CN104017572B - A kind of extreme acidic pH fluorescent probe and its preparation method and application - Google Patents
A kind of extreme acidic pH fluorescent probe and its preparation method and application Download PDFInfo
- Publication number
- CN104017572B CN104017572B CN201410262408.6A CN201410262408A CN104017572B CN 104017572 B CN104017572 B CN 104017572B CN 201410262408 A CN201410262408 A CN 201410262408A CN 104017572 B CN104017572 B CN 104017572B
- Authority
- CN
- China
- Prior art keywords
- probe
- indoles
- fluorescent probe
- value
- extreme acidic
- 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.)
- Expired - Fee Related
Links
- 0 CC(C)(C1CCC=C[C@@](C2C(CC3)CCC3F)[C@@](C(*)**)C3=C2C=CCC3)c(c(cccc2)c2cc2)c2[N+]1[O-] Chemical compound CC(C)(C1CCC=C[C@@](C2C(CC3)CCC3F)[C@@](C(*)**)C3=C2C=CCC3)c(c(cccc2)c2cc2)c2[N+]1[O-] 0.000 description 1
Landscapes
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
PH fluorescent probe that the invention discloses a kind of extreme acidic and its preparation method and application.This probe utilizes divinyl bridging indoles or derivatives thereof and 3-[3-(4-fluorophenyl)-1-alkyl-1H-indoles builds the pH fluorescent probe based on Intramolecular electron transfer effect.Its preparation method: under protection of inert gas; by 2,3,3-tri-methyl indole or 1; 1; 2-trimethylammonium-1H-benzo [e] indoles and 3-[3-(4-fluorophenyl)-1-alkyl-1H-indoles-2-base]-propenal are dissolved in DMF, add KOH solid; stirring at room temperature; spend the night, concentrating under reduced pressure obtains crude product, is separated obtained sterling through silicagel column.This probe has lower pKa value, to H
+there are higher sensitivity, good selectivity and large Stocks displacement.Because this probe is in conjunction with H
+front and back colour-change is comparatively obvious, can conveniently by bore hole identification.This probe can permeates cell membranes, is applicable to detection and the cell imaging of intracellular ph value.
Description
Technical field
The present invention relates to fluorescent probe, specifically belong to a kind of extreme acidic pH fluorescent probe and preparation method thereof, and the application of this probe in cell imaging.
Background technology
PH value not only has obvious impact to the physico-chemical property of material and reactivity worth thereof, and many important physiological processs of cell and crganelle etc. are also all closely related with pH value.Acid and alkalescence is crossed and the heart, tuberculosis can be caused by force to become or neural class disease, even can be in peril of one's life time serious.Under normal physiological conditions, H in enchylema
+concentration be about 40nmol/L (pH=7.40), its amplitude of fluctuation is generally at 0.1 ~ 0.2 pH unit.Also have a pH value range to be acid range in cell, if lysosomal pH value range is 4.5-5.5, the pH value range of endosome is 5.4-6.2, is called acidic organelles.Extreme acidic's environment (pH<4) is fatal to most of organism.The substantial connection of intracellular ph value and cell function means can for studying the little physiology to individual cells device and pathological process provides key message to the accurate measurement of intracellular ph value.So the dynamic change of monitoring intracellular ph value has important effect for the mechanism understanding many physiological functions in cell.
Life system requires small pH value change to be detected.Measure pH value and generally use glass electrode, but be unsuitable for live body pH owing to there is the defects such as electrochemistry interference, possible physical abuse and monitor.Also have and adopt colorimetric method for determining pH value, but colorimetry also also exists obvious shortcoming, cannot be applied to life system and carry out active somatic cell radiography.In addition, the sensitivity of colorimetry is lower.The advantages such as fluorescent probe has quick real-time response, high and selectivity is single-minded to cell not damaged, response sensitivity, utilize fluorescent probe to detect to intracellular ph value change the broad interest causing people.Many pH fluorescent probes of current report concentrate on detect in cell neutral, i.e. the pH value of enchylema and the acidic organelles such as lysosome and endosome pH value; Measure extreme environmental, it is considerably less that the fluorescent probe as pH<4 or pH>9 is reported.Therefore, design and synthesis has large Stokes displacement, to H
+ion is extremely sensitive, selectivity good, extreme environmental can be measured under the fluorescent probe of pH in biological and chemical etc., all there is potential using value.
Summary of the invention
An object of the present invention be to provide a kind of there is large Stocks displacement, extremely sensitive to pH, selectivity is high, extreme acidic's condition can be measured under the fluorescent probe of pH value; Two of object is to provide the preparation method of this probe, and the method technique is simple, with low cost; Three of object is to provide the purposes of this probe, and namely this probe detects intracellular ph value as fluorescent indicator.
The pH fluorescent probe of a kind of extreme acidic provided by the invention, its structural formula is:
Wherein, R
1, R
2for hydrogen atom or alkyl.
The synthetic route of the pH fluorescent probe of extreme acidic provided by the invention is as follows:
Its preparation method, comprises the steps:
(1) under protection of inert gas, by 2,3,3-tri-methyl indole or 1,1,2-trimethylammonium-1H-benzo [e] indoles and 3-[3-(4-fluorophenyl)-1-alkyl-1H-indoles-2-base]-propenal in molar ratio 1 ︰ 1.0-1.5 are dissolved in N, dinethylformamide, by 2,3,3-tri-methyl indole or 1,1,2-trimethylammonium-1H-benzo [e] indoles meter, adds the KOH solid of 5 times of mole numbers, stirring at room temperature, reaction is spent the night;
(2) excessively filter KOH solid, in reaction solution, add the H of 1-2 times of volume
2o, uses CH
2cl
2extract 2-3 time; Merge organic phase, use anhydrous Na
2sO
4after drying, underpressure distillation obtains crude product;
(3) crude product is separated through silicagel column, obtains sterling.
Probe of the present invention has good cell membrane penetration, can be used for detection and the cell imaging of intracellular ph value.
Compared with existing pH fluorescent probe, the probe of the present invention's synthesis has the following advantages: the Stokes displacement that (1) is large, is conducive to the interference reducing exciting light in angiographic procedure; (2) low pKa value, can detect the pH value under extreme acidic's condition; (3) good selectivity, this probe is to H
+response by the interference of common metal ion; (4) this probe is based on Intramolecular electron transfer (ICT) principle design, and change with Absorption and emission spectra in the process of deprotonation protonated, under natural light, solution colour obviously changes, by visual inspection.(5) this probe has good cell membrane penetration, utilizes laser confocal imaging technology can carry out detection and the cell imaging of internal pH; (6) synthesis step of this probe is simple, with low cost, has larger practical value.
Accompanying drawing explanation
Fig. 1. the uv absorption spectra that the embodiment of the present invention 1 middle probe changes with pH value.
Fig. 2. the embodiment of the present invention 1 middle probe under natural light in conjunction with H
+front and back colour-change, color becomes a piece redness from glassy yellow.
Fig. 3. the fluorescence emission spectrogram that the embodiment of the present invention 1 middle probe changes with pH value.
Fig. 4. the sigmoidal matched curve that the fluorescence intensity of the embodiment of the present invention 1 middle probe changes with pH value.
Fig. 5. the embodiment of the present invention 1 middle probe is to the response condition of common metal ion.
Fig. 6. the embodiment of the present invention 1 middle probe and human renal carcinoma cell 7860 jointly hatch the laser confocal imaging figure of 30min under the condition of pH value 7.4, and cell sends bright green glow (a) and ruddiness (b).
Fig. 7. the embodiment of the present invention 1 middle probe and human renal carcinoma cell 7860 jointly hatch the laser confocal imaging figure of 30min under the condition of pH value 4.2, and the fluorescence of cell obviously weakens when comparatively pH value is 7.4.
