CN104447774A - Rhodamine B-based fluorescence sensor and preparation - Google Patents
Rhodamine B-based fluorescence sensor and preparation Download PDFInfo
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- CN104447774A CN104447774A CN201410637758.6A CN201410637758A CN104447774A CN 104447774 A CN104447774 A CN 104447774A CN 201410637758 A CN201410637758 A CN 201410637758A CN 104447774 A CN104447774 A CN 104447774A
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- C—CHEMISTRY; METALLURGY
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- C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
- C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
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- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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Abstract
The invention discloses a rhodamine B-based fluorescence sensor and preparation. The preparation comprises the following steps: dissolving 3-(3',6'-bi(diethyl amino)-3-oxaspiro[isoindoline-1,-1,9'-xanthene]-2-yl) propanal (A) in methyl alcohol, adding 6-aminocaproic acid (B), and stirring at room temperature; dissolving sodium triacetoxyborohydride in tetrahydrofuran, adding to a reaction liquid in the step 1, and further carrying out stirring reaction at room temperature; dropwise adding a saturated sodium bicarbonate solution to the reaction liquid, and further stirring; and removing a solvent by virtue of a rotary evaporator, and separating by using a thin layer chromatography, so as to obtain a target product. The rhodamine B-based fluorescence sensor is relatively simple in synthesis step; resources are saved; and nearly all the reported fluorescence probes can only be applied to an organic solvent or an organic water solution, thus the application of the fluorescence sensor in the fields of actual water sample analysis and life is limited.
Description
Technical field
The present invention relates to a kind of fluorescent optical sensor based on rhodamine B and preparation, belong to bioluminescence field of sensor preparation.
Background technology
Tin is one of requisite trace element in human body, and it carries out various physiological activity to human body and safeguards that HUMAN HEALTH has material impact, and its main physiological function shows antitumor and promotes the synthesis of protein and nucleic acid.Research finds that in tumor tissues, Theil indices is organized normally lower than other.Lack the metabolic disturbance that tin can cause protein and nucleic acid in human body, but if accumulate too much tin in human body, the ill symptoms such as will occur dizziness, diarrhoea, feel sick.Chromium is also trace element required in human body, and it is that normal growth is grown and regulates the important element of blood sugar.The heavy metal ion content such as tin and chromium is too low or too highly all can cause detrimentally affect to health.Therefore, how effectively the heavy metal cation in testing environment and organism has great meaning for environment measuring, medical diagnosis on disease and health assessment etc.
Detection people up to now for these heavy metal ion have developed multiple method, as atomic absorption spectrum, atomic emissions spectrum etc.But these methods have a lot of shortcoming, as sample need pre-treatment, detect not fast, detect expensive etc.Therefore, people in the urgent need to fast, accurately, the method for these heavy metals of analyzing and testing at low cost.
The advantages such as fluorescent optical sensor is fast with highly sensitive, detection speed, easy handling enjoy people to pay close attention to.Puhui Xie (Journal of Luminescence 140 (2013) 45 – 50) reports a kind of specificity based on rhodamine and detects Cr
3+fluorescent optical sensor, the chromium ion in the aqueous solution and viable cell can be detected, to environment measuring and medical diagnosis on disease significant.Han OuYang (Tetrahedron Letters (2013), 54 (23), 2964-2966) reports a kind of to Fe
3+the fluorescent optical sensor that specificity detects.
What Li seminar reported connects rhodamine B acylhydrazone and the compound containing benzene-naphthalene diimide or ferrocene units structure by quinoline, this compound and Cr
3+after selective coordination, the open loop of induction rhodamine B volution, after the optical excitation of 405nm place, Resonance energy transfer occurs, and make its fluorescence emission wavelengths red shift to 594nm, fluorescence color also becomes redness from yellow, can realize bore hole observation.Structural formula of compound is as follows:
Summary of the invention
The object of the present invention is to provide a kind of fluorescent optical sensor based on rhodamine and preparation thereof, described fluorescent optical sensor is to Sn
2+, Cr
3+have good fluorescence response, have potential application detecting this two heavy metal species ions in organism, medical diagnosis on disease, health assessment in have good application prospect.
