectivity, the test paper of preparation has can be portable, and convenient use is cheap to wait many merits, is with a wide range of applications.
Description
Technical field
The present invention relates to the Ratio-type fluorescence of technical field of analytical chemistry, more particularly to a kind of pair of rare earth ion doping
Probe and the probe are detecting the purposes of bacillus anthracis biomarker concentration.
Background technology
Bacillus anthracis is the aerobic bacteria of Gram-positive formation gemma, and anthrax is a kind of main disease of herbivore
Disease, is contacted in soil, the gemma on fur can cause infection.Gemma can be contacted by respiratory tract, alimentary canal and skin to be felt
The mankind are contaminated, it is most common with localized anthrax.Because the resistance of anthrax spore in the environment is extremely strong, in military aspect, always by
It is No.1 biological warfare agent to be classified as, and is used as the main constituents of a Bacillus anthracis shell, 2,6 pyridinedicarboxylic acids（DPA）
About 5-15% dry anthrax bacillus gemma quality is occupied, therefore can be by quantitative detection DPA content come indirect detection anthrax
Bacillus.
Between the past few decades, domestic and international scientist has taken up quantitative detection of the various methods implementations to DPA,
Including surface-enhanced Raman method, electrochemical detection method etc..However, these methods require that longer time is spent mostly, operation
Complicated instrument and expensive reagent.Therefore, a kind of simple cheap is developed, sensitive portable detection method is for preventing anthrax bar
It is significant for the harm of bacterium biomarker.
Before instrument analytical method appearance, just there is the analysis method that analyte detection to be measured is carried out using colorimetric analysis principle.
Chemical colorimetry determines various concentrations using the color distortion of solution, with easy to be quick, the advantage of mass detection.But
It is that this method, which has background light, influences larger, and color judges subjective, it is impossible to realize that quantitative analysis automation etc. lacks
Point.And the development of image procossing and colour recognition technology make it that color shades quantify.By color of image identification technology
A kind of new qualitative and quantitative analysis method will be brought by being combined with chemical colorimetric detection.At present, the color mode of image photograph is main
With RGB（RGB）It is by red based on color standard pattern（R）, it is green（G）, it is blue（B）The change of 3 Color Channels and it
Between be overlapped mutually to obtain shades of colour, it is all that this 3 Standard Colors almost include that human eyesight can perceive
Color, is that any one color can be recorded and expressed by one group of rgb value with one of most wide color system.
The content of the invention
A kind of Ratio-type for rare earth ion doping that the purpose of the present invention is in view of the shortcomings of the prior art and provided
Fluorescence probe and the probe are used for early-stage preparations complicated in the application for detecting DPA, the method to overcome existing detection DPA and held high
Expensive large-scale instrument is used, it is difficult to realize the defect of accurate, simple, portable quick detection.
Realizing the object of the invention concrete technical scheme is：
A kind of Ratio-type rare-earth fluorescent probe, feature is：By rare earth terbium ion solution (Tb3+) and pyridinedicarboxylic acid（DPA）Pass through
The synthesis of reverse microemulsion method obtains structural formula for Tb/DPA@SiO2Presoma；It is 0.5 ~ 1 through centrifuge washing compound concentration scope
After the mg/ml aqueous solution add equimolar than rare-earth europium ion solution E u3+With guanosine ribonucleoside acid solution (GMP), through shake
Swing after reaction, centrifuge washing obtains structural formula for Tb/DPA@SiO2- Eu/GMP fluorescence probe；Utilize Tb3+Fluorescence intensity
Value and Eu3+Both fluorescence intensity levels ratio component ratio type rare-earth fluorescent probe Tb/DPA@SiO2-Eu/GMP。
The reverse microemulsion composition is that volume ratio is：It is molten that Zheng Ji Chun ︰ Qulas lead to X-100 ︰ Huan Ji Wan ︰ tetraethyl orthosilicates
The ︰ 17 of 200 ︰ of liquid (TEOS)=200 ︰ 800.
