CN104592983A

CN104592983A - Fluorescent probes based on pyrene, and preparation method and application thereof

Info

Publication number: CN104592983A
Application number: CN201410725553.3A
Authority: CN
Inventors: 钟克利; