CN107505302A - It is prepared by a kind of unmarked biological thiol fluorescence detection method and its application and probe based on the lobate nitrogen-doped carbon nanobelt of rice - Google Patents
It is prepared by a kind of unmarked biological thiol fluorescence detection method and its application and probe based on the lobate nitrogen-doped carbon nanobelt of rice Download PDFInfo
- Publication number
- CN107505302A CN107505302A CN201710857274.6A CN201710857274A CN107505302A CN 107505302 A CN107505302 A CN 107505302A CN 201710857274 A CN201710857274 A CN 201710857274A CN 107505302 A CN107505302 A CN 107505302A
- Authority
- CN
- China
- Prior art keywords
- fluorescence
- biological thiol
- rice
- nrpcnrs
- nitrogen
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- 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"
-
- 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
- G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Optics & Photonics (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The invention discloses a kind of unmarked biological thiol fluorescence detection method and its application, detecting step based on the lobate nitrogen-doped carbon nanobelt of rice to include：1. the lobate nitrogen-doped carbon nanobelt fluorescence probe solution of phosphate buffered saline solution, rice and silver ion solution will be sequentially added in container；2. being grouped and adding the standard liquid of the biological thiol containing various concentrations and the testing sample of the biological thiol containing unknown concentration, 3. dilution incubates after mixing；4. determine the change of each group solution fluorescence intensity；5. for the biological thiol concentration added using standard liquid group as transverse axis, standard liquid fluorescence recovery extent is the longitudinal axis, standard curve is drawn；6. being contrasted by the fluorescence recovery extent and the standard curve of testing sample, the biological thiol concentration of sample is judged.It utilizes NRPCNRs Ag+Reversible complexing, without fluorescence labeling, just can realize that fluorescence recovers and real-time and quantification analysis in biological thiol.
Description
Technical field
The present invention relates to trace detection field, and in particular to a kind of unmarked life based on the lobate nitrogen-doped carbon nanobelt of rice
Thing mercaptan fluorescence detection method and its preparation and application.
Background technology
Biological thiol class sulfhydryl compound, generally comprise cysteine (Cys), homocysteine (Hcy), glutathione
(GSH).Because these molecules play important physiological action in organism, more and more researchers are caused at present
Concern.Numerous studies prove that sulfhydryl compound is unbalance closely related with many diseases in organism.For example, half in human body
The excessive concentration of cystine can cause neuron excitotoxicity, and cysteine shortage will cause body edema, decreased growth, leucocyte
Reduction, epilation and other physical function disorders.Homotype semicystinol concentration investigating rise also with many diseases, such as heart
Vascular diseases, osteoporosis, senile dementia etc. are closely related.If in addition, human body GSH-PX activity horizontal abnormality, it will
Suppress body immune system function, accelerate aging, and it is related to some diseases such as diabetes, tuberculosis, AIDS.Due to leading to
The level of these compounds in detection such as blood plasma, living cells or urine biological sample is crossed, can examining for clinically relevant disease
It is disconnected that diagnosis basis is provided.Therefore, the development effective detection method of biological thiol has very important realistic meaning.
At present, the detection that many methods are used for biological thiol, including HPLC MS, area has been developed
Band capillary electrophoresis, electrochemical process and inductively coupled plasma mass spectrometry method (abbreviation ICP-MS) etc..But these are examined
The shortcomings that some are inevitable, such as expensive equipment and heaviness, operation sequence complexity, time of measuring length often be present in survey technology,
It is and high to the skill set requirements of operator, it is necessary to dedicated technician people's operation etc..In order to overcome the above-mentioned subject matter being related to,
Technology based on fluorescence probe detection biological thiol receives more and more attention, and has also obtained extensive development.However, although
These detection techniques have higher selectivity and sensitivity to biological thiol, but these methods often relate to complexity and held high
The mark of the modification of expensive molecular beacon probe and fluorogen and fluorescent quenching group, thus such sensor often take and
Need cumbersome pretreatment process.Therefore, need badly and develop the good fluorescence sense inspection of a kind of simple and quick, high sensitivity, selectivity
The analysis that survey technology is applied to biological thiol detects.
