CN110483386A - A kind of superoxide anion probe and its preparation method and application - Google Patents
A kind of superoxide anion probe and its preparation method and application Download PDFInfo
- Publication number
- CN110483386A CN110483386A CN201910837565.8A CN201910837565A CN110483386A CN 110483386 A CN110483386 A CN 110483386A CN 201910837565 A CN201910837565 A CN 201910837565A CN 110483386 A CN110483386 A CN 110483386A
- Authority
- CN
- China
- Prior art keywords
- superoxide anion
- probe
- follows
- anion probe
- application
- 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
- OUUQCZGPVNCOIJ-UHFFFAOYSA-M Superoxide Chemical compound [O-][O] OUUQCZGPVNCOIJ-UHFFFAOYSA-M 0.000 title claims abstract description 71
- 239000000523 sample Substances 0.000 title claims abstract description 44
- 238000002360 preparation method Methods 0.000 title claims abstract description 9
- YQUVCSBJEUQKSH-UHFFFAOYSA-N 3,4-dihydroxybenzoic acid Chemical compound OC(=O)C1=CC=C(O)C(O)=C1 YQUVCSBJEUQKSH-UHFFFAOYSA-N 0.000 claims abstract description 14
- 238000002059 diagnostic imaging Methods 0.000 claims abstract description 7
- 150000001875 compounds Chemical class 0.000 claims abstract description 6
- 239000002994 raw material Substances 0.000 claims abstract description 5
- WQGWDDDVZFFDIG-UHFFFAOYSA-N pyrogallol Chemical compound OC1=CC=CC(O)=C1O WQGWDDDVZFFDIG-UHFFFAOYSA-N 0.000 claims description 38
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims description 24
- 229940079877 pyrogallol Drugs 0.000 claims description 19
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 12
- ICSNLGPSRYBMBD-UHFFFAOYSA-N 2-aminopyridine Chemical compound NC1=CC=CC=N1 ICSNLGPSRYBMBD-UHFFFAOYSA-N 0.000 claims description 10
- 239000001301 oxygen Substances 0.000 claims description 9
- 229910052760 oxygen Inorganic materials 0.000 claims description 9
- 238000003756 stirring Methods 0.000 claims description 8
- 238000001514 detection method Methods 0.000 claims description 7
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 6
- CSDSSGBPEUDDEE-UHFFFAOYSA-N 2-formylpyridine Chemical compound O=CC1=CC=CC=N1 CSDSSGBPEUDDEE-UHFFFAOYSA-N 0.000 claims description 5
- 229960000935 dehydrated alcohol Drugs 0.000 claims description 4
- 239000000706 filtrate Substances 0.000 claims description 4
- 239000012216 imaging agent Substances 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 4
- 239000002904 solvent Substances 0.000 claims description 4
- 238000010025 steaming Methods 0.000 claims description 4
- 230000005778 DNA damage Effects 0.000 claims description 3
- 231100000277 DNA damage Toxicity 0.000 claims description 3
- 230000008030 elimination Effects 0.000 claims description 3
- 238000003379 elimination reaction Methods 0.000 claims description 3
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 3
- 239000003795 chemical substances by application Substances 0.000 claims description 2
- 239000013066 combination product Substances 0.000 claims description 2
- 229940127555 combination product Drugs 0.000 claims description 2
- 238000000034 method Methods 0.000 abstract description 20
- JUJWROOIHBZHMG-UHFFFAOYSA-N pyridine Substances C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 abstract description 11
- 238000006243 chemical reaction Methods 0.000 abstract description 10
- 238000010791 quenching Methods 0.000 abstract description 7
- 230000000171 quenching effect Effects 0.000 abstract description 7
- 239000011159 matrix material Substances 0.000 abstract description 4
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 abstract description 4
- 230000035945 sensitivity Effects 0.000 abstract description 4
- 210000001541 thymus gland Anatomy 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 23
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 14
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 10
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 9
- 229960005070 ascorbic acid Drugs 0.000 description 7
- 239000000872 buffer Substances 0.000 description 6
- 230000008859 change Effects 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 230000005284 excitation Effects 0.000 description 6
- 239000002211 L-ascorbic acid Substances 0.000 description 5
- 235000000069 L-ascorbic acid Nutrition 0.000 description 5
- 238000004435 EPR spectroscopy Methods 0.000 description 4
- 229960004756 ethanol Drugs 0.000 description 4
- UHOVQNZJYSORNB-UHFFFAOYSA-N monobenzene Natural products C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 4
- 239000010453 quartz Substances 0.000 description 4
- 239000002516 radical scavenger Substances 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 230000032683 aging Effects 0.000 description 3
- 239000007853 buffer solution Substances 0.000 description 3
- 230000006378 damage Effects 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 150000003254 radicals Chemical class 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 3
- 238000004566 IR spectroscopy Methods 0.000 description 2
- 239000007983 Tris buffer Substances 0.000 description 2
- 235000010323 ascorbic acid Nutrition 0.000 description 2
- 239000011668 ascorbic acid Substances 0.000 description 2
- 238000006701 autoxidation reaction Methods 0.000 description 2
- 230000003139 buffering effect Effects 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 239000012488 sample solution Substances 0.000 description 2
- 230000002000 scavenging effect Effects 0.000 description 2
- PFTAWBLQPZVEMU-DZGCQCFKSA-N (+)-catechin Chemical compound C1([C@H]2OC3=CC(O)=CC(O)=C3C[C@@H]2O)=CC=C(O)C(O)=C1 PFTAWBLQPZVEMU-DZGCQCFKSA-N 0.000 description 1
- CQOZJDNCADWEKH-UHFFFAOYSA-N 2-[3,3-bis(2-hydroxyphenyl)propyl]phenol Chemical compound OC1=CC=CC=C1CCC(C=1C(=CC=CC=1)O)C1=CC=CC=C1O CQOZJDNCADWEKH-UHFFFAOYSA-N 0.000 description 1
- 208000024827 Alzheimer disease Diseases 0.000 description 1
- 244000080767 Areca catechu Species 0.000 description 1
- 235000006226 Areca catechu Nutrition 0.000 description 1
- 201000001320 Atherosclerosis Diseases 0.000 description 1
- ZGTMUACCHSMWAC-UHFFFAOYSA-L EDTA disodium salt (anhydrous) Chemical compound [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 241000007401 Magnolia odora Species 0.000 description 1
- 238000005481 NMR spectroscopy Methods 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 206010039966 Senile dementia Diseases 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 230000009429 distress Effects 0.000 description 1
- 238000002848 electrochemical method Methods 0.000 description 1
- 238000000921 elemental analysis Methods 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000000269 nucleophilic effect Effects 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 238000011897 real-time detection Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 210000002345 respiratory system Anatomy 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 239000000341 volatile oil Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
- A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
- C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
- C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D213/72—Nitrogen atoms
- C07D213/76—Nitrogen atoms to which a second hetero atom is attached
-
- 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
-
- 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
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1029—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Epidemiology (AREA)
- Biomedical Technology (AREA)
- Materials Engineering (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The invention discloses a kind of superoxide anion probes and its preparation method and application, and in particular to medical imaging field, including a kind of superoxide anion probe, structural formula are as follows:Its raw material compound structural formula is respectively as follows:WithThe present invention passes through using 2- (2 '-pyridine imine methyl) pyridine (2-APC) as fluorescence probe, mouse thymus cells system is as superoxide anion source, it is reacted using 2-APC with superoxide anion generation fluorescent quenching and makes the significantly reduced principle of reaction system fluorescence intensity, establish the measuring method of superoxide anion and protocatechuic acid relative concentration and superoxide anion clearance rate, easy to operate, high sensitivity, stability are good, superoxide anion can be preferably removed, the superoxide anion mitigated in superoxide anion probe is damaged caused by patient's matrix.
Description
Technical field
The present invention relates to medical imaging technical field, it is more particularly related to a kind of superoxide anion probe and
Preparation method and application.
Background technique
Have document report everyone there are about the intake oxygen of 1%-3% to be changed into superoxide anion and its reactive derivative daily.
The active oxygen of proper level can maintain body normal physiological function, and can be to machine when accumulated active oxygen is to a certain concentration in body
Body biomolecule causes to damage, so as to cause a variety of diseases.Some document reports claim active oxygen and hypertension, Atherosclerosis
Close relationship compared with change, respiratory tract Distress syndrome, senile dementia, tumour, diabetes have with aging.In recent years, freely
The research of base context of detection is paid close attention to by more and more researchers.The detection method of free radical mainly has at present: electron paramagnetic
Resonance method (electron paramagnetic resonance, EPR), high performance liquid chromatography (high-performance
Liquid chromatographic, HPLC), electrochemical process (electrochemical method), chemoluminescence method
(chemiluminescence method) and fluorescence probe method (fluorescence probe method).Wherein EPR method is
To the unique method that free radical is directly detected, but expensive equipment is unsuitable for conventional analysis.
In recent years, fluorescence probe method was widely used in the real-time detection of free radical in cell.The emphasis of such detection method
To synthesize highly selective, highly sensitive and with good fluorescence spectral characteristic novel fluorescence probe.The research is taken based on nucleophilic
For reaction principle, using 2-aminopyridine and pyridine-2-formaldehyde as a kind of novel fluorescence probe of Material synthesis: (2 '-pyridines are sub- by 2-
Amine methyl) pyridine (2-APC), pass through fusing point test, elemental analysis, infrared spectroscopy (IR) and hydrogen nuclear magnetic resonance wave spectrum (1H-NMR)
The methods of synthetic product is characterized.And establish the ultra-oxygen anion free radical fluoremetry side reacted based on fluorescent quenching
Method, while being applied to the evaluation of L-AA (L-ascorbic acid) to superoxide anion Scavenging activity.
The compound of existing fluorescence probe label is in use process, although can be good at playing fine to medical imaging
Fiting effect and disease treatment effect, be difficult to be purged it after imaging, although it is existing medically utilize Tsoongiodendron odorum
Blade volatile oil inhibit superoxide anion generation and remove its activity, damage of the superoxide anion to cell DNA can be reduced,
But its inhibitory effect is poor, can not comprehensively clear up superoxide anion, so that it is partially left at patient's body, endanger patient's
Health.
Summary of the invention
In order to overcome the drawbacks described above of the prior art, the embodiment of the present invention provides a kind of superoxide anion probe and its system
Preparation Method and application, by using 2- (2 '-pyridine imine methyl) pyridine (2-APC) as fluorescence probe, mouse thymus cells
System is reacted with superoxide anion generation fluorescent quenching using 2-APC as superoxide anion source and makes reaction system fluorescence
The significantly reduced principle of intensity, establishes the measurement of superoxide anion and protocatechuic acid relative concentration and superoxide anion clearance rate
Method, easy to operate, high sensitivity, stability are good, can preferably remove superoxide anion, mitigate in superoxide anion probe
Superoxide anion damaged caused by patient's matrix.
To achieve the above object, the invention provides the following technical scheme: a kind of superoxide anion probe, structural formula is such as
Under:
Its raw material compound structural formula is respectively as follows:
The superoxide anion probe the preparation method is as follows:
A) 20mL toluene, 4.71g 2-aminopyridine are added in 200mL round-bottomed flask；
B) mixed liquor of 5.36g pyridine-2-formaldehyde and 40mL toluene is added after stirring 30min at room temperature；
C) filter: Buchner funnel filters after stirring 10h at room temperature；
D) distill: filtrate is placed in Rotary Evaporators and is evaporated under reduced pressure；
E) it crystallizes: after steaming most of solvent, being recrystallized with dehydrated alcohol；
F) dry: dry the pale yellow powder in vacuum freeze drier；
The application of superoxide anion probe:
Superoxide anion probe can be used as the detection that medical imaging agent applies to medical conditions, needed after the completion of to be detected using
Remover is removed.
In a preferred embodiment, the product after the superoxide anion probe is combined with hydroxyl (- OH) can be led
Cell DNA damage is caused, human body function is destroyed, needs to dispel superoxide anion using pyrogallol after being detected.
In a preferred embodiment, protocatechuic acid can also be used in the remover of the superoxide anion.
Technical effect and advantage of the invention:
1, the present invention is by using 2- (2 '-pyridine imine methyl) pyridine (2-APC) as fluorescence probe, and pyrogallol is from oxygen
Change system is reacted with superoxide anion generation fluorescent quenching using 2-APC as superoxide anion source and keeps reaction system glimmering
The significantly reduced principle of luminous intensity establishes the measuring method of superoxide anion relative concentration Yu superoxide anion clearance rate, behaviour
It is good to make simplicity, high sensitivity, stability, can preferably remove superoxide anion；
2, removing of the present invention by the scavenger using protocatechuic acid as ultra-oxygen anion free radical, relative to aging method
Agent can effectively promote the elimination effect to superoxide anion, mitigate the superoxide anion in superoxide anion probe to patient
It is damaged caused by matrix.
Detailed description of the invention
Fig. 1 is superoxide anion probe synthetic route chart of the invention.
Fig. 2 is the fluorescence intensity change curve of pyrogallol of the invention as remover when excitation wavelength is 295nm
Figure.
Fig. 3 is the fluorescence intensity change curve of pyrogallol of the invention as remover when excitation wavelength is 365nm
Figure.
Fig. 4 is for pyrogallol of the invention as remover pH value to fluorescence intensity change curve graph.
Fig. 5 is for protocatechuic acid of the invention as remover to superoxide anion Scavenging activity line chart.
Specific embodiment
Below in conjunction with the embodiment in the present invention, technical solution in the embodiment of the present invention is carried out clearly and completely
Description, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.Based on this hair
Embodiment in bright, every other implementation obtained by those of ordinary skill in the art without making creative efforts
Example, shall fall within the protection scope of the present invention.
Embodiment 1:
The present invention provides a kind of superoxide anion probe, structural formula is as follows:
Its raw material compound structural formula is respectively as follows:
The superoxide anion probe the preparation method is as follows:
A) 20mL toluene, 4.71g 2-aminopyridine are added in 200mL round-bottomed flask；
B) mixed liquor of 5.36g pyridine-2-formaldehyde and 40mL toluene is added after stirring 30min at room temperature；
C) filter: Buchner funnel filters after stirring 10h at room temperature；
D) distill: filtrate is placed in Rotary Evaporators and is evaporated under reduced pressure；
E) it crystallizes: after steaming most of solvent, being recrystallized with dehydrated alcohol；
F) dry: dry the pale yellow powder in vacuum freeze drier；
The application of superoxide anion probe:
Superoxide anion probe can be used as the detection that medical imaging agent applies to medical conditions, needed after the completion of to be detected using
Remover is removed；
As shown in Figs 1-4, specific in the present embodiment: after the superoxide anion probe and hydroxyl (- OH) combination
Product will lead to cell DNA damage, destroy human body function, be needed superoxide anion after being detected using pyrogallol
It dispels.
The superoxide anion generated by the research using pyrogallol alkaline condition autoxidation system is measure object, measurement
Principle are as follows: superoxide anion can destroy 2-APC-C=N- group and reduce 2-APC fluorescence intensity by (fluorescent quenching phenomenon),
And superoxide anion content and 2-APC fluorescence intensity decreasing value (Δ F) are proportional；2.00mL is sequentially added in 10mL colorimetric cylinder
Tris-HCl buffer solution (pH=8.20,0.4mol/L), 0.3mL 2-APC ethanol solution (4.0 × 10-4Mol/L), 2.0mL
Pyrogallol solution (4.0 × 10-4Mol/L), ultrapure water is settled to scale, shakes up, the water-bath 40min in 40 DEG C of water-baths；Choosing
With 1cm quartz colorimetric utensil, λ ex=295nm, λ em=365nm are set, and excitation and transmite slit width are that 5nm measures its fluorescence
Intensity is denoted as F；It does pyrogallol blank test simultaneously and measures its fluorescence intensity and be denoted as F0, then Δ F=F0-F；Every group of test 3
It is a parallel；Different volumes L-AA solution (2.0 × 10 is added in measurement system-3Mol/L), its fluorescence intensity note is measured
For FVc；And the fluorescence intensity for measuring the system for being not added with ascorbic acid is denoted as F0；Not plus scavenger ascorbic acid and adjacent benzene is not added
The blank system of triphenol, fluorescence intensity F；Its fluorescence intensity is reduced by (note since L-AA can be reacted with 2-APC
For Fj), therefore this influence, calculation formula of the L-AA to superoxide anion clearance rate Q should be deducted when calculating clearance rate are as follows:
Q (%)=(FVc+Fj-F0)/(F-F0)×100；
The determination of maximum excitation wavelength and launch wavelength: 1cm quartz colorimetric utensil, slit width 5nm is selected to carry out 2-APC
Maximum excitation wavelength (λ ex) and launch wavelength (λ em) scanning (result is as shown in Figure 2)；(note: A, 3 curve reagent compositions in B
For 1,0.3mL 2-APC ethanol solution (4.0 × 10-4Mol/L), 2.00mL Tris-HCl buffer solution (pH=8.20,4.0 ×
10-4mol/L)；2,0.3mL 2-APC ethanol solution (4.0 × 10-4Mol/L), 2.00mL Tris-HCl buffer solution (pH=
8.20,4.0×10-4Mol/L), 1.5mL pyrogallol solution (4.0 × 10-4mol/L)；3,2.00mL Tris-HCl buffering is molten
Liquid (pH=8.20,4.0 × 10-4Mol/L), 1.5mL pyrogallol solution (4.0 × 14.0 × 10-4))；2-APC as shown in Figure 2
Maximum excitation wavelength X ex=295nm；The maximum emission wavelength λ em=365nm of 2-APC as shown in Figure 3；And by Fig. 21,2
It is had occurred after pyrogallol solution is added in 2-APC solution known to fluorescence intensity difference shown in two curves more apparent
Fluorescent quenching reaction；PH value measures the relative intensity of fluorescence (Δ F) of different pH reaction systems, (result such as Fig. 4 to the influence of Δ F
It is shown)；(note: reagent set becomes 0.3mL 2-APC ethanol solution (4.0 × 10-4Mol/L), 2.00mL Tris-HCl buffering is molten
Liquid (4.0 × 10-4) and 2.0mL pyrogallol solution (4.0 × 10 mol/L-4Mol/L), T=40 DEG C of bath temperature, water bath time
T=40min, λ ex=295nm, λ em=365nm, slit width 5nm))；As pH < 8.2, with the increase of pH, Δ F becomes rapidly
Greatly；As pH=8.2, Δ F reaches maximum value；And as pH > 8.2, Δ F is gradually decreased with the increase of pH；This is because adjacent benzene three
Phenol autoxidation generates O2 -Reaction rate improved with the increase of pH, as pH > 8.2, Δ F reduce the reason of may be with pH
Increase, 2-APC and O2 -Reactivity relative reduction；Therefore, the research select reaction system pH be 8.2, using 2-APC with
Superoxide anion occurs fluorescent quenching reaction and makes the significantly reduced principle of reaction system fluorescence intensity, establishes superoxide anion
The measuring method of relative concentration and superoxide anion clearance rate, easy to operate, high sensitivity, stability are good, can preferably go
Except superoxide anion.
Embodiment 2:
Its structural formula is as follows:
Its raw material compound structural formula is respectively as follows:
The superoxide anion probe the preparation method is as follows:
A) 20mL toluene, 4.71g 2-aminopyridine are added in 200mL round-bottomed flask；
B) mixed liquor of 5.36g pyridine-2-formaldehyde and 40mL toluene is added after stirring 30min at room temperature；
C) filter: Buchner funnel filters after stirring 10h at room temperature；
D) distill: filtrate is placed in Rotary Evaporators and is evaporated under reduced pressure；
E) it crystallizes: after steaming most of solvent, being recrystallized with dehydrated alcohol；
F) dry: dry the pale yellow powder in vacuum freeze drier；
The application of superoxide anion probe:
Superoxide anion probe can be used as the detection that medical imaging agent applies to medical conditions, needed after the completion of to be detected using
Remover is removed.
As described in figures 1 and 5, specific in the present embodiment: former catechu can also be used in the remover of the superoxide anion
Acid.
Solution prepare specific method: Tris solution (0.1mol/L), 1.21gTris (trishydroxymethylaminomethane,
M.W.121.1)+100mL distilled water, HCl solution (0.1mol/L): taking 0.1mL concentrated hydrochloric acid, add distilled water be diluted to 6mL,
Tris-HCl buffer (0.05mol/L, pH7.4, Na containing 1mmol/L2EDTA) 40mL0.1 mol/L Tris solution+x
ML0.1 mol/L HCl solution+15.2mg Na2EDTA, mixing, is diluted to 80mL, is measured with pH meter, pH should be 7.4, with palm fibre
Color bottle is stored in refrigerator (at most save three days), and (the above are the dosages of a sample) is with preceding slightly hot to room temperature, then surveys pH value,
It meets the requirements；60mmol/L pyrogallol solution (being dissolved in 1mmol/L hydrochloric acid) takes 20 μ L of 0.1mol/L HCl solution, uses
Distilled water is diluted to 2mL, obtains 1mmol/L hydrochloric acid solution (being measured with pH meter, pH=2.5-3.0)；Add pyrogallol inward again
14.6mg(M.W.126.1) to get (same day is effective, and the above are the dosages of 1 sample)；Pyrogallol solution: 2950 μ L are taken
Tris-HCl buffer is added in quartz colorimetric utensil, then plus about 50 μ L pyrogallol solution, rapidly mix (overturning formula), start
Timing, every A value (325nm) of 30 seconds readings, when 300 seconds (5min) until.(blank reference: Tris-HCl buffer)
Δ A=A325nm,300s-A325nm,30s.O is generated since Δ A value reflects2 -Initial concentration, so, for it is same a batch experiment and
Speech, Δ A value at this time must be equal, and Δ A at this time is Δ A0；Sample solution: x μ L sample solution is taken to be added to big quartz cuvette
In ware, then plus (2950-x) μ L Tris-HCl buffer, then plus 50 μ L pyrogallol solution, rapidly mix (overturning formula), start
Timing read primary (A value, 325nm) every 30 seconds, when 300 seconds until.(blank reference: Tris-HCl buffer) Δ A
=A325nm,300s-A325nm,30s；Δ A at this time is Δ ASample；Calculation formula:
Clearance rate=(Δ A0-ΔASample)/ΔA0*100；
Due to rosette like growth react it is very sensitive to temperature and pH value, and pH value by temperature fluctuation and change,
Therefore, strict temperature control, buffer is preferably more, cuvette 3.5mL specification, more stable, the experimental result of such data
(referring to Fig. 5), can be effective relative to the scavenger of aging method as the scavenger of ultra-oxygen anion free radical using protocatechuic acid
Promotion to the elimination effect of superoxide anion, mitigate the superoxide anion in superoxide anion probe caused by patient's matrix
Damage.
Last: the foregoing is only a preferred embodiment of the present invention, is not intended to restrict the invention, all in the present invention
Spirit and principle within, any modification, equivalent replacement, improvement and so on, should be included in protection scope of the present invention it
It is interior.
Claims (3)
1. a kind of superoxide anion probe, it is characterised in that: its structural formula is as follows:
Its raw material compound structural formula is respectively as follows:
The superoxide anion probe the preparation method is as follows:
A) 20mL toluene, 4.71g 2-aminopyridine are added in 200mL round-bottomed flask；
B) mixed liquor of 5.36g pyridine-2-formaldehyde and 40mL toluene is added after stirring 30min at room temperature；
C) filter: Buchner funnel filters after stirring 10h at room temperature；
D) distill: filtrate is placed in Rotary Evaporators and is evaporated under reduced pressure；
E) it crystallizes: after steaming most of solvent, being recrystallized with dehydrated alcohol；
F) dry: dry the pale yellow powder in vacuum freeze drier；
The application of superoxide anion probe:
Superoxide anion probe can be used as the detection that medical imaging agent applies to medical conditions, need after the completion of to be detected using elimination
Agent is removed.
2. a kind of application of superoxide anion probe, it is characterised in that: after the superoxide anion probe and hydroxyl (- OH) combination
Product will lead to cell DNA damage, destroy human body function, needed after being detected using pyrogallol by super oxygen yin from
Son is dispelled.
3. a kind of application of superoxide anion probe according to claim 2, it is characterised in that: the superoxide anion
Protocatechuic acid can also be used in remover.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910837565.8A CN110483386B (en) | 2019-09-05 | 2019-09-05 | Superoxide anion probe and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910837565.8A CN110483386B (en) | 2019-09-05 | 2019-09-05 | Superoxide anion probe and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110483386A true CN110483386A (en) | 2019-11-22 |
| CN110483386B CN110483386B (en) | 2022-03-15 |
Family
ID=68556668
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910837565.8A Active CN110483386B (en) | 2019-09-05 | 2019-09-05 | Superoxide anion probe and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110483386B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114456079A (en) * | 2021-12-23 | 2022-05-10 | 山东师范大学 | Fluorescent probe compound, preparation method and application of fluorescent probe compound as superoxide anion indicator |
-
2019
- 2019-09-05 CN CN201910837565.8A patent/CN110483386B/en active Active
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114456079A (en) * | 2021-12-23 | 2022-05-10 | 山东师范大学 | Fluorescent probe compound, preparation method and application of fluorescent probe compound as superoxide anion indicator |
| CN114456079B (en) * | 2021-12-23 | 2023-06-23 | 山东师范大学 | Fluorescent probe compound, preparation method and application of fluorescent probe compound as superoxide anion indicator |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110483386B (en) | 2022-03-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104610955B (en) | A kind of Ratio-type detects the synthesis and application of fluorine ion and inferior sulfate radical fluorescent molecular probe simultaneously | |
| Drujan et al. | The differential determination of catecholamines in urine | |
| CN111233880B (en) | Preparation method of hypochlorite fluorescent probe | |
| CN106946902B (en) | A kind of sulfur dioxide near-infrared-two-photon ratio fluorescent probe and preparation method thereof | |
| Peng et al. | Fluorescent nanoprobe for in-vivo ratiometric imaging of endogenous hydrogen peroxide resulted from drug-induced organ damages | |
| CN107556305B (en) | Fluorescent probe for detecting aluminum ions, preparation method and application | |
| CN105842235B (en) | It is highly sensitive, can open hole detection effumability organic amine fluorescent test paper and preparation | |
| CN106749359B (en) | A kind of synthesis and application for detecting hydrogen peroxide novel fluorescence probe | |
| JP2003509442A (en) | Glucose sensing molecules with selected fluorescent properties | |
| Yoe et al. | The colorimetric determination of palladium with p-nitrosodiphenylamine | |
| CN107216270A (en) | A kind of application for detecting hydrogen sulfide high selectivity response type fluorescence probe | |
| CN110746321B (en) | Malononitrile Schiff base hypochlorous acid fluorescent probe and preparation method thereof | |
| CN110015992A (en) | Fluorescence probe and its preparation method and application of the one kind for the detection of sulfur dioxide/sulfurous acid (hydrogen) salt | |
| Hu et al. | A ratiometric fluorescent probe for hyaluronidase detection via hyaluronan-induced formation of red-light emitting excimers | |
| CN110483386A (en) | A kind of superoxide anion probe and its preparation method and application | |
| CN107383078A (en) | Phenylboric acid ester compounds and the benzoyl peroxide detection kit comprising the compound | |
| CN113121580B (en) | Hydrogen peroxide fluorescent probe and preparation method and application thereof | |
| CN110878085B (en) | Rapid high-selectivity hypobromous acid fluorescent probe, preparation method and application | |
| CN107033078B (en) | Iron ion sensor molecule and its synthesis and application containing Hydroxynaphthaldehyde structure | |
| CN110981857B (en) | Ultrasensitive ferrous ion fluorescent probe, preparation method and application | |
| CN105820183B (en) | Fluoroboropyrrole compound containing α -unsaturated ketone and application thereof in sulfite detection | |
| CN111548304A (en) | Derivative based on triphenylamine and preparation method and application thereof | |
| CN108148056B (en) | Ratio-type near-infrared cysteine fluorescence probe | |
| CN108191848B (en) | Prepare the method for detecting the kit of cysteine | |
| CN112341366B (en) | Triarylamine derivative fluorescent 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 |