CN110887819B - Preparation and application of high-selectivity fluorescent probe for detecting phenylalanine - Google Patents
Preparation and application of high-selectivity fluorescent probe for detecting phenylalanine Download PDFInfo
- Publication number
- CN110887819B CN110887819B CN201910797922.2A CN201910797922A CN110887819B CN 110887819 B CN110887819 B CN 110887819B CN 201910797922 A CN201910797922 A CN 201910797922A CN 110887819 B CN110887819 B CN 110887819B
- Authority
- CN
- China
- Prior art keywords
- water
- probe
- phenylalanine
- solution
- detecting
- 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.)
- Active
Links
Images
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"
- G01N21/643—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" non-biological material
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
Abstract
The invention discloses preparation and application of a high-selectivity fluorescent probe for detecting phenylalanine. The molecular formula of the probe is C 48 H 48 N 32 O 16 @C 15 H 16 ClN3S· 0.5 ZnCl 2 Is composed of eight-membered cucurbituril (Q [8 ])]) With Toluidine Blue (TB) to form said eight-membered cucurbituril Q [8]]Molar ratio to Toluidine Blue (TB) 1:2. the invention can detect the L-phenylalanine in water, realize the highly selective detection of amino acid by a single probe, has low detection cost and high detection efficiency, and is beneficial to the analysis of a complex microscopic system. In addition, when the probe is used for detection, the probe also has the characteristics of higher sensitivity, simple sample treatment, convenient operation, quick determination and real-time detection.
Description
Technical Field
The invention relates to a fluorescent probe, in particular to preparation and application of a high-selectivity fluorescent probe for detecting phenylalanine.
Background
Amino acids play an extremely important role in the growth and development of organisms: 1. is a constituent of human tissue; 2. various substances constituting the human body; 3. supplying heat; 4. immune regulation; 5. as an important carrier in vivo, plays a role in transportation; 6. and (4) oxidation function. Amino acid is the basic composition substance of protein, the ingested amino acid is the only way for human body to obtain nitrogen source, after human body absorbs amino acid, one part is directly used for synthesizing protein, one part is oxidized and decomposed, wherein the nitrogen-containing part is used for synthesizing other necessary amino acid, and one part is decomposed as energy and is discharged out of body in the form of urea. Therefore, the method is particularly important for detecting the content and the type of amino acid in food, and the fluorescent probe is a novel detection reagent, so that the fluorescent probe is favored by people due to the advantages of higher sensitivity, lower detection cost, simple sample treatment, convenient operation, quick determination and real-time detection. It is necessary to develop a novel fluorescent probe for detecting L-amino acid in water and a novel fluorescent probe detection method.
Disclosure of Invention
The invention aims to provide preparation and application of a high-selectivity fluorescent probe for detecting phenylalanine. The invention can detect the L-phenylalanine in water, realize the highly selective detection of amino acid by a single probe, has low detection cost and high detection efficiency, and is beneficial to the analysis of a complex microscopic system. In addition, when the probe is used for detection, the probe also has the characteristics of higher sensitivity, simple sample treatment, convenient operation, quick determination and real-time detection.
The technical scheme of the invention is as follows: a high-selectivity fluorescent probe for detecting phenylalanine has a molecular formula of C 48 H 48 N 32 O 16 @C 15 H 16 ClN 3 S•0.5ZnCl 2 。
The preparation of the high selectivity fluorescent probe for detecting phenylalanine is prepared from eight-membered cucurbituril (Q8) and Toluidine Blue (TB), wherein the molar ratio of the eight-membered cucurbituril Q8 to the Toluidine Blue (TB) is 1:2, the method specifically comprises the following steps:
(1) Dissolving eight-element cucurbituril (Q8) and eight-element cucurbituril (Q8) in water to obtain solution A;
(2) Dissolving Toluidine Blue (TB) in water to obtain a solution B;
(3) And mixing the solution A and the solution B to obtain the probe.
In the preparation of the fluorescent probe for detecting phenylalanine with high selectivity, in the step (1), the solution A contains eight-membered cucurbituril (Q [8 ])]) Has a concentration of 1.0X 10 -3 mol/L。
In the preparation of the fluorescent probe for detecting phenylalanine with high selectivity, in the step (2), the concentration of Toluidine Blue (TB) in the solution B is 1.0 × 10 -3 mol/L。
In the preparation of the fluorescent probe with high selectivity for detecting phenylalanine, in the step (3), the solution A and the solution B are mixed according to a molar ratio of eight-membered cucurbituril (Q8) to Toluidine Blue (TB) of 1:2, and reacting for 10 seconds at normal temperature to obtain the probe.
In the preparation of the fluorescent probe for detecting phenylalanine with high selectivity, the water added in the steps (1) and (2) is secondary water with pH = 7.
The application of the high-selectivity fluorescent probe for detecting phenylalanine is used for detecting various L-amino acids in water.
In the application of the fluorescent probe for detecting high selectivity of phenylalanine, the method for detecting various L-amino acids in water is used for detecting L-phenylalanine in water.
In the application of the fluorescent probe for detecting phenylalanine with high selectivity, when the fluorescent probe is used for detecting L-phenylalanine in water, the specific detection method comprises the following steps:
1) Diluting the fluorescent probe with pH 7 water to 2.0 × 10 -5 Obtaining a probe standard solution by using a mol/L solution;
2) Adding water to be detected into the prepared probe standard solution, standing for 10S, then carrying out fluorescence emission spectrum measurement at a fixed excitation wavelength of 630 nm, and drawing a change curve of the fluorescence intensity at the excited laser wavelength;
3) And (3) calculating the fluorescence emission spectrum intensity change value under the corresponding 660nm before and after the water to be detected is added into the fluorescent probe solution according to the curve of the step 2), and detecting the L-phenylalanine in the water.
In the application of the high-selectivity fluorescent probe for detecting phenylalanine, in the step 3), when the fluorescence emission spectrum intensity of the probe consisting of the eight-membered cucurbituril (Q8) and the Toluidine Blue (TB) is enhanced corresponding to 660nm before and after the probe is added into the water to be detected, the fact indicates that the water to be detected contains L-phenylalanine; and when the fluorescence emission spectrum intensity corresponding to 660nm before and after the water to be detected is added does not change or is not subjected to blue shift, indicating that the water to be detected does not contain L-phenylalanine.
In order to verify the beneficial effects of the invention, the inventor carries out a great deal of experimental research, and the experimental process and the results are as follows: [ note: eight membered cucurbituril (Q8) abbreviated as Q8, toluidine Blue (TB) abbreviated as TB
Experimental example 1: exploring for the appropriate molar ratio of probe formed by Q8 to TB;
in order to explore the suitable molar ratio of the probe formed by Q8 and TB, the interaction between the host and the object was investigated by fluorescence spectroscopy and ultraviolet absorption spectroscopy. And (3) determining the ultraviolet absorption spectrum and the fluorescence spectrum and fluorescence spectrum data among the systems by a molar ratio method.
The specific method comprises the following steps: mixing TB and Q [8]]Respectively preparing into 1.0 mmol/L and 0.1 mmol/L aqueous solutions, wherein the concentration of immobilized object TB is 0.02mmol/L, change Q [8]]Concentration of (2), configuration N Q[8] /N TB An aqueous solution having a molar ratio of 0,0.1,0.2,0.3,0.5, 0.6,0.7,0.8,0.9,1.0,1.1,1.2,1.3,1.4,1.5, and the ultraviolet-visible absorption spectrum of the solution is measured at room temperature; measuring the fluorescence emission spectrum of the solution under the conditions of an excitation wavelength of 630 nm, an excitation slit of 5 nm, an emission slit of 5 nm and a voltage of 600V, measuring the ultraviolet absorption spectrum and the fluorescence spectrum between systems by adopting an equimolar continuous variation method (JOB method) as shown in figure 1, keeping the total concentration of the immobilized host and the immobilized object constant at 4.0 mmol/L, and preparing a series of different molar ratios N by continuously changing the ratio of the amount of substances between the host and the object Q[8] /( N Q[8] +N G ) Measuring the solution to be measured of 0.1, 0.2. 0.8,0.9,1.0 by the method, and measuring the ultraviolet absorption spectrum and the fluorescence spectrum when N is Q[8] / N TB The molar ratio is 1:2, the fluorescence intensity reaches a minimum value and N, determined by the JOB method Q[8] /( N Q[8] +N TB ) The difference in fluorescence intensity between TB and the same concentration was the greatest at a molar ratio of 0.33, thus giving TB and de-Q [8]]In a molar ratio of 2:1.
experimental example 2: carrying out quantitative analysis;
the concentration of the product prepared by the invention is 2.0X 10 -5 The L-Phe-containing solutions with different volume fractions were added to the mol/L fluorescent probe standard solution (prepared according to example 1) for detection, and the detection results are shown in FIG. 2, it can be seen that the L-Phe concentrations in the standard solutions are different after the addition of different volume fractions, the L-Phe concentrations can cause the fluorescent probe solution to generate different degrees of fluorescence sensitization, and the linear range of the L-Phe response is (1.0-20.0) × 10 × -5 mol/L, detection limit of 1.6581X 10 -6 mol/L (as shown in FIG. 3).
Experimental example 3 investigated the mechanism of action of the probe formed by (Q8) and (TB) and L-phenylalanine
In the invention, in the process of forming the probe by Q8 and TB, as can be seen from a nuclear magnetic spectrum shown in figure 4, after the probe is formed by Q8 and TB, L-phenylalanine is dripped into the probe, the proton peak moves to a low field, which indicates that after the probe formed by Q8 and TB, TB is wrapped in the cavity of Q8, and when phenylalanine is dripped, the phenylalanine ejects TB in the citrullus ring.
Compared with the prior art, the invention has the following beneficial effects:
1. the fluorescent probe can be used for simultaneously detecting the L-phenylalanine in the water, only one amino acid is detected, the specificity and pertinence of the probe are reflected, and the fluorescent probe has the advantages of wide detection range and specificity. The detection cost is low, the detection efficiency is high, and the analysis of a complex microscopic system can be facilitated.
2. The present invention utilizes Q8 and TB to form supermolecular complex, and when the above-mentioned amino acid is detected, phenylalanine and probe can be formed into new complex, so that the fluorescence of probe can produce sensitization action, so that it can simply, quickly and sensitively detect L-phenylalanine in water.
3. Compared with the traditional detection method, the detection cost is greatly reduced, the operation is convenient, and the real-time detection can be realized. Therefore, the probe can detect the L-phenylalanine in the water, the two probes detect the same amino acid, the specificity detection of the probe on the amino acid can be realized, the amino acid can be identified in a targeted manner, the detection cost is low, the detection efficiency is high, and the analysis on a complex microscopic system is facilitated. In addition, when the probe is used for detection, the probe also has the characteristics of high sensitivity, simple sample treatment, convenient operation, quick determination and real-time detection.
In conclusion, the method can detect the L-phenylalanine in the water, can realize the highly selective detection of the amino acid by the single probe, has low detection cost and high detection efficiency, and is favorable for the analysis of a complex microscopic system. In addition, when the probe is used for detection, the probe also has the advantages of higher sensitivity, simple sample treatment, convenient operation, quick determination and real-time detection.
Drawings
FIG. 1 is a graph showing fluorescence spectra of TB solutions with different Q8 solutions;
FIG. 2 is a graph showing fluorescence spectra of Q8 and TB in the case of adding a probe standard solution to a solution containing different L-amino acids;
FIG. 3 is a graph showing fluorescence spectra of Q8 and TB for a probe standard solution when a solution containing L-Phe at different concentrations was added;
FIG. 4 is a diagram showing nuclear magnetic titration of a probe standard solution into different concentrations of a solution containing L-Phe; wherein, the first and the second end of the pipe are connected with each other,
（A）TB:Q[8]=2 ：1
（B）L-Phe；
(C)TB:Q[8]:L-Phe=1 ：1 ：7.15
(D)TB:Q[8]:L-Phe=1 ：3 : 1
(E)TB:Q[8]:L-Phe=1：3 : 2
(F)TB:Q[8]:L-Phe=2:1:2。
Detailed Description
The invention is further described with reference to the following figures and examples, which are not to be construed as limiting the invention.
Example 1: a high-selectivity fluorescent probe for detecting phenylalanine is disclosed, wherein the molecular formula of the probe is C 48 H 48 N 32 O 16 @C 15 H 16 ClN3S• 0.5 ZnCl 2 Is composed of eight-membered cucurbituril (Q [8 ])]) With Toluidine Blue (TB) to form said eight-membered cucurbituril Q [8]]Molar ratio to Toluidine Blue (TB) 1:2, the method specifically comprises the following steps:
(1) Taking eight-membered cucurbituril (Q8)]) Eight yuan melon ring (Q8)]) Dissolving in water to obtain solution A containing eight-membered cucurbituril (Q8)]) Has a concentration of 1.0X 10 -3 mol/L；
(2) Dissolving Toluidine Blue (TB) in water to obtain solution B, wherein the concentration of Toluidine Blue (TB) in solution B is 1.0 × 10 -3 mol/L
(3) Mixing solution A and solution B at a molar ratio of octatomic cucurbituril (Q8) to Toluidine Blue (TB) of 1:2, and reacting for 10S at normal temperature to obtain the probe.
The method for detecting the L-phenylalanine in the water by using the fluorescent probe comprises the following steps:
1) Taking the fluorescent probe, adding water with pH of 7 to diluteThe release concentration is 2.0 × 10 -5 Obtaining a probe standard solution by using a mol/L solution;
2) Adding water to be detected into the prepared probe standard solution, standing for 10S, then performing fluorescence emission spectrometry at a fixed excitation wavelength of 630 nm, and drawing a variation curve of the fluorescence intensity at the excited laser wavelength;
3) And 3) calculating the intensity change value of the fluorescence emission spectrum corresponding to 660nm before and after the water to be detected is added into the fluorescent probe solution according to the curve in the step 2), and detecting the L-phenylalanine in the water.
When the fluorescence emission spectrum intensity of 660nm is increased before and after the probe composed of Q8 and TB is added into the water to be detected, it indicates that the water to be detected contains L-phenylalanine; and when the fluorescence emission spectrum intensity under 660nm does not change or blue shift before and after the water to be detected is added, indicating that the water to be detected does not contain L-phenylalanine.
Claims (1)
1. The application of the high-selectivity fluorescent probe for detecting phenylalanine is characterized in that:
the probe is used for detecting L-phenylalanine in water;
the molecular formula of the probe is C 48 H 48 N 32 O 16 @C 15 H 16 ClN 3 S•0.5ZnCl 2 Is prepared from eight-membered cucurbituril and toluidine blue,
the molar ratio of the eight-membered cucurbituril to the toluidine blue is 1:2, the method specifically comprises the following steps:
(1) Dissolving the eight-element cucurbituril in water, wherein the water is secondary water with pH =7, and the concentration of the eight-element cucurbituril is 1.0 × 10 - 3 A solution A of mol/L;
(2) Dissolving toluidine blue in water (pH = 7) twice to obtain toluidine blue with concentration of 1.0 × 10 -3 A mol/L solution B;
(3) And (3) mixing the solution A and the solution B according to the molar ratio of the eight-membered cucurbituril to the toluidine blue of 1:2, reacting at normal temperature for 10 seconds to obtain the probe;
when the probe is used for detecting the L-phenylalanine in the water, the specific detection method comprises the following steps:
1) Diluting the fluorescent probe with water of pH 7 to 2.0 × 10 -5 Obtaining a probe standard solution by using a mol/L solution;
2) Adding water to be detected into the prepared probe standard solution, standing for 10S, then carrying out fluorescence emission spectrometry at a fixed excitation wavelength of 630 nm, and drawing a change curve of the excited fluorescence intensity;
3) Calculating the fluorescence emission spectrum intensity change under 660nm before and after the water to be detected is added into the fluorescent probe solution according to the curve of the step 2), and detecting the L-phenylalanine in the water;
when the fluorescence emission spectrum intensity under 660nm is enhanced before and after the probe consisting of the eight-membered cucurbituril and the toluidine blue is added into the water to be detected, the fact that the water to be detected contains the L-phenylalanine is indicated; and when the fluorescence emission spectrum intensity under the corresponding 660nm does not change before and after the water to be detected is added, indicating that the water to be detected does not contain L-phenylalanine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910797922.2A CN110887819B (en) | 2019-08-27 | 2019-08-27 | Preparation and application of high-selectivity fluorescent probe for detecting phenylalanine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910797922.2A CN110887819B (en) | 2019-08-27 | 2019-08-27 | Preparation and application of high-selectivity fluorescent probe for detecting phenylalanine |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110887819A CN110887819A (en) | 2020-03-17 |
| CN110887819B true CN110887819B (en) | 2022-10-28 |
Family
ID=69745876
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910797922.2A Active CN110887819B (en) | 2019-08-27 | 2019-08-27 | Preparation and application of high-selectivity fluorescent probe for detecting phenylalanine |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110887819B (en) |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109232589B (en) * | 2018-10-19 | 2021-04-06 | 贵州大学 | Fluorescent probe and preparation and application thereof |
| CN109652062B (en) * | 2018-12-25 | 2021-07-13 | 贵州大学 | Fluorescent probe T and preparation and application thereof |
-
2019
- 2019-08-27 CN CN201910797922.2A patent/CN110887819B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN110887819A (en) | 2020-03-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109232589B (en) | Fluorescent probe and preparation and application thereof | |
| Wu et al. | Aptamer-functionalized magnetic nanoparticle-based bioassay for the detection of ochratoxin a using upconversion nanoparticles as labels | |
| CN103264165B (en) | A kind of method of synthesizing silver nanoclusters taking single stranded DNA as template | |
| Zhou et al. | Selective detection of zwitterionic arginine with a new Zn (II)-terpyridine complex: potential application in protein labeling and determination | |
| CN111687408A (en) | Fluorescent copper nanocluster, preparation method and application thereof | |
| CN113913182A (en) | Fluorescent probe for cosmetic deterioration viscosity detection and preparation method and application thereof | |
| CN113340860A (en) | Manganese-doped carbon dot and Mn-CDs solution for detecting Fe3+, test paper, preparation method of test paper and detection method of test paper | |
| CN110669026B (en) | Fluorescent probe molecule for detecting nitrite and preparation method thereof | |
| Robrish et al. | The use of the o-phthalaldehyde reaction as a sensitive assay for protein and to determine protein in bacterial cells and dental plaque | |
| CN110412000B (en) | Fluorescent probe for detecting L-tryptophan based on ten-element cucurbituril and detection method thereof | |
| CN110018146B (en) | Method for detecting palladium ions based on fluorescent carbon quantum dots | |
| CN102435587B (en) | Method for rapidly determining nitrite in water through nanogold resonance scattering spectrometry | |
| CN110887819B (en) | Preparation and application of high-selectivity fluorescent probe for detecting phenylalanine | |
| CN110346338A (en) | A kind of fluorescence probe and preparation method thereof, biosensor and its construction method and application | |
| CN110376172B (en) | Method for identifying L-phenylalanine | |
| CN110411999B (en) | Fluorescent probe for detecting L-phenylalanine based on eight-membered cucurbituril and detection method thereof | |
| CN110527506A (en) | A kind of highly selective fluorescence probe and its application for detecting phenylalanine | |
| CN112362628A (en) | Fluorescent probe for detecting L-tryptophan | |
| CN110596056A (en) | Fluorescent probe for detecting L-phenylalanine based on seven-element cucurbituril and detection method thereof | |
| CN112251218B (en) | Preparation method of ethylenediamine functionalized carbon quantum dots and application of ethylenediamine functionalized carbon quantum dots in catechol detection | |
| CN110698390B (en) | Fluorescent probe for identifying bisulfite and preparation method and detection method thereof | |
| CN113933280A (en) | Fluorescent and resonant Rayleigh scattering light binary signal probe for detecting tetracycline and preparation method and application thereof | |
| CN113075179A (en) | Method for detecting ketamine by using supramolecular fluorescent probe | |
| CN116554181A (en) | Preparation method and application of fluorescent probe for detecting amino acid based on G5 pyridine dye molecule/methyl cucurbit [6] uril | |
| Zhu | A novel chemiluminescent flow injection analysis of trace amounts of rutin by its inhibition of the luminol–hydrogen peroxide reaction catalyzed by tetrasulfonated colbalt phthalocyanine |
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 |