The rare-earth europium ion solution E u3+End reaction concentration with guanosine ribonucleoside acid solution (GMP) is 0.5 ~ 2
mM。
A kind of sketch-based user interface type rare-earth fluorescent probe is used for bacillus anthracis biomarker DPA detection method, and feature is：
Tb/DPA@SiO are added in the solution of the buffer solution containing Tris-HCl2- Eu/GMP fluorescence probes, are added to be measured after solution-stabilized
DPA solution is reacted, and occurs Enhancement of Fluorescence, and passage time resolved fluorometric analysis method determines DPA concentration；Its
In：The fluorescence probe is dissolved in the Tb/DPA@SiO that water formation concentration is 0.1 ~ 0.2 mg/ml2- Eu/GMP the aqueous solution；It is described slow
The solution concentration of fliud flushing is 20 ~ 50 mM, and pH value is 4~9；The reaction temperature is 5~35 DEG C, the DPA solution to be measured
Concentration is 0.025~2 μM.
The fluorescence probe lowest detection line is 7.3 nM DPA solution, and high sensitivity, the DPA of high selectivity can be achieved
Concentration Testing.
A kind of test paper of sketch-based user interface type rare-earth fluorescent probe, feature is that ordinary filter paper is cut into a diameter of 5 ~ 8 mm's
Circular paper, is immersed in concentration after 10 ~ 30 minutes, to be dried in its natural state in 1 ~ 5 mg/ml fluorescence probe solution, makes
Must have the test paper of fluorescence probe.
A kind of application of above-mentioned test paper in Visual retrieval DPA concentration, feature is：The test paper is adding various concentrations
DPA after have different color responses, as DPA concentration gradually increases, test paper color gradually changes, according to the intensity of color
Value quantifies DPA concentration, can to DPA concentration real-time online, Intelligent Measurement.
The test paper is in green under ultra violet lamp, and it is still green to add test paper after low concentration DPA, and adds high concentration
Test paper is changed into red after DPA, passes through colour recognition, red R ed, the green Green of scanning analysis test paper color, blue Blue numbers
Value, adds the red R of test paper color after low concentration DPA：Green G intensity rates (R/G) are small, add test paper color after high concentration DPA
Red R/G values are big, and DPA concentration is determined according to R/G value sizes.
Compared with prior art, beneficial effect of the present invention includes as follows：
1. the present invention changes detection DPA conventional at present detection pattern, without complicated early stage preparation of samples, it is not necessary to
Using expensive large-scale instrument, cost has been saved；
2. the present invention is the Ratiometric fluorescent probe that report is adulterated using a kind of pair of rare earth ion current for the first time and visual
Change test paper approach, propose to recognize cell phone software using color of image first to realize that the quantitative analysis to DPA is detected；
3. detection method consumption is few, whole reaction system only needs trace level, has saved the usage amount of probe,
Realize micro inexpensive detection.Raw material obtain simple cheap in the fluorescence probe, are applied to the inspection of DPA in blood
Survey practical；
4. detection method can effectively reduce the influence of environmental factor, using rare earth terbium ion green fluorescence as
Reference signal, the red fluorescence of rare-earth europium ion is as response signal, and the fluorescence probe has longer fluorescence lifetime（Reach
To ms grades）, so as to using the method for Time-resolved fluorescence assay, can effectively eliminate background fluorescence in detection process and
The influence of fluorescence is scattered, so as to significantly improve sensitivity and signal to noise ratio；
5. present invention detection fast response time, is directly sensitized the fluorescence probe using test substance DPA, whole detection DPA processes
No more than 1 minute.The inventive method determines DPA 0.025~2 μM of concentration range, and minimal detectable concentration is 7.3 nM, is had
Higher sensitivity and selectivity.
Brief description of the drawings
Fig. 1 is that fluorescence probe of the present invention is prepared and detection principle diagram；
Fig. 2 is the phenogram of fluorescence probe prepared by the present invention；
Fig. 3 is the experimental result picture that the present invention is applied to detection DPA；
Fig. 4 is that the test paper prepared by the present invention is used for the experimental result picture of Visual retrieval DPA concentration.
Embodiment
With reference to specific examples below and accompanying drawing, the present invention is described in further detail, protection content of the invention
It is not limited to following examples.Under the spirit and scope without departing substantially from inventive concept, those skilled in the art it is conceivable that change
Change and advantage is all included in the present invention, and using appending claims as protection domain.The process of the implementation present invention,
Condition, reagent, experimental method etc., in addition to the following content specially referred to, are the universal knowledege and common knowledge of this area,
Content is not particularly limited in the present invention.
As shown in figure 1, the Ratio-type rare-earth fluorescent probe of the present invention is the Ratio-type of a kind of pair of rare earth ion doping
Fluorescence probe：By rare earth terbium ion solution (Tb3+) and 2,6 pyridinedicarboxylic acids（DPA）Tied by the synthesis of reverse microemulsion method
Structure formula is Tb/DPA@SiO2Presoma；After being formulated as solution through centrifuge washing add equimolar than rare-earth europium ion solution
Eu3+With guanosine ribonucleoside acid solution (GMP), after concussion reaction, centrifuge washing obtains structural formula for Tb/DPA@SiO2-
Eu/GMP fluorescence probe, utilizes Tb3+Fluorescence intensity level and Eu3+Both fluorescence intensity levels ratio component ratio type
Rare-earth fluorescent probe Tb/DPA@SiO2- Eu/GMP, the fluorescence probe can be used for detection DPA.In the probe, Tb/DPA@SiO2
It is wrapped in the network structure for entering and being formed by Eu/GMP polymer, forms double rear-earth-doped polymer Tb/DPA@SiO2-
Eu/GMP.After the probe is acted on DPA, Tb/DPA@SiO2Due to the green that the protective effect transmitting of silica shell is stable
Fluorescence is as reference signal, and DPA is sensitized Eu/GMP as antenna ligand and produces response of the red fluorescent as determinand
Signal, so as to realize the Ratio-type fluoroscopic examination to DPA.
The present invention comprises the following steps：
(1) method for preparing fluorescence probe：Tb/DPA SiO are prepared by reverse microemulsion method2- Eu/GMP Ratio-types fluorescence is visited
The green fluorescence of pin, wherein Tb makees response fluorescence as reference fluorescent, Eu red fluorescence；
(2) passage time resolved fluorometric detection method realizes the detection to DPA：Take the Tb/DPA@SiO that 10 μ L are obtained2-
Eu/GMP Ratiometric fluorescent probes simultaneously add DPA standard liquids to be measured or blood serum sample is reacted.The DPA marks to be measured
The concentration of quasi- solution is 0.025~2 μM.In a particular embodiment, blood serum sample is rat blood serum sample；
(3) visualization test paper is prepared：By the way that filter paper is immersed in fluorescence probe solution, test paper of the system with fluorescence probe enters
One step, which develops a kind of portable test paper, is used for Visual retrieval DPA；
(4) visualization test paper detection DPA：By the cell phone software of colour recognition, test paper after addition various concentrations DPA is scanned
The rgb value of color, realizes DPA quantitative detection.
The reaction volume of the present invention can be controlled in a microlitre rank, and volume is 100 μ L.Optimal Experimental scheme can be expressed as follows：
10 μ L solution to be measured are taken to be added in 90 μ L fluorescence probe solution, passage time resolved fluorometric analytic approach is detected.
In the fluorescence probe, the concentration of the fluorescence probe is 0.01 ~ 1 mg/ml, it is preferable that be 0.1 mg/ml.
The reaction density of the DPA solution to be measured is 0.025~2 μM.
The buffer solution is Tris-HCl, and its concentration is 40 mM, and pH value is 7.0~9.0；Preferably, pH=7.4.
The reaction temperature is 4 ~ 40 DEG C, it is preferable that under room temperature condition.
The reaction time is 0 ~ 2 minute, it is preferable that the reaction time is 1 minute.
The inventive method is Sparklet testing method.In a particular embodiment, minimal detectable concentration of the invention is 7.3
NM DPA solution.
Fluorescence spectrum is detected：Multi-function microplate reader（Infinite M200 pro, TECAN, Switzerland）Or the same sex
Can spectrometer, excitation wavelength：272 nm, time delay：50 μs；The time of integration：2000 μs；Scanning wavelength scope：450~
750 nm, using 384 micropore blackboards, take 100 μ L reaction solutions to be measured.
Embodiment 1
Fluorescence probe is prepared first；Absolute ethyl alcohols of the 500 μ L containing 9.0 mg EDC and 2.8 mg NHS is added 600 μ L's
20 mM DPA solution, and stir 40 minutes.Then, 100 μ L APTES are added and are reacted 100 minutes.Then, 200 μ L are added
20 mM Tb(NO3)3Solution is into mixed solution.It regard resulting mixture as precursor.Then, reverse micro emulsion legal system is used
Standby Tb/DPA@SiO2.Using 1mL n-hexyl alcohols, 1mL triton x-100s and 4 mL hexamethylenes are made microemulsion and are added to 300 μ
Before L in prepared Tb/DPA solution, after lasting stirring in 40 minutes, 25 μ L ammonia spirits (28%) and 85 are added in the solution
μ L TEOS's.Stirring reaction continues 24 hours.Equal-volume acetone is used for the separating nano-particles from microemulsion, and uses second
Alcohol and water rinses centrifugation 3 times.In order to prepare amino modified Tb/DPA@SiO2-NH2, by Tb/DPA@SiO2It is suspended in above-mentioned
Microemulsion.Then add in 20 μ L APTES solution and stirring reaction 2 hours.Finally, collected using method same as described above
Nano particle.By resulting Tb/DPA@SiO2-NH2Dried in 40 °C of drying boxes.
Embodiment 2
By the mM GMP solution of 500 μ L 2 and the mg/mL of 1.0 mL 0.5 Tb/DPA@SiO2-NH2Solution is mixed, and is then shaken
30 minutes.Then, by the mM Eu (NO of 500 μ L 23)3It is added in mixed solution and shakes 100 minutes at room temperature.Then,
5 minutes are centrifuged under 10000 rpm to collect the product, and with milli-Q water 3 times, to remove unreacted reagent.Finally,
By Tb/DPA@SiO2- Eu/GMP mixtures are stored in 2 mL H2It is for further use in O.(prepared material characterization is as schemed
Shown in 2, Fig. 2 (A) is Tb/DPA@SiO2-NH2Transmission scanning electron microscope phenogram, Fig. 2 (B) is Tb/DPA@SiO2-NH2It is saturating
ESEM phenogram is penetrated, Fig. 2 (C) is Tb/DPA@SiO2-NH2, Tb/DPA@SiO2-Eu/GMP, Tb/DPA@SiO2-Eu/
GMP/DPA infrared spectrum characterization figure, Fig. 2 (D) is Tb/DPA@SiO2- Eu/GMP/DPA fluorescence excitation EX and emission spectrum
Scheme EM).
Embodiment 3
DPA is detected with the fluorescence probe of preparation and foregoing testing conditions.By 80 μ L Tris-HCl buffer solutions（pH
7.4,50 mM）It is added to the Tb/DPA@SiO prepared by 10 μ L2In-Eu/GMP solution.Afterwards, by 10 μ L various concentrations
DPA（0.25-20 µM）It is added in above-mentioned solution.Tris-HCl ultimate density is 40 mM, and DPA ultimate density is
In 0.025-2.0 μM of scope.In order to probe into the selectivity for the Ratiometric fluorescent probe for detecting DPA, different materials are selected to carry out
Interference is tested, including phenoxyacetic acid（POA）, terephthalic acid (TPA)（p - PA）, benzoic acid（BA）, cysteine（Cys）, paddy
Amino acid（Glu）, the ultimate density of these materials is 50 μM.Finally, lower record emission spectrum is excited in 272 nm wavelength.It is dense
When spending 0.025-2.0 μM, fluorescence intensity is linearly related to DPA concentration：Y=1.225X+0.033, R2=0.9911, based on sky
White sample three times standard deviation, obtained DPA minimal detectable concentration is 7.3 nM（As shown in figure 3, Fig. 3 (A) is to be separately added into
Fluorogram after the DPA of 0.025 ~ 2 μM of concentration, Fig. 3 (B) is adds after the DPA of 0.025 ~ 2 μM of concentration, in 618 nm
At wavelength and 548nm wavelength fluorescence intensity F618/F548The linear graph that ratio changes with the change of DPA concentration, Fig. 3 (C)
It is the selective lab diagram of fluorescence probe, Fig. 3 (D) is the competitive experimental result picture of sample mixing）.
Embodiment 4
In order to further probe into whether the probe can be used for biological actual sample detection, detected using the probe in blood serum sample
DPA contents.A series of blood serum samples containing various concentrations DPA are added in detection architecture by Standard entertion method.
Table 1 is the experimental result that the present invention is used in blood detect DPA, as shown in table 1, the rate of recovery of blood serum sample in 95.70 % extremely
Between 104.65 %, relative standard deviation（RSD, n=3）Below 5%, it was demonstrated that Tb/DPA@SiO2- Eu/GMP time resolutions ratio
Rate type fluorescence probe is applicable to the analysis detection of DPA in complicated actual sample.
Table 1
Embodiment 5
Develop a kind of easy visualization test paper：By the way that filter paper is immersed in fluorescence probe solution, it is made and has fluorescence probe
Test paper, for quick detection DPA contents.As shown in Fig. 4 (A), test paper is in green when test paper is added without DPA under ultra violet lamp
Color, gradually increase adds DPA concentration, and test paper is changed into red from green, and is taken on a red color after addition high concentration DPA test paper, such as Fig. 4
(B) shown in, by colour recognition cell phone software, red R ed, the green Green of scanning analysis color, blue Blue numerical value, plus
Enter the red R of test paper color after low concentration DPA：Green G intensity rates (R/G) are small, add the red R/G of test paper color after high concentration DPA
Value is big, determines DPA concentration, the minimum detectable concentration of test paper indirectly according to the linear relationship of R/G ratios and DPA concentration
（LDC）About 1 μM.Therefore, the Tb/DPA@SiO determined for DPA2Method associated with-Eu/GMP test paper and cell phone software has
There is a rapid and handy, cheap advantage and there is huge application potential in actual applications.
To sum up result, based on Tb/DPA@SiO2The time resolution ratio of double rare earth ions doping of-Eu/GMP polymer
Rate type fluorescence probe is synthesized first, wherein Tb/DPA@SiO2As stable interior reference, Eu/GMP CP are used as the spirit of determinand
Quick signal response.Time resolution Ratiometric fluorescent probe is shown detects higher sensitivity and selectivity to DPA；Detection is limited to
7.3 nM, the range of linearity is 0.025~2 μM, linearly dependent coefficient 0.9911.With in standard addition method detection rat blood serum
Other concurrents in DPA, blood are not detected to it to be impacted.The practicality and biocompatibility of the present invention is strong, can
It is prevented effectively from interference of the concurrent to detection in actual sample.In addition, using being fixed with Tb/DPA@SiO2- Eu/GMP filter paper
The simple test paper determined for DPA is successfully devised, and can be under uv lamps by direct visual perception from green to red
The color conversion of color.The novel simple test paper meets quick and convenient detection DPA demand, recognizes that mobile phone is soft by color of image
Part can determine DPA concentration indirectly according to R/G numerical value, it is to avoid complicated operation and the use of expensive instrument, only use hand
Machine software just can realize quick detection.Therefore, the time resolution ratio fluorescent probe be it is credible, accurately, sensitively, and will be in reality
More flexible easy sensing strategy is provided in the application of border, application of the image recognition technology in chemical analysis context of detection is promoted.
It is only presently preferred embodiments of the present invention in summary, not for limiting the scope of the invention.It is all according to the present invention
Equivalence changes and modification that content is made, all should be protection category of the present invention.
Claims (7)
1. a kind of Ratio-type rare-earth fluorescent probe, it is characterised in that by rare earth terbium ion solution and 2,6 pyridinedicarboxylic acids pass through anti-
Structural formula is obtained for Tb/DPA@SiO to microemulsion method synthesis2Presoma；It is 0.5 ~ 1 through centrifuge washing compound concentration scope
After the mg/ml aqueous solution add equimolar than rare-earth europium ion solution E u3+With guanosine ribonucleoside acid solution, through concussion reaction
Afterwards, centrifuge washing obtains structural formula for Tb/DPA@SiO2- Eu/GMP fluorescence probe；Utilize Tb3+Fluorescence intensity level and Eu3+
Both fluorescence intensity levels ratio component ratio type rare-earth fluorescent probe Tb/DPA@SiO2-Eu/GMP。
2. fluorescence probe as claimed in claim 1, it is characterised in that the reverse microemulsion composition is that volume ratio is：Just
Ji Chun ︰ Qulas lead to X-100 ︰ Huan Ji Wan ︰ teos solutions=︰ 17 of 200 ︰, 200 ︰ 800.
3. fluorescence probe as claimed in claim 1, it is characterised in that the rare-earth europium ion solution E u3+With guanine ribose
The end reaction concentration of nucleotide solution is 0.5 ~ 2 mM.
4. a kind of sketch-based user interface type rare-earth fluorescent probe is used for bacillus anthracis biomarker DPA detection method, its feature exists
In the addition Tb/DPA@SiO in the solution of the buffer solution containing Tris-HCl2- Eu/GMP fluorescence probes, are added after solution-stabilized
DPA solution to be measured is reacted, and occurs Enhancement of Fluorescence, and passage time resolved fluorometric analysis method determines DPA concentration；
Wherein：The fluorescence probe is dissolved in the Tb/DPA@SiO that water formation concentration is 0.1 ~ 0.2 mg/ml2- Eu/GMP the aqueous solution；It is described
The solution concentration of buffer solution is 20 ~ 50 mM, and pH value is 4~9；The reaction temperature is 5~35 DEG C, the DPA solution to be measured
Concentration be 0.025~2 μM.
5. detection method as claimed in claim 4, it is characterised in that the fluorescence probe lowest detection line is 7.3 nM's
DPA solution.
6. a kind of test paper of sketch-based user interface type rare-earth fluorescent probe, it is characterised in that ordinary filter paper is cut into a diameter of 5 ~ 8 mm
Circular paper, concentration is immersed in after 10 ~ 30 minutes, test paper to be taken out, in natural shape in 1 ~ 5 mg/ml fluorescence probe solution
Dried under state, the test paper with fluorescence probe is made.
7. application of the test paper described in a kind of claim 6 in Visual retrieval DPA concentration, it is characterised in that the test paper exists
Adding has different color responses after the DPA of various concentrations, as DPA concentration gradually increases, test paper color gradually changes, root
Quantify DPA concentration according to the intensity level of color, can be to DPA concentration real-time online, Intelligent Measurement.
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| CN115109584A (en) * | 2022-07-11 | 2022-09-27 | 北京石油化工学院 | Method for detecting dipicolinic acid and Cu 2+ Fluorescent nano probe composite material and preparation method and application thereof |
| WO2023185543A1 (en) * | 2022-03-28 | 2023-10-05 | 河南理工大学 | Preparation and detection methods for dual-channel visualized multicolor fluorescent probe |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101486903A (en) * | 2009-02-23 | 2009-07-22 | 东南大学 | Preparation of rare earth luminous nanoparticle based on pyridine dicarboxylic acid |
| CN101705084A (en) * | 2009-11-09 | 2010-05-12 | 东南大学 | Multicolour rare earth nano particle and preparation method thereof |
| CN105424661A (en) * | 2015-11-13 | 2016-03-23 | 贵州大学 | Probe method for detecting a trace amount of F- through ratio fluorescence and ratio absorption or visual observation |
-
2017
- 2017-03-09 CN CN201710137454.7A patent/CN106959290A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101486903A (en) * | 2009-02-23 | 2009-07-22 | 东南大学 | Preparation of rare earth luminous nanoparticle based on pyridine dicarboxylic acid |
| CN101705084A (en) * | 2009-11-09 | 2010-05-12 | 东南大学 | Multicolour rare earth nano particle and preparation method thereof |
| CN105424661A (en) * | 2015-11-13 | 2016-03-23 | 贵州大学 | Probe method for detecting a trace amount of F- through ratio fluorescence and ratio absorption or visual observation |
Non-Patent Citations (6)
| Title |
|---|
| HONGLIANG TAN ET AL: "Nanoscaled lanthanide/nucleotide coordination polymer for detection of an anthrax biomarker", 《SENSORS AND ACTUATORS B》 * |
| MINGJING LIU ET AL: "Development of a ratiometric time-resolved luminescence sensor for pH based on lanthanide complexes", 《ANALYTICA CHIMICA ACTA》 * |
| RUI ZHOU ET. AL.: "Europium-decorated ZnO quantum dots as a fluorescent sensor for the detection of an anthrax biomarker", 《JOURNAL OF MATERIALS CHEMISTRY C》 * |
| RYUHEI NISHIYABU ET. AL.: "Nanoparticles of Adaptive Supramolecular Networks Self-Assembled from Nucleotides and Lanthanide Ions", 《JACS》 * |
| TIAN-YING SUN ET AL: "Dual-emitting nanocomposites derived from rare-earth compound nanotubes for ratiometric fluorescence sensing applications", 《NANOSCALE》 * |
| YIHE ZHANG ET AL: "Rapid andfacileratiometricdetectionofan anthrax biomarker by regulatingenergytransferprocessinbio-metal-organicframework", 《BIOSENSORS AND BIOELECTRONICS》 * |
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