CN107817230B - Detection method of tetracycline antibiotics based on composite system of graphene quantum dots and europium ions - Google Patents
Detection method of tetracycline antibiotics based on composite system of graphene quantum dots and europium ions Download PDFInfo
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- 229910052693 Europium Inorganic materials 0.000 title claims abstract description 31
- -1 europium ions Chemical class 0.000 title claims abstract description 31
- 239000002131 composite material Substances 0.000 title claims abstract description 24
- 229940072172 tetracycline antibiotic Drugs 0.000 title claims abstract description 21
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 title claims abstract description 11
- 238000001514 detection method Methods 0.000 title claims description 18
- 239000002096 quantum dot Substances 0.000 claims abstract description 37
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 36
- 229910021389 graphene Inorganic materials 0.000 claims abstract description 35
- XMEVHPAGJVLHIG-FMZCEJRJSA-N chembl454950 Chemical compound [Cl-].C1=CC=C2[C@](O)(C)[C@H]3C[C@H]4[C@H]([NH+](C)C)C(O)=C(C(N)=O)C(=O)[C@@]4(O)C(O)=C3C(=O)C2=C1O XMEVHPAGJVLHIG-FMZCEJRJSA-N 0.000 claims abstract description 28
- 229960004989 tetracycline hydrochloride Drugs 0.000 claims abstract description 28
- 238000012360 testing method Methods 0.000 claims abstract description 22
- 238000012921 fluorescence analysis Methods 0.000 claims abstract description 12
- 239000000243 solution Substances 0.000 claims description 76
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 48
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 39
- 239000007853 buffer solution Substances 0.000 claims description 11
- 239000012153 distilled water Substances 0.000 claims description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 11
- NNMXSTWQJRPBJZ-UHFFFAOYSA-K europium(iii) chloride Chemical compound Cl[Eu](Cl)Cl NNMXSTWQJRPBJZ-UHFFFAOYSA-K 0.000 claims description 9
- 238000010438 heat treatment Methods 0.000 claims description 5
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical group Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 claims description 4
- AWDWVTKHJOZOBQ-UHFFFAOYSA-K europium(3+);trichloride;hexahydrate Chemical compound O.O.O.O.O.O.[Cl-].[Cl-].[Cl-].[Eu+3] AWDWVTKHJOZOBQ-UHFFFAOYSA-K 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 3
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- 150000001336 alkenes Chemical class 0.000 claims 1
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- 239000010439 graphite Substances 0.000 claims 1
- 238000000034 method Methods 0.000 abstract description 15
- 230000035945 sensitivity Effects 0.000 abstract description 6
- 239000004098 Tetracycline Substances 0.000 description 15
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- 238000002189 fluorescence spectrum Methods 0.000 description 6
- 230000005284 excitation Effects 0.000 description 5
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- 238000004458 analytical method Methods 0.000 description 2
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- 238000001000 micrograph Methods 0.000 description 2
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- XIYOPDCBBDCGOE-IWVLMIASSA-N (4s,4ar,5s,5ar,12ar)-4-(dimethylamino)-1,5,10,11,12a-pentahydroxy-6-methylidene-3,12-dioxo-4,4a,5,5a-tetrahydrotetracene-2-carboxamide Chemical class C=C1C2=CC=CC(O)=C2C(O)=C2[C@@H]1[C@H](O)[C@H]1[C@H](N(C)C)C(=O)C(C(N)=O)=C(O)[C@@]1(O)C2=O XIYOPDCBBDCGOE-IWVLMIASSA-N 0.000 description 1
- 241000186361 Actinobacteria <class> Species 0.000 description 1
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- 239000004100 Oxytetracycline Substances 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- IOYNQIMAUDJVEI-BMVIKAAMSA-N Tepraloxydim Chemical group C1C(=O)C(C(=N/OC\C=C\Cl)/CC)=C(O)CC1C1CCOCC1 IOYNQIMAUDJVEI-BMVIKAAMSA-N 0.000 description 1
- XBDYBAVJXHJMNQ-UHFFFAOYSA-N Tetrahydroanthracene Natural products C1=CC=C2C=C(CCCC3)C3=CC2=C1 XBDYBAVJXHJMNQ-UHFFFAOYSA-N 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 229960003022 amoxicillin Drugs 0.000 description 1
- LSQZJLSUYDQPKJ-NJBDSQKTSA-N amoxicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=C(O)C=C1 LSQZJLSUYDQPKJ-NJBDSQKTSA-N 0.000 description 1
- 229960004099 azithromycin Drugs 0.000 description 1
- MQTOSJVFKKJCRP-BICOPXKESA-N azithromycin Chemical compound O([C@@H]1[C@@H](C)C(=O)O[C@@H]([C@@]([C@H](O)[C@@H](C)N(C)C[C@H](C)C[C@@](C)(O)[C@H](O[C@H]2[C@@H]([C@H](C[C@@H](C)O2)N(C)C)O)[C@H]1C)(C)O)CC)[C@H]1C[C@@](C)(OC)[C@@H](O)[C@H](C)O1 MQTOSJVFKKJCRP-BICOPXKESA-N 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 238000005251 capillar electrophoresis Methods 0.000 description 1
- 239000011852 carbon nanoparticle Substances 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 229940124587 cephalosporin Drugs 0.000 description 1
- 150000001780 cephalosporins Chemical class 0.000 description 1
- CYDMQBQPVICBEU-UHFFFAOYSA-N chlorotetracycline Natural products C1=CC(Cl)=C2C(O)(C)C3CC4C(N(C)C)C(O)=C(C(N)=O)C(=O)C4(O)C(O)=C3C(=O)C2=C1O CYDMQBQPVICBEU-UHFFFAOYSA-N 0.000 description 1
- 229960004475 chlortetracycline Drugs 0.000 description 1
- CYDMQBQPVICBEU-XRNKAMNCSA-N chlortetracycline Chemical compound C1=CC(Cl)=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 CYDMQBQPVICBEU-XRNKAMNCSA-N 0.000 description 1
- 235000019365 chlortetracycline Nutrition 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000003271 compound fluorescence assay Methods 0.000 description 1
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- 230000002708 enhancing effect Effects 0.000 description 1
- OGPBJKLSAFTDLK-UHFFFAOYSA-N europium atom Chemical compound [Eu] OGPBJKLSAFTDLK-UHFFFAOYSA-N 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
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- 230000004048 modification Effects 0.000 description 1
- 239000002086 nanomaterial Substances 0.000 description 1
- 229960000625 oxytetracycline Drugs 0.000 description 1
- IWVCMVBTMGNXQD-PXOLEDIWSA-N oxytetracycline Chemical compound C1=CC=C2[C@](O)(C)[C@H]3[C@H](O)[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 IWVCMVBTMGNXQD-PXOLEDIWSA-N 0.000 description 1
- 235000019366 oxytetracycline Nutrition 0.000 description 1
- LSQZJLSUYDQPKJ-UHFFFAOYSA-N p-Hydroxyampicillin Natural products O=C1N2C(C(O)=O)C(C)(C)SC2C1NC(=O)C(N)C1=CC=C(O)C=C1 LSQZJLSUYDQPKJ-UHFFFAOYSA-N 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 238000010791 quenching Methods 0.000 description 1
- 230000000171 quenching effect Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- IWVCMVBTMGNXQD-UHFFFAOYSA-N terramycin dehydrate Natural products C1=CC=C2C(O)(C)C3C(O)C4C(N(C)C)C(O)=C(C(N)=O)C(=O)C4(O)C(O)=C3C(=O)C2=C1O IWVCMVBTMGNXQD-UHFFFAOYSA-N 0.000 description 1
- 150000003518 tetracenes Chemical class 0.000 description 1
- 229940040944 tetracyclines Drugs 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- 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
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N2021/6432—Quenching
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- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Optics & Photonics (AREA)
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- Analytical Chemistry (AREA)
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- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
本发明提供了一种基于石墨烯量子点与铕离子复合体系的四环素类抗生素检测方法，其特征在于，包括以下步骤：A.制备石墨烯量子点溶液；B.制备铕离子溶液；C.制备不同浓度的盐酸四环素溶液；D.将石墨烯量子点溶液和铕离子溶液混合得到复合体系溶液；E.将不同浓度的盐酸四环素溶液依次与复合体系溶液混合，通过荧光分析法绘制盐酸四环素浓度的标准曲线；F.将未知浓度的盐酸四环素溶液与复合体系溶液混合，通过荧光测试得到荧光强度值，通过与标准曲线的比对得到浓度数据。本发明的信号输出方式为比率荧光型，并具有操作简便、耗时短、灵敏度高以及干扰小等优点。