The technical solution realizing the object of the invention is: a kind of fluorescent optical sensor based on rhodamine B, and described fluorescent optical sensor has following structure:
The preparation of the above-mentioned fluorescent optical sensor based on rhodamine B, comprises the steps:
Step 1, by 3-(3', two (diethylamino)-3-oxaspiro [isoindoline-1 ,-1,9'-the xanthene]-2-base of 6'-) propionic aldehyde (A) is dissolved in methyl alcohol, then add 6-aminocaprolc acid (B), stir at normal temperatures;
Step 2, Sodium triacetoxyborohydride is dissolved in tetrahydrofuran (THF), joins in step 1 reaction solution, continue stirring reaction at normal temperatures;
Step 3, dropping saturated sodium bicarbonate solution, in above-mentioned reaction solution, continue to stir;
Step 4, with Rotary Evaporators except desolventizing, be separated by tlc, obtain target product.
3-(two (diethylamino)-3-oxaspiro [isoindoline-1 ,-1,9'-the xanthene]-2-base of 3', 6'-) propionic aldehyde described in step 1 and 6-aminocaprolc acid mol ratio are 1:3, and under normal temperature, churning time is more than 2h.
The mol ratio of the Sodium triacetoxyborohydride described in step 2 and 6-aminocaprolc acid is 2:3, and under normal temperature, churning time is more than 3h.
Compared with prior art, advantage of the present invention is:
Synthesis step is simpler, economize on resources, the fluorescent probe of report nearly all can only be applied to organic solvent or organic-aqueous solution, which limits its application in actual water sample analysis and life field, thus design one structure is simple, selectivity good, highly sensitive water-soluble fluorescent probe molecule seems particularly important.
Accompanying drawing explanation
Fig. 1 is the fluorescence response figure of fluorescent optical sensor to different ions solution, and wherein X-coordinate is numbering (1, the none of different ions; 2, Sn
2+; 3, Cr
3+; 4, Ca
2+; 5, Ag
+; 6, Mg
2+; 7, K
+; 8, Ba
2+; 9, Co
2+; 10, Zn
2+; 11, Li
+; 12, Cd
2+; 13, Fe
2+; 14, Cu
2+; 15, Mn
2+; 16, Pb
2+; 17, Na
+), ordinate zou is fluorescent value.
Fig. 2 is the Sn of fluorescent optical sensor to different concns
2+the fluorogram of solution, wherein X-coordinate is wavelength, unit: nm; Ordinate zou is fluorescent value.
Fig. 3 is the Cr of fluorescent optical sensor to different concns
2+the fluorogram of solution, wherein X-coordinate is wavelength, unit: nm; Ordinate zou is fluorescent value.
Embodiment
Synthetic route of the present invention is as follows:
One, the synthesis of target compound
1. by 100mg 3-(3', two (the diethylamino)-3-oxaspiro [isoindoline-1 of 6'-,-1,9' – xanthene]-2-base) propane (see article Design and synthesis of a novel Rhodamine B [2] rotaxane) is dissolved in 5mL methyl alcohol, then add 79mg 6-aminocaprolc acid, stir 2h at normal temperatures;
2. take 85mg Sodium triacetoxyborohydride, be dissolved in 1ml tetrahydrofuran (THF), joined in above-mentioned reaction solution with liquid-transfering gun, continue to stir at normal temperatures, by TLC detection reaction progress, within 3 hours, react completely;
3. drip 1mL saturated sodium bicarbonate solution in above-mentioned reaction solution, continue to stir 10min, saturated sodium bicarbonate is in order to quencher adds Sodium triacetoxyborohydride wherein;
4. remove desolventizing with Rotary Evaporators, be separated by tlc, obtain target product.
