CN103232358A - Vesicle probe used for detecting neomycin, and application and preparation method thereof - Google Patents
Vesicle probe used for detecting neomycin, and application and preparation method thereof Download PDFInfo
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- 239000000523 sample Substances 0.000 title claims abstract description 64
- 238000002360 preparation method Methods 0.000 title abstract description 8
- 229930193140 Neomycin Natural products 0.000 title abstract description 5
- 229960004927 neomycin Drugs 0.000 title abstract description 5
- 238000006243 chemical reaction Methods 0.000 claims abstract description 38
- 150000001875 compounds Chemical class 0.000 claims abstract description 25
- 239000002253 acid Substances 0.000 claims abstract description 15
- 238000001514 detection method Methods 0.000 claims abstract description 14
- DBLXOVFQHHSKRC-UHFFFAOYSA-N ethanesulfonic acid;2-piperazin-1-ylethanol Chemical compound CCS(O)(=O)=O.OCCN1CCNCC1 DBLXOVFQHHSKRC-UHFFFAOYSA-N 0.000 claims abstract description 10
- 238000006116 polymerization reaction Methods 0.000 claims abstract description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 28
- 238000000034 method Methods 0.000 claims description 21
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 18
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 15
- 239000002243 precursor Substances 0.000 claims description 14
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 12
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 claims description 10
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 9
- 238000013016 damping Methods 0.000 claims description 9
- 239000012530 fluid Substances 0.000 claims description 9
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 9
- 229910052757 nitrogen Inorganic materials 0.000 claims description 8
- 238000003756 stirring Methods 0.000 claims description 8
- 239000012043 crude product Substances 0.000 claims description 6
- 238000004821 distillation Methods 0.000 claims description 6
- 239000002904 solvent Substances 0.000 claims description 6
- CWLKGDAVCFYWJK-UHFFFAOYSA-N 3-aminophenol Chemical compound NC1=CC=CC(O)=C1 CWLKGDAVCFYWJK-UHFFFAOYSA-N 0.000 claims description 5
- 238000004587 chromatography analysis Methods 0.000 claims description 5
- HOGDNTQCSIKEEV-UHFFFAOYSA-N n'-hydroxybutanediamide Chemical compound NC(=O)CCC(=O)NO HOGDNTQCSIKEEV-UHFFFAOYSA-N 0.000 claims description 5
- 238000001816 cooling Methods 0.000 claims description 4
- 150000001408 amides Chemical class 0.000 claims description 2
- 238000004090 dissolution Methods 0.000 claims description 2
- 238000010926 purge Methods 0.000 claims description 2
- 238000009281 ultraviolet germicidal irradiation Methods 0.000 claims description 2
- 230000035945 sensitivity Effects 0.000 abstract description 2
- 239000007853 buffer solution Substances 0.000 abstract 1
- 239000000243 solution Substances 0.000 abstract 1
- 239000007788 liquid Substances 0.000 description 7
- 229930182555 Penicillin Natural products 0.000 description 4
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 4
- 150000001345 alkine derivatives Chemical class 0.000 description 4
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 4
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 description 4
- 238000002189 fluorescence spectrum Methods 0.000 description 4
- 238000005457 optimization Methods 0.000 description 4
- 229940049954 penicillin Drugs 0.000 description 4
- 239000010453 quartz Substances 0.000 description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 4
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 4
- 238000000862 absorption spectrum Methods 0.000 description 3
- 239000003242 anti bacterial agent Substances 0.000 description 3
- 229940088710 antibiotic agent Drugs 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 230000003287 optical effect Effects 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 2
- 206010033109 Ototoxicity Diseases 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 229960000723 ampicillin Drugs 0.000 description 2
- 244000052616 bacterial pathogen Species 0.000 description 2
- 229960003669 carbenicillin Drugs 0.000 description 2
- FPPNZSSZRUTDAP-UWFZAAFLSA-N carbenicillin Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)C(C(O)=O)C1=CC=CC=C1 FPPNZSSZRUTDAP-UWFZAAFLSA-N 0.000 description 2
- RTYJTGSCYUUYAL-YCAHSCEMSA-L carbenicillin disodium Chemical compound [Na+].[Na+].N([C@H]1[C@H]2SC([C@@H](N2C1=O)C([O-])=O)(C)C)C(=O)C(C([O-])=O)C1=CC=CC=C1 RTYJTGSCYUUYAL-YCAHSCEMSA-L 0.000 description 2
- 229960005091 chloramphenicol Drugs 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 231100000262 ototoxicity Toxicity 0.000 description 2
- 229940049547 paraxin Drugs 0.000 description 2
- 229960005322 streptomycin Drugs 0.000 description 2
- QTENRWWVYAAPBI-YCRXJPFRSA-N streptomycin sulfate Chemical compound OS(O)(=O)=O.OS(O)(=O)=O.OS(O)(=O)=O.CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](N=C(N)N)[C@H](O)[C@@H](N=C(N)N)[C@H](O)[C@H]1O.CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](N=C(N)N)[C@H](O)[C@@H](N=C(N)N)[C@H](O)[C@H]1O QTENRWWVYAAPBI-YCRXJPFRSA-N 0.000 description 2
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 1
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- 206010049712 Dysacusis Diseases 0.000 description 1
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- 241000588724 Escherichia coli Species 0.000 description 1
- 206010017964 Gastrointestinal infection Diseases 0.000 description 1
- 208000016621 Hearing disease Diseases 0.000 description 1
- 241000588748 Klebsiella Species 0.000 description 1
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 description 1
- 208000009205 Tinnitus Diseases 0.000 description 1
- 229940126575 aminoglycoside Drugs 0.000 description 1
- 239000002647 aminoglycoside antibiotic agent Substances 0.000 description 1
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- 239000007850 fluorescent dye Substances 0.000 description 1
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Images
Abstract
The invention relates to a vesicle probe used for detecting neomycin, and an application and a preparation method thereof. The probe is obtained by polymerization by using 10,12-diyne pentacosoic acid and a compound represented by the formula I, according to a weight ratio of 9:1. The vesicle probe provided by the invention is blue and non-fluorescent in a 4-hydroxyethylpiperazine ethanesulfonic acid buffer solution. When the vesicle probe contact neomycin and is subjected to a reaction, the vesicle probe turns orange and fluorescent, wherein a fluorescence emission wavelength is 550nm. The probe has high selectivity and high sensitivity. A probe detection method adopting the probe has good application prospect in the field of biomedicine.
Description
Technical field
The present invention relates to a kind of vesica probe, purposes and preparation method for detection of Xin Meisu.
Background technology
Microbiotic pollutes and is considered to threaten one of the most serious arch-criminal of human health at present, and wherein, abuse of antibiotics has three harm greatly: endanger one, use microbiotic can bring strong toxic side effect in a large number, directly injure health, especially to children's hearing; Harm two, microbiotic excessively use can make bacterium develop immunity to drugs, and make the effects of antibiotics variation or directly do not have effect; Harm three, the too much mistake of microbiotic are used excessively, can kill a large amount of normal bacteria in the body, thereby pathogenic bacterium are availed oneself of the opportunity to get in, and cause human body death.
Xin Meisu is a kind of microbiotic, belongs to aminoglycoside antibiotics, and Xin Meisu has anti-microbial effect to many gram negative bacillis such as escherichia coli, klebsiella spp etc., is mainly used in treating class diseases such as the gastro-intestinal infection of poultry and stomatitis, mazoitis.The excessive use of Xin Meisu can cause serious pathology, for example dysacusis, tinnitus or ear's satiety (ototoxicity); Lurch, dizzy or staggered (ototoxicity, influence vestibular); Urine amount or number of micturitions significantly reduce, or extreme thirsty (renal toxicity).
In aminoglycosides antibiotics, the neuromuscular junction restraining effect of Xin Meisu is the strongest, once has many patients that serious respiration inhibition even death take place.Therefore, the detection by quantitative of Xin Meisu content is significant in biological chemistry and clinical chemistry.
The technique means that detects Xin Meisu at present mainly is: enzyme-linked immunosorbent assay, fluorescence polarization immunoassay and high performance liquid chromatography etc., but these technique means are consuming time, sensitivity is low and need complicated experimental assembly and operating process.
Summary of the invention
Technical problem to be solved by this invention is: overcome the problem that prior art exists, a kind of vesica probe for detection of Xin Meisu, purposes are provided, reach the preparation method.
The technology of the present invention design is as follows: had the detection method of fluorescent probe in the prior art, these class methods are quick, sensitive, accurate, therefore produce the probe of fluorescence as long as prepare to react with Xin Meisu.
The technical scheme that the present invention solves its technical problem is as follows:
A kind of probe precursor for detection of Xin Meisu of the present invention is characterized in that, has suc as formula the structure shown in the I:
A kind of method for preparing above-mentioned probe precursor of the present invention is characterized in that, may further comprise the steps:
The first step, with 10,12-diine neocerotic acid and N-hydroxy-succinamide, N, N '-dicyclohexylcarbodiimide reaction obtains formula II compound;
Second step, formula II compound and meta-aminophenol reacted obtain probe precursor shown in the formula I.
Further improve and be, the detailed process of the first step is: with 10,12-diine neocerotic acid is dissolved in and contains N-hydroxy-succinamide and N, in the dichloromethane solution of N'-dicyclohexylcarbodiimide, obtain reaction solution, continues to stir this reaction solution and finish until reaction under nitrogen protection; Then, solvent is removed in underpressure distillation, namely gets formula II compound.
Further improve and be, the detailed process in second step is: earlier formula II compound dissolution is obtained formula II compound solution in tetrahydrofuran (THF), add to formula II compound solution in the tetrahydrofuran solution that contains meta-aminophenol again, obtain reaction solution, under nitrogen protection, continue to stir this reaction solution and finish until reaction; Then, underpressure distillation is removed solvent and is obtained crude product, and with crude product with the chromatography column purifying, namely get probe precursor shown in the formula I.
Further improve and be: in second step, adopt the methylene dichloride of volume ratio 100:0.5 in the chromatography column purge process: methyl alcohol is elutriant.
A kind of vesica probe for detection of Xin Meisu of the present invention is characterized in that, is 10 of 9:1 by weight ratio, and 12-diine neocerotic acid and formula I compound polymerization form.
A kind of method for preparing above-mentioned vesica probe of the present invention is characterized in that, may further comprise the steps:
With 10,12-diine neocerotic acid and formula I compound are dissolved in dimethyl sulfoxide (DMSO) by aforementioned ratio and obtain DMSO solution, DMSO solution is heated at least 80 ℃ and add in the 4-hydroxyethyl piperazine ethanesulfonic acid damping fluid and obtain reaction solution, the cooling reaction solution, carry out polyreaction with the UV-irradiation reaction solution, namely get the vesica probe.
Further improve and be, before the cooling reaction solution, that reaction solution is ultrasonic and filter with filter.
Further improve is to adopt the 254nm ultraviolet lamp as ultraviolet source.
In addition, the present invention also provides the purposes of aforementioned vesica probe for detection of Xin Meisu.
Vesica probe of the present invention is blue and does not have fluorescence in 4-hydroxyethyl piperazine ethanesulfonic acid damping fluid, meet to become orange after Xin Meisu reacts and send fluorescence (fluorescent emission wavelength 550nm), and selectivity is strong, and is highly sensitive.
Compare with existing detection method, adopt the fluorescence detection method speed of response of vesica probe of the present invention fast, can develop the color in 5 minutes; Highly sensitive, can reach 10 to the detectability of Xin Meisu
-6The M level; Only under the Xin Meisu existence condition, react, have very strong selectivity and specificity; Has suitable fluorescent emission wavelength (550nm); Can realize detection by quantitative to micro-Xin Meisu at aqueous phase.Therefore, the present invention has great application prospect at biomedicine field.
Description of drawings
Fig. 1 is the embodiment of the invention 2 vesica probe optimizations figure as a result.
Fig. 2 is the embodiment of the invention 2 Xin Meisu concentration optimum result figure.
Fig. 3 is the embodiment of the invention 2 vesica probe selectivity tests figure as a result.
Fig. 4 is the fluorescence emission spectrum of the embodiment of the invention 3.
Fig. 5 is the UV, visible light optical absorption spectra of the embodiment of the invention 3.
Fig. 6 is the result schematic diagram of the embodiment of the invention 4.
Embodiment
Describe in further detail below in conjunction with the present invention of embodiment.But the invention is not restricted to given example.
The experiment material that relates in the following content and reagent then are commercially available product as not specifying.
The preparation of embodiment 1 formula I probe precursor, vesica probe
One, preparation formula I probe precursor
The first step, with 0.375g (1mmol) 10,12-diine neocerotic acid is dissolved in and contains 0.15g (1.3mmol) N-hydroxy-succinamide (English name: N-Hydroxysuccinimide, abbreviation: NHS) and 0.3g (1.45mmol) N, N'-dicyclohexylcarbodiimide (English name: N, N '-Dicyclohexylcarbodiimide, abbreviation: in 20ml dichloromethane solution DCC), obtain reaction solution, under nitrogen protection, continue to stir this reaction solution and finish (be generally to stir and spend the night) until reaction; Then, solvent is removed in underpressure distillation, namely gets the formula II compound that is colorless oil.
Second step, elder generation are dissolved in the first step gained formula II compound and obtain formula II compound solution in a small amount of tetrahydrofuran (THF), drop to formula II compound solution in the 20ml tetrahydrofuran solution that contains the 0.15g meta-aminophenol again, obtain reaction solution, under nitrogen protection, continue to stir this reaction solution and finish (be generally to stir and spend the night) until reaction; Then, underpressure distillation is removed solvent and is obtained crude product, and with crude product with chromatography column purifying (adopt the methylene dichloride of volume ratio 100:0.5: methyl alcohol is elutriant), namely get probe precursor shown in the formula I, weight 120.8mg, the solid that is white in color, molecular weight are 465.
Formula I probe precursor
1H NMR (DMSO, 500MHz) δ (ppm): δ 0.83-0.86 (t, 3H, J=13.55Hz), 1.24 (s, 26H), and 1.40-1.45 (t, 4H, J=25.85Hz), 1.70-1.73 (t, 2H, J=15.7Hz), 2.24-2.27 (t, 6H, J=17.65Hz), 6.39-6.41 (d, 1H), 6.91-6.93 (d, 1H), 7.0-7.041 (t, 1H, J=15.7Hz), 7.17 (s, 1H), 9.26 (s, 1H), 9.65 (s, 1H).
Formula I probe precursor
13C NMR (DMSO, 500MHz) δ (ppm): 13.85,18.23,22.03,24.41,25.07,27.65,27.68,28.11,28.16,28.25,28.30,28.33,28.45,28.54,28.59,28.62,28.64,28.80,28.88,28.94,31.24,33.61,36.40,65.30,77.85,106.21,109.71,129.10,140.31,157.48,174.37.
HRMS (EI) m/z 465.3533 (C of formula I probe precursor
31H
47NO
2).
Operational path is as follows:
Two, preparation vesica probe
With 10,12-diine neocerotic acid and formula I compound are dissolved in the 1ml dimethyl sulfoxide (DMSO) by weight 9:1 and obtain DMSO solution, DMSO solution is heated to 80 ℃, and in 5 minutes, drop in 4-hydroxyethyl piperazine ethanesulfonic acid (HEPES) damping fluid (10mM pH=7.4) with syringe and to obtain reaction solution, this moment 10, the ultimate density of 12-diine neocerotic acid is 1mM; Reaction solution was descended ultrasonic 25 minutes at 80 ℃, and filter with 0.8 μ m filter, remove insoluble particle; Reaction solution is put 4 ℃ to be cooled off 12 hours down; Adopt 254nm ultraviolet lamp (1mW/cm
2) irradiation reaction solution carried out polyreaction in 25 minutes, namely get blue vesica probe.
The optimization of embodiment 2 vesica probes, Xin Meisu concentration, and vesica probe selectivity test
One, the optimization of vesica probe
Two, the optimization of Xin Meisu concentration
Get six part of 100 μ l vesica probe respectively and put into 96 orifice plates, it is 500 μ M that adding 4-hydroxyethyl piperazine ethanesulfonic acid (HEPES) damping fluid (10mM pH=7.4) makes each vesica probe ultimate density, adds 0.1 μ l respectively in each hole, 0.2 μ l, 0.4 μ l, 1 μ l, the Xin Meisu of 2 μ l 10mM, make the Xin Meisu ultimate density be respectively 0 μ M, 5 μ M, 10 μ M, 20 μ M, 50 μ M, 100 μ M; React and observe colour-change after 5 minutes.The result increases with Xin Meisu concentration as shown in Figure 2, and liquid is become orangely gradually in each hole by blueness, and wherein, the Xin Meisu colour-change of 50 μ M is comparatively obvious, and the Xin Meisu colour-change of 100 μ M is the most obvious, becomes orangely fully, and naked eyes can observe directly.
Three, vesica probe selectivity test
Get 18 part of 100 μ l vesica probe respectively and put into 96 orifice plates, it is 500 μ M that adding 4-hydroxyethyl piperazine ethanesulfonic acid (HEPES) damping fluid (10mM pH=7.4) makes each vesica probe ultimate density; 100 each analyte of μ M that add equal volume (as 1 μ l) by the hole order to each hole respectively: penicillin (penicillin), Mg
2+, NH
4+, K
+, Ca
2+, Na
+, SO4
2-, Cl
-, HCO
3-, NO
3-, PO
4 3-, NH
3H
2O, Streptomycin sulphate (streptomycin), penbritin (ampicillin), paraxin (chloramphenicol), Pyocianil (carbenicillin), Xin Meisu (neomycin); React after 5 minutes, observe each hole liquid color and change.As shown in Figure 3, except hole, Xin Meisu place liquid become orange, all the other each hole liquid colors do not have considerable change.
More than three experimental results show:
1) have only 10,12-diine neocerotic acid and formula I compound by weight the vesica probe that is polymerized for 9:1 could with Xin Meisu generation color reaction;
2) adding Xin Meisu that concentration is at least 50 μ M to 500 μ M vesica probes can produce obvious color and change;
3) the vesica probe has selectivity and the specificity of height to Xin Meisu, can tell Xin Meisu with visual inspection from experimental result.
The spectral quality of embodiment 3 vesica probes and Xin Meisu reaction
Get 150 μ l 1mM vesica probes respectively and add in a series of quartz containeres, add 4-hydroxyethyl piperazine ethanesulfonic acid (HEPES) damping fluid (10mM pH=7.4) then respectively and dilute and be settled to 3ml; Then, add the Xin Meisu (0 μ M, 1 μ M, 2 μ M, 4 μ M, 8 μ M, 10 μ M) of equal volume (as 1 μ l), different concns respectively, react and measure fluorescence emission spectrum and UV, visible light optical absorption spectra after 5 minutes respectively.Excite with 492nm when fluorescence emission spectrum is measured, exciting with the slit width of launching is 5nm.
Fluorescence emission spectrum as shown in Figure 4, Xin Meisu concentration is in 0-10 μ M scope the time, the fluorescence intensity at 550nm place strengthens with the increase of Xin Meisu concentration, when Xin Meisu concentration was 8 μ M, fluorescence intensity was increased to maximum value.
The UV, visible light optical absorption spectra as shown in Figure 5, Xin Meisu concentration is in 0-10 μ M scope the time, with the increase of Xin Meisu concentration, the absorbancy at 620nm place reduces gradually, the absorbancy at 541nm place increases gradually; As calculated, when Xin Meisu concentration was 8 μ M, it is maximum that its CR value reaches, i.e. the colour-change maximum.
This experiment shows that the fluorescence intensity that liquid sends in each quartz container strengthens with the increase of Xin Meisu concentration, and its color response value is increasing with the increase of Xin Meisu concentration.
The color response value (CR) of embodiment 4 vesica probes and different analytes
Get 150 μ l 1mM vesica probes respectively and add in a series of quartz containeres, add 4-hydroxyethyl piperazine ethanesulfonic acid (HEPES) damping fluid (10mM pH=7.4) then respectively and dilute and be settled to 3ml; Then, the different analytes of 8 μ M that add equal volume (as 10 μ l) respectively: penicillin (penicillin), Mg
2+, NH
4+, K
+, Ca
2+, Na
+, SO4
2-, Cl
-, HCO
3-, NO
3-, PO
4 3-, NH
3H
2O, Streptomycin sulphate (streptomycin), penbritin (ampicillin), paraxin (chloramphenicol), Pyocianil (carbenicillin), Xin Meisu (neomycin); Reacted 5 minutes, and before and after reaction, measured respectively that liquid calculated each CR value according to following formula then in the absorbancy at 630nm and 530nm place in each quartz container:
CR=(PB
0-PB
1)/PB
0* 100%, PB wherein
0Or PB
1=A
Blue/ (A
Blue+ A
Red) * 100%,
Wherein, A
BlueRefer to the absorbancy (the two alkynes of red gathering are to the absorption intensity of blue ripple) at 530nm place, A
RedRefer to the absorbancy (the blue absorption intensity of two alkynes to red ripple of meeting) at 630nm place, PB
0Poly-two alkynes absorb shared percentage ratio, PB to red ripple before referring to react
1Refer to react the poly-two alkynes in back red ripple is absorbed shared percentage ratio.The CR value is more big, shows that the variable color degree of system is more big.
The result show the vesica probe to the color response value maximum of Xin Meisu, so this probe can detect to Xin Meisu optionally as shown in Figure 6.
Claims (10)
1. the probe precursor for detection of Xin Meisu is characterized in that, has suc as formula the structure shown in the I:
2. a method for preparing the described probe precursor of claim 1 is characterized in that, may further comprise the steps:
The first step, with 10,12-diine neocerotic acid and N-hydroxy-succinamide, N, N '-dicyclohexylcarbodiimide reaction obtains formula II compound;
Second step, formula II compound and meta-aminophenol reacted obtain probe precursor shown in the formula I.
3. according to the described method of claim 2, it is characterized in that, the detailed process of the first step is: with 10,12-diine neocerotic acid is dissolved in and contains N-hydroxy-succinamide and N, in the dichloromethane solution of N '-dicyclohexylcarbodiimide, obtain reaction solution, under nitrogen protection, continue to stir this reaction solution and finish until reaction; Then, solvent is removed in underpressure distillation, namely gets formula II compound.
4. according to the described method of claim 2, it is characterized in that, the detailed process in second step is: earlier formula II compound dissolution is obtained formula II compound solution in tetrahydrofuran (THF), add to formula II compound solution in the tetrahydrofuran solution that contains meta-aminophenol again, obtain reaction solution, under nitrogen protection, continue to stir this reaction solution and finish until reaction; Then, underpressure distillation is removed solvent and is obtained crude product, and with crude product with the chromatography column purifying, namely get probe precursor shown in the formula I.
5. according to the described method of claim 4, it is characterized in that in second step, adopt the methylene dichloride of volume ratio 100:0.5 in the chromatography column purge process: methyl alcohol is elutriant.
6. the vesica probe for detection of Xin Meisu is characterized in that, is 10 of 9:1 by weight ratio, and 12-diine neocerotic acid and formula I compound polymerization form.
7. a method for preparing the described vesica probe of claim 6 is characterized in that, may further comprise the steps:
With 10,12-diine neocerotic acid and formula I compound are dissolved in dimethyl sulfoxide (DMSO) by weight 9:1 and obtain DMSO solution, DMSO solution is heated at least 80 ℃ and add in the 4-hydroxyethyl piperazine ethanesulfonic acid damping fluid and obtain reaction solution, the cooling reaction solution, carry out polyreaction with the UV-irradiation reaction solution, namely get the vesica probe.
8. according to the described method of claim 7, it is characterized in that, before the cooling reaction solution, that reaction solution is ultrasonic and filter with filter.
9. described method according to Claim 8 is characterized in that, adopts the 254nm ultraviolet lamp as ultraviolet source.
10. the described vesica probe of claim 6 is for detection of the purposes of Xin Meisu.
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