The invention provides a method for detecting tetracycline antibiotics based on a composite system of graphene quantum dots and europium ions, which is characterized by comprising the following steps: A. preparing a graphene quantum dot solution; B. preparing a europium ion solution; C. preparing Tetracycline hydrochloride solutions of different concentrations; D. The graphene quantum dot solution and the europium ion solution are mixed to obtain a composite system solution; E. The tetracycline hydrochloride solutions of different concentrations are mixed with the composite system solution in turn, and the concentration of tetracycline hydrochloride is drawn by fluorescence analysis. Standard curve; F. Mix the tetracycline hydrochloride solution of unknown concentration with the composite system solution, obtain the fluorescence intensity value through the fluorescence test, and obtain the concentration data through the comparison with the standard curve. The signal output method of the present invention is a ratiometric fluorescence type, and has the advantages of simple operation, short time-consuming, high sensitivity and little interference.
Description
Technical Field
The invention belongs to the field of chemical detection, and particularly relates to a tetracycline antibiotic detection method based on a graphene quantum dot and europium ion composite system.
Background
The tetracycline antibiotics are a broad-spectrum antibiotics produced by actinomycetes, and include aureomycin, oxytetracycline, tetracycline and semisynthetic derivatives of methacycline, doxycycline, dimethylamino tetracycline, and the like, and the structures of the antibiotics all contain the basic skeleton of tetracene. As a broad spectrum antibiotic, contamination by abuse of tetracycline antibiotics has attracted attention.
The existing common analysis and detection methods of tetracycline antibiotics include liquid chromatography, liquid chromatography-mass spectrometry, capillary electrophoresis, colorimetry, fluorescence analysis and the like. However, these methods generally have some disadvantages, such as complicated operation of chromatography and electrophoresis, long analysis time, and insufficient sensitivity of colorimetry. The fluorescence analysis method is simple to operate and high in sensitivity, so that the development of the fluorescence analysis method of the tetracycline antibiotics is of great significance. Existing fluorescence analysis methods for antibiotics are roughly divided into two types, One is established by quenching fluorescent nanomaterials (such as fluorescent carbon dots) with tetracycline antibiotics (x. Yang, y. Luo, s. Zhu, y. Feng, y. Zhuo, y. Dou, One-potyntheses of high fluorescence carbon nanoparticles and the above applications for detection of tetracyclines, biosen. bioelectrode.56 (2014) 6-11.), and the other is established by forming luminescent complexes with tetracycline using Europium ions (l.d.s. Teixeira, a.n. Grasso, a.m. Monteiro, a.m.f. Neto, n.d. vireira jr., m. Giundl.c. coupled, and the same as 20. c.e. 20 and 19. the same of the same type), and the other is established by using Europium ions and tetracycline to form luminescent complexes with tetracycline. The first kind of fluorescence analysis method is a fluorescence reduction type, and the second kind of fluorescence analysis method needs to search and add a substance capable of enhancing the fluorescence of the europium luminescent complex into the system because the luminescent complex formed by europium ions and tetracycline has weak fluorescence intensity.
The present invention has been made in view of the above problems.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides a tetracycline antibiotic detection method based on a graphene quantum dot and europium ion composite system, and the detection method is solved by the following technical scheme.
The tetracycline antibiotic detection method based on the graphene quantum dot and europium ion composite system is characterized by comprising the following steps of: A. preparing a graphene quantum dot solution; B. preparing a europium ion solution; C. preparing tetracycline hydrochloride solutions with different concentrations; D. mixing the graphene quantum dot solution and the europium ion solution to obtain a composite system solution; E. sequentially mixing tetracycline hydrochloride solutions with different concentrations with the composite system solution, and drawing a standard curve of the tetracycline hydrochloride concentration by a fluorescence analysis method; F. and mixing the tetracycline hydrochloride solution with unknown concentration with the composite system solution, obtaining a fluorescence intensity value through fluorescence test, and obtaining concentration data through comparison with a standard curve.
Preferably, in the step a, the preparation of the graphene quantum dot solution comprises the following steps: A1. putting solid citric acid into a beaker, putting the beaker into a heating sleeve, and heating at 200 ℃ until the citric acid becomes liquid and the color of the citric acid changes from colorless to light yellow; A2. and mixing the molten citric acid with a sodium hydroxide solution to obtain a graphene quantum dot solution.
Preferably, in the step a2, the mixing operation of citric acid and sodium hydroxide solution is as follows: and placing the beaker together with the molten citric acid into a container containing 10mg/mL sodium hydroxide solution, inclining the beaker, and slowly introducing the sodium hydroxide solution in the container into the beaker to obtain the graphene quantum dot solution.
Preferably, in the step B, the preparation of the europium ion solution comprises the following steps: europium chloride hexahydrate is taken and added into distilled water to obtain a europium chloride solution.
Preferably, in step E, F, the buffer solution used in the fluorescence assay test is a Tris-HCl buffer solution, the concentration of the Tris-HCl buffer solution is 9-11 mM, and the pH value is 8-10.
Preferably, in the step E, F, the concentration of the graphene quantum dot solution used in the fluorescence analysis test is 10 to 11 μ g/mL, and the concentration of the europium ion solution is 19 to 21 μ M.
Compared with the prior art, the invention provides a method for detecting tetracycline antibiotics by using a graphene quantum dot and europium ion composite system, wherein the graphene quantum dot has two functions: on one hand, the fluorescence of the graphene quantum dot can be quenched by tetracycline antibiotics, and on the other hand, the surface of the graphene quantum dot prepared by the method is rich in carboxyl, and can be coordinated with europium ions, so that the intensity of a luminescent complex formed by the europium ions and the tetracycline can be enhanced; therefore, the signal output mode of the invention is a ratio fluorescence type, and has the advantages of simple operation, short time consumption, high sensitivity, small interference and the like.
Drawings
Fig. 1 shows an electron microscope image of the graphene quantum dot prepared in step 1 in example 1 of the present invention.
Fig. 2 shows a fluorescence spectrum of a graphene quantum dot and europium ion composite system obtained in step 4 in example 1 of the present invention under the condition of adding tetracycline hydrochloride of different concentrations.
FIG. 3 shows a standard curve of fluorescence intensity ratio F616/F460 and tetracycline hydrochloride concentration plotted in step 4 of example 1 of the present invention.
FIG. 4 shows a test curve of the addition of tetracycline hydrochloride at a concentration of 20. mu.M in example 1 of the present invention.
FIG. 5 shows a test curve of example 2 of the present invention in which tetracycline hydrochloride was added at a concentration of 1 mM.
FIG. 6 shows a standard curve of fluorescence intensity ratio F616/F460 and doxycycline concentration plotted in step 4 of example 3 of the present invention.
FIG. 7 shows a test curve of the addition of tetracycline hydrochloride at a concentration of 250. mu.M in example 3 of the present invention.
Detailed Description
The invention is described in further detail below with reference to specific embodiments and with reference to the following drawings.
Example 1: preparing tetracycline hydrochloride solution with the concentration of 20 mu M as a solution to be detected, and comprising the following steps:
(1) 2.1 g of citric acid was placed in a 25 mL beaker, which was then heated in a digital 200 ℃ heating mantle. After about 5 minutes the citric acid became liquid and changed its color from colorless to light yellow. Then, placing the small beaker in a large beaker filled with 100 mL of 10mg/mL sodium hydroxide by using a pair of tweezers, and inclining the small beaker to enable the sodium hydroxide solution to slowly enter the small beaker to obtain a solution-state graphene quantum dot aqueous solution, wherein an electron microscope image of the aqueous solution is shown in fig. 1, and the graphene quantum dots are uniformly distributed.
(2) And (2) taking 5 mL of the graphene quantum dot solution obtained in the step (1), and adding 5 mL of distilled water to obtain the graphene quantum dot solution with the mass concentration of 10.5 mg/mL.
(3) 0.0258 g of europium chloride hexahydrate was weighed out and 10 mL of distilled water was added to obtain a 10 mM europium chloride solution.
(4) 2 mu L of graphene quantum dot solution with the concentration of 10.5mg/mL, 4 mu L of europium chloride solution with the concentration of 10 mM, 200 mu L of buffer solution with the concentration of 100 mM and the pH value of 9.0, tetracycline hydrochloride solution and distilled water are added into a cuvette, so that the total test system is 2 mL (the concentration of tetracycline in the test system is 0, 0.1, 0.2, 0.5, 1, 2, 3, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22 and 24 mu M), and after uniform mixing, waiting for one minute, the dispersion is uniform. The fluorescence spectrum of the system at 400-650 nm is recorded with 360 nm as the excitation wavelength, as shown in FIG. 2. A standard curve for tetracycline hydrochloride detection is drawn by taking the tetracycline concentration as the abscissa and the fluorescence intensity ratio F616/F460 at 616 nm to 460 nm as the ordinate, as shown in FIG. 3.
(5) Adding 2 mu L of graphene quantum dot solution with the concentration of 10.5mg/mL, 4 mu L of europium chloride solution with the concentration of 10 mM, 200 mu L of buffer solution with the concentration of 100 mM and the pH value of 9.0, 20 mu L of tetracycline hydrochloride solution to be detected and distilled water into a cuvette to enable the total test system to be 2 mL, uniformly mixing, and waiting for one minute. The fluorescence spectrum of the system at 400-650 nm was recorded with 360 nm as the excitation wavelength, as shown in FIG. 4. The fluorescence intensity at 616 nm and 460 nm was read and the ratio of F616/F460 was calculated. Substituting the ratio into the standard curve to obtain the concentration of tetracycline in the test system of 196.16 nM, and further to obtain the concentration of tetracycline in the solution to be tested of 19.16 μ M with error of 4.2%.
Example 2: preparing tetracycline hydrochloride solution with the concentration of 1mM as a solution to be detected, and comprising the following steps:
(1) the standard curve for tetracycline hydrochloride detection was the same as in example 1.
(2) Adding 2 mu L of graphene quantum dot solution with the concentration of 10.5mg/mL, 4 mu L of europium chloride solution with the concentration of 10 mM, 200 mu L of buffer solution with the concentration of 100 mM and the pH value of 9.0, 20 mu L of tetracycline hydrochloride solution to be detected and distilled water into a cuvette to enable the total test system to be 2 mL, uniformly mixing, and waiting for one minute. The fluorescence spectrum of the system at 400-650 nm was recorded with 360 nm as the excitation wavelength, as shown in FIG. 5. The fluorescence intensity at 616 nm and 460 nm was read and the ratio of F616/F460 was calculated. Substituting the ratio into the standard curve to obtain the concentration of tetracycline in the test system of 9.794 μ M, and further obtaining the concentration of tetracycline in the solution to be tested of 979.4 μ M with an error of 2.06%.
Example 3: preparing a doxycycline solution with the concentration of 250 mu M as a solution to be detected, and comprising the following steps:
(1) 2.1 g of citric acid was placed in a 25 mL beaker, which was then heated in a digital 200 ℃ heating mantle. After about 5 minutes the citric acid became liquid and changed its color from colorless to light yellow. And then placing the small beaker into a large beaker filled with 100 mL of 10mg/mL sodium hydroxide by using forceps, and inclining the small beaker to ensure that the sodium hydroxide solution slowly enters the small beaker to obtain a solution-state graphene quantum dot aqueous solution.
(2) And (2) taking 5 mL of the quantum dot solution obtained in the step (1), and adding 5 mL of distilled water to obtain a graphene quantum dot solution with the mass concentration of 10.5 mg/mL.
(3) 0.0258 g of europium chloride hexahydrate was weighed out and 10 mL of distilled water was added to obtain a 10 mM europium chloride solution.
(4) 2 μ L of graphene quantum dot solution with a concentration of 10.5mg/mL, 4 μ L of europium chloride solution with a concentration of 10 mM, 200 μ L of buffer solution with a concentration of 100 mM and a pH value of 9.0, doxycycline solution and distilled water were added to a cuvette, so that the total test system was 2 mL (the concentration of tetracycline in the test system was 0, 0.1, 0.2, 0.5, 1, 2, 3, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24 μ M), and the fluorescence spectrum of the system at 400-650 nm was recorded with 360 nm as an excitation wavelength. A doxycycline standard curve is drawn by using the doxycycline concentration as the abscissa and using the ratio of the fluorescence intensity at 616 nm to that at 460 nm, F616/F460, as the ordinate, as shown in FIG. 6.
(5) 2 mu L of graphene quantum dot solution with the concentration of 10.5mg/mL, 4 mu L of europium chloride solution with the concentration of 10 mM, 200 mu L of buffer solution with the concentration of 100 mM and the pH value of 9.0, 20 mu L of doxycycline solution to be tested and distilled water are added into a cuvette, so that the total test system is 2 mL, mixed evenly and waited for one minute. The fluorescence spectrum of the system at 400-650 nm was recorded with 360 nm as the excitation wavelength, as shown in FIG. 7. The fluorescence intensity at 616 nm and 460 nm was read and the ratio of F616/F460 was calculated. Substituting the ratio into the standard curve to obtain the doxycycline concentration of 2.435 μ M in the test system, and further to obtain the doxycycline concentration of 243.5 μ M in the solution to be tested with an error of 2.6%.
The method for detecting the tetracycline antibiotics can be used for detecting the tetracycline hydrochloride only by simple mixing, is simple to operate and high in speed, and is high in sensitivity, the linear detection range is 0-20 mu M, and the sensitivity is 8.2 nM.
In the detection method, the fluorescence intensity changes obviously along with different tetracycline hydrochloride concentrations, the fluorescence intensity at 616 nm is obviously increased along with the increase of the tetracycline hydrochloride concentration, and the fluorescence intensity at 460 nm is reduced along with the increase of the tetracycline hydrochloride concentration, which are consistent with two effects of the addition of the graphene quantum dot solution.
In addition, in the invention, the fluorescence signal is in a ratio type, is less influenced by a light source and test conditions, and can effectively reduce test errors; the used graphene quantum dot material is simple to prepare, small in dosage and easy to store for a long time, so that the method is low in cost. The method has good selectivity, obvious fluorescent signal change can be generated when tetracycline antibiotics are added into a system of the graphene quantum dots and europium ions, and no fluorescent signal change is generated when several other common antibiotics (such as streptomycin, penicillin, azithromycin, amoxicillin and cephalosporins) are added.
The scope of the present invention includes, but is not limited to, the above embodiments, and the present invention is defined by the appended claims, and any alterations, modifications, and improvements that may occur to those skilled in the art are all within the scope of the present invention.
Claims (6)
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Non-Patent Citations (4)
| Title |
|---|
| Europium-decorated graphene quantum dots as a fluorescent probe for label-free, rapid and sensitive detection of Cu2+ and l-cysteine;Liping Lin et.al;《Analytica Chimica Acta》;elsevier;20150822;第891卷;261-268 * |
| Graphene Quantum Dots Combined with Europium Ions as Photoluminescent Probes for Phosphate Sensing;Jiang-Mei Bai et.al;《Chemistry A European Journal》;Wiley;20130218;第19卷;3822-3826 * |
| 基于KGdF4:Tb3+纳米材料检测四环素的生物传感新方法;段诺等;《食品与生物技术学报》;20141231;第33卷(第11期);1160-1167 * |
| 荧光共振能量转移技术在食品分析检测中的应用;袁云霞等;《食品安全质量检测学报》;20170228;第8卷(第2期);434-437 * |
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