CN118225746A - Method for rapidly, simply, conveniently and highly sensitively quantitatively detecting tetracycline in water sample - Google Patents
Method for rapidly, simply, conveniently and highly sensitively quantitatively detecting tetracycline in water sample Download PDFInfo
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- CN118225746A CN118225746A CN202410305450.5A CN202410305450A CN118225746A CN 118225746 A CN118225746 A CN 118225746A CN 202410305450 A CN202410305450 A CN 202410305450A CN 118225746 A CN118225746 A CN 118225746A
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- 239000004098 Tetracycline Substances 0.000 title claims abstract description 81
- 235000019364 tetracycline Nutrition 0.000 title claims abstract description 81
- 150000003522 tetracyclines Chemical class 0.000 title claims abstract description 76
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 title claims abstract description 22
- 238000000034 method Methods 0.000 title claims abstract description 19
- 229960002180 tetracycline Drugs 0.000 title claims abstract description 14
- 229930101283 tetracycline Natural products 0.000 title claims abstract description 14
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 claims abstract description 28
- 238000001514 detection method Methods 0.000 claims abstract description 27
- 239000007853 buffer solution Substances 0.000 claims abstract description 18
- 239000011259 mixed solution Substances 0.000 claims abstract description 6
- 238000002189 fluorescence spectrum Methods 0.000 claims description 10
- 239000000243 solution Substances 0.000 claims description 10
- 238000005259 measurement Methods 0.000 claims description 9
- 238000006243 chemical reaction Methods 0.000 claims description 4
- DBLXOVFQHHSKRC-UHFFFAOYSA-N ethanesulfonic acid;2-piperazin-1-ylethanol Chemical compound CCS(O)(=O)=O.OCCN1CCNCC1 DBLXOVFQHHSKRC-UHFFFAOYSA-N 0.000 claims description 4
- 239000011541 reaction mixture Substances 0.000 claims description 2
- 230000000694 effects Effects 0.000 abstract description 10
- 230000008859 change Effects 0.000 abstract description 7
- 230000035945 sensitivity Effects 0.000 abstract description 5
- 238000004020 luminiscence type Methods 0.000 abstract description 3
- 238000004445 quantitative analysis Methods 0.000 abstract description 3
- OFVLGDICTFRJMM-WESIUVDSSA-N tetracycline Chemical compound C1=CC=C2[C@](O)(C)[C@H]3C[C@H]4[C@H](N(C)C)C(O)=C(C(N)=O)C(=O)[C@@]4(O)C(O)=C3C(=O)C2=C1O OFVLGDICTFRJMM-WESIUVDSSA-N 0.000 abstract 5
- 229940040944 tetracyclines Drugs 0.000 description 67
- 230000004044 response Effects 0.000 description 5
- 238000005457 optimization Methods 0.000 description 4
- 230000035484 reaction time Effects 0.000 description 4
- 230000005284 excitation Effects 0.000 description 3
- 238000000695 excitation spectrum Methods 0.000 description 3
- 239000012621 metal-organic framework Substances 0.000 description 3
- 229910052693 Europium Inorganic materials 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 238000000862 absorption spectrum Methods 0.000 description 2
- 238000000295 emission spectrum Methods 0.000 description 2
- 238000001506 fluorescence spectroscopy Methods 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 239000008399 tap water Substances 0.000 description 2
- 235000020679 tap water Nutrition 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 229910002538 Eu(NO3)3·6H2O Inorganic materials 0.000 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- 238000003917 TEM image Methods 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 235000019730 animal feed additive Nutrition 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000002924 anti-infective effect Effects 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000003899 bactericide agent Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000012742 biochemical analysis Methods 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000004993 emission spectroscopy Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- JVYYYCWKSSSCEI-UHFFFAOYSA-N europium(3+);trinitrate;hexahydrate Chemical compound O.O.O.O.O.O.[Eu+3].[O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O JVYYYCWKSSSCEI-UHFFFAOYSA-N 0.000 description 1
- 238000012921 fluorescence analysis Methods 0.000 description 1
- 230000007160 gastrointestinal dysfunction Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 238000012417 linear regression Methods 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000002086 nanomaterial Substances 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000002352 surface water Substances 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 238000004809 thin layer chromatography Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
Landscapes
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The invention discloses a method for rapidly, simply and conveniently quantitatively detecting Tetracycline (TC) in a water sample with high sensitivity, wherein the TC can generate Eu MOF which emits red fluorescence when being added into a mixed solution containing Eu3+ (1 mM) and Hepes buffer solution. The addition of TC can generate an antenna effect to obviously enhance the luminescence of Eu3+, the measured fluorescence signal of Eu MOF is closely related to the concentration of TC, and quantitative analysis and detection of TC are realized through the change of the fluorescence signal of Eu MOF.