The content of the invention
For above-mentioned problem of the prior art, it is an object of the invention to provide one kind to be based on the lobate nitrogen-doped carbon nanometer of rice
The unmarked biological thiol fluorescence detection method of band, utilizes NRPCNRs-Ag+Reversible complexing, without fluorescence labeling, just can
Realize that fluorescence recovers and real-time and quantification is analyzed in biological thiol.
A kind of unmarked biological thiol fluorescence detection method based on the lobate nitrogen-doped carbon nanobelt of rice, detecting step are provided
Including：1. phosphate buffered saline solution (Phosphate Buffered Saline, abbreviation PBS), rice leaf will be sequentially added in container
Shape nitrogen-doped carbon nanobelt (abbreviation NRPCNRs) fluorescence probe solution and silver ion solution；2. it is grouped and adds containing various concentrations
The standard liquid of biological thiol and the testing sample of the biological thiol containing unknown concentration, 3. dilution incubate after mixing；4. determine each group
The change of solution fluorescence intensity；5. the biological thiol concentration added using standard liquid group is transverse axis, standard liquid fluorescence recovers journey
Spend [(FL-FL0)]/FL0For the longitudinal axis, standard curve is drawn；6. fluorescence recovery extent and the standard curve by testing sample
Contrast, judge the biological thiol concentration of sample.
Preferably, step 1. in, the mass ratio of the lobate nitrogen-doped carbon nanobelt fluorescence probe of the rice and silver ion is
(2-3):1。
Preferably, step 3. in, the lobate nitrogen-doped carbon nanobelt fluorescence probe concentration of rice is 10-25 μ g/ml, silver
Ion concentration is 70-90 μM.
Preferably, step 4. in, the N doping content of the lobate nitrogen-doped carbon nanobelt of rice is (35-37) wt%.
Preferably, fluorescence exciting wavelength 355nm, to excite wide with transmite slit be 3nm.Biological thiol includes cysteine and paddy Guang
At least one of sweet peptide, it is furthermore preferred that the testing sample is biological sample, contain at least one of serum and cell.
Preferably, step 5. and 6. in, draw the standard curve and calculate biological thiol content in testing sample
Method, using standard addition method.
A kind of application of rice shape leaf nitrogen-doped carbon nanobelt in biological thiol detection, by above-mentioned detection method, with reference to
NRPCNRs-Ag+Unmarked fluoroscopic examination is carried out to testing sample.
Preferably, fluorescence probe profile is that rice is lobate, particle diameter 100-120nm, N element doping (35-37) wt%, is swashed
Hair wavelength is 355nm.
Further, the fluorescence probe of the rice shape leaf nitrogen-doped carbon nanobelt is prepared by hydrothermal synthesis method, preparation process
Including：
By 1.1g uric acid (Uric Acid), 25mL absolute ethyl alcohols and the mixing of 25mL deionized waters, mixed under ultrasound condition
For suspension；The 25mL suspension of preparation is transferred in polytetrafluoroethylene (PTFE) pyroreaction kettle, and 4.5h is kept at 180 DEG C；
Room temperature is naturally cooled to, products therefrom is extracted with dichloromethane, and extraction gained aqueous phase solution is refrigerated 5-10 days, stands and removes
Length is more than 1 μm of large scale NRPCNRs；Then, 15min then by the solution being collected into 8000rpm is centrifuged, takes upper strata bright orange
The color NRPCNRs aqueous solution, and gained bright yellow solution is stored for future use at 4 DEG C.
