CN107383078A - Phenylboric acid ester compounds and the benzoyl peroxide detection kit comprising the compound - Google Patents
Phenylboric acid ester compounds and the benzoyl peroxide detection kit comprising the compound Download PDFInfo
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- CN107383078A CN107383078A CN201710654441.7A CN201710654441A CN107383078A CN 107383078 A CN107383078 A CN 107383078A CN 201710654441 A CN201710654441 A CN 201710654441A CN 107383078 A CN107383078 A CN 107383078A
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- C07F5/02—Boron compounds
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- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
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- 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
- G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
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- 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
- C09K2211/1096—Heterocyclic compounds characterised by ligands containing other heteroatoms
Abstract
The present invention relates to benzoyl peroxide detection technique field, and in particular to a kind of phenylboric acid ester compounds and the benzoyl peroxide detection kit comprising the compound.Benzoyl peroxide detection kit includes reagent stock liquid a and reagent stock liquid b；The reagent stock liquid a is the phosphate buffer for the ethanol that pH value is 6~8, and the reagent stock liquid b is the organic solvent solution of phenylboric acid ester compounds.Reagent storing solution b fluorescence itself is weaker in the benzoyl peroxide detection kit of the present invention；And then fluorescence intensity can be remarkably reinforced after being reacted with benzoyl peroxide, strong absorption is produced at 690nm~750nm, and fluorescence significantly increases at 706nm, the fluorescent emission bands are near-infrared region, ambient interferences are few, therefore have higher accuracy and sensitivity；And the fluorescence probe has specificity to benzoyl peroxide, do not disturbed by other common inorganic ions and oxide species.
Description
Technical field
The invention belongs to benzoyl peroxide detection technique field, and in particular to phenylboric acid ester compounds and include the change
The benzoyl peroxide detection kit of compound.
Background technology
In recent years, as the improvement of people's living standards, food-safety problem also has been to be concerned by more and more people.Ensure
Food security is to build Health China, promote the important content of the well-being of the people, is the specific body of the development thought centered on the people
It is existing.Food security sampling observation situation is classified as the key content of food security job rating by " 13 " state food safety program,
Food security sampling observation covering all foods classification, kind.Food additives are the main reason for causing food problems, numerous
In food additives, benzoyl peroxide is that important food brightens antistaling agent, is widely used in food, chemical industry and medicine neck
Domain.In field of food, it is often used as the bleaching agent of wheat flour, and it such as can also play antibacterial, accelerate the ripening at effect in addition.
However, benzoyl peroxide (benzoyl peroxide) solve our daily lifes, production take it is hiding
Huge potential safety hazard.Excessive addition as the benzoyl peroxide of active oxygen species can cause the nutritional ingredient in food
The degraded of such as VitAVitE and vitamin B1, benzoyl peroxide can also aggravate burden of liver, inspire canceration, right
Peripheral blood can produce heredity toxic side effect.In addition, the benzoyl peroxide in food can produce in food heating process
Benzene radical, and then the noxious materials such as benzene, phenol, biphenyl can be formed, there is huge damage to human body, free radical accelerates human body
Cell ageing, cause atherosclerosis, or even induce a variety of diseases.Based on this, many countries and regions are all kept under strict control peroxidating
The use of benzoyl.China's formally addition of disabling benzoyl peroxide in flour in 1 day May in 2011.But can be small
Used in flour and bactericide.
The method for being presently used for benzoyl peroxide detection has high performance liquid chromatography (HPLC), gas-chromatography (GC), mass spectrum
With chromatograph joint used, chromatography of ions, chemoluminescence method, electrochemistry etc..The illegal addition of benzoyl peroxide occurs mainly in food
In, food is the product that must disappear of national product life, even sampling observation also has substantial amounts of sample, it is therefore desirable to more convenient quickly
Detection method.
China application CN201110084232.6 discloses a kind of violated additive benzoyl peroxide quick detection of flour
Kit, it is made up of reaction reagent, developer, positive control agent storing solution and standard color comparison card；Reaction reagent is concentration
100mg/L to 300mg/L ferrous ion acidic aqueous solution；Developer is concentration 0.5mol/L to 0.75mol/L thiocyanic acid
The root aqueous solution；Positive control agent storing solution is concentration 500mg/L to 1500mg/L benzoyl peroxide ethanol solution；Standard
Colorimetric card includes some small color lumps, and each color lump corresponds to benzoyl peroxide concentration.Its Cleaning Principle is to utilize benzoyl peroxide first
The strong oxidizing property of acyl, by the ferrous ions in reaction reagent into ferric ion, ferric ion is again in an acidic solution
Chromogenic reaction occurs with the thiocyanate radical in developer, generates orange red ferric rhodanate complex, orange red complex color
The depth carries out half-quantitative detection with Determination of benzoyl peroxide in wheat in correlation, but this method has larger limitation, shows
Colour response is in visible light wave range, and many sample compositions are complex, and the pigment of itself has very big do for chromogenic reaction
Immunity, especially for the detection of biological tissue or biological tissue, this detection kit can not be applicable completely.
China application CN201611163372.1 discloses a kind of benzoyl peroxide quick determination method, its Cleaning Principle
Can form the material that can develop the color of visible light wave range with detection reagent also with the benzoyl peroxide in sample, using with mark
Quasi- colorimetric card carries out color and compares progress semi-quantitative analysis.In above-mentioned common method, sample pre-treatments are cumbersome time-consuming, it is necessary to sample
Product are extracted, and can just carry out chromogenic reaction；And need to carry out colorimetric twice and judge that cost is high, is not easy to benzoyl peroxide first
The quick detection of acyl, it is not easy to promote the use of.
Therefore, a kind of new method of the sensitive quick detection Determination of benzoyl peroxide in wheat of energy is established, is carried for basic unit's quality testing department
For technical support, the safe handling for realizing perbenzoic acid is significantly.
Optical probe is that one kind can react with target substance and cause optics (including luminous, fluorescence, extinction) property
Change., can be with the strong of qualitative or even quantitative environmental stimuli by means of means such as such as XRF, fluorescence imaging microscopes
It is weak, so as to realize the purpose of analysis detection.Fluorescence probe is mainly detection means by fluorescence signal, generally there is the increasing of fluorescence
By force, it is quenched or the change of emission wavelength.The change of luminescence phenomenon and absorption spectrum that fluorescence probe passes through molecule detects point
Interaction between son, its advantage may be summarized to be the following aspects：(1) it is convenient and swift, there is very high sensitivity；(2)
Single celled detection in real time can be realized using optical fiber technology；(3) if having bigger change, Ke Yi on absorption spectrum
In the case of without the help of any instrument, the purpose of detection is directly reached by the change of color.
Fluorescent spectrometry high sensitivity, selectivity are good, and the information of acquisition is directly perceived, accurate, can the complicated sample of Scientific Expression explanation
Structure, distribution, content and physiological function of product etc., therefore, obtain analyzing the concern of detection field scientific research personnel.But fluorescence
The foundation of spectroscopic methodology many organisms and its is organized under the exciting of visible ray and itself can launch fluorescence there is also some difficult point,
The fluoroscopic examination of severe jamming biological sample and radiography, as haemocyanin in blood plasma fluorescent wavelength ranges for 325nm~
The fluorescent wavelength ranges of 350nm, NADH phosphatase (NADPH) and bilirubin be 430nm~
470nm, so that the sensitivity of fluorescence analysis and accuracy receive very big influence.Launch wavelength is in the glimmering of visible light wave range
Light probe when applied to biological living or biological sample analysis sensitivity and accuracy deficiency, it is necessary to develop it is possible to prevente effectively from
The novel probe compound of biological tissue fluorescence interference itself.
The maximum absorption wavelength and launch wavelength of near infrared fluorescent probe are 600nm~900nm, and light infiltrating tissues are up to number
Centimetre, excellent biological tissue's penetration capacity is detected in biological sample analysis with resistance ambient interferences ability, near-infrared fluorescent
Have shown that obvious superiority.In consideration of it, exploitation launch wavelength belongs to the fluorescence detection reagent kit of near-infrared region, for biology
The detection of benzoyl peroxide has important value in sample.
The content of the invention
In order to solve Detrmination of Benzoyl Peroxide poor selectivity in the prior art, sample pre-treatments are cumbersome, and colour developing is special
The interference that property belongs to the chromogenic reaction of visible ray section, sensitivity and accuracy by biological specimen in itself can be greatly reduced, because
And the defects of being not suitable for biology sample detection, on the one hand, it is of the invention using tricarbocyanine IR-780 skeleton derivatives as fluorogen, with
As recognition group, design had synthesized one kind to be detected near infrared emission wave-length coverage the phenyl boronate of ortho position substitution
The compound of BP, on the other hand, present invention also offers a kind of benzoyl peroxide based on fluorescence probe method
Detection kit and its detection method.
Phenylboric acid ester compounds shown in offer formula (I) of the present invention,
The preparation method of phenylboric acid ester compounds shown in offer formula (I) of the present invention, comprises the following steps：
In the presence of a base, compound shown in formula (IV) and the compound shown in formula (V) are substituted point step (A)
Rearrangement reaction is solved, obtains compound shown in formula (II)；
In the presence of a base, with the compound shown in formula (III) parent occurs step (B) for the compound shown in formula (II)
Core substitution reaction, obtain phenylboric acid ester compounds shown in formula (I)；
Further, in the preparation method of above-mentioned phenylboric acid ester compounds, in step (A) and step (B), the alkali is equal
Selected from organic base or inorganic base；Wherein, the organic base in triethylamine, diisopropyl ethyl amine or pyridine at least one
Kind；The inorganic base is selected from least one of potassium carbonate, sodium carbonate, sodium hydroxide or sodium acid carbonate.
Further, in the preparation method of above-mentioned phenylboric acid ester compounds, in step (A), the chemical combination shown in formula (IV)
The molar ratio of compound and alkali shown in thing, formula (V) is 1：1~5：0.5~5；In some embodiments, feed intake mole
Than for 1：2.5：2.5, reaction time of step (A) according to inventory is not all 1 hour~24 hours, inventory increase, instead
Accordingly extend between seasonable.Reaction is preferably in inert gas, as reacted in nitrogen or argon gas.
In step (B), the molar ratio of compound shown in compound, formula (III) and alkali shown in formula (II) is 1：1
~1.5：1.5~2, in some embodiments, molar ratio 1：1.5：1.5；When step (B) need to strictly control reaction
Between, every when carry out thin-layer chromatography TLC detection reaction process, in some embodiments, reaction time of the step (B) according to
Inventory is not all 2 hours~4 hours, and inventory increase, the reaction time accordingly extends.Reaction is preferably in inert gas, such as
Reacted in nitrogen or argon gas.
Further, in the preparation method of above-mentioned phenylboric acid ester compounds, in step (A), reaction temperature be 30 DEG C~
60 DEG C, reaction dissolvent is selected from least one of DMF, dichloromethane, acetonitrile, in some embodiments,
The reaction temperature of reaction is 40 DEG C~50 DEG C；In step (B), reaction temperature is 20 DEG C~60 DEG C, and reaction dissolvent is selected from N, N- bis-
One kind in NMF, acetonitrile, in some embodiments, reaction temperature are 40 DEG C~50 DEG C.In step (A) and step
(B) in, the dosage of reaction dissolvent is defined by being completely dissolved reactant, without being particularly limited to its dosage.
The preparation method of phenylboric acid ester compounds (I) of the present invention, simple and easy to do, post processing is simple, is easy to scale metaplasia
Production.
The present invention also provides a kind of benzoyl peroxide detection kit, includes reagent stock liquid a and reagent stock liquid b；
The reagent stock liquid a is the phosphate buffer for the ethanol that pH value is 6~8, and the reagent stock liquid b is benzene shown in formula (I)
The organic solvent solution of ylboronic acid ester compounds, the organic solvent are selected from dimethyl sulfoxide (DMSO), DMF, acetonitrile
At least one of.
Further, in benzoyl peroxide detection kit of the present invention, the phosphoric acid in the reagent stock liquid a
Salt is selected from Na2HPO4、NaH2PO4And KH2PO4At least one of, the concentration expressed in percentage by volume of ethanol is in reagent stock liquid a
10%, the phosphatic molar concentration is 5mM~15mM；Phenyl boronate chemical combination shown in formula (I) in the reagent stock liquid b
The concentration of thing is 0.01mM~1mM.In some embodiments, the reagent stock liquid a in the kit and reagent storage
Standby liquid b volume ratio is 200/1.
Further, in benzoyl peroxide detection kit of the present invention, the pH value of the reagent stock liquid a is
7.4, wherein phosphatic molar concentration is 10mM；Phenylboric acid ester compounds shown in formula (I) is dense in the reagent stock liquid b
Spend for 0.01mM~1mM.
The Cleaning Principle of benzoyl peroxide detection kit of the present invention：
Phenylboric acid ester compounds (I) in reagent stock liquid b are used as probe, and the fluorescence intensity of itself is relatively low, peroxide
Changing benzoyl can cause phenyl boric acid which ester of frequency oxidation in compound (I), electronics to be reset, finally again release have it is glimmering by force
The compound (II) of light blob, the fluorescence of the compound significantly increase at 690nm~750nm, have at 706nm most strong glimmering
Light, therefore this property can be detected for benzoyl peroxide；The change of fluorescence intensity and the concentration of benzoyl peroxide be in than
Example, the change by determining relevant fluorescence signal can determine the content of benzoyl peroxide.Course of reaction is as follows：
In yet a further aspect, the present invention provides the peroxidating in described benzoyl peroxide detection kit determination sample
The detection method of benzoyl content, it is comprised the steps of：
(1) standard curve is prepared
Using 635nm~690nm as excitation wavelength, a series of benzoyl peroxide standard solution of various concentrations is determined
Fluorescence intensity at launch wavelength 690nm~750nm is designated as F, and measure reagent blank is in the fluorescence intensity of transmitted wave strong point, note
For F0, fluorescence intensity difference DELTA F=F-F0, using the concentration C of benzoyl peroxide as abscissa, fluorescence intensity difference DELTA F is sat to be vertical
Mark, standard curve is drawn, wherein, the benzoyl peroxide standard solution of the various concentrations is stored up by the reagent in kit
Standby liquid a, reagent stock liquid b and benzoyl peroxide Standard Stock solutions prepare what is obtained；
(2) concentration of benzoyl peroxide in sample is detected
Benzoyl peroxide standard solution prepared by step (1) is replaced with into testing sample, adds reagent storage thereto
Standby liquid b, testing sample is detected in the fluorescence intensity with step (1) identical transmitted wave strong point, note according to step (1) methods described
For F ', the F ' is substituted into standard curve obtained by step (1), and then obtain the concentration of benzoyl peroxide in testing sample.
When in some embodiments, using described kit, wherein, a series of benzoyl peroxide of various concentrations
Formyl standard solution is matched somebody with somebody by reagent stock liquid a, reagent stock liquid b and the benzoyl peroxide Standard Stock solutions in kit
Make and obtain, in the benzoyl peroxide Standard Stock solutions, reagent stock liquid b additions are equal, and solvent is mainly reagent
Storing solution a；A series of solvent of the benzoyl peroxide standard solution of various concentrations is reagent stock liquid a, and volume is equal
For 2mL, concentration be followed successively by 0 μM, 0.5 μM, 1.0 μM, 1.5 μM, 2.0 μM, 2.5 μM, 3.0 μM, 3.5 μM, 4.0 μM, 4.5 μM, 5.0
μM, 5.5 μM and 6.0 μM.After fluorescence intensity, find benzoyl peroxide concentration when between 0.5 μM~4 μM, concentration with
Linear between fluorescence intensity, degree of correlation R reaches more than 0.99, can carry out quantitative detection.
During using heretofore described kit, the optional 630nm~690nm wavelength periods of excitation wavelength, wherein in some realities
Apply in scheme, selective exitation wavelength 635nm；In other embodiments, selective exitation wavelength 670nm；Launch wavelength is optional
690nm~750nm wavelength periods, wherein in some embodiments, selecting launch wavelength 706nm；In other embodiments,
Select launch wavelength 700nm.
In yet a further aspect, described benzoyl peroxide detection kit is used to determine present invention also offers another
The detection method of benzoyl peroxide in sample, it is comprised the steps of：
(i) fluorescence intensity or fluorescence imaging of blank control sample are determined
Using 635nm~690nm as excitation wavelength, finite concentration is added into the control sample without benzoyl peroxide
Reagent stock liquid b, cleaned after certain time with reagent stock liquid a to remove unnecessary reagent stock liquid b, then in transmitted wave
The fluorescence intensity of system is determined at a length of 690nm~750nm with luminoscope, or fluorescence imaging is carried out with laser co-focusing；
(ii) testing sample fluorescence intensity or fluorescence imaging are determined
The reagent stock liquid b with step (i) same concentrations is added into testing sample, reagent stock liquid is used after certain time
A is cleaned, measure system with the fluorescence intensity of step (i) identical transmitted wave strong point or shooting fluorescence imaging, and and step
(i) fluorescence intensity or the fluorescence imaging contrast measured；
When the fluorescence intensity of testing sample is more than the fluorescence intensity or testing sample fluorescence imaging brightness ratio of control sample
When control sample fluorescence imaging is strong, then illustrate to contain benzoyl peroxide in testing sample.
Compared with prior art, benzoyl peroxide detection kit of the invention, has the characteristics that：
1st, reagent storing solution b fluorescence itself is weaker in benzoyl peroxide detection kit of the invention；And and peroxidating
Then fluorescence intensity can be remarkably reinforced after benzoyl reaction, strong absorption be produced at 690nm~750nm, and fluorescence shows at 706nm
Enhancing is write, the fluorescent emission bands are near-infrared region, and ambient interferences are few, therefore have higher accuracy and sensitivity.
2nd, benzoyl peroxide detection kit of the invention, fluorescence generation reaction speed is fast, can be developed the color in 20 minutes
It is stable.
3rd, benzoyl peroxide detection kit of the invention, detection sensitivity is high, high sensitivity, and benzoyl peroxide is dense
Degree has Fluorescence Increasing signal when more than 0.5 μM, and fluorescence intensity strengthens with the increase of benzoyl peroxide concentration, in peroxide
Change benzoyl concentration when between 0.5 μM~4 μM, fluorescence intensity and benzoyl peroxide concentration, between it is linear, it is related
Degree R reaches more than 0.99.
4th, benzoyl peroxide detection kit of the invention, intensified response of the benzoyl peroxide to fluorescence have special
Property, not by other common inorganic salts such as calcium chloride (CaCl2), magnesium chloride (MgCl2), copper chloride (CuCl2), frerrous chloride
(FeCl2), zinc sulfate (ZnSO4), iron chloride (FeCl3), hypochlorite (ClO–), hydroxyl (OH–) or vitamin B6, dimension
Raw plain C, glucose, arginine, serine, singlet oxygen (1O2), nitric oxide (NO), nitrite anions (NO2 –), the tertiary fourth of peroxide
Alcohol (TBHP), potassium permanganate, potassium bichromate, potassium chlorate, sodium chlorate, sodium perchlorate, Potassiumiodate, sodium hypochlorite, hydrogen peroxide etc.
The interference of typical oxidation active specy.
5th, benzoyl peroxide detection kit of the invention, has near-infrared fluorescent launch wavelength, can use fluorescence spectrum
Method detects, and background signal is relatively low, and background noise is few, is adapted to detect for the biological sample of complicated component, is also adapted to monitor live body sample
The distribution of benzoyl peroxide in this.
Brief description of the drawings
Fig. 1 is the fluorescence emission spectrum of the benzoyl peroxide reaction of kit and various concentrations.
Fig. 2 is standard curve when kit is used to detect benzoyl peroxide concentration.
Fig. 3 is the fluorescence emission spectrum that kit is used for the reaction of various interfering materials.
Fig. 4 is the fluorescence intensity change that kit detects various concentrations benzoyl peroxide in Hela cells.
Fig. 5 is that the zebra fish fluorescence intensity change after benzoyl peroxide is bred in kit detection.
Embodiment
Further detailed description is done to the present invention with reference to specific embodiment, but embodiments of the present invention are not limited to
This.Those skilled in the art will realize that：Chemical reaction described in the invention can be used for suitably preparing many
Other compounds of invention, for example, some conventional modifications are made according to reaction condition of the present invention, for preparing the chemical combination of the present invention
Other methods of thing are considered as within the scope of the present invention.
The structure of compound be by nuclear magnetic resonance (1H-NMR、13C-NMR) determine.1H-NMR、13C-NMR chemical potentials
(δ) is moved to provide with the unit of hundred a ten thousandths (ppm).1H-NMR、13C-NMR measure is to use Bruker Ultrashield-
400 nuclear magnetic resonance spectrometers and the nuclear magnetic resonance spectrometers of Bruker Avance III HD 600, measure solvent are deuterated methanol
(CD3OD).Reference standard is used as by the use of TMS (0ppm) or chloroform (7.25ppm).When there is multiplet, it will use following
Abbreviation：S (singlet, unimodal), d (doublet, bimodal), t (triplet, triplet), m (multiplet, multiplet),
Br (broadened, broad peak), dd (doublet of doublets, double doublet), and brs (broadened singlet, it is wide
It is unimodal).Coupling constant, represented with hertz (Hz).
Column chromatography is carrier typically using the mesh silica gel of the mesh of Qingdao Haiyang chemical industry 200~300.
Experimental method described in following embodiments, it is conventional method unless otherwise specified；The reagent such as IR-780 iodine
For thing and biomaterial, unless otherwise specified, commercially obtain, can either use or according to known in the art
Method synthesize.
Carried out under nitrogen atmosphere without specified otherwise, reaction in embodiment；
Blanket of nitrogen refers to that reaction bulb connects the nitrogen balloon or steel kettle of an about 1L volume；
Without specified otherwise in specification, room temperature is 20 DEG C~30 DEG C, and the temperature error described in embodiment is ± 5 DEG C.
The monitoring of reaction process in embodiment uses the system of solvent used in thin-layered chromatography (TLC) reaction
Have：Petroleum ether and ethyl acetate system, the volume ratio of solvent is different according to the polarity of compound and is adjusted.
Embodiment 1：The preparation of formula (I) phenylboric acid ester compounds
The preparation of (1-1) compound (III)
Compound (III) can directly be commercially available or prepare in accordance with the following methods：
30mL toluene is added into a there-necked flask equipped with Dean-Stark water knockout drums, then adds 2- methylphenylboronic acids
(0.21g, 1.55mmol), pinacol (pinacol-H2O, 0.22g, 1.86mmol), gained mixture is heated to reflux two small
When.After reaction terminates, (30mL × 3) are washed with water in mixture, remove unreacted pinacol and phenyl boric acid, then have what is separated
Removing toluene solvant is mutually evaporated under reduced pressure in machine.Gained crude product dichloromethane (50mL) is dissolved, after washing (30mL) again, to
Add anhydrous sodium sulfate in gained organic phase to be dried, dried solution decompression evaporates to obtain crude product 0.32g.
Take gained crude product (0.32g, 1.47mmol), NBS (N-bromosuccinimide, 0.39g, 2.2mmol), AIBN
In the acetonitrile (10mL) that (azodiisobutyronitrile, 0.003g, 0.018mmol) is dissolved into.Gained reaction solution flows back at 90 DEG C
Two hours.After cooling down at room temperature, remove solvent under reduced pressure, obtain crude product.Gained crude product silica gel column chromatography separating purification, with
Petroleum ether (60~90 DEG C)/ethyl acetate (v/v)=1：1 makees eluent, and it is white solid (0.53g, yield to obtain product
80%).
The preparation of (1-2) compound (II)
Resorcinol (86.0mg, 0.78mmol) and potassium carbonate (102mg, 0.78mmol) are dissolved in acetonitrile (20mL)
In, in room temperature and stirred under nitrogen atmosphere 10 minutes, compound (IV) (the i.e. IR-780 that then will be dissolved in acetonitrile (1mL)
Iodo thing, 207mg, 0.31mmol) it is added in system.Reactant mixture is reacted 4 hours at 50 DEG C, reaction finishes, and subtracts
Solvent is evaporated off in pressure, obtains crude product.With silica gel column chromatography purifying crude product, with petroleum ether (60~90 DEG C)/ethyl acetate (1：1, v/
V) eluant, eluent is made, it is cyan powders (67.3mg, yield 52.6%) to obtain product.
The structural characterization data result of compound (II) is as follows：
Hydrogen is composed：1H NMR(400MHz,CDCl3):δ (ppm) 8.03 (d, 1H, J=14.2Hz), 7.28-7.23 (m, 3H),
7.19 (d, 1H, J=9.2Hz), 7.04 (t, 1H, J=7.6Hz), 6.81 (d, 1H, J=7.6Hz), 6.74 (d, 1H, J=
8.8Hz), 6.51 (s, 1H), 5.61 (d, 1H, J=14.2Hz), 3.76 (t, 2H, J=7.2Hz), 2.67 (t, 2H, J=
6.0Hz), 2.61 (t, 2H, J=6.0Hz), 1.91-1.78 (m, 4H), 1.66 (s, 6H), 1.06 (t, 3H, J=7.6Hz)
Carbon is composed：13C NMR(400MHz,CH3OD):δ(ppm)174.5,173.7,163.8,158.5,144.0,142.2,
141.6,140.3,131.1,129.9,126.1,123.5,123.2,121.9,116.2,115.9,112.0,103.6,
100.1,50.4,46.6,29.6,29.0,25.3,22.1,21.8,11.9.
Preparing for compound shown in formula (II) can also be carried out according to the experiment condition of table 1 below.
Table 1：
According to the experiment condition of upper table 1, compound (II) can be obtained.
The preparation of (1-3) compound (I)
Compound (II) (40mg, 1.0mmol) and potassium carbonate (20mg, 1.5mmol) are dissolved in acetonitrile (4mL),
Room temperature and stirred under nitrogen atmosphere 10 minutes, the compound (III) (20mg, 1.0mmol) that then will be dissolved in acetonitrile (1mL)
It is added in mixed solution, reactant mixture is reacted 2.5 hours at 50 DEG C.Reaction finishes, and it is dilute to add dichloromethane (20mL)
Mixture is released, is washed successively with water (5mL) and saturated aqueous sodium carbonate (5mL).Then anhydrous sodium sulfate drying organic phase is used,
Remove solvent under reduced pressure, obtain crude product.Crude product, methylene chloride/methanol=10 are purified with silica gel column chromatography：1, product is obtained as deeply
Blue solid (25mg, 36%).
The structural characterization data result of compound (I) is as follows：
Hydrogen is composed：1H NMR (600MHz, MeOD) δ (ppm) 8.72 (d, J=14.9Hz, 1H), 7.84 (dd, J=7.4,
0.9Hz, 1H), 7.64 (d, J=7.4Hz, 1H), 7.58-7.43 (m, 6H), 7.39-7.34 (m, 2H), 7.03 (d, J=
2.1Hz, 1H), 7.00 (dd, J=8.5,2.3Hz, 1H), 6.51 (d, J=14.9Hz, 1H), 5.42 (s, 2H), 4.33 (t, J=
7.4Hz, 2H), 2.81-2.75 (m, 2H), 2.72 (t, J=6.0Hz, 2H), 1.98-1.91 (m, 4H), 1.81 (s, 6H), 1.28
(s, 12H), 1.08 (t, J=7.4Hz, 3H)；
Carbon is composed：13C NMR(151MHz,MeOD)δ(ppm)179.19,164.13,163.07,155.83,146.95,
143.42,143.36,143.08,137.18,135.21,132.32,130.29,130.15,129.58,128.78,128.56,
128.44,123.80,117.24,115.77,115.20,114.05,104.88,102.50,85.16,71.83,52.00,
47.66,30.11,28.53,25.32,25.16,22.31,21.67,11.68.
Preparing for phenylboric acid ester compounds shown in formula (I) can also be carried out according to the experiment condition of table 2 below.
Table 2：
According to the experiment condition of upper table 2, compound (II) can be obtained.
Reaction temperature is 40 DEG C~50 DEG C, and the compound shown in compound, formula (III) and alkali shown in formula (II) feed intake
Mol ratio is 1：1.5：1.5, combined reaction best results.
Embodiment 2：Fluorescence probe shown in formula (I) and the spectral quality of various concentrations benzoyl peroxide reaction
The phosphate buffer (PBS) of the 10mM PH=7.4 containing ethanol is first prepared, according to this area conventional practices
Preparation phosphate concn is 10mM, and pH is 7.4 phosphate buffer containing ethanol, and the concentration expressed in percentage by volume of wherein ethanol is
10%, PBS are prepared and are used NaHPO4-KH2PO4System, therefore phosphate concn refers to NaHPO4And KH2PO4Total concentration, so
Afterwards, the solution prepared is as reagent stock liquid a.The phenylboric acid ester compounds (I) that embodiment 1 is prepared are configured to dense
It is acetonitrile to spend the solution for 1mM as reagent stock liquid b, selected solvent, obtains reagent stock liquid b.Empirical tests, for
The species of reagent stock liquid b organic solvents processed does not influence Detection results, uses dimethyl sulfoxide (DMSO) instead or DMF is
Effect is the same during solvent.The volume ratio of reagent stock liquid a and reagent stock liquid b in kit are 200/1.
Take reagent stock liquid b (1mM, 20 μ L) to be dissolved in reagent stock liquid a, arrived with reagent stock liquid a or ethanol constant volume
2mL.Now, phenylboric acid ester compounds (I) are that concentration and probe concentration is 10 μM.Then, by adding the benzoyl peroxides of different volumes
Formyl Standard Stock solutions (1mM), final prepare obtain a series of benzoyl peroxide standard solution of various concentrations, wherein
The concentration of benzoyl peroxide be followed successively by 0 μM, 0.5 μM, 1.0 μM, 1.5 μM, 2.0 μM, 2.5 μM, 3.0 μM, 3.5 μM, 4.0 μ
M, 4.5 μM, 5.0 μM, 5.5 μM and 6.0 μM.A series of volume of the reaction solution of benzoyl peroxide standard items containing various concentrations
It is 2mL.The mixed solution of gained reacts 20min at 37 DEG C, and luminoscope F-4600 measures its fluorescence excitation spectrum and fluorescence
Emission spectrum, the slit width for exciting and launching is 10nm.Using 670nm as excitation wavelength when fluorescence emission spectrum determines, obtain
The emission spectrum of the benzoyl peroxide standard solution of various concentrations is as shown in Figure 1.
Using 670nm as excitation wavelength when fluorescence emission spectrum determines, a series of benzoyl peroxide of various concentrations is determined
The solution of standard items is the fluorescence intensity at 706nm in launch wavelength, is designated as F；It is 706nm that reagent blank, which is determined, in launch wavelength
The fluorescence intensity at place, is designated as F0, fluorescence intensity difference DELTA F=F-F0, using the concentration C of benzoyl peroxide as abscissa, fluorescence is strong
Degree difference DELTA F is ordinate, draws standard curve, it is as shown in Figure 2 to obtain standard curve.
Equation of linear regression is corresponding to standard curve：Δ F=676.3 × [BPO] (μM) -100.1, (R2=0.993),
From standard curve, benzoyl peroxide concentration is linearly closed when between 0.5 μM~4 μM between concentration and fluorescence intensity
System, degree of correlation R reach more than 0.99.In linear scope, reagent stock liquid b is added into testing sample, detects testing sample
In the fluorescence intensity of transmitted wave strong point, the fluorescence intensity is substituted into standard curve, so that it may obtain benzoyl peroxide first in testing sample
The concentration of acyl.Wherein, the concentration unit of benzoyl peroxide [BPO] is μM.
According to this area conventional methods, the test limit (S/N=3) that the probe is drawn after 11 repetitions are tested is
47nM。
Phosphate buffer solution in the present embodiment is prepared according to the conventional PBS compound methods in this area, reagent stock liquid a
In phosphate be selected from Na2HPO4、NaH2PO4And KH2PO4At least one of, phosphatic molar concentration can be 5mM~
15mM；The concentration of fluorescence probe shown in formula (I) can be 0.01mM~1mM in reagent stock liquid b.Conventional reagent stock liquid a
It is concentration 1mM solution for concentration 10mM, pH 7.4, the PBS containing ethanol 10%, reagent stock liquid b.
It is above-mentioned test result indicates that, the present invention in fluorescence probe phenylboric acid ester compounds (I) have the characteristics that：
1st, fluorescence signal is extremely low in the solution for fluorescence probe phenylboric acid ester compounds (I), and background signal is relatively low；But with
The addition of benzoyl peroxide, compound (I) and benzoyl peroxide react after then fluorescence intensity can be remarkably reinforced, in 690nm
Produced at~750nm it is strong absorb, and fluorescence is most strong at 706nm, and the fluorescent emission bands are near-infrared region, ambient interferences
Few, this probe has higher accuracy and sensitivity.
2nd, fluorescence probe phenylboric acid ester compounds (I) fluorescence generation reaction speed is fast, can color stability in 20 minutes.
3rd, fluorescence probe phenylboric acid ester compounds (I) have fluorescence in benzoyl peroxide concentration when more than 0.5 μM
Strengthen signal, fluorescence intensity strengthens with the increase of benzoyl peroxide concentration, in benzoyl peroxide concentration in 0.5 μM~4 μ
When between M, fluorescence intensity and benzoyl peroxide concentration are linear, can carry out the quantitative detection of benzoyl peroxide.
Embodiment 3：The response situation (selection Journal of Sex Research) of kit containing fluorescence probe (I) and other ions
The reagent stock liquid b of 1mM concentration is dissolved in be configured in reagent stock liquid a (10mM, pH 7.4) it is glimmering at 10 μM
Light probe solution, isometric various common inorganic salts, Biomedia, activating oxide and typical oxygen are separately added into thereto
Compound, such as 30 μM of calcium chloride, magnesium chloride, copper chloride, frerrous chloride, zinc sulfate, iron chloride；1.5mM vitamin B6, dimension life
Plain C, glucose, arginine, serine；30 μM of hypochlorite, hydroxyl, singlet oxygen, nitric oxide, nitrite anions, height
Potassium manganate, potassium bichromate, potassium chlorate, sodium chlorate, sodium perchlorate, Potassiumiodate, sodium hypochlorite, hydrogen peroxide and 6 μM of peroxidating
Benzoyl (as a control group).Interfering material is 10mM reagent stock liquid a as solvent using concentration, and it is molten to obtain one group of mixing
Liquid, after reacting 20min at 37 DEG C, its fluorescence emission spectrum is measured respectively using luminoscope F-6000.Fluorescence emission spectrum
With 670nm deexcitations during measure；The slit width for exciting and launching is 10nm.
Experimental result is as shown in figure 3, Fig. 3 is the fluorescence emission spectrum that kit is used for the reaction of various interfering materials.
Test result indicates that only benzoyl peroxide can cause fluorescence probe to produce obvious optical signal significant changes sound
Should, it was demonstrated that the fluorescence probe has the selectivity of height to benzoyl peroxide；Other common inorganic salts, oxidation activity species
With typical oxidation thing interference effect unobvious.
Embodiment 4：Various concentrations in benzoyl peroxide detection kit detection Hela cells containing fluorescence probe (I)
The fluorescence intensity change of benzoyl peroxide
Hela cells adherent growth in glass culture dish；Nutrient solution is DMEM nutrient solutions, wherein containing 10% hyclone,
100mg/mL penicillin and 100mg/mL streptomysins；Condition of culture is 37 DEG C, 5%CO2；Incubation time is 12~24 hours.
Before the fluorescence imaging for carrying out intracellular peroxidation benzoyl, one group of cell is as blank control, remaining four groups of Hela cells difference
It is incubated with the DMEM of the hyclone of the benzoyl peroxide (0 μM, 2 μM, 4 μM, 6 μM) containing various concentrations at 37 DEG C
10min, cleaned three times with reagent stock liquid a (concentration 10mM, pH 7.4).Again respectively with 10 μM of probe solution (by 1mM
Reagent stock liquid b be diluted with reagent stock liquid a form) respectively with four groups of experimental group cell effect 20min, with same
Reagent stock liquid a is cleaned three times.Fluorescence reaction intensity is big after reagent stock liquid b reacts with benzoyl peroxide, in biological sample
During detection, obvious Detection results are can reach using low concentration, therefore, 10 μM or so of concentration is typically diluted to and uses,
Reagent dosage adds according to this area conventional amount used.
With laser confocal microscope TCS SP5 under 635nm excitation wavelength fluorescence imaging.Experimental result such as Fig. 4 institutes
Show, Fig. 4 is the fluorescence intensity change that kit detects various concentrations benzoyl peroxide in Hela cells.
4A is the fluorescence imaging for the blank control that unused reagent stock liquid b is treated in Fig. 4, and 4B is without addition peroxidating
Benzoyl is only respectively to use 2 μM, 4 μM, 6 μM of concentration with the fluorescence imaging of the cell after reagent stock liquid b processing, 4C, 4D and 4E
The treated cell sample of benzoyl peroxide, the fluorescence imaging added after reagent stock liquid b.Fig. 4 F~4J are Fig. 4 A~4E
Corresponding light field cytological map.
Contrasted from the 4A in Fig. 4 and 4B：Hela cell itself unstressed configurations, it is individually added into Hela cells after fluorescence probe
Less fluorescence.From 4C, 4D and 4E in Fig. 4, Hela cell fluorescence intensities compared with 4B have it is different degrees of significantly increase, say
It is bright that benzoyl peroxide has different degrees of humidification to the fluorescence of probe in cell sample, and with benzoyl peroxide first
The increase of acyl concentration, Fluorescence Increasing；Fluorescence intensity change is obvious, can judge the dense of benzoyl peroxide by fluorescence intensity
Spend difference.By Fig. 4 F~4J it can also be seen that with benzoyl peroxide increasing concentrations, cell fluorescence intensity gradually strengthens.
Embodiment 5：Various concentrations benzoyl peroxide in benzoyl peroxide detection zebra fish containing fluorescence probe (I)
Fluorescence intensity change
Zebra fish is incubated in E3 embryo mediums (15mM sodium chloride, 0.5mM potassium chloride, 1mM magnesium sulfate, 1mM chlorinations
Calcium, 0.15mM sodium dihydrogen phosphates, 0.05mM disodium hydrogen phosphates, 0.7mM sodium acid carbonates, 5~10% methylenum careuleum, pH 7.5), use
The zebra fish of growth 1 day carries out fluorescence imaging.Carrying out benzoyl peroxide before processing, one group of zebra fish as blank control,
Remaining four groups of zebra fish cultivates 5min with (0 μM, 2 μM, 4 μM, 6 μM) of the benzoyl peroxide solution containing various concentrations respectively,
Then cleaned three times with reagent stock liquid a (concentration 10mM, pH 7.4)；Afterwards with 10 μM of reagent stock liquid b (by 1mM examination
Agent storing solution b is diluted with reagent stock liquid a and formed), 20min is cultivated, then cleaned three times with same reagent stock liquid a,
To wash excess probes off.Zebra fish after the completion of processing carries out fluorescence imaging, excitation wavelength 635nm, hair with living imaging instrument
A length of 690nm~the 750nm of ejected wave.
Experimental result is as shown in figure 5, Fig. 5 is with benzoyl peroxide change in concentration, zebra fish afterbody (A1~E1) and ovum
The change of yellow capsule portion (A2~E2) fluorescence intensity.Fig. 5 A1 and Fig. 5 A2 are the blank control that unused reagent stock liquid b is treated
Fluorescence imaging, Fig. 5 B1 and Fig. 5 B2 are only with the work of the zebra fish after reagent stock liquid b processing without addition benzoyl peroxide
Body fluorescence imaging, Fig. 5 C1/5C2,5D1/5D2,5E1/5E2 are respectively to use 2 μM of concentration, 4 μM, 6 μM of benzoyl peroxide processing
The fluorescence imaging that the zebra fish biopsy samples crossed are added after reagent stock liquid b.As a comparison, Fig. 5 F1~5J1 are Fig. 5 A1~5E1
Original photo under non-fluorescence state, Fig. 5 F2~5J2 are original photos of Fig. 5 A2~5E2 under non-fluorescence state.
From Fig. 5 A1 and 5A2, under the conditions of same test, the afterbody of zebra fish and yolk bag portion do not have fluorescence in itself, by
Fig. 5 B1 and 5B2 understand that after individually adding fluorescence probe in zebra fish live body, the afterbody and yolk bag portion fluorescence of zebra fish are micro-
It is weak.Understood to carry out the zebra fish afterbody of various concentrations benzoyl peroxide pre-treatment by Fig. 5 C1/5C2,5D1/5D2,5E1/5E2
Strengthen with the fluorescence intensity in yolk bag portion, and the increase of fluorescence intensity and benzoyl peroxide concentration are proportional.
Meanwhile the change for observation fluorescence intensity directly perceived carries out quantization detection to the fluorescence intensity of zebra fish live body afterbody,
And Numerical analysis is carried out to fluorescence intensity level.Zebra fish (5B1) fluorescence intensity for adding 0 μM of benzoyl peroxide processing is denoted as
1.0；Compared to this, add treatment group (5C1~5E1) fluorescence intensity of 2,4,6 μM of benzoyl peroxides to add 0.77 respectively,
1.11 and 1.36 multiple, illustrate that the probe compound in this kit is suitable for detecting the benzoyl peroxide inside live body
Distribution.
Above example 4 and embodiment 5 demonstrate well, using the phenyl boronate shown in containing formula (I) of the present invention
Whether the kit of compound can intuitively judge biological sample to be measured by way of contrasting fluorescence intensity or fluorescence imaging
Containing benzoyl peroxide, common biological specimen (cell) is applicable not only to, it is equally applicable for complex biological biopsy sample, and
And in terms of benzoyl peroxide is detected, kit provided by the present invention is swift in response, sensitivity level is high, simple to operate, fits
In extensive use.
In summary, benzoyl peroxide kit of the invention is a kind of function admirable, benzoyl peroxide easy to use
Formyl detection device, there is huge application prospect in the fields such as food, medicine, living body biological imaging.Above content is knot
Close specific preferred embodiment further description made for the present invention, it is impossible to assert the specific implementation office of the present invention
It is limited to these explanations.For general technical staff of the technical field of the invention, before present inventive concept is not departed from
Put, some simple deduction or replace can also be made, should all be considered as belonging to protection scope of the present invention.
Claims (10)
1. the phenylboric acid ester compounds shown in formula (I),
2. the preparation method of the phenylboric acid ester compounds described in claim 1, comprises the following steps：
In the presence of a base, the compound shown in formula (IV) is substituted decomposition weight to step (A) with the compound shown in formula (V)
Row's reaction, obtains compound shown in formula (II)；
In the presence of a base, the compound shown in formula (II) occurs nucleophilic with the compound shown in formula (III) and taken step (B)
Generation reaction, obtains phenylboric acid ester compounds shown in formula (I)；
3. preparation method according to claim 2, it is characterised in that in step (A) and step (B), the alkali is selected from
Organic base or inorganic base；
Wherein, the organic base is selected from least one of triethylamine, diisopropyl ethyl amine or pyridine；
The inorganic base is selected from least one of potassium carbonate, sodium carbonate, sodium hydroxide or sodium acid carbonate.
4. the preparation method according to Claims 2 or 3, it is characterised in that in step (A), compound shown in formula (IV),
The molar ratio of compound and alkali shown in formula (V) is 1：1~5：0.5~5；
In step (B), the molar ratio of compound shown in compound, formula (III) and alkali shown in formula (II) is 1：1~
1.5：1.5~2.
5. the preparation method according to Claims 2 or 3, it is characterised in that in step (A), reaction temperature is 30 DEG C~60
℃；Reaction dissolvent is selected from least one of N,N-dimethylformamide, dichloromethane, acetonitrile；
In step (B), reaction temperature is 20 DEG C~60 DEG C；The one kind of reaction dissolvent in N,N-dimethylformamide, acetonitrile.
6. a kind of benzoyl peroxide detection kit, include reagent stock liquid a and reagent stock liquid b, the reagent stock liquid a
It is the phosphate buffer for the ethanol that pH value is 6~8, the reagent stock liquid b is phenylboric acid ester compounds shown in formula (I)
Organic solvent solution, the organic solvent are selected from least one of dimethyl sulfoxide (DMSO), DMF, acetonitrile.
7. benzoyl peroxide detection kit according to claim 6, it is characterised in that in the reagent stock liquid a
Phosphate be selected from Na2HPO4、NaH2PO4And KH2PO4At least one of, the concentration expressed in percentage by volume of ethanol in reagent stock liquid a
For 10%, the phosphatic molar concentration is 5mM~15mM；
The concentration of phenylboric acid ester compounds shown in formula (I) is 0.01mM~1mM in the reagent stock liquid b.
8. benzoyl peroxide detection kit according to claim 7, it is characterised in that the reagent stock liquid a's
PH value is 7.4, wherein phosphatic molar concentration is 10mM；
The concentration of phenylboric acid ester compounds shown in formula (I) is 0.01mM~1mM in the reagent stock liquid b.
9. usage right requires the benzoyl peroxide in the benzoyl peroxide detection kit determination sample described in 6~8 any one
The detection method of formyl content, it is comprised the steps of：
(1) standard curve is prepared
Using 635nm~690nm as excitation wavelength, a series of benzoyl peroxide standard solution for determining various concentrations is being sent out
Fluorescence intensity at the long 690nm~750nm of ejected wave is designated as F, and measure reagent blank is strong for the fluorescence at 706nm in launch wavelength
Degree, is designated as F0, fluorescence intensity difference DELTA F=F-F0, using the concentration C of benzoyl peroxide as abscissa, fluorescence intensity difference DELTA F
For ordinate, standard curve is drawn, wherein, the benzoyl peroxide standard solution of the various concentrations is by kit
Reagent stock liquid a, reagent stock liquid b and benzoyl peroxide Standard Stock solutions prepare what is obtained；
(2) concentration of benzoyl peroxide in sample is detected
Benzoyl peroxide standard solution in step (1) is replaced with into testing sample, adds reagent stock liquid b thereto,
According to step (1) methods described detection testing sample in the fluorescence intensity with step (1) identical transmitted wave strong point, F ' is designated as,
The F ' is substituted into standard curve obtained by step (1), and then obtains the concentration of benzoyl peroxide in testing sample.
10. usage right requires the benzoyl peroxide in the benzoyl peroxide detection kit determination sample described in 6~8 any one
The detection method of formyl, it is comprised the steps of：
(i) fluorescence intensity or fluorescence imaging of blank control sample are determined
Using 635nm~690nm as excitation wavelength, certain density examination is added into the control sample without benzoyl peroxide
Agent storing solution b, cleaned after certain time with reagent stock liquid a, to remove unnecessary reagent stock liquid b, then in launch wavelength
To determine the fluorescence intensity of system at 690nm~750nm with luminoscope, or with laser co-focusing carry out fluorescence imaging；
(ii) testing sample fluorescence intensity or fluorescence imaging are determined
The reagent stock liquid b with step (I) same concentrations is added into testing sample, it is clear with reagent stock liquid a after certain time
Wash, to remove unnecessary reagent stock liquid b, measure system with the fluorescence intensity of step (I) identical transmitted wave strong point or
Fluorescence imaging is shot, and fluorescence intensity or the fluorescence imaging contrast measured with step (I)；
When the fluorescence intensity of testing sample is more than fluorescence intensity or the control of testing sample fluorescence imaging brightness ratio of control sample
When fluorescent imaging is strong, then illustrate to contain benzoyl peroxide in testing sample.
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| CN109320537A (en) * | 2018-12-04 | 2019-02-12 | 湖南工业大学 | A kind of soluble two-photon fluorescence probe and its preparation method and application of for flour and in vivo benzoyl peroxide detection |
| CN110745781A (en) * | 2019-10-30 | 2020-02-04 | 汕头大学 | Novel method for generating singlet oxygen by exciting small-molecule anthraquinone charge transfer state by blue light or near infrared light |
| CN113552099A (en) * | 2020-04-24 | 2021-10-26 | 中国科学院宁波工业技术研究院慈溪生物医学工程研究所 | Fluorescent diagnosis kit and application thereof |
| CN115029132A (en) * | 2022-05-27 | 2022-09-09 | 重庆师范大学 | Preparation method of novel dopamine functionalized fluorescent carbon dots, product and application thereof |
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| CN102532177A (en) * | 2011-12-22 | 2012-07-04 | 中国科学院化学研究所 | Rapid detection reagent kit for benzoyl peroxide |
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| CN102532177A (en) * | 2011-12-22 | 2012-07-04 | 中国科学院化学研究所 | Rapid detection reagent kit for benzoyl peroxide |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109320537A (en) * | 2018-12-04 | 2019-02-12 | 湖南工业大学 | A kind of soluble two-photon fluorescence probe and its preparation method and application of for flour and in vivo benzoyl peroxide detection |
| CN110745781A (en) * | 2019-10-30 | 2020-02-04 | 汕头大学 | Novel method for generating singlet oxygen by exciting small-molecule anthraquinone charge transfer state by blue light or near infrared light |
| CN113552099A (en) * | 2020-04-24 | 2021-10-26 | 中国科学院宁波工业技术研究院慈溪生物医学工程研究所 | Fluorescent diagnosis kit and application thereof |
| CN113552099B (en) * | 2020-04-24 | 2024-04-16 | 中国科学院宁波工业技术研究院慈溪生物医学工程研究所 | Fluorescent diagnostic kit and application thereof |
| CN115029132A (en) * | 2022-05-27 | 2022-09-09 | 重庆师范大学 | Preparation method of novel dopamine functionalized fluorescent carbon dots, product and application thereof |
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