CN109180707A - Fluorescence probe and its preparation method and application for detecting xanthine - Google Patents
Fluorescence probe and its preparation method and application for detecting xanthine Download PDFInfo
- Publication number
- CN109180707A CN109180707A CN201811168452.5A CN201811168452A CN109180707A CN 109180707 A CN109180707 A CN 109180707A CN 201811168452 A CN201811168452 A CN 201811168452A CN 109180707 A CN109180707 A CN 109180707A
- Authority
- CN
- China
- Prior art keywords
- xanthine
- dtpa
- fluorescence
- fluorescence probe
- 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.)
- Granted
Links
- LRFVTYWOQMYALW-UHFFFAOYSA-N 9H-xanthine Chemical compound O=C1NC(=O)NC2=C1NC=N2 LRFVTYWOQMYALW-UHFFFAOYSA-N 0.000 title claims abstract description 254
- 229940075420 xanthine Drugs 0.000 title claims abstract description 127
- 239000000523 sample Substances 0.000 title claims abstract description 70
- 238000002360 preparation method Methods 0.000 title claims abstract description 12
- 210000002700 urine Anatomy 0.000 claims abstract description 45
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims abstract description 36
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 claims abstract description 27
- 238000000034 method Methods 0.000 claims abstract description 23
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 claims abstract description 18
- 229930024421 Adenine Natural products 0.000 claims abstract description 17
- 229960000643 adenine Drugs 0.000 claims abstract description 17
- RAZLJUXJEOEYAM-UHFFFAOYSA-N 2-[bis[2-(2,6-dioxomorpholin-4-yl)ethyl]azaniumyl]acetate Chemical compound C1C(=O)OC(=O)CN1CCN(CC(=O)O)CCN1CC(=O)OC(=O)C1 RAZLJUXJEOEYAM-UHFFFAOYSA-N 0.000 claims abstract description 14
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 claims abstract description 12
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims abstract description 8
- 238000010438 heat treatment Methods 0.000 claims abstract description 8
- 230000006837 decompression Effects 0.000 claims abstract description 7
- 239000013067 intermediate product Substances 0.000 claims abstract description 7
- QPCDCPDFJACHGM-UHFFFAOYSA-N N,N-bis{2-[bis(carboxymethyl)amino]ethyl}glycine Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(=O)O)CCN(CC(O)=O)CC(O)=O QPCDCPDFJACHGM-UHFFFAOYSA-N 0.000 claims abstract description 6
- 229960003330 pentetic acid Drugs 0.000 claims abstract description 6
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 claims abstract description 6
- 238000010992 reflux Methods 0.000 claims abstract description 6
- 238000001816 cooling Methods 0.000 claims abstract description 4
- 238000001514 detection method Methods 0.000 claims description 21
- 238000002189 fluorescence spectrum Methods 0.000 claims description 19
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 15
- 239000008367 deionised water Substances 0.000 claims description 12
- 229910021641 deionized water Inorganic materials 0.000 claims description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 12
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 8
- 238000002390 rotary evaporation Methods 0.000 claims description 5
- 239000007864 aqueous solution Substances 0.000 claims description 4
- 238000001917 fluorescence detection Methods 0.000 claims description 4
- KJHJAABRDASKAF-UHFFFAOYSA-N 3,7-dihydropurine-2,6-dione Chemical compound OC1=NC(O)=C2N=CNC2=N1.O=C1NC(=O)NC2=C1NC=N2 KJHJAABRDASKAF-UHFFFAOYSA-N 0.000 claims description 3
- 238000007689 inspection Methods 0.000 claims description 2
- 238000003756 stirring Methods 0.000 abstract description 5
- 239000000047 product Substances 0.000 abstract description 3
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 24
- 239000000243 solution Substances 0.000 description 19
- 239000011550 stock solution Substances 0.000 description 14
- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 13
- 239000011668 ascorbic acid Substances 0.000 description 13
- 230000005284 excitation Effects 0.000 description 13
- 229960005070 ascorbic acid Drugs 0.000 description 12
- 235000010323 ascorbic acid Nutrition 0.000 description 12
- 238000010521 absorption reaction Methods 0.000 description 9
- 238000000862 absorption spectrum Methods 0.000 description 9
- DDRJAANPRJIHGJ-UHFFFAOYSA-N creatinine Chemical compound CN1CC(=O)NC1=N DDRJAANPRJIHGJ-UHFFFAOYSA-N 0.000 description 9
- 239000000126 substance Substances 0.000 description 9
- 230000015572 biosynthetic process Effects 0.000 description 7
- 239000011259 mixed solution Substances 0.000 description 7
- KDCGOANMDULRCW-UHFFFAOYSA-N 7H-purine Chemical compound N1=CNC2=NC=NC2=C1 KDCGOANMDULRCW-UHFFFAOYSA-N 0.000 description 6
- RPNUMPOLZDHAAY-UHFFFAOYSA-N Diethylenetriamine Chemical compound NCCNCCN RPNUMPOLZDHAAY-UHFFFAOYSA-N 0.000 description 6
- QIAFMBKCNZACKA-UHFFFAOYSA-N N-benzoylglycine Chemical compound OC(=O)CNC(=O)C1=CC=CC=C1 QIAFMBKCNZACKA-UHFFFAOYSA-N 0.000 description 6
- 230000008859 change Effects 0.000 description 6
- 238000003786 synthesis reaction Methods 0.000 description 6
- 238000001035 drying Methods 0.000 description 5
- RYYVLZVUVIJVGH-UHFFFAOYSA-N caffeine Chemical compound CN1C(=O)N(C)C(=O)C2=C1N=CN2C RYYVLZVUVIJVGH-UHFFFAOYSA-N 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 241000251468 Actinopterygii Species 0.000 description 3
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 3
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 description 3
- 239000002253 acid Substances 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- -1 terbium-diethylenetriamine Chemical compound 0.000 description 3
- LPHGQDQBBGAPDZ-UHFFFAOYSA-N Isocaffeine Natural products CN1C(=O)N(C)C(=O)C2=C1N(C)C=N2 LPHGQDQBBGAPDZ-UHFFFAOYSA-N 0.000 description 2
- 108010093894 Xanthine oxidase Proteins 0.000 description 2
- 102100033220 Xanthine oxidase Human genes 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- GTDPSWPPOUPBNX-UHFFFAOYSA-N ac1mqpva Chemical compound CC12C(=O)OC(=O)C1(C)C1(C)C2(C)C(=O)OC1=O GTDPSWPPOUPBNX-UHFFFAOYSA-N 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 229960001948 caffeine Drugs 0.000 description 2
- VJEONQKOZGKCAK-UHFFFAOYSA-N caffeine Natural products CN1C(=O)N(C)C(=O)C2=C1C=CN2C VJEONQKOZGKCAK-UHFFFAOYSA-N 0.000 description 2
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Chemical compound NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 description 2
- 235000005911 diet Nutrition 0.000 description 2
- 230000037213 diet Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- ZFXYFBGIUFBOJW-UHFFFAOYSA-N theophylline Chemical compound O=C1N(C)C(=O)N(C)C2=C1NC=N2 ZFXYFBGIUFBOJW-UHFFFAOYSA-N 0.000 description 2
- 201000002928 xanthinuria Diseases 0.000 description 2
- XDIYNQZUNSSENW-UUBOPVPUSA-N (2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanal Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O.OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O XDIYNQZUNSSENW-UUBOPVPUSA-N 0.000 description 1
- 206010013786 Dry skin Diseases 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 238000004566 IR spectroscopy Methods 0.000 description 1
- 208000007101 Muscle Cramp Diseases 0.000 description 1
- 208000000112 Myalgia Diseases 0.000 description 1
- 244000269722 Thea sinensis Species 0.000 description 1
- LEHOTFFKMJEONL-UHFFFAOYSA-N Uric Acid Chemical compound N1C(=O)NC(=O)C2=C1NC(=O)N2 LEHOTFFKMJEONL-UHFFFAOYSA-N 0.000 description 1
- TVWHNULVHGKJHS-UHFFFAOYSA-N Uric acid Natural products N1C(=O)NC(=O)C2NC(=O)NC21 TVWHNULVHGKJHS-UHFFFAOYSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 238000003965 capillary gas chromatography Methods 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 230000000536 complexating effect Effects 0.000 description 1
- 208000029078 coronary artery disease Diseases 0.000 description 1
- 229940104302 cytosine Drugs 0.000 description 1
- 238000005034 decoration Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- 125000004177 diethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 230000005672 electromagnetic field Effects 0.000 description 1
- FPVGTPBMTFTMRT-NSKUCRDLSA-L fast yellow Chemical compound [Na+].[Na+].C1=C(S([O-])(=O)=O)C(N)=CC=C1\N=N\C1=CC=C(S([O-])(=O)=O)C=C1 FPVGTPBMTFTMRT-NSKUCRDLSA-L 0.000 description 1
- 206010016256 fatigue Diseases 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 230000001631 hypertensive effect Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 230000027939 micturition Effects 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 230000004144 purine metabolism Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000011896 sensitive detection Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 238000011895 specific detection Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 238000000967 suction filtration Methods 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 229960000278 theophylline Drugs 0.000 description 1
- 229940116269 uric acid Drugs 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F5/00—Compounds containing elements of Groups 3 or 13 of the Periodic Table
- C07F5/003—Compounds containing elements of Groups 3 or 13 of the Periodic Table without C-Metal linkages
-
- 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
-
- 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
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/18—Metal complexes
- C09K2211/182—Metal complexes of the rare earth metals, i.e. Sc, Y or lanthanide
-
- 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
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Physics & Mathematics (AREA)
- Optics & Photonics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Molecular Biology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Engineering & Computer Science (AREA)
- Materials Engineering (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The present invention discloses the fluorescence probe and its preparation method and application for detecting xanthine.Diethylenetriamine pentaacetic acid, acetic anhydride and pyridine are taken, is stirred at reflux, cooling, decompression filters, and washs, dry；Obtained diethylenetriamine pentaacetic acid dianhydride is stirred at reflux with triethylamine, dimethylformamide, adenine and/or cytimidine, it is cooling, it rotates, washs, it is dry, by obtained intermediate product and Tb (NO3)3·6H2O obtains target product in 60 DEG C of heating stirring 2h.By TbШ- dtpa-2A, TbШ- dtpa-2C and TbШ- dtpa-AC detects xanthine respectively as probe combination fluorescent method.The method of the present invention is simple and novel, at low cost, high-efficient, and can be applicable in practical urine sample.
Description
Technical field
The invention belongs to the synthesis of analytical chemistry field more particularly to a kind of novel fluorescence probe and its in true urine
Detect the application of xanthine.
Background technique
Xanthine (Xanthine) is the purine base in a kind of organ and body fluid for being distributed widely in people and other organisms,
Its normal physiological concentration is 0.5~2.5 μm of ol/L in serum, is 40~160 μm of ol/L in urine.Meanwhile xanthine
It is also the product of purine metabolism, uric acid is converted under the action of xanthine oxidase, is finally excreted with urine.When internal
When lacking xanthine oxidase, xanthine cannot be converted but be accumulated in vivo.Lead to xanthine concentration in blood and urine
It steeply rises, is eventually developed to xanthinuria.Patient with xanthinuria is it is possible that such as fatigability myalgia
Bitterly, the symptoms such as muscle cramp or muscle spasmus frequent micturition.
Containing the mild excitant being largely derived by xanthine in tea and coffee, such as caffeine and theophylline.However,
Caffeine in coffee may induce coronary heart disease, this will increase the blood pressure and heart rate of hypertensive patient.In addition, food industry is to meat
The freshness of class requires very high.After fish is dead, intracorporal ATP is degraded to xanthine, content with storage time increasing
Add and increases.Therefore, in the flesh of fish content of xanthine can also be used for assessment the flesh of fish freshness.To sum up, xanthine
It detects either still all extremely important in medical field in field of food.
The method for being commonly used for detection xanthine includes colorimetric method, electrochemical process, high performance liquid chromatography and capillary column
Gas chromatography.However, these methods are complicated, time-consuming and sensitivity is low.Moreover, in these methods, pre-treatment step is complicated
And instrument and equipment is expensive.It is well known that fluorescence probe detection method is because its equipment is simple, it is easy to operate, analysis speed is fast etc.
Advantage is widely used in many fields.The especially usual high sensitivity of fluorescence probe method, selectivity is good and detection limit is low.
Therefore, developing a kind of fluorescence probe that can fast and accurately detect xanthine is very important.
Summary of the invention
An object of the present invention is the novel fluorescence probe that design synthesis can be used for effectively detecting xanthine.
It is a kind of easy to operate the second object of the present invention is to provide, it is at low cost, it is sensitive quick, and the detection Huang that selectivity is good
The method of purine.
The technical solution adopted by the present invention is that: for detecting the fluorescence probe of xanthine, preparation method includes the following steps:
1) diethylenetriamine pentaacetic acid, acetic anhydride and pyridine are uniformly mixed, are stirred at reflux at 65 DEG C for 24 hours, are cooled to room
Temperature, decompression are filtered, are successively washed with acetic anhydride and anhydrous ether, dry at 60 DEG C, obtain diethylenetriamine pentaacetic acid dianhydride
(dtpaa).Preferably, in molar ratio, diethylenetriamine pentaacetic acid: acetic anhydride: pyridine=1:4:6.
2) diethylenetriamine pentaacetic acid dianhydride, triethylamine, anhydrous DMF, adenine and/or cytimidine are uniformly mixed, 100
It is stirred at reflux 24-48h at DEG C, is cooled to room temperature, rotary evaporation is successively washed with acetonitrile and anhydrous ether, and decompression filters, in 50
DEG C drying, obtains intermediate product.Preferably, in molar ratio, diethylenetriamine pentaacetic acid dianhydride: triethylamine: adenine=1:3:2；Two
Second pentaacetic acid dianhydride: triethylamine: cytimidine=1:3:2；Diethylenetriamine pentaacetic acid dianhydride: triethylamine: adenine: cytimidine
=1:3:1:1.
3) intermediate product and Tb (NO for taking step 2) to obtain3)3·6H2O adds deionized water dissolving, in 60 DEG C of agitating and heatings
2-3h, it is cooling, obtain the fluorescence probe for detecting xanthine.Preferably, in molar ratio, intermediate product: Tb (NO3)3·6H2O
=1:1.
Above-mentioned application of the fluorescence probe in qualitative and quantitative detection xanthine for detecting xanthine.
Preferably, for detecting the application of fluorescence probe xanthine in qualitative and quantitative detection urine of xanthine.Side
Method is as follows:
The method of xanthine in qualitative detection urine: taking urine, is added above-mentioned for detecting the fluorescence probe of xanthine
Aqueous solution is sufficiently mixed, and is carried out fluorescence detection, is observed the variation of fluorescence spectrum.
The method of xanthine in quantitative detection urine: taking 50 μ L concentration is 5.0 × 10-3Mol/L's is above-mentioned for detecting Huang
The aqueous solution of the fluorescence probe of purine is settled to 5mL in 10mL colorimetric cylinder, with urine, carries out fluorescence detection.
The beneficial effects of the present invention are:
1. the present invention is directed to the design feature of detected material xanthine, dtpa is repaired using base pair complementarity principle
Decorations, design have synthesized three kinds of novel fluorescence probes.
2. by means of the present invention, three kinds of probes can carry out sensitive and specific detection to xanthine.With it is other
The method of detection xanthine is compared, and is had simply, and quickly, at low cost, selectivity is good, the advantages that influence by external electromagnetic field.
Detailed description of the invention
Fig. 1 is fluorescence probe TbШ- dtpa-2A, TbШ- dtpa-2C and TbШThe synthetic route chart of-dtpa-AC.
Fig. 2 a is Fourier transform infrared spectroscopy (FT-IR) figure of dtpa.
Fig. 2 b is Fourier transform infrared spectroscopy (FT-IR) figure of adenine (adenine).
Fig. 2 c is Fourier transform infrared spectroscopy (FT-IR) figure of cytimidine (cytosine).
Fig. 2 d is Fourier transform infrared spectroscopy (FT-IR) figure of dtpa-2A.
Fig. 2 e is Fourier transform infrared spectroscopy (FT-IR) figure of dtpa-2C.
Fig. 2 f is Fourier transform infrared spectroscopy (FT-IR) figure of dtpa-AC.
Fig. 3 a is Tb3+, dtpa-2A, TbШ- dtpa-2A and TbШThe uv absorption spectra of-dtpa-2A+ xanthine.
Fig. 3 b is Tb3+, dtpa-2C, TbШ- dtpa-2C and TbШThe uv absorption spectra of-dtpa-2C+ xanthine.
Fig. 3 c is Tb3+, dtpa-AC, TbШ- dtpa-AC and TbШThe uv absorption spectra of-dtpa-AC+ xanthine.
Fig. 4 a-1 is fluorescence probe TbШThe fluorescence spectra that-dtpa-2A detects xanthine.
Fig. 4 a-2 is fluorescence probe TbШ- dtpa-2A compares histogram to the fluorescence spectrum that xanthine detects.
Fig. 4 b-1 is fluorescence probe TbШThe fluorescence spectra that-dtpa-2C detects xanthine.
Fig. 4 b-2 is fluorescence probe TbШ- dtpa-2C compares histogram to the fluorescence spectra that xanthine detects.
Fig. 4 c-1 is fluorescence probe TbШThe fluorescence spectra that-dtpa-AC detects xanthine.
Fig. 4 c-2 is fluorescence probe TbШ- dtpa-AC compares histogram to the fluorescence spectra that xanthine detects.
Fig. 5 a is fluorescence probe TbШ- dtpa-2A compares the interference fluorescence spectrum that xanthine is mixed with different material respectively
Figure.
Fig. 5 b is fluorescence probe TbШ- dtpa-2C compares the interference fluorescence spectrum that xanthine is mixed with different material respectively
Figure.
Fig. 5 c is fluorescence probe TbШ- dtpa-AC compares the interference fluorescence spectrum that xanthine is mixed with different material respectively
Figure.
Fig. 6 a is fluorescence probe TbШThe fluorescence spectrum comparison diagram that-dtpa-2A detects xanthine in practical urine sample (Ur).
Fig. 6 b is fluorescence probe TbШThe fluorescence spectrum comparison diagram that-dtpa-2C detects xanthine in practical urine sample (Ur).
Fig. 6 c is fluorescence probe TbШThe fluorescence spectrum comparison diagram that-dtpa-AC detects xanthine in practical urine sample (Ur).
Specific embodiment
Embodiment 1
The preparation of diethylenetriamine pentaacetic acid dianhydride (dtpaa): 7.8670g (0.02mol) diethylenetriamine pentaacetic acid is weighed
(dtpa), acetic anhydride 16.0mL (0.08mol), pyridine 10.0mL (0.12mol) are placed in three neck round bottom, are delayed at 65 DEG C
Slow agitating and heating is condensed back for 24 hours.Stop heating and stirring, product at reduced pressure is filtered after being cooled to room temperature, successively uses acetic anhydride
Wash (3 × 10mL) three times respectively with anhydrous ether, and depressurize suction filtration, by product in drying box 60 DEG C of dryings to get diethyl
Pentaacetic acid dianhydride (dtpaa).
(1) for detecting fluorescence probe --- bis- (the adenine) (Tb of terbium-diethylenetriamine pentaacetic acid-of xanthineШ-
dtpa-2A)
1, the preparation of diethylenetriamine pentaacetic acid-bis- (adenines) (dtpa-2A): diethylenetriamine pentaacetic acid dianhydride is taken
(dtpaa) triethylamine (16.5mmol) of 1.9635g (5.5mmol), 2.334mL, anhydrous DMF (50mL), adenine (A)
1.4864g (11mmol), in three neck round bottom.It under the conditions of 100 DEG C of constant temperature, quickly stirs, is condensed back for 24 hours.It has reacted
Stood after complete, after being cooled to room temperature, rotary evaporation removes solvent, obtains Off-white solid substance, and decompression filters, successively with acetonitrile and
Anhydrous ether washs (3 × 10mL) three times respectively.Drying is bis- (adenine) to get diethylenetriamine pentaacetic acid-under the conditions of 50 DEG C
(dtpa-2A)。
2、TbШThe preparation of-dtpa-2A: it is bis- (adenine) to weigh 0.7838g (1.25mmol) diethylenetriamine pentaacetic acid-
(dtpa-2A) in three neck round bottom, add 30mL deionized water dissolving.Tb (the NO of 0.5663g is weighed simultaneously3)3·6H2O
(1.25mmol) is placed in a beaker, and is transferred in round-bottomed flask after adding 30mL deionized water dissolving, in 60 DEG C of agitating and heating 2h.It will
Solution after reaction is cooled to room temperature, and obtains TbШ-dtpa-2A.Synthesis process is as shown in Figure 1.
(2) for detecting fluorescence probe --- bis- (the cytimidine) (Tb of terbium-diethylenetriamine pentaacetic acid-of xanthineШ-
dtpa-2C)
1, the preparation of diethylenetriamine pentaacetic acid-bis- (cytimidines) (dtpa-2C): diethylenetriamine pentaacetic acid dianhydride is taken
(dtpaa) triethylamine (16.5mmol) of 1.9635g (5.5mmol), 2.334mL, anhydrous DMF (50mL), cytimidine (C)
1.2210g (11mmol), in three neck round bottom.It under the conditions of 100 DEG C of constant temperature, quickly stirs, is condensed back for 24 hours.It has reacted
Stood after complete, after being cooled to room temperature, rotary evaporation removes solvent, obtains light yellow solid substance, and decompression filters, successively with acetonitrile and
Anhydrous ether washs (3 × 10mL) three times respectively.Drying is bis- (cytimidine) to get diethylenetriamine pentaacetic acid-under the conditions of 50 DEG C
(dtpa-2C)。
2、TbШThe preparation of-dtpa-2C: it is bis- (cytimidine) to weigh 0.7288g (1.25mmol) diethylenetriamine pentaacetic acid-
(dtpa-2C) in three neck round bottom, add 30mL deionized water dissolving.Tb (the NO of 0.5663g is weighed simultaneously3)3·6H2O
(1.25mmol) is placed in a beaker, and is transferred in round-bottomed flask after adding 30mL deionized water dissolving, in 60 DEG C of agitating and heating 2h.It will
Solution after reaction is cooled to room temperature, and obtains TbШ-dtpa-2C.Synthesis process is as shown in Figure 1.
(3) for detecting fluorescence probe --- the terbium-adenine-diethylenetriamine pentaacetic acid-cytimidine (Tb of xanthineШ-
dtpa-AC)
1, adenine-diethylenetriamine pentaacetic acid-cytimidine (dtpa-AC) preparation: diethylenetriamine pentaacetic acid dianhydride is taken
(dtpaa) triethylamine (16.5mmol) of 1.9635g (5.5mmol), 2.334mL, anhydrous DMF (50mL), adenine (A)
0.7432g (5.5mmol), in three neck round bottom.It under the conditions of 100 DEG C of constant temperature, quickly stirs, is condensed back for 24 hours.To above-mentioned
It is added in reaction solution cytimidine 0.6105g (5.5mmol), continuation is quickly stirred under the conditions of 100 DEG C of constant temperature, is condensed back
24h.It is stood after fully reacting, after being cooled to room temperature, rotary evaporation removes solvent, obtains Off-white solid substance, and decompression filters, according to
It is secondary to wash (3 × 10mL) three times respectively with acetonitrile and anhydrous ether.It dries under the conditions of 50 DEG C to get adenine-Diethylenetriamine
Pentaacetic acid-cytimidine (dtpa-AC).
2、TbШThe preparation of-dtpa-AC: 0.7563g (1.25mmol) adenine-diethylenetriamine pentaacetic acid-cytimidine is weighed
(dtpa-AC) in three neck round bottom, add 30mL deionized water dissolving.Tb (the NO of 0.5663g is weighed simultaneously3)3·6H2O
(1.25mmol) is placed in a beaker, and is transferred in round-bottomed flask after adding 30mL deionized water dissolving, in 60 DEG C of agitating and heating 2h.It will
Solution after reaction is cooled to room temperature, and obtains TbШ-dtpa-AC.Synthesis process is as shown in Figure 1.
(4) it detects
1, FT-IR figure such as Fig. 2 a, 2b, the 2c of dtpa, adenine, cytimidine, dtpa-2A, dtpa-2C and dtpa-AC,
Shown in 2d, 2e, 2f.With dtpa, adenine is compared with the infrared spectroscopy of cytimidine, dtpa-2A, and dtpa-2C and dtpa-AC's is red
The position at respective absorption peak shows apparent variation in external spectrum.Especially dtpa-2A, dtpa-2C and dtpa-AC's is infrared
The C=O absorption peak of amido bond respectively appears in 1638cm in spectrum-1, 1648cm-1And 1638cm-1Place.With 1752cm in dtpa-1
The absorption peak of the carboxyl C=O at place is compared, they move 114cm respectively-1, 104cm-1And 114cm-1.In addition, dtpa-2A,
The NH stretching vibration peak of amido bond respectively appears in 3395cm in dtpa-2C and dtpa-AC-1And 2933cm-1(dtpa-2A),
3378cm-1And 2928cm-1(dtpa-2C) and 3378cm-1And 2931cm-1(dtpa-AC) at.With in adenine and cytimidine-
NH23294cm-1, 3117cm-1And 3382cm-1, 3169cm-1It compares, it can be seen that apparent variation has occurred.These features
The movement at peak all illustrates the formation of amido bond, i.e. three kinds of fluorescence probes are successfully synthesized.
Embodiment 2 is used to detect application of the fluorescence probe of xanthine in detection xanthine
Taking a certain amount of xanthine to be configured to concentration with deionized water is 5.0 × 10-3The solution of mol/L, as xanthine
Stock solution.
Tb prepared by Example 1 respectivelyШ-dtpa-2A、TbШ- dtpa-2C and TbШ- dtpa-AC is matched with deionized water
Being set to concentration is 5.0 × 10-3The Tb of mol/LШ-dtpa-2A、TbШ- dtpa-2C and TbШThe stock solution of-dtpa-AC.
(1) ultra-violet absorption spectrum of fluorescence probe
1, method
9 colorimetric cylinders are taken to be divided into TbШ- dtpa-2A group, TbШ- dtpa-2C group and TbШTotally three groups of-dtpa-AC group, carries out
Ultra-violet absorption spectrum detection.
TbШ- dtpa-2A component are as follows: dtpa-2A, TbШ- dtpa-2A and TbШ- dtpa-2A+ xanthine (TbШ-dtpa-2A
+Xanthine)；
TbШ- dtpa-2C component are as follows: dtpa-2C, TbШ- dtpa-2C and TbШ- dtpa-2C+ xanthine (TbШ-dtpa-2C
+Xanthine)；
TbШ- dtpa-AC component are as follows: dtpa-AC, TbШ- dtpa-AC and TbШ- dtpa-AC+ xanthine (TbШ-dtpa-
AC+Xanthine)；
2, it detects
Dtpa-2A, dtpa-2C, dtpa-AC, TbШ- dtpa-2A, TbШ- dtpa-2C, TbШ- dtpa-AC and TbШ-
Dtpa-2A+ xanthine (TbШ- dtpa-2A+Xanthine), TbШ- dtpa-2C+ xanthine (TbШ-dtpa-2C+
Xanthine), TbШ- dtpa-AC+ xanthine (TbШ- dtpa-AC+Xanthine) uv absorption spectra such as Fig. 3 a, 3b,
Shown in 3c.
As can be seen that Tb from Fig. 3 a3+There is weak absorption at 219nm.The dtpa-2A of synthesis is in 219nm and 264nm
There are two absorption peaks at place.As dtpa-2A and Tb3+It is complexed and forms TbIIITwo suctions when-dtpa-2A, at 219nm and 264nm
The intensity for receiving peak slightly enhances.However, again to TbIIIAfter xanthine is added in-dtpa-2A solution, TbIIITwo of-dtpa-2A
The significant reduction of the absorbance of absorption peak.This shows that the addition of xanthine can substantially change TbIIIThe absorption spectrum of-dtpa-2A.
Simultaneously it may be speculated that after xanthine is added, TbIIIThe fluorescence spectrum of-dtpa-2A can change a lot.Show TbIII-
Dtpa-2A has the potentiality as fluorescence probe detection xanthine.
From Fig. 3 b can be seen that dtpa-2C respectively at 221nm and 270nm tool there are two absorption peak, but absorbance compared with
It is weak.When with Tb3+Complexing forms TbIIIWhen-dtpa-2C, have almost no change compared with the absorption spectrum of dtpa-2C.However,
After xanthine is added, TbIIITwo absorption peaks of-dtpa-2C are remarkably reinforced.Show TbIII- dtpa-2C equally has as fluorescence
Probe detects the possibility of xanthine.
It can be seen that dtpa-AC and Tb from Fig. 3 cIIIThe absorption spectrum of-dtpa-AC is almost the same.However, will be yellow fast
Tb is added in purineIIIAfter-dtpa-AC solution, two absorption peak is obviously increased.This also indicates that TbIII- dtpa-AC has conduct
The potentiality of fluorescence probe detection xanthine.
(2) fluorescence spectrum that fluorescence probe detects xanthine
Method: 9 10mL colorimetric cylinders are taken to be divided into TbШ- dtpa-2A group, TbШ- dtpa-2C group and TbШ- dtpa-AC group is altogether
Three groups.Every group of the 1st 150 μ L xanthine stock solutions of addition.2nd 50 μ L concentration of addition are 5.0 × 10-3The fluorescence of mol/L is visited
Needle stock solution.3rd 50 μ L concentration of addition are 5.0 × 10-3The fluorescence probe stock solution of mol/L and 150 μ L xanthine stock solutions.
Then every colorimetric cylinder is all settled to 5mL with deionized water.It is observed under the excitation of 340nm wavelength light in 280nm, 330nm respectively
The variation of fluorescence spectrum.As a result shown in such as Fig. 4 a-1,4a-2,4b-1,4b-2,4c-1 and 4c-2.
From in Fig. 4 a-1 as can be seen that under the excitation of 280nm wavelength light, TbIII- dtpa-2A launches at 320nm
Hyperfluorescence, and xanthine hardly issues fluorescence at 320nm.When xanthine is added to TbIIIAfter in-dtpa-2A solution,
TbIIIThe fluorescence of-dtpa-2A is obviously quenched.Fig. 4 a-2 can more intuitively compare the difference of fluorescence intensity at 320nm.
From in Fig. 4 b-1 as can be seen that under the excitation of 330nm wavelength light, TbIII- dtpa-2C launches at 372nm
Compared with hypofluorescence, and xanthine also launches certain fluorescence at 372nm.When xanthine is added to TbIII- dtpa-2C solution
In after, TbIIIThe fluorescence intensity of-dtpa-2C obviously increases.Fluorescence intensity from 372nm can more intuitively be compared from Fig. 4 b-2
Difference.
From in Fig. 4 c-1 as can be seen that under the excitation of 340nm wavelength light, TbIII- dtpa-AC launches at 381nm
The fluorescence of moderate strength, and xanthine launches weak fluorescence at 381nm.When xanthine is added to TbIII- dtpa-AC is molten
After in liquid, TbIIIThe fluorescence intensity of-dtpa-AC is slightly increased.It should be apparent that fluorescence intensity from 381nm from Fig. 4 c-2
Difference.
(3) different coexisting substances are mixed with xanthine to fluorescence probe TbШ- dtpa-2A, TbШ- dtpa-2C and TbШ-
The influence of dtpa-AC detection
Method: 15 colorimetric cylinders are taken to be divided into TbШ- dtpa-2A group, TbШ- dtpa-2C group and TbШ- dtpa-AC group totally three
Group.It is 5.0 × 10 that 1.5mL concentration is separately added into every group of 5 colorimetric cylinders-4The hippuric acid (Hipa) of mol/L, ascorbic acid
(AA), histidine (His), kreatinin (Cre) and glucose (Glucose) solution.Then TbШ- dtpa-2A organizes every colorimetric cylinder
Add 50 μ LTb respectively againШ- dtpa-2A stock solution and 150 μ L xanthine stock solutions, TbШ- dtpa-2C organizes every colorimetric cylinder difference
Add 50 μ LTbШ- dtpa-2C stock solution and 150 μ L xanthine stock solutions, TbШ- dtpa-AC organizes every colorimetric cylinder and adds 50 μ respectively
LTbШ- dtpa-AC stock solution and 150 μ L xanthine stock solutions.Finally 5mL is settled to deionized water respectively.At this time in solution
The concentration of probe is 5.0 × 10-5The concentration of mol/L, xanthine and other coexisting substances solution is 1.5 × 10-4mol/L.?
The variation of fluorescence spectrum is observed under the excitation of 280nm, 330nm, 340nm wavelength light.As a result such as Fig. 5 a, 5b, 5c and table 1-3 institute
Show.
It can be seen from Fig. 5a that under the excitation of 280nm wavelength light, TbШ- dtpa-2A solution issues by force at 320nm
Fluorescence, after xanthine is added, TbШThe fluorescence of-dtpa-2A is obviously quenched.However work as hippuric acid, and ascorbic acid, histidine,
Kreatinin, the coexisting substances such as glucose are separately added into TbШAfter in the mixed solution of-dtpa-2A and xanthine, in addition to ascorbic acid
Except other substances have little influence on the fluorescence of mixed solution.This illustrates that other are total with xanthine in addition to ascorbic acid in urine
The substance deposited will not interfere detection of the probe to xanthine.Seem that the presence of ascorbic acid may will affect the inspection of xanthine
It surveys, still, in fact, the content of ascorbic acid is related with the diet of preceding a couple of days in urine.That is, control can be passed through
Diet reduces the content of ascorbic acid in urine, so that making the detection of xanthine will not be interfered.It can be more from table 1
The clearly variation of observation fluorescence intensity.
Table 1
It can be seen from figure 5b that under the excitation of 330nm wavelength light, TbIII- dtpa-2C solution emits at 372nm
Hypofluorescence out.Xanthine is added to TbIIIAfter-dtpa-2C solution, TbIIIThe fluorescence intensity of-dtpa-2C obviously increases.When point
Not in TbIIIHippuric acid, ascorbic acid, histidine, kreatinin, Portugal are further added in the mixed solution of-dtpa-2C+ xanthine
When grape sugar, in addition to ascorbic acid, other substances have little influence on the fluorescence of mixed solution.Ascorbic acid makes TbIII-dtpa-2C
The fluorescence intensity of+xanthine mixed solution slightly reduces, but Tb in the presence of ascorbic acidIII- dtpa-2C+ xanthine is mixed
The fluorescence intensity for closing solution is still higher than TbIII-dtpa-2C.It can be considered that due to xanthine there are TbIII-dtpa-2C
Fluorescence intensity dramatically increase.From in table 2 can it is more intuitive from fluorescence intensity variation.
Table 2
From in Fig. 5 c as can be seen that under the excitation of 340nm wavelength light, TbIII- dtpa-AC emits medium at 381nm
The fluorescence of intensity.After xanthine is added, TbIIIThe fluorescence intensity of-dtpa-AC is increased slightly.Although the increase of fluorescence intensity is unknown
It is aobvious, but it can be seen that the variation of fluorescence intensity before and after xanthine is added.In TbIIIIn the mixed solution of-dtpa-AC+ xanthine
Further it is added hippuric acid, ascorbic acid, histidine, kreatinin, after glucose, the fluorescence intensity of mixed solution is almost without change
Change.From in table 3 can it is clearer from fluorescence intensity variation.
Table 3
It is above-mentioned the experiment results show that Tb of the inventionШ- dtpa-2A, TbШ- dtpa-2C and TbШ- dtpa-AC is used as fluorescence
Probe in detecting xanthine has good anti-interference ability.
(4) fluorescence probe TbШ- dtpa-2A, TbШ- dtpa-2C, TbШ- dtpa-AC detects xanthine in practical urine sample
Method: 18 10mL colorimetric cylinders are taken to be divided into TbШ- dtpa-2A group, TbШ- dtpa-2C group and TbШ- dtpa-AC group is altogether
Three groups.Every group the 1st is blank urine sample, the 2nd 150 μ L xanthine stock solutions of addition, the 3rd 50 μ L fluorescence probe deposits of addition
Liquid, 4-6 branch are being separately added into 50 μ L, the xanthine storage of 100 μ L, 150 μ L after being separately added into 50 μ L fluorescence probe stock solutions
Standby liquid, is then settled to 5mL with urine sample.Respectively in 280nm, 330nm, fluorescence spectrum is observed under the excitation of 340nm wavelength light
Variation.As a result such as Fig. 6 a, 6b, shown in 6c.
From as can be seen that under the excitation of 280nm wavelength light, pure urine issues relatively strong glimmering at 390nm in Fig. 6 a
Light.When xanthine (ultimate density is 150 μm of ol/L) is added into urine, the fluorescence intensity of urine is significantly reduced.This shows
When containing a large amount of xanthine in urine, the fluorescence intensity of urine may be substantially reduced.But this method is for judging urine
It is inaccurate that a large amount of xanthine whether are generated in liquid.When by TbIII- dtpa-2A is added in urine (final concentration of 50 μm of ol/L)
When, it launches hyperfluorescence at 320nm.Since the fluorescence of pure urine is weaker at 320nm, therefore, it is considered that the fluorescence of urine is not
Tb can be interferedIIIThe fluorescence of-dtpa-2A.Then, xanthine is added gradually to TbIIIIn-dtpa-2A solution, with xanthine
The increase of concentration, TbIIIThe fluorescence intensity of-dtpa-2A gradually decreases.This shows to use TbIII- dtpa-2A can as fluorescence probe
Xanthine in accurate detection urine.From in table 4 can more clearly from fluorescence intensity variation.
Table 4
From being can be seen that under the light excitation of 330nm wavelength in Fig. 6 b, pure urine is launched very weak at 372nm
Fluorescence.When xanthine is added in urine (ultimate density is 150 μm of ol/L), the fluorescence intensity of urine is slightly increased.It will
TbIIIAfter-dtpa-2C is added in urine, it can be seen that TbIII- dtpa-2C has at 372nm apparent glimmering in urine
Light emitting.Then, it states further up and xanthine is added in solution, with the increase of xanthine concentration, TbIII- dtpa-2C's
Fluorescence intensity also gradually increases.As can be seen that the presence of xanthine can obviously increase TbIIIThe fluorescence intensity of-dtpa-2C.Together
When show TbIII- dtpa-2C can also be with the xanthine in accurate detection urine as fluorescence probe.More intuitive fluorescence
Strength Changes can be observed from table 5.
Table 5
It can be seen that under the excitation of 340nm wavelength light from Fig. 6 c, pure urine launches faint fluorescence.To urine
After middle addition xanthine (ultimate density is 150 μm of ol/L), the fluorescence intensity of urine is increased slightly.By TbIII- dtpa-AC is added
When in urine, under the light excitation of 340nm wavelength, TbIII- dtpa-AC issues relatively strong fluorescence at 381nm.Further plus
Enter xanthine, TbIIIThe fluorescence intensity of-dtpa-AC is gradually increased with the increase of xanthine concentration.This shows TbIII-
Dtpa-AC can also serve as fluorescence probe for detecting the xanthine in urine.Fluorescence intensity change can be from table 6
Out.
Table 6
It is above-mentioned the experiment results show that Tb of the inventionШ- dtpa-2A, TbШ- dtpa-2C and TbШ- dtpa-AC is used as fluorescence
Probe quantitative detects xanthine.
Claims (8)
1. the fluorescence probe for detecting xanthine, which is characterized in that preparation method includes the following steps:
1) diethylenetriamine pentaacetic acid, acetic anhydride and pyridine are uniformly mixed, are stirred at reflux for 24 hours, are cooled to room temperature at 65 DEG C, subtracts
Pressure filters, and is successively washed with acetic anhydride and anhydrous ether, dry at 60 DEG C, obtains diethylenetriamine pentaacetic acid dianhydride；
2) diethylenetriamine pentaacetic acid dianhydride, triethylamine, anhydrous DMF, adenine and/or cytimidine are uniformly mixed, at 100 DEG C
It is stirred at reflux 24-48h, is cooled to room temperature, rotary evaporation is successively washed with acetonitrile and anhydrous ether, and decompression filters, dry in 50 DEG C
It is dry, obtain intermediate product；
3) intermediate product and Tb (NO for taking step 2) to obtain3)3·6H2O adds deionized water dissolving, in 60 DEG C of agitating and heating 2-
3h, it is cooling, obtain the fluorescence probe for detecting xanthine.
2. as described in claim 1 for detecting the fluorescence probe of xanthine, which is characterized in that in step 1), in molar ratio,
Diethylenetriamine pentaacetic acid: acetic anhydride: pyridine=1:4:6.
3. as described in claim 1 for detecting the fluorescence probe of xanthine, which is characterized in that in step 2), in molar ratio,
Diethylenetriamine pentaacetic acid dianhydride: triethylamine: adenine=1:3:2；Diethylenetriamine pentaacetic acid dianhydride: triethylamine: cytimidine=1:
3:2；Diethylenetriamine pentaacetic acid dianhydride: triethylamine: adenine: cytimidine=1:3:1:1.
4. as described in claim 1 for detecting the fluorescence probe of xanthine, which is characterized in that in step 3), in molar ratio,
Intermediate product: Tb (NO3)3·6H2O=1:1.
5. the described in any item fluorescence probes for detecting xanthine of claim 1-4 are in qualitative and quantitative detection xanthine
Application.
6. application as claimed in claim 5, which is characterized in that for detecting the fluorescence probe of xanthine in qualitative and quantitative inspection
Survey the application of xanthine in urine.
7. application as claimed in claim 6, which is characterized in that in the fluorescence probe qualitative detection urine for detecting xanthine
Xanthine, the method is as follows: take urine, the aqueous solution of the above-mentioned fluorescence probe for being used to detect xanthine is added, is sufficiently mixed, carries out
The variation of fluorescence spectrum is observed in fluorescence detection.
8. application as claimed in claim 6, which is characterized in that in the fluorescence probe quantitative detection urine for detecting xanthine
Xanthine, the method is as follows: taking 50 μ L concentration is 5.0 × 10-3Mol/L for detect xanthine fluorescence probe aqueous solution in
In 10mL colorimetric cylinder, it is settled to 5mL with urine, carries out fluorescence detection.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811168452.5A CN109180707B (en) | 2018-10-08 | 2018-10-08 | Fluorescent probe for detecting xanthine and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811168452.5A CN109180707B (en) | 2018-10-08 | 2018-10-08 | Fluorescent probe for detecting xanthine and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109180707A true CN109180707A (en) | 2019-01-11 |
| CN109180707B CN109180707B (en) | 2020-05-12 |
Family
ID=64946818
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811168452.5A Active CN109180707B (en) | 2018-10-08 | 2018-10-08 | Fluorescent probe for detecting xanthine and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109180707B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106674134A (en) * | 2016-11-30 | 2017-05-17 | 辽宁大学 | Novel fluorescent probe, preparation method thereof and application of novel fluorescent probe to detection of 6-thioguanine |
| CN108358956A (en) * | 2018-03-22 | 2018-08-03 | 辽宁大学 | Fluorescence probe EuШ- dtpa-bis (adenine) and its application in detecting urine in orotic acid |
-
2018
- 2018-10-08 CN CN201811168452.5A patent/CN109180707B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106674134A (en) * | 2016-11-30 | 2017-05-17 | 辽宁大学 | Novel fluorescent probe, preparation method thereof and application of novel fluorescent probe to detection of 6-thioguanine |
| CN108358956A (en) * | 2018-03-22 | 2018-08-03 | 辽宁大学 | Fluorescence probe EuШ- dtpa-bis (adenine) and its application in detecting urine in orotic acid |
Non-Patent Citations (1)
| Title |
|---|
| YANG FAN等: "Design and synthesis of a novel lanthanide fluorescent probe (Tb-III-dtpa-bis(2,6-diaminopurine)) and its application to the detection of uric acid in urine sample", 《SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109180707B (en) | 2020-05-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zhou et al. | Design principles of spectroscopic probes for biological applications | |
| CN106970061A (en) | The preparation method of carbon point/copper nano-cluster compound ratio fluorescent dopamine probe | |
| CN107699234B (en) | A kind of Schiff bases Multifunction fluorescent probe and its preparation method and application | |
| CN107021953A (en) | A kind of coumarin fluorescent probe and preparation method and its application on detection hypochlorite ion | |
| CN109535147A (en) | A kind of formaldehyde fluorescence probe of quick response and its preparation method and application | |
| CN108358956B (en) | Fluorescent probe Eu III-dtpa-bis (adenine) and application thereof in detection of orotic acid in urine | |
| CN110964044B (en) | Peroxynitrite fluorescent probe based on dicoumarin derivative, preparation method and application | |
| CN111518066B (en) | Bifunctional fluorescent probe for identifying hypochlorite and bisulfite and preparation method and application thereof | |
| CN105777607B (en) | A kind of double indoles salt compounded of iodine of triphenylamine and its synthetic method and application | |
| CN111777598A (en) | Detect Cu2+And can utilize Cu2+Detection of HPO42–Fluorescent probe and preparation method and application thereof | |
| CN109180707A (en) | Fluorescence probe and its preparation method and application for detecting xanthine | |
| CN113736091B (en) | Method for detecting quercetin by using fluorescent micrometer probe and application | |
| CN114790202B (en) | Preparation and application of biological thiol activated precipitation dye type efficient diagnosis and treatment integrated probe | |
| CN112557355B (en) | Method for detecting bismuth ions in biological fluid by rare earth fluorescent probe | |
| CN110590762B (en) | Ratio type fluorescent probe for detecting bisulfite and preparation method and application thereof | |
| CN111732574B (en) | Naphthalimide-hydrazine pyridine derivative fluorescent probe and preparation method and application thereof | |
| CN114965417A (en) | Method for rapidly detecting methyl mercury by surface enhanced Raman scattering | |
| CN108896523B (en) | Fluorescence enhancement type hypochlorite detection method and application | |
| CN113563592A (en) | Fluorescent microsphere, fluorescent probe and method for detecting tetracycline | |
| CN106674048A (en) | CN<-> detection reagent and synthesis method and application thereof | |
| CN108693157B (en) | A method based on 2, 2': preparation and application method of fluorescent chemical sensor of 6 ', 2' -terpyridine derivative | |
| CN112500382A (en) | Ratiometric fluorescent probe for reversible detection of sulfur dioxide/formaldehyde, preparation method and application | |
| CN110563609A (en) | Preparation method and application of near-infrared fluorescent probe for detecting selenious acid roots | |
| CN107098890B (en) | Colorimetric fluorescent probe for detecting copper ions with high selectivity and ultra-sensitivity | |
| CN112760090B (en) | Palladium ion probe and preparation method and application thereof |
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 |