Resultant effect caused by the present invention：
1st, the simple and quick hydro-thermal method synthesizing water-solubility of present invention use is good, is uniformly dispersed, has fluorescent characteristic, fluorescence steady
The lobate nitrogen-doped carbon nanobelt of fixed rice, and build highly sensitive, high selectivity detection life using the NRPCNRs as fluorescence probe
The fluorescence nano sensor of thing mercaptan.
2nd, because the N atomic interactions on Ag+ ions and NRPCNRs can induce NRPCNR that fluorescent quenching, shape occurs
Into non-fluorescence co-ordination complex, and once in NRPCNRs-Ag+Biological thiol is added in reaction system, due to Ag+Ion is easier
Stable Ag-S keys are formed with the S atom of biological thiol intramolecular, so that being incorporated in the Ag on NRPCNRs+Discharge,
NRPCNRs fluorescence intensity will be recovered, and the recovery extent of NRPCNRs fluorescence intensities is biological with being added in reaction system
The amount of mercaptan is in certain correlation.
3rd, the unmarked biological thiol new detecting method based on NRPCNRs of the invention can be effectively realized in human serum
Biological thiol is highly sensitive, high selectivity detection, and method is simple, quick, linear detection range is wide, is efficient detection biology sample
Biological thiol in product provides effective way, has potential application value.
Scheme explanation
Fig. 1 is the solution fluorescence spectrum and fluorescent brightness schematic diagram of the embodiment of the present invention；
Wherein, a.NRPCNRs (20 μ g/mL), b.NRPCNRs (20 μ g/mL)+Ag+(80μM),c.NRPCNRs(20μg/
mL)+Ag+(80μM)+Cys(100μM),d.NRPCNRs(20μg/mL)+Cys(100μM)；Interior illustration is from left to right to be different molten
The fluorescence photo of liquid:1.NRPCNRs, 2.NRPCNRs+Ag+, 3.NRPCNRs+Ag++ Cys, 4.NRPCNRs+Cys.
Fig. 2 is that NRPCNRs ultraviolet spectrogram and NRPCNRs excite in 355nm in detection method of the embodiment of the present invention
Under launching light spectrogram；Interior illustration is pictures of the NRPCNRs under 1. natural lights and 2. uviol lamp 365nm irradiations.
Fig. 3 is transmission electron microscope (TEM) figure of solution in detection method of the embodiment of the present invention；Wherein A.NRPCNRs,
B.NRPCNRs-Ag+And C.NRPCNRs-Ag+-Cys。
Fig. 4 is the fluorescence intensity spectrum that detection method of the embodiment of the present invention obtains NRPCNRs at maximum emission wavelength 420nm
Figure；Wherein, testing conditions variable is A.Ag+Concentration, B.PBS ph value of buffer solution.
Fig. 5 is the spectrogram that detection method of the embodiment of the present invention detects Cys and GSH using NRPCNRs：A.NRPCNRs-Ag+
Solution, in Cys concentration from the fluorescence spectrum under 0 to 200 μM；The relative intensity of fluorescence [(FL- of B.NRPCNRs fluorescence probes
FL0)/FL0], the canonical plotting of Cys concentration is quantitatively detected with Cys change in concentration；C.NRPCNRs-Ag+Solution, in GSH concentration
Fluorescence spectrum under 0 to 200 μM；[(the FL-FL of D.NRPCNRs fluorescence probes0)/FL0], quantitatively detected with GSH change in concentration
The canonical plotting of GSH concentration.
Fig. 6 is that detection method of the embodiment of the present invention is NRPCNRs-Ag+Detect the [(FL-FL of different biological samples0)/FL0]
Intensity contrast figure, NRPCNRs is investigated in detection method to Cys selectivity.
Embodiment
Explanation is further explained to the present invention with reference to specific embodiment and figure, it should be appreciated that the scope of the present invention
Not limited to this.In the present embodiment, spectrophotometer selects the XRF of Hitachi Hitachis, model F-7000.It is fixed
Amount detection solution biological thiol content carries out data processing using standard addition method of the prior art.Specifically, with reference to country
Standard addition method and US Gov Env Protection Agency described in the national standard GB18582 of State Administration of Quality Supervision, Inspection and Quarantine's issue《METHOD OF
STANDARD ADDITIONS AND EFFECTS OF DILUTION》Described in standard addition method implemented.