1H NMR(500MHz,CDCl
3)δ7.94–7.81(m,1H),7.52–7.33(m,2H),7.05(tq,J=7.1,3.6Hz,1H),6.41(d,J=8.9Hz,2H),6.38(t,J=6.2Hz,2H),6.26(dd,J=8.9,2.6Hz,2H),3.38–3.27(m,9H),3.18(dd,J=16.3,9.3Hz,2H),3.12(dd,J=15.9,8.3Hz,4H),2.52–2.33(m,2H),1.71–1.59(m,3H),1.59–1.48(m,3H),1.48–1.42(m,1H),1.35(dd,J=16.2,9.0Hz,2H),1.16(d,J=7.1Hz,11H)。
Two, target compound fluorescence property test
1. the fluorescence response test of pair different heavy metal ion
The target product of gained is mixed with 10 μm of ol/L methanol-waters mixing solutions (4:6, V:V), gets 3000 μ L and be placed in liquid cell use, detect its initial fluorescence value.The MgCl prepared is measured with microsyringe
26H
2o, SnCl
2h
2o, CrCl
36H
2o, AgNO
3, CaCl
2, NaCl, PbCl
2, KCl, MnCl
24H
2o, ZnCl
2, CuCl
22H
2o, CdCl
22.5H
2o, LiClH
2o, Ba (NO
3)
2, FeCl
24H
2o, CoCl
26H
2the solion of O, observes fluorogram change and record.We find to add Sn
2+, Cr
3+after, fluorescence intensity changes to some extent, sees Fig. 1.
2. the fluorescence response change of different concns tin ion solution
To in the 10 μm of ol/L solution title compound be mixed with, add mother liquor respectively and tin ion mol ratio is that 1:0.1,1:0.2,1:0.3,1:0.4,1:0.5,1:1,1:2,1:3 are until the Sn of 1:130
2+, corresponding Sn
2+concentration is increased to 1.3mmol/L from 0.1 μm of ol/L.We find within the specific limits, and present linear relationship, when mol ratio is 1:50, fluorescence intensity reaches capacity, and sees Fig. 2.
3. the fluorescence response change of different concns chromium ion solution
Detect mother liquor according to the method same with step 2 and the fluorescence response of chromium ion mol ratio from 1:0.1 to 1:130 changes, find within the specific limits, present linear relationship, when mol ratio is 1:50, fluorescence intensity reaches capacity, and sees Fig. 3 always.
Claims (4)
1. based on a fluorescent optical sensor for rhodamine B, it is characterized in that, described fluorescent optical sensor has following structure:
2. based on a preparation for the fluorescent optical sensor of rhodamine B, it is characterized in that, comprise the steps:
Step 1,3-(3', 6'-two (diethylamino)-3-oxaspiro [isoindoline-1 ,-1,9'-xanthene]-2-base) propionic aldehyde is dissolved in methyl alcohol, then adds 6-aminocaprolc acid, stir at normal temperatures;
Step 2, Sodium triacetoxyborohydride is dissolved in tetrahydrofuran (THF), joins in step 1 reaction solution, continue stirring reaction at normal temperatures;
Step 3, dropping saturated sodium bicarbonate solution, in above-mentioned reaction solution, continue to stir;
Step 4, with Rotary Evaporators except desolventizing, be separated by tlc, obtain target product.
3. as claimed in claim 2 based on the preparation of the fluorescent optical sensor of rhodamine B, it is characterized in that, 3-(3' described in step 1, two (the diethylamino)-3-oxaspiro [isoindoline-1 of 6'-,-1,9'-xanthene]-2-base) propionic aldehyde and 6-aminocaprolc acid mol ratio be 1:3, under normal temperature, churning time is more than 2h.
4., as claimed in claim 2 based on the preparation of the fluorescent optical sensor of rhodamine B, it is characterized in that, the mol ratio of the Sodium triacetoxyborohydride described in step 2 and 6-aminocaprolc acid is 2:3, and under normal temperature, churning time is more than 3h.
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Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105331358A (en) * | 2015-12-07 | 2016-02-17 | 河南省农业科学院 | Dirhodamine-based Sn4+ fluorescent probe molecule and preparation method and application thereof |
CN105505376A (en) * | 2015-12-07 | 2016-04-20 | 河南省农业科学院 | Hg<2+> fluorescence probe containing rhodamine with double-carbon-sulfur-bond structure as well as preparation method and application of Hg<2+> fluorescence probe |
CN105907387A (en) * | 2016-04-22 | 2016-08-31 | 东华大学 | Method for detecting stannous ions by using rhodamine fluorescence probe |
CN107129503A (en) * | 2017-05-06 | 2017-09-05 | 渤海大学 | A kind of quick detection Cr in aqueous3+The preparation method and applications of the enhanced probe of ion fluorescence |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103242327A (en) * | 2013-04-11 | 2013-08-14 | 天津师范大学 | P-N-methyl cyclopentaldehyde rhodamine 6G pH fluorescence molecular probe as well as preparation method and use thereof |
CN103540312A (en) * | 2013-10-09 | 2014-01-29 | 大连理工大学 | Rhodamine fluorescent probe with pseudo nucleic acid base as recognition site and preparation thereof and application to nucleotide image |
CN104017569A (en) * | 2014-05-23 | 2014-09-03 | 苏州科技学院 | Rhodamine-containing lactam group micromolecule pH fluorescent probe and synthetic method |
-
2014
- 2014-11-12 CN CN201410637758.6A patent/CN104447774B/en not_active Expired - Fee Related
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103242327A (en) * | 2013-04-11 | 2013-08-14 | 天津师范大学 | P-N-methyl cyclopentaldehyde rhodamine 6G pH fluorescence molecular probe as well as preparation method and use thereof |
CN103540312A (en) * | 2013-10-09 | 2014-01-29 | 大连理工大学 | Rhodamine fluorescent probe with pseudo nucleic acid base as recognition site and preparation thereof and application to nucleotide image |
CN104017569A (en) * | 2014-05-23 | 2014-09-03 | 苏州科技学院 | Rhodamine-containing lactam group micromolecule pH fluorescent probe and synthetic method |
Non-Patent Citations (1)
Title |
---|
XIAOFENG BAO ET AL.: "Design and synthesis of a novel Rhodamine B[2]rotaxane", 《RSC ADV.》 * |
Cited By (8)
Publication number | Priority date | Publication date | Assignee | Title |
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CN105331358A (en) * | 2015-12-07 | 2016-02-17 | 河南省农业科学院 | Dirhodamine-based Sn4+ fluorescent probe molecule and preparation method and application thereof |
CN105505376A (en) * | 2015-12-07 | 2016-04-20 | 河南省农业科学院 | Hg<2+> fluorescence probe containing rhodamine with double-carbon-sulfur-bond structure as well as preparation method and application of Hg<2+> fluorescence probe |
CN105331358B (en) * | 2015-12-07 | 2017-04-05 | 河南省农业科学院 | A kind of Sn based on double rhodamines4+Fluorescent probe molecule and preparation method and application |
CN105505376B (en) * | 2015-12-07 | 2017-05-31 | 河南省农业科学院 | A kind of Hg containing double carbon-sulfur bond structure rhodamines2+Fluorescence probe and preparation method and application |
CN105907387A (en) * | 2016-04-22 | 2016-08-31 | 东华大学 | Method for detecting stannous ions by using rhodamine fluorescence probe |
CN105907387B (en) * | 2016-04-22 | 2018-01-19 | 东华大学 | A kind of method that stannous ion is detected using rhodamine fluorescence probe |
CN107129503A (en) * | 2017-05-06 | 2017-09-05 | 渤海大学 | A kind of quick detection Cr in aqueous3+The preparation method and applications of the enhanced probe of ion fluorescence |
CN107129503B (en) * | 2017-05-06 | 2019-05-31 | 渤海大学 | A kind of detection Cr quick in aqueous solution3+The preparation method and applications of the enhanced probe of ion fluorescence |
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