Embodiment
Embodiment 1
A kind of preparation of the extreme acidic pH fluorescent probe based on benzindole:
(1) under protection of inert gas; by 0.283g (1.35mmol) 1; 1; 2-trimethylammonium-1H-benzo [e] indoles and 0.454g (1.48mmol) 3-[3-(4-fluorophenyl)-1-sec.-propyl-1H-indoles-2-base]-propenal are dissolved in 6mLDMF; add 0.378g (6.75mmol) KOH solid; stirring at room temperature, reaction is spent the night.
(2) excessively filter KOH solid, in reaction solution, add H
2o (6mL), mixing solutions CH
2cl
2extraction (10mL × 3).The organic phase merged is through anhydrous Na
2sO
4after drying, underpressure distillation obtains crude product.
(3) product crude product is separated through silicagel column, and methylene dichloride is eluent, obtains orange/yellow solid.
1HNMR(300MHz,DMSO-d
6),δ:1.556(s,6H),1.648-1.671(d,6H),5.009-5.055(m,1H),6.607-6.658(m,2H),7.037-7.085(m,1H),7.178-7.258(m,2H),7.309-7.364(t,2H),7.414-7.570(m,4H),7.596-7.691(m,1H),7.691-7.755(t,3H),7.917-7.946(d,1H),7.999-8.025(d,1H),8.125-8.153(d,1H).
Embodiment 2
Concentration and probe concentration in embodiment 1 is remained on 5 μm of ol/L, by the HCl adjust ph of 1mol/L in ethanol/water (volume ratio is 2:1) system, record its absorption spectrum (Fig. 1).Along with the reduction of pH value, the absorption peak at short wavelength 426nm place declines gradually, and the absorption peak at long wavelength 546nm place significantly strengthens, and in 450nm place existence isobestic point.The color of solution has also become piece redness (Fig. 2) from original glassy yellow.
Embodiment 3
Concentration and probe concentration in embodiment 1 is remained on 5 μm of ol/L, by the HCl adjust ph of 1mol/L in ethanol/water (volume ratio is 2:1) system, be excitation wavelength with 426nm, record its fluorescence emission spectrum (Fig. 3).Along with the reduction of pH value, the fluorescence peak at 571nm place weakens gradually.Utilize the fluorescence intensity at 571nm place to map to pH value, and utilize sigmoidal to carry out matching (Fig. 4), the pKa value obtaining the probe in embodiment 1 is 2.89.
Embodiment 4
Concentration and probe concentration in embodiment 1 is remained on 5 μm of ol/L, investigate the response condition of this probe to common metal ion respectively.As shown in Figure 5, this probe does not almost respond metal ion, proves that this probe is to H
+the selectivity had.In Fig. 5, the order of material and concentration are followed successively by: 1. probe; 2.Na
+(30mM); 3.K
+(30mM); 4.Mg
2+(25mM); 5.Ca
2+(25mM); 6.Ba
2+(25mM); 7.Fe
3+(60 μMs); 8.Fe
2+(1mM); 9.Al
3+(1mM); 10.Cu
2+(1mM); 11.Zn
2+(25mM); 12.Mn
2+(25mM); 13.Ni
2+(15mM); 14.Co
2+(4mM); 15.Cr
3+(1 μM); 16.Pb
2+(0.5mM); 17.Cd
2+(0.1mM); 18.Hg
2+(0.1mM); 19.Ag
+(0.1mM).
Embodiment 5
By the probe in cultured human renal carcinoma cell 7860 and embodiment 1 respectively under pH value 7.4 and 4.2 condition, at 37 DEG C, 5%CO
2incubator in jointly hatch 30min, then wash 3 times with the phosphate buffer soln of corresponding pH value, observe under laser confocal microscope, fixing excitation wavelength is 426nm, respectively at the fluorescence that green glow and red channel collecting cell send.Experimental result shows, the cell of pH value 7.4 send comparatively bright green (in Fig. 6 a) and ruddiness (b) fluorescence in Fig. 6, and cell green fluorescence during pH4.2 (in Fig. 7 a) and red fluorescence (b) all obviously weaken in Fig. 7.