Embodiment
(1) hydrothermal synthesis method prepares NRPCNRs fluorescent nano probes
By 1.1g uric acid, 25mL absolute ethyl alcohols and the mixing of 25mL deionized waters, it is suspension to be mixed under ultrasound condition；Will
The 25mL suspension of preparation is transferred in polytetrafluoroethylene (PTFE) pyroreaction kettle, and keeps 4.5h at 180 DEG C；Naturally cool to room
Temperature, products therefrom is extracted with dichloromethane, and gained aqueous phase solution will be extracted and refrigerated 5-10 days, stood and remove length more than 1 μm
Large scale NRPCNRs；Then, 15min then by the solution being collected into 8000rpm is centrifuged, takes upper strata glassy yellow NRPCNRs water
Solution, and gained bright yellow solution is stored for future use at 4 DEG C.
(2) the lobate nitrogen-doped carbon nanobelt of rice of the present embodiment based on above-mentioned synthesis is built high as unmarked fluorescence probe
Sensitive, high selectivity detection biological thiol fluorescence nano sensor, for the detection of biological thiol, detecting step includes：
1. it is 50mM to sequentially add 50 μ L concentration into 1.5mL centrifuge tubes first, pH is 7.4 PBS cushioning liquid, 5 μ L
Concentration is 2mg/mL NRPCNRs fluorescence probe solution, and 40 μ L concentration are 1mM Ag+ solions；2. it is grouped and adds containing not
With the standard liquid of concentration biological thiol and the testing sample of the biological thiol containing unknown concentration, deionized water diluting reaction is 3. used again
Reactant mixture is incubated 20min by liquor capacity at room temperature on turbine mixer to 500 μ L after abundant mixing；4. survey
Determine the change of each group solution fluorescence intensity；5. the biological thiol concentration added using standard liquid group is transverse axis, standard liquid fluorescence
Recovery extent [(FL-FL0)]/FL0For the longitudinal axis, standard curve is drawn；6. by the fluorescence recovery extent and the institute that test testing sample
Standard curve contrast is stated, judges the biological thiol concentration of sample.
Step 1. in, the mass ratio of the lobate nitrogen-doped carbon nanobelt fluorescence probe of the rice and silver ion is 2.3:1.
Step 3. in, the lobate nitrogen-doped carbon nanobelt fluorescence probe concentration of rice is 20 μ g/ml, and concentration of silver ions is
80μM。
Step 4. in, the N doping content of the lobate nitrogen-doped carbon nanobelt of rice is 35-37wt%.Fluorescence excitation ripple
A length of 355nm, to excite wide with transmite slit be 3nm.Biological thiol includes cysteine and glutathione, the testing sample
For biological sample, contain serum.
Step 5. and 6. in, draw the standard curve and the method for calculating biological thiol content in testing sample, adopt
Use standard addition method.
The preparation method of described NRPCNRs fluorescence probes, is prepared using hydrothermal synthesis method, and fluorescence probe profile is rice leaf
Shape, particle diameter 150-180nm, N element doping (35-37) wt%, excitation wavelength 355nm.
Detect mechanism：Due to Ag+It can induce NRPCNR that fluorescent quenching occurs with the N atomic interactions on NRPCNRs,
Non-fluorescence co-ordination complex is formed, and once in NRPCNRs-Ag+Biological thiol is added in reaction system, due to Ag+Ion more holds
Stable Ag-S keys are easily formed with the S atom of biological thiol intramolecular, so that being incorporated in the Ag on NRPCNRs+Discharge,
NRPCNRs fluorescence intensity will be recovered, and the recovery extent of NRPCNRs fluorescence intensities is biological with being added in reaction system
The amount of mercaptan is in certain correlation.
(3) rice shape leaf nitrogen-doped carbon nanobelt is applied, biological thiol is detected by the present embodiment method, with reference to NRPCNRs-
Ag+Unmarked fluoroscopic examination is carried out to testing sample, sample detection and probe characterization result are as shown in figures 1 to 6：
As a result show, the present invention is designed to effectively realize that biological thiol is highly sensitive, high selectivity detection.
1) as shown in figure 1, the NRPCNRs that the present invention is prepared using hydrothermal synthesis method has good fluorescence property (figure
1a), when the Ag+ ions of appropriate amount are added in NRPCNRs solution, NRPCNRs fluorescence is by Ag+Ion quenching (figure
1b), and when adding biological thiol (Cys) in being further continued for its solution, NRPCNRs 90% fluorescence is restored (Fig. 1 c),
It also demonstrate that Cys will not have an impact (Fig. 1 d) to NRPCNRs fluorescence simultaneously.Illustrate to visit NRPCNRs as fluorescence
Pin establishes the nano fluorescent sensing detection new method of biological thiol.
Before NRPCNRs is detected as fluorescence probe, ultraviolet sign is carried out to it first and TEM is characterized, has characterized knot
Fruit is as shown in Figures 2 and 3.
2) from Fig. 2, it can be seen that as shown in Figure 2, NRPCNRs ultra-violet absorption spectrum exists respectively WithPlace shows two characteristic peaks (curve a), and using 355nm as maximum excitation wavelength, can excite NRPCNRs
Send fluorescence spectrum (420nm) (curve b), meanwhile, the interior illustration in Fig. 2 has further showed that NRPCNRs fluorescent characteristic.
The above results show that prepared NRPCNRs can act as fluorescent nano probe, the quantitative analysis for biological thiol.
3) Fig. 3 is different solutions TEM figures.As shown in Figure 3A, NRPCNRs fluorescence probes profile is lobate similar to rice, and particle diameter is
150-180nm；Due to Ag+Ag-N keys can be formed between the nitrogen-atoms in NRPCNs, make NRPCNRs that aggregation (figure occur
3B), so as to causing NRPCNRs fluorescent quenching.And when in NRPCNRs-Ag+Reaction system in add Cys after, NRPCNRs
Obvious clustering phenomena (Fig. 3 C) does not occur, this result shows, Ag+Ion is easier former with the S of biological thiol intramolecular
Son forms stable Ag-S keys, so that the Ag+ being incorporated on NRPCNRs is discharged, NRPCNRs fluorescence intensity is recovered.
4) as shown in figure 4, in order to preferably realize the effective detection of biological thiol, the present invention have studied Ag+It is dense
Influences of the pH of degree and PBS cushioning liquid to testing result.Optimum results show, the optimal Ag of detection architecture+Concentration be 80 μM,
The optimal acid-base value of PBS cushioning liquid is pH=7.4.
5) Cys and GSH detection.Under optimal conditions (50mM, pH 7.4PBS, 20 μ g/mL NRPCNRs, 80 μM
Ag+), using the Cys and GSH of a series of various concentrations of present invention method measure, as a result as shown in figure 5, Fig. 5 A are shown
The changes of Cys NRPCNRs fluorescence intensities in 0-200 μM of concentration range.In 80 μM of Ag+In the presence of, with Cys concentration
Increase, fluorescence intensities of the NRPCNRs at 420nm also increase (Fig. 5 B), and presents in 0.05-10 μM of concentration range good
Good linear relationship.Fig. 5 C and 5D are GSH testing result figures.As a result show, with the increase of GSH concentration, NRPCNRs exists
Fluorescence intensity at 420nm also increases (Fig. 5 D), and good linear relationship, the above are presented in 0.5-30 μM of concentration range
As a result absolutely proved that the fluorescent optical sensor of the invention designed has good response to biological thiol.