Description
Technical Field
The invention belongs to the fields of nano materials, fluorescence sensing technology and biochemical analysis and detection, and in particular relates to a fluorescence sensing platform based on a metal organic framework, which can rapidly, simply and conveniently quantitatively detect tetracycline with high sensitivity.
Background
Tetracyclines (TCs) are one of the spectral antibiotics found in the forty of the twentieth century to treat human and animal infections caused by gram-positive and negative bacteria. Due to its high anti-infective effectiveness and low cost, it has been widely used in bactericides, medicine and animal feed additives. However, improper use or abuse may cause excessive or residual amounts of TC, which is difficult to degrade, may accumulate in food, soil, and surface water, and may ultimately cause serious damage to animal and human health, such as allergic reactions, gastrointestinal dysfunction, drug resistance, and damage to organs such as the liver. Therefore, it is necessary to establish a rapid, simple and highly sensitive quantitative detection method of TC.
The traditional analysis methods for detecting TC, such as an enzyme-linked immunosorbent assay, a thin layer chromatography, a high performance liquid chromatography, a lateral chromatography detection method, a chromatography-mass spectrometry combination method and the like, have the advantages of high accuracy, low detection limit and the like, and become conventional methods for detecting TC. However, these methods often require cumbersome sample pretreatment, specialized handling skills, and complex and expensive instrumentation, etc., which are somewhat limited in TC detection applications. The fluorescence analysis method has the advantages of simple operation, low cost, high response speed, good selectivity, high sensitivity and the like, and is paid attention to. To our knowledge, there is no method for adding TC, hepes and Eu 3+
The reaction generates luminescent EuMOF nano particles to carry out quantitative analysis and detection of TC.
Disclosure of Invention
Aiming at the problems of the prior art, the invention aims to provide a method for rapidly, simply and conveniently quantitatively detecting the tetracycline in a water sample with high sensitivity.
In order to achieve the above purpose, the technical scheme and steps adopted by the invention are as follows:
TC of different concentrations was added to a buffer solution containing Eu 3+ (final concentration 50. Mu.M) of 4-hydroxyethyl piperazine ethane sulfonic acid (Hepes, final concentration 25 mM), the final volume was 100. Mu.L, and the mixture was mixed, incubated at room temperature for 2min, and a red-fluorescence-emitting EuMOF was synthesized, and fluorescence spectrum measurement was performed.
The invention quantitatively detects TC principle in water: when the TC solution is mixed with a Hepes buffer solution containing Eu 3+ (1 mM), eu MOF is generated. When no TC is added, eu 3+ in the solution is spin-forbidden according to Laplat rule f-f, and fluorescence emission intensity is weak. After adding TC, the TC can effectively transfer the absorbed photon energy to Eu 3+ through antenna effect to sensitize the luminescence of Eu 3+, so that the luminescence of Eu 3+ is obviously enhanced, a fluorescence signal detected in the solution is closely related to the concentration of TC, and quantitative analysis and detection of TC are realized through the change of the fluorescence signal of Eu MOF.
The beneficial effects are that: according to the invention, a novel method for quantitatively analyzing and detecting TC in a water sample is constructed by adding different fluorescence signals of red light Eu MOF synthesized by TC with different concentrations. Compared with the traditional TC detection method, the method has the advantages of simplicity in operation, low cost, rapid response, high sensitivity and good selectivity. The whole detection process is simple, environment-friendly and safe.
Drawings
FIG. 1A is a graph showing fluorescence spectra of the components of example 1 alone and in combination. As shown, the characteristic emission peak of Eu MOF at 615nm is strongest only when TC and Eu 3+, hepes buffer solutions are co-present. FIG. 1B is a graph of the UV-visible absorption spectrum of example 1 when the components are present alone and in combination. As shown, the Eu MOF exhibits a characteristic absorption peak at 397nm only when TC and Eu 3+, hepes buffer solutions are present together.
FIG. 2A is the excitation and emission spectra of the EuMOF synthesized after TC addition, with the excitation spectra being strongest at 397nm and the characteristic fluorescence emission wavelengths being strongest at 615 nm. Fig. 2B is a transmission electron micrograph of the Tb MOF prepared in example 1, with particles approximately circular and uniformly dispersed.
FIG. 3A is a graph of the optimization of the reaction time for detecting TC in example 1. As shown, the ratio of fluorescence intensities was saturated when the reaction time reached 2min.
FIG. 3B is a chart of optimization of Hepes buffer concentration for TC detection in example 1. As shown, hepes concentration was 25mM with optimal signal response.
FIG. 3C is a solution pH optimization graph for TC detection in example 1. As shown, the pH was 7.0 with the best signal response.
FIG. 3D is a graph showing the Eu 3+ concentration optimization for TC detection in example 1. As shown, eu 3+ has an optimal signal response at a concentration of 50. Mu.M.
FIG. 4A is a graph showing the fluorescence change spectrum of TC concentration and Eu MOF formation in example 1. As shown, the fluorescence of EuMOF gradually increases with increasing TC concentration.
FIG. 4B is a graph showing the correlation between TC concentration and fluorescence intensity value at 615nm of Eu MOF generated in example 1; as shown, the Eu MOF increases in fluorescence intensity value at 615nm gradually with increasing TC concentration, and exhibits a good linear relationship, and the linear regression equation is y=0.1986 [ TC ] +0.0781, and the correlation coefficient r 2 =1.000.
Detailed Description
Tetracyclines (TC), 4-hydroxyethylpiperazine ethanesulfonic acid (Hepes) (biotechnology company); europium nitrate hexahydrate (Eu (NO 3)3·6H2 O) (hadamard reagent limited).
Example 1 feasibility verification of Eu MOF-based fluorescent nanoprobe to detect TC, the procedure is as follows:
To a certain amount of sterilized water, hepes buffer solution of pH7.0, eu 3+, TC, hepes buffer solution of pH 7.0+Eu 3+, hepes buffer solution of pH 7.0+TC, eu 3+ +TC, hepes buffer solution of pH 7.0+Eu 3+ +TC were added, respectively, to give a sample volume of 100. Mu.L, eu 3+ was 50. Mu.M, TC was 150. Mu.M, and Hepes was 25mM. All samples were mixed and incubated at room temperature for 2min for fluorescence spectroscopy and UV-visible absorbance spectroscopy measurements. The fluorescence and UV-visible absorption spectra are shown in FIGS. 1A and 1B, respectively.
Example 2 excitation and emission spectrometry of Eu MOF and particle morphology characterization, the procedure is as follows:
To a certain amount of sterilized water, hepes buffer solution of pH 7.0, eu 3+ and TC were added in this order, respectively, with a final sample volume of 100. Mu.L, eu 3+ final concentration of 50. Mu.M, TC final concentration of 150. Mu.M, hepes final concentration of 25mM. The samples were mixed well and incubated at room temperature for 2min for fluorescence spectroscopy. The fluorescence excitation and emission spectra are shown in FIG. 2A, and the transmission electron microscope is shown in FIG. 2B.
Example 3A the effect of Hepes concentration on TC detection was examined as follows:
Hepes buffer solution, eu 3+ and TC of different concentrations are sequentially added into a certain amount of sterilized water respectively, the final volume of the sample is 100 mu L, the final concentration of Eu 3+ is 50 mu M, and the final concentration of TC is 150 mu M. The samples were mixed and subjected to fluorescence spectrum measurement at room temperature for different times, the change in the fluorescence intensity at 615nm was recorded, and the effect of the solution pH on TC detection was examined, and the results are shown in FIG. 3A.
Example 3B examines the effect of Eu 3+ concentration on TC detection as follows:
To a certain amount of sterilized water, hepes buffer solution of pH7.0, eu 3+ of different concentrations, TC of 100. Mu.L in final sample volume, TC of 150. Mu.M in final Hepes concentration of 25mM were sequentially added, respectively. The samples were mixed and reacted at room temperature for 2min, the fluorescence spectrum was measured, the change in the fluorescence intensity at 615nm was recorded, and the effect of Eu 3+ concentration on TC detection was examined, and the result was shown in FIG. 3B.
Example 3C the effect of solution pH on TC detection was examined as follows:
To a certain amount of sterilized water, hepes buffer solution of different pH, eu 3+ and TC were sequentially added, the final volume of the sample was 100. Mu.L, eu 3+ was 50. Mu.M, TC was 150. Mu.M, and Hepes was 25mM. The samples were mixed and subjected to fluorescence spectrum measurements at room temperature for different times of reaction, the change in the value of fluorescence intensity at 615nm was recorded, and the effect of the solution pH on TC detection was examined, and the results are shown in FIG. 3C.
Example 3D the effect of reaction time on TC detection was examined as follows:
To a certain amount of sterilized water, hepes buffer solution of pH7.0, eu 3+ and TC were sequentially added, the final volume of the sample was 100. Mu.L, eu 3+ was 50. Mu.M, TC was 150. Mu.M, and Hepes was 25mM. The samples were mixed and subjected to fluorescence spectrum measurement at room temperature for different times of reaction, the numerical change of fluorescence intensity at 615nm was recorded, and the effect of reaction time on TC detection was examined, and the results are shown in FIG. 3D.
Example 4TC detection, the procedure is as follows:
TC solutions (10. Mu.L) of different concentrations were added to a buffer solution containing Eu 3+ (final concentration 50. Mu.M) of 4-hydroxyethylpiperazine ethanesulfonic acid (Hepes, final concentration 25 mM). Mixing uniformly, incubating for 2min at room temperature, synthesizing Eu MOF which emits red fluorescence, and performing fluorescence spectrum measurement. Wherein the final volume of the reaction mixture solution is 100. Mu.L, the final Eu 3+ concentration is 50. Mu.M, and the final Hepes concentration is 25mM.
Example 5 detection of TC in tap water, the procedure is as follows:
Tap water filtered by a 0.22 mu m filter membrane and TC with different concentrations are added into a mixed solution containing a pH7.0hepes buffer solution and Eu 3+, all samples are uniformly mixed, incubated for 2min at room temperature, 90 mu L of the mixed solution is taken for fluorescence spectrum measurement, and the content of TC in lake water is calculated according to a standard curve. The TC contents in the lake water detected by the five samples in the examples are shown in table 1, respectively, and each detection result is within the error range.
TABLE 1
Example 6 detection of TC in lake water the following steps are followed:
Adding lake water filtered by a 0.22 mu m filter membrane and TC with different concentrations into a mixed solution containing a pH7.0hepes buffer solution and Eu 3+, uniformly mixing all samples, incubating for 2min at room temperature, taking 90 mu L for fluorescence spectrum measurement, and calculating according to a standard curve to obtain the content of TC in the lake water. The TC contents in the lake water detected by the five samples in the examples are shown in table 2, respectively, and each detection result is within the error range.
TABLE 2
Claims (4)
1. A method for rapidly, simply, conveniently and highly sensitively quantitatively detecting tetracycline in a water sample comprises the following steps:
TC (10. Mu.L) with different concentrations is added into a mixed solution containing Eu 3+ and 4-hydroxyethyl piperazine ethane sulfonic acid (Hepes) buffer solution, the mixed solution is uniformly mixed, and incubated for 2min at room temperature, eu MOF which emits red fluorescence is synthesized, and fluorescence spectrum measurement is carried out. Wherein the final volume of the reaction mixture solution is 100. Mu.L, the final Eu 3+ concentration is 50. Mu.M, and the final Hepes concentration is 25mM.
2. The method for quantitatively detecting tetracycline in a water sample of claim 1, wherein the Eu MOF emits red light at a wavelength of 615 nm.
3. The method for quantitatively detecting tetracycline in a water sample of claim 1, wherein the reaction conditions are room temperature for 2min.
4. The method for quantitatively detecting tetracycline in a water sample of claim 1, wherein the fluorescent detection is by a conventional microplate reader.
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