Claims (4)
1. an extreme acidic's pH fluorescent probe, is characterized in that structural formula is:
Wherein, R
1, R
2for hydrogen atom or alkyl; Described extreme acidic is pH<4.
2. the preparation method of the pH fluorescent probe of a kind of extreme acidic as claimed in claim 1, is characterized in that, comprise the steps:
(1) under protection of inert gas, by 2,3,3-tri-methyl indole or 1,1,2-trimethylammonium-1H-benzo [e] indoles and 3-[3-(4-fluorophenyl)-1-alkyl-1H-indoles-2-base]-propenal in molar ratio 1 ︰ 1.0-1.5 are dissolved in N, dinethylformamide, by 2,3,3-tri-methyl indole or 1,1,2-trimethylammonium-1H-benzo [e] indoles meter adds the KOH solid of 5 times of mole numbers, stirring at room temperature, and reaction is spent the night;
(2) excessively filter KOH solid, in reaction solution, add the H of 1-2 times of volume
2o, uses CH
2cl
2extract 2-3 time; Merge organic phase, use anhydrous Na
2sO
4after drying, underpressure distillation obtains crude product;
(3) crude product is separated through silicagel column, obtains sterling.
3. the pH fluorescent probe of a kind of extreme acidic as claimed in claim 1 is used as the indicator detecting intracellular ph value.
4. the application of pH fluorescent probe in cell imaging of a kind of extreme acidic as claimed in claim 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410262408.6A CN104017572B (en) | 2014-06-12 | 2014-06-12 | A kind of extreme acidic pH fluorescent probe and its preparation method and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410262408.6A CN104017572B (en) | 2014-06-12 | 2014-06-12 | A kind of extreme acidic pH fluorescent probe and its preparation method and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104017572A CN104017572A (en) | 2014-09-03 |
| CN104017572B true CN104017572B (en) | 2016-03-30 |
Family
ID=51434584
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410262408.6A Expired - Fee Related CN104017572B (en) | 2014-06-12 | 2014-06-12 | A kind of extreme acidic pH fluorescent probe and its preparation method and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104017572B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104535552B (en) * | 2015-01-21 | 2017-04-19 | 山西大学 | Novel application of acraldehyde derivative |
| CN105001141B (en) * | 2015-08-12 | 2017-05-24 | 中国广州分析测试中心 | Fluoro phenylindole compound, application of fluoro phenylindole compound as red organic luminescent material, and preparation method of fluoro phenylindole compound |
| CN111398269B (en) * | 2020-04-09 | 2023-03-03 | 湖北科技学院 | Method for preparing PH test paper by indole pyridinium derivative |
| CN113461588B (en) * | 2021-06-29 | 2022-09-02 | 山东省科学院海洋仪器仪表研究所 | Fluorescent probe for monitoring gastric acid and preparation method and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0525398B1 (en) * | 1991-07-29 | 1996-06-12 | Toa Medical Electronics Co., Ltd. | Method of preparing specimen for classifying and counting leukocytes |
| CN101805526A (en) * | 2010-04-10 | 2010-08-18 | 大连理工大学 | Synthesis and application of indole hemicyanine dye |
| KR20120041001A (en) * | 2010-10-20 | 2012-04-30 | 경북대학교 산학협력단 | A stimuli-responsive charge transfer dye and a method of preparing the same |
| CN103756669A (en) * | 2014-01-22 | 2014-04-30 | 山西大学 | Indole pH fluorescence probe, and preparation method and application thereof |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9410186B2 (en) * | 2012-07-11 | 2016-08-09 | Nubad Llc | Methods and compositions related to nucleic acid binding assays |
-
2014
- 2014-06-12 CN CN201410262408.6A patent/CN104017572B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0525398B1 (en) * | 1991-07-29 | 1996-06-12 | Toa Medical Electronics Co., Ltd. | Method of preparing specimen for classifying and counting leukocytes |
| CN101805526A (en) * | 2010-04-10 | 2010-08-18 | 大连理工大学 | Synthesis and application of indole hemicyanine dye |