6) method choice is investigated.The present invention has investigated the selectivity of detecting system, to possible in the biosystem of complexity
Existing non-thiol amino acid interfering material includes Pro, Val, Tyr, Ser, His, Try, Arg, Glu, Thr, Phe, Lys, Ala
Investigated with Gly.As shown in fig. 6, in containing different non-thiol amino acid complex biological system environment, detecting system
There is very high selectivity to Cys and GSH.Illustrate that the fluorescent optical sensor that the present invention is built is biological in detection of complex biosystem
Mercaptan has preferably selectivity.
7) actual sample is analyzed.The present invention verifies that the present invention is set by taking Cys in Standard Addition Method for Determination normal human serum as an example
Practicality of the fluorescent optical sensor of meter in actual sample analysis detection.As a result as shown in appendix 1, the Cys rate of recovery exists
Between 96.2%-104.8%, and relative standard deviation is less than 5.0%, and it is preferably reliable to illustrate that inventive embodiments method has
Property.Meanwhile the content of Cys in normal human serum is determined also in normal range (NR).The above results illustrate, the present invention design based on
NRPCNRs unmarked biological thiol new detecting method can effectively realize that biological thiol in human serum is highly sensitive, high selection
Property detection, and method is simple, quick, linear detection range is wide, is provided for the biological thiol in efficient detection biological sample
Effective way, there is potential application value.
The NRCNRs of table 1 detects the Cys in testing sample as fluorescence probe
Although the present invention has been described in detail, it will be understood by those skilled in the art that in spirit and scope of the invention
Modification will be apparent.However, it should be understood that each side of the invention recorded, different embodiments
Each several part and the various features enumerated can be combined or all or part of exchange.In above-mentioned each embodiment, that
A little embodiments with reference to another embodiment can be combined suitably with other embodiment, and this is by by this area skill
Art personnel are to understand.In addition, it will be understood to those of skill in the art that description above is only the mode of example, not purport
In the limitation present invention.
Claims (10)
- A kind of 1. unmarked biological thiol fluorescence detection method based on the lobate nitrogen-doped carbon nanobelt of rice, it is characterised in that inspection Surveying step includes：1. the lobate nitrogen-doped carbon nanobelt fluorescence probe solution of phosphate buffered saline solution, rice will be sequentially added in container And silver ion solution；2. it is grouped and adds the standard liquid of the biological thiol containing various concentrations and treating for the biological thiol containing unknown concentration Test sample product, 3. dilution incubate after mixing；4. determine the change of each group solution fluorescence intensity；5. the biology added with standard liquid group Concentrations of mercaptans is transverse axis, and standard liquid fluorescence recovery extent is the longitudinal axis, draws standard curve；It is 6. extensive by the fluorescence of testing sample Multiple degree contrasts with the standard curve, judges the biological thiol concentration of sample.
- 2. according to the method for claim 1, it is characterised in that step 1. in, the lobate nitrogen-doped carbon nanobelt of rice The mass ratio of fluorescence probe and silver ion is (2-3):1.
- 3. according to the method for claim 2, it is characterised in that step 3. in, the lobate nitrogen-doped carbon nanobelt of rice Fluorescence probe concentration is 10-25 μ g/ml, and concentration of silver ions is 70-90 μM.
- 4. according to the method for claim 1, it is characterised in that step 4. in, the lobate nitrogen-doped carbon nanobelt of rice N doping content be (35-37) wt%.
- 5. according to the method for claim 3, it is characterised in that fluorescence exciting wavelength 355nm, excite it is wide with transmite slit It is 3nm.
- 6. according to the method for claim 3, it is characterised in that biological thiol is included in cysteine and glutathione extremely Few one kind, it is preferred that the testing sample is biological sample, contains at least one of serum and cell.
- 7. according to the method for claim 3, it is characterised in that step 5. and 6. in, draw the standard curve and meter The method for calculating biological thiol content in testing sample, using standard addition method.
- 8. application of a kind of rice shape leaf nitrogen-doped carbon nanobelt in biological thiol detection, it is characterised in that will by such as right Method any one of 1-6 is sought, with reference to NRPCNRs-Ag+Unmarked fluoroscopic examination is carried out to testing sample.
- 9. application according to claim 8, it is characterised in that fluorescence probe profile is that rice is lobate, particle diameter 150- 180nm, N element doping (35-37) wt%, excitation wavelength 355nm.
- 10. application according to claim 9, it is characterised in that the fluorescence probe of the rice shape leaf nitrogen-doped carbon nanobelt Prepared by hydrothermal synthesis method, preparation process preferably includes：By 1.1g uric acid, 25mL absolute ethyl alcohols and the mixing of 25mL deionized waters, it is suspension to be mixed under ultrasound condition；It will prepare 25mL suspension be transferred in polytetrafluoroethylene (PTFE) pyroreaction kettle, and keep 4.5h at 180 DEG C；Naturally cool to room temperature, Products therefrom is extracted with dichloromethane, and extraction gained aqueous phase solution is refrigerated 5-10 days, it is big more than 1 μm to stand removing length Size NRPCNRs；Then, 15min then by the solution being collected into 8000rpm is centrifuged, takes upper strata glassy yellow NRPCNRs water-soluble Liquid, and gained bright yellow solution is stored at 4 DEG C.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710857274.6A CN107505302B (en) | 2017-09-21 | 2017-09-21 | Application of rice-shaped leaf nitrogen-doped carbon nanoribbon in biological thiol detection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710857274.6A CN107505302B (en) | 2017-09-21 | 2017-09-21 | Application of rice-shaped leaf nitrogen-doped carbon nanoribbon in biological thiol detection |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107505302A true CN107505302A (en) | 2017-12-22 |
| CN107505302B CN107505302B (en) | 2020-07-07 |
Family
ID=60698310
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710857274.6A Active CN107505302B (en) | 2017-09-21 | 2017-09-21 | Application of rice-shaped leaf nitrogen-doped carbon nanoribbon in biological thiol detection |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107505302B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108760706A (en) * | 2018-06-08 | 2018-11-06 | 农业部环境保护科研监测所 | A kind of method of quick screening low cadmium-accumulation rice varieties |
| CN113916856A (en) * | 2021-10-09 | 2022-01-11 | 江南大学 | fluorescence-Raman dual-mode nano sensor, preparation method thereof and application thereof in mercaptan detection |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105588823A (en) * | 2016-01-19 | 2016-05-18 | 济南大学 | Preparation and application of fluorescent switch type sensor for detecting biological thiols |
| CN105842209A (en) * | 2016-03-18 | 2016-08-10 | 中国科学院合肥物质科学研究院 | Paper sensor for rapid on-site detection of fluorine ions in water and preparation method thereof |
| CN106645035A (en) * | 2016-12-16 | 2017-05-10 | 盐城工学院 | Method for detecting content of heavy metal silver ions based on carbon-spot light scattering sensing |
-
2017
- 2017-09-21 CN CN201710857274.6A patent/CN107505302B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105588823A (en) * | 2016-01-19 | 2016-05-18 | 济南大学 | Preparation and application of fluorescent switch type sensor for detecting biological thiols |
| CN105842209A (en) * | 2016-03-18 | 2016-08-10 | 中国科学院合肥物质科学研究院 | Paper sensor for rapid on-site detection of fluorine ions in water and preparation method thereof |
| CN106645035A (en) * | 2016-12-16 | 2017-05-10 | 盐城工学院 | Method for detecting content of heavy metal silver ions based on carbon-spot light scattering sensing |
Non-Patent Citations (1)
| Title |
|---|
| VIVEK BORSE ET.AL: "N-doped multi-fluorescent carbon dots for ‘turn off-on’ silver-biothioldual sensing and mammalian cell imaging application", 《SENSORS AND ACTUATORS B》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108760706A (en) * | 2018-06-08 | 2018-11-06 | 农业部环境保护科研监测所 | A kind of method of quick screening low cadmium-accumulation rice varieties |
| CN108760706B (en) * | 2018-06-08 | 2021-08-06 | 农业部环境保护科研监测所 | Method for rapidly screening rice varieties with low cadmium accumulation |
| CN113916856A (en) * | 2021-10-09 | 2022-01-11 | 江南大学 | fluorescence-Raman dual-mode nano sensor, preparation method thereof and application thereof in mercaptan detection |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107505302B (en) | 2020-07-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Lou et al. | A fluorescent probe for rapid detection of thiols and imaging of thiols reducing repair and H 2 O 2 oxidative stress cycles in living cells | |
| Zeng et al. | A colorimetric and ratiometric fluorescent probe for quantitative detection of GSH at physiologically relevant levels | |
| Yang et al. | A selective coumarin-based “turn-on” fluorescent sensor for the detection of cysteine and its applications for bioimaging | |
| Hu et al. | A rapid and sensitive turn-on fluorescent probe for ascorbic acid detection based on carbon dots–MnO 2 nanocomposites | |
| CN107389636A (en) | A kind of preparation and application of the water soluble fluorescence sensor that endogenous glutathione can be detected in cancer cell | |
| Zheng et al. | A rhodol-derived probe for intracellular biothiols imaging and rapid labelling of sulfhydryl-containing proteins | |
| CN107941774A (en) | Phosphatic method is detected based on metal organic frame composite material ratio fluorescent | |
| JPH0241708B2 (en) | ||
| Wang et al. | A coumarin-based dual optical probe for homocysteine with rapid response time, high sensitivity and selectivity | |
| CN109406487A (en) | A kind of Raman biosensor and its preparation method and application detecting Alzheimer's disease marker | |
| Wang et al. | A one-pot synthesis of fluorescent N, P-codoped carbon dots for vitamin B 12 determination and bioimaging application | |
| CN107936035A (en) | A kind of cysteine-modifying graphene quantum dot GQCY and preparation method are with preparing the application on dopamine luciferase assay reagent | |
| CN107253961A (en) | It is a kind of can ratio test cysteine water soluble fluorescence sensor preparation and application | |
| Chen et al. | Imidazo [1, 5-α] pyridine-derived fluorescent turn-on probe for cellular thiols imaging with a large Stokes shift | |
| CN110878049B (en) | Preparation and application of fluorescent probe for specifically analyzing hydrogen sulfide in Golgi apparatus | |
| Niu et al. | A novel fluorescent probe for detection of Glutathione dynamics during ROS-induced redox imbalance | |
| CN107505302A (en) | It is prepared by a kind of unmarked biological thiol fluorescence detection method and its application and probe based on the lobate nitrogen-doped carbon nanobelt of rice | |
| CN109320490A (en) | A kind of fluorescence probe of near-infrared specific detection cysteine | |
| CN110387230A (en) | A kind of highly selective fluorescent nano probe, preparation method and its application on detection ascorbic acid | |
| CN108689933A (en) | A kind of hypochlorous fluorescence probe of quick high-selectivity analysis | |
| Li et al. | Rapid pattern recognition of different types of sulphur-containing species as well as serum and bacteria discrimination using Au NCs-Cu2+ complexes | |
| CN107253932A (en) | A kind of quick high-selectivity hypersensitive nickel ion ratio fluorescent probe and preparation method thereof | |
| CN105503768A (en) | Preparation method of alpha-oxoglutarate fluorescent/ultraviolet molecular probe and application of alpha-oxoglutarate fluorescent/ultraviolet molecular probe to biological samples | |
| CN114790202B (en) | Preparation and application of biological thiol activated precipitation dye type efficient diagnosis and treatment integrated probe | |
| CN110669503A (en) | Preparation and application of carbon monoxide near-infrared fluorescent probe |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |