CN109776369A - A kind of hypersensitive is highly selective to analyze hypochlorous fluorescence probe in real time - Google Patents
A kind of hypersensitive is highly selective to analyze hypochlorous fluorescence probe in real time Download PDFInfo
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- CN109776369A CN109776369A CN201910193501.9A CN201910193501A CN109776369A CN 109776369 A CN109776369 A CN 109776369A CN 201910193501 A CN201910193501 A CN 201910193501A CN 109776369 A CN109776369 A CN 109776369A
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- hypochlorous acid
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Abstract
Hypochlorous fluorescence probe is analyzed in real time the present invention relates to a kind of hypersensitive is highly selective.Specifically, probe of the invention is a kind of indanone compounds, can be used as hypochlorous acid fluorescence probe for hypochlorous detection.This kind of probe can realize following at least one of technical effect: identify hypochlorous acid with high selectivity；Immediately hypochlorous acid can be realized and is responded；It may be implemented to analyze hypochlorous hypersensitive；Property is stablized, and can be used with long-term preservation.
Description
Technical field
The present invention relates to indanone compounds as hypochlorous acid fluorescence probe, can be rapidly to the highly selective oversoul of hypochlorous acid
Quick identification or its can measure hypochlorous concentration in actual water sample immediately.
Background technique
Signaling molecule of the active oxygen as life system, it is closely related with physiology and pathologic process.In high response activity
In oxygen, hypochlorous acid is to be generated under peroxide enzyme effect by hydrogen peroxide and chloride ion, it is passed in the signal of biosystem
It leads and is played a key role in innate immune response, had received widespread attention in biological and medical field.Although hypochlorous acid
There is certain protective effect to human health, still, since hypochlorous acid has very strong oxidability, mistake to various biomolecule
The hypochlorous acid of amount will lead to cell and tissue damage.More and more evidences also indicate that improper hypochlorous acid regulation is that induction is various
The key factor of disease, such as cancer, neurodegenerative disease and diseases associated with inflammation.Therefore, it is suitable to find in biosystem
Chemical tools be of great significance to track the variation of hypochlorous acid concentration.
Due to its highly sensitive and significant spatial and temporal resolution, fluorescence probe is as a kind of Noninvasive tool application in life
Analyte determination in object imaging.Up to the present, it has devised and has been largely used to detect hypochlorous fluorescence spy in biosystem
Needle.But there are still some defects for the hypochlorous acid fluorescence probe reported at present, including selectivity is not good enough, sensitivity is not high enough,
Poorly water-soluble, synthesis are complicated.In addition, being explored a kind of high since oxidation time is short in physiological conditions, concentration is low for hypochlorous acid
The hypochlorous acid imitated in fluorescence probe real-time detection living body and living cells is still a hot issue.In addition, for detecting time chlorine
The fluorescence probe that good water solubility, the quantum efficiency of acid are high is also relatively fewer.Therefore, a kind of high quantum production is found in biosystem
Rate, highly selective, hypersensitive, the hypochlorous tool of real-time detection are still vital.
Summary of the invention
This field urgent need prepares a kind of quick high-selectivity hypochlorous acid fluorescence probe of high quantum production rate, so as to effective
Detect hypochlorous acid.For this purpose, the present invention has synthesized a kind of novel hypochlorous acid fluorescence probe, quantum yield is high, selectivity is good, clever
Sensitivity is high, can quickly identify hypochlorous acid.Specifically, being indone class the present invention provides a kind of hypochlorous acid fluorescence probe
Compound, structure are as follows:
Preferably, fluorescence probe of the invention is:
It is by the way that probe of the invention will be corresponded to the present invention also provides the preparation method of hypochlorous acid fluorescence probe
Corresponding indanone compounds and the thio formyl chloride of dimethylamino flow back 48h in dichloromethane solution and synthesize obtained.
The present invention also provides for detecting the detection preparation of hypochlorous acid concentration in sample (such as swimming-pool water), it includes
Probe of the invention.Preferably, detection preparation of the invention or kit also include the operation instructions of product.Preferably, originally
The detection preparation of invention also includes the buffer for measuring the hypochlorous acid concentration in sample.
The present invention also provides the methods of hypochlorous acid concentration in detection sample (such as swimming-pool water) comprising will be of the invention
Probe and sample to be tested the step of contacting.
It is dense for detecting hypochlorous acid in sample (such as swimming-pool water) in preparation that the present invention also provides probes of the invention
Purposes in the preparation of degree.
Fluorescence probe of the invention can be acted on hypochlorous acid, generate the variation of fluorescence spectrum, to realize to secondary chlorine
The quantitative detection of acid.
Specifically, hypochlorous acid fluorescence probe of the invention respectively with potassium ion, calcium ion, zinc ion, sodium ion, copper from
Son, magnesium ion, ferrous iron, ferric iron, chloride ion, nitrate anion, nitrite anions, sulfate radical, hydrogen peroxide, potassium superoxide etc. other
Substance, which is acted on, cannot lead to substantially changeing for fluorescence spectrum, to realize to hypochlorous Selective recognition, Jin Erke
Optionally for exclusion these potassium ions, calcium ion, zinc ion, sodium ion, copper ion, magnesium ion, ferrous iron, ferric iron, chlorine
The presence pair of other ions in the substances such as ion, nitrate anion, nitrite anions, sulfate radical, hydrogen peroxide, potassium superoxide and human body
The interference of hypochlorous quantitative determination.
Fluorescence probe of the invention reacts very sensitive with hypochlorous acid, to be conducive to hypochlorous rapid detection.
Selectively, the stability of hypochlorous acid fluorescence probe of the invention is good, and then being capable of long-term preservation use.
Further, hypochlorous acid fluorescence probe of the invention is the glimmering color probe of quick high-selectivity hypochlorous acid, and synthesizes letter
It is single, be conducive to commercialized popularization and application.
Detailed description of the invention
Fig. 1 is (5 μM) of the probe response times being added after hypochlorous acid (1 μM)；
Fig. 2 is (5 μM) of the probe fluorescence spectrums being added before and after hypochlorous acid (0-5 μM)；
Fig. 3 is that various concentration hypochlorous acid (0-1 μM) meets good linear relationship to probe (5 μM).
Fig. 4 is influence of the substance common in human body to the fluorescence intensity of probe (5 μM).Wherein number 1-28 is respectively sky
White, potassium ion, calcium ion, zinc ion, sodium ion, copper ion, magnesium ion, ferrous iron, ferric iron, chloride ion, nitrate anion, nitrous
Acid group, inferior sulfate radical, sulfate radical, vulcanized sodium, glutathione (5mM), homocysteine (500 μM), cysteine (500 μ
M), hydrogen peroxide, hydroxyl radical free radical, tertbutanol peroxide, tertbutanol peroxide free radical, potassium superoxide, singlet oxygen, an oxygen
Change nitrogen, peroxynitrite, sodium hypochlorite (5 μM) (in addition to special indicate, other analyte concentrations are 100 μM).Column
Scheme fluorescence intensity level of the probe at 505nm in the presence of being different analytes represented.
Specific embodiment:
The present invention provides synthetic route, method and its light of above-mentioned quick high-selectivity hypersensitive hypochlorous acid fluorescence probe
Spectrality energy.
Hypochlorous acid fluorescence probe of the invention is a kind of indanone compounds, has following structure general formula
In above formula: R1, R2, R3, R4, R5,R6, R7, R8, R9And R10, it is hydrogen atom, linear or branched alkyl group, linear chain or branched chain
Alkoxy, sulfonic group, ester group, carboxyl；R1, R2, R3, R4, R5, R6, R7, R8And R9And R10, can be identical or different.
The synthetic route and method of such hypochlorous acid fluorescent needle are as follows:
Specifically, fluorescence probe of the invention can be prepared via a method which, by certain mol proportion (such as 1:2~1:
6) indanone compounds and the thio formyl chloride of dimethylamino are dissolved in methylene chloride, be then added DIPEA (such as 1:2~1:
6) it, is heated to reflux a period of time (such as 48h), is then rotated to obtain crude product using Rotary Evaporators, if to obtain
The mixed system (such as v/v, 2:1) of filtrate methylene chloride and petroleum ether can be carried out pillar layer separation by purer product
Obtain sterling.
Therefore, the present invention also provides the thio formyl chlorides of dimethylamino to prepare for detecting in hypochlorous fluorescence probe
Purposes.
The present invention also provides preparing indanone compounds for detecting the purposes in hypochlorous fluorescence probe.
Hypochlorous acid fluorescence probe of the invention be noteworthy characterized by can quick high-selectivity hypersensitive identification hypochlorous acid with
And quantitative analysis accurately can be carried out to hypochlorous acid in the presence of other intracorporal ions of people.
It below will be by the way that the present invention be described in more detail by following embodiment.Following embodiment is merely illustrative,
It should be understood that the present invention is not limited by following embodiments.
Embodiment 1
(scheme 1) 472mg (2mmol) 2- benzylidene -5- hydroxide radical-1-indenone is dissolved in 10mL methylene chloride, is added
Then the thio formyl chloride of 496mg (4mmol) dimethylamino is added DIPEA516mg (4mmol) reflux 10h, then utilizes rotation
Evaporimeter is rotated, and crude product is obtained.If obtaining purer product, the mixture of methylene chloride and petroleum ether can be used
It is that (such as v/v, 2:1) carries out the isolated sterling of chromatographic column, obtains pure product 460mg, yield 71.2%.
472mg (2mmol) 2- benzylidene -5- hydroxide radical-1-indenone is dissolved in 10mL methylene chloride by (scheme 2), then plus
Enter the thio formyl chloride of 744mg (6mmol) dimethylamino, DIPEA 774mg (6mmol) reflux 16h is then added, then utilizes rotation
Turn evaporimeter to be rotated to obtain crude product.If obtaining purer product, the mixing of methylene chloride and petroleum ether can be used
System (such as v/v, 2:1) carries out chromatographic column and purifies to obtain sterling, obtains pure product 500mg, yield 77.4%.
472mg (2mmol) 2- benzylidene -5- hydroxide radical-1-indenone is dissolved in 10mL methylene chloride by (scheme 3), then plus
Enter the thio formyl chloride of 992mg (8mmol) dimethylamino, DIPEA 1032mg (8mmol) reflux is then added and for 24 hours, then utilizes
Rotary Evaporators are rotated to obtain crude product.It, can be mixed with methylene chloride and petroleum ether if obtaining purer product
Zoarium system (such as v/v, 2:1) carries out the isolated sterling of chromatographic column, obtains pure product 540mg, yield 83.6%.
472mg (2mmol) 2- benzylidene -5- hydroxide radical-1-indenone is dissolved in 10mL methylene chloride by (scheme 4), then plus
Enter the thio formyl chloride of 1488mg (12mmol) dimethylamino, DIPEA 1548mg (12mmol) reflux 48h is then added, it is then sharp
It is filtered with high-pressure pump, obtains filtrate, filtrate becomes our crude product.If obtaining purer product, can will filter
The mixed system (such as v/v, 2:1) of liquid methylene chloride and petroleum ether carries out the isolated sterling of chromatographic column.It is pure to obtain yellow
Net products 580mg, yield 89.8%.
472mg (2mmol) 2- benzylidene -5- hydroxide radical-1-indenone is dissolved in 10mL methylene chloride by (scheme 5), then plus
Enter the thio formyl chloride of 330mg (12mmol) dimethylamino, DIPEA 3096mg (12mmol) reflux is then added for 24 hours, it is then sharp
It is filtered with high-pressure pump, obtains filtrate, filtrate becomes our crude product.If obtaining purer product, can will filter
The mixed system (such as v/v, 2:1) of liquid methylene chloride and methanol carries out the isolated sterling of chromatographic column.It is pure to obtain yellow
Product 550mg, yield 85.1%.
Embodiment 2
Fig. 1 is (5 μM) of the probe response times being added after hypochlorous acid (1 μM).50 μ L are taken out from probe mother liquor to be placed in
Then 1 μM of hypochlorous acid is added in test system by the test system of 10mL, surveyed immediately with Fluorescence Spectrometer after rocking uniformly
Try its fluorescence intensity change.Said determination is carried out in pure water (5mM PBS, pH 7.4), and used probe is to implement
Prepared probe in example 1, and all spectrum tests are measured at 25 DEG C.
By figure it will be clear that after hypochlorous acid is added, fluorescence intensity reaches maximum value and keeps in detection 3s
Constant, this illustrates that the probe is swift in response with hypochlorous acid, can provide quick analysis method for hypochlorous measurement.
Embodiment 3
Fig. 2 is (5 μM) of the hypochlorous acid fluorescence probe fluorescence spectrum variation diagrams that hypochlorous acid (0-5 μM) is added.Fig. 3 is different dense
Hypochlorous acid (0-1 μM) is spent to the linear relationship chart of probe (5 μM).
The parallel sample that multiple concentration and probe concentrations are 5 μM is configured in 10mL colorimetric cylinder, then by the hypochlorous acid of various concentration
It is added in test system, stands 1 minute after rocking uniformly.Said determination is carried out in pure water (5mM PBS, pH 7.4)
, used probe is probe prepared in embodiment 1, and all spectrum tests are measured at 25 DEG C.
Its fluorescence intensity change is tested with Fluorescence Spectrometer, can clearly be found out from Fig. 2, with addition hypochlorous acid concentration
Increase, the fluorescence intensity at 505nm gradually increases.Also, as seen from Figure 3, at 505nm, hypochlorous acid fluorescence probe
Fluorescence intensity presents good linear relationship after (5 μM) addition hypochlorous acid (0-1 μM), this is proved by means of the fluorescence probe
Quantitative analysis can be carried out to hypochlorous acid.
Embodiment 4
Fig. 4 is fluorescence intensity of the different ions analyte to probe (5 μM).Analyte be respectively blank, potassium ion, calcium from
Son, zinc ion, sodium ion, copper ion, magnesium ion, ferrous iron, ferric iron, chloride ion, nitrate anion, nitrite anions, inferior sulfate radical,
Bisulfite, sulfate radical, vulcanized sodium, glutathione (5mM), homocysteine (500 μM), cysteine (500 μM), mistake
Hydrogen oxide, hydroxyl radical free radical, tertbutanol peroxide, tertbutanol peroxide free radical, ultra-oxygen anion free radical, singlet oxygen,
Nitric oxide, peroxynitrite, sodium hypochlorite (5 μM) (in addition to special indicate, other analyte concentrations are 100 μM).
Fluorescence intensity level of the probe at 505nm in the presence of being different analytes that histogram represents.Said determination is in pure water (5mM
PBS, pH 7.4) in carry out, used probe is probe prepared in embodiment 1, and all spectrum tests be all
It is measured at 25 DEG C.Specifically, the parallel sample that multiple concentration and probe concentrations are 5 μM is configured in 10mL colorimetric cylinder, is then added one
Quantitative analyte, shakes up, and measures after 1 minute.
From fig. 4, it can be seen that will not to significantly interfere with probe strong to hypochlorous fluorescence for existing common ion in organism
Degree, therefore probe has good selectivity.
Although with above embodiments describe the present invention, it should be appreciated that before without departing substantially from spirit of the invention
It puts, the present invention further can be modified and be changed, and these modifications and variation all belong to the scope of protection of the present invention it
It is interior.
Claims (10)
1. compound has following structure
Wherein: R1, R2, R3, R4, R5, R6, R7, R8, R9And R10For independently selected from by hydrogen atom, linear or branched alkyl group, straight chain
Or the group of branched alkoxy, sulfonic group, ester group and hydroxyl composition；And R therein1, R2, R3, R4, R5, R6, R7, R8, R9And R10It can
With identical or different.
2. compound according to claim 1 is the compound such as flowering structure:
3. the preparation for detecting hypochlorous acid content in sample, it includes the compounds having following structure:
Wherein: R1, R2, R3, R4, R5, R6, R7, R8, R9And R10For independently selected from by hydrogen atom, linear or branched alkyl group, straight chain
Or the group of branched alkoxy, sulfonic group, ester group and hydroxyl composition；And R therein1, R2, R3, R4, R5, R6, R7, R8, R9And R10It can
With identical or different.
4. preparation according to claim 3, wherein the compound is:
5. preparation according to claim 3, wherein the sample is tissue or water (such as swimming-pool water).
6. the method for preparing the compound of claims 1 or 2 comprising following steps:
2- benzylidene -5- the hydroxide radical-1-indenone of certain mol proportion is dissolved in 10mL methylene chloride, is added a certain proportion of two
The thio formyl chloride of methylamino and DIPEA, are then refluxed for a period of time, and revolving obtains crude product；Optionally, by crude product dichloro
The mixed system of methane and petroleum ether carries out the isolated sterling of chromatographic column.
7. method according to claim 6, wherein the 2- benzylidene -5- hydroxide radical-1-indenone and the thio formyl of dimethylamino
The molar ratio of chlorine is 1:2~1:6.
8. method according to claim 6, wherein the return time is 10~48 hours.
9. the compound of claims 1 or 2 is in preparation for for detecting hypochlorous preparation in sample or the use in kit
On the way.
10. purposes according to claim 9, wherein the sample is tissue or water (such as swimming-pool water).
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| CN109503443A (en) * | 2018-12-25 | 2019-03-22 | 姜懿珊 | A kind of hypersensitive is highly selective to analyze hypochlorous fluorescence probe in real time |
| CN110372632A (en) * | 2019-07-26 | 2019-10-25 | 广东轻工职业技术学院 | A kind of fluorescent probe molecule and its preparation method and application of quick identification hypochlorite ion |
| CN115073338A (en) * | 2022-07-14 | 2022-09-20 | 济南大学 | High-selectivity mercury ion recognition fluorescent probe, preparation method and application |
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| CN107746406A (en) * | 2017-11-22 | 2018-03-02 | 济南大学 | A kind of preparation and application of hypersensitive high selectivity hypochlorous acid fluorescence probe |
| CN108409697A (en) * | 2018-04-23 | 2018-08-17 | 济南大学 | A kind of highly selective hypochlorous probe of detection of hypersensitive |
| CN108690040A (en) * | 2018-05-07 | 2018-10-23 | 济南大学 | A kind of highly selective hypochlorous colorimetric fluorescence probe of detection of hypersensitive |
| CN109503443A (en) * | 2018-12-25 | 2019-03-22 | 姜懿珊 | A kind of hypersensitive is highly selective to analyze hypochlorous fluorescence probe in real time |
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| CN107266397A (en) * | 2017-06-26 | 2017-10-20 | 闫语 | A kind of preparation and application of thiocarbamates hypochlorous acid fluorescence probe |
| CN107746406A (en) * | 2017-11-22 | 2018-03-02 | 济南大学 | A kind of preparation and application of hypersensitive high selectivity hypochlorous acid fluorescence probe |
| CN108409697A (en) * | 2018-04-23 | 2018-08-17 | 济南大学 | A kind of highly selective hypochlorous probe of detection of hypersensitive |
| CN108690040A (en) * | 2018-05-07 | 2018-10-23 | 济南大学 | A kind of highly selective hypochlorous colorimetric fluorescence probe of detection of hypersensitive |
| CN109503443A (en) * | 2018-12-25 | 2019-03-22 | 姜懿珊 | A kind of hypersensitive is highly selective to analyze hypochlorous fluorescence probe in real time |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109503443A (en) * | 2018-12-25 | 2019-03-22 | 姜懿珊 | A kind of hypersensitive is highly selective to analyze hypochlorous fluorescence probe in real time |
| CN110372632A (en) * | 2019-07-26 | 2019-10-25 | 广东轻工职业技术学院 | A kind of fluorescent probe molecule and its preparation method and application of quick identification hypochlorite ion |
| CN110372632B (en) * | 2019-07-26 | 2021-04-06 | 广东轻工职业技术学院 | Fluorescent probe molecule for rapidly identifying hypochlorite ions and preparation method and application thereof |
| CN115073338A (en) * | 2022-07-14 | 2022-09-20 | 济南大学 | High-selectivity mercury ion recognition fluorescent probe, preparation method and application |
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Application publication date: 20190521 |