| KR20120041001A (en) * | 2010-10-20 | 2012-04-30 | 경북대학교 산학협력단 | A stimuli-responsive charge transfer dye and a method of preparing the same |
| CN103756669A (en) * | 2014-01-22 | 2014-04-30 | 山西大学 | Indole pH fluorescence probe, and preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| Design, synthesis and evaluation of near-infrared fluorescent pH indicators in a physiologically relevant range;Zongren Zhang等;《Chem. Commun.》;20051231;5887–5889 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104017572A (en) | 2014-09-03 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Jiang et al. | High-performance near-infrared fluorescence probe for fast and specific visualization of harmful sulfite in food, living cells, and zebrafish | |
| Brewer et al. | A 2-aza-Cope reactivity-based platform for ratiometric fluorescence imaging of formaldehyde in living cells | |
| CN104017572B (en) | A kind of extreme acidic pH fluorescent probe and its preparation method and application | |
| CN105524079B (en) | Ratio-type pH fluorescence probes, preparation method, application and test method for water-soluble positioning lysosome | |
| Mao et al. | A photostable Si-rhodamine-based near-infrared fluorescent probe for monitoring lysosomal pH during heat stroke | |
| CN103436253B (en) | Rhodamine fluorescent probe for detecting ferrous ion, and preparation method thereof | |
| CN103756669B (en) | Indole pH fluorescence probe, and preparation method and application thereof | |
| Zhu et al. | A highly selective ratiometric fluorescent probe for hydrogen peroxide displaying a large emission shift | |
| CN105017196B (en) | A kind of near-infrared ratio fluorescent probe of detection hydrazine and its application | |
| CN109593078B (en) | N-butyl-4-hydroxy-1, 8-naphthalimide-3-formaldehyde- (2-pyridine) hydrazone and application thereof | |
| Liang et al. | A coumarin-based TICT fluorescent probe for real-time fluorescence lifetime imaging of mitochondrial viscosity and systemic inflammation in vivo | |
| CN104830315A (en) | Bivalent copper ion fluorescent probe and preparation method and application thereof | |
| Zhou et al. | Ratiometric fluorescent Zn2+ chemosensor constructed by appending a pair of carboxamidoquinoline on 1, 2-diaminocyclohexane scaffold | |
| Li et al. | A julolidine-chalcone-based fluorescent probe for detection of Al3+ in real water sample and cell imaging | |
| CN107417671A (en) | A kind of coumarin derivative of the substitution containing quinoline and preparation method thereof and the application on Ratio-type pH fluorescence probes | |
| CN106544007B (en) | Hypochlorous fluorescence probe and its application in a kind of detection biosystem | |
| CN111004246B (en) | Rhodamine pH fluorescent probe for monitoring mitochondrial autophagy, preparation and application thereof | |
| CN104949946A (en) | Application of fluorescent probe to hydrogen peroxide molecule detection | |
| CN103242327A (en) | P-N-methyl cyclopentaldehyde rhodamine 6G pH fluorescence molecular probe as well as preparation method and use thereof | |
| CN102095724A (en) | Method for determining copper content in water by biquinolyl spectrophotometry | |
| CN105838352B (en) | A kind of synthetic method of probe of the Visual retrieval mercury ion based on chromene nitrile and application | |
| Perinchery et al. | The potential of autofluorescence spectroscopy to detect human urinary tract infection | |
| Zhou et al. | A novel turn-on fluorescent probe for detection of pH in extremely acidic environment and its application | |
| CN107698557A (en) | Fluorescence probe based on pyridine connection pyrazoles acyl hydrazone derivative and its preparation method and application | |
| CN104132920A (en) | Method for measuring Ag<+> or F<-> through fluorescence quenching |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20160330 Termination date: 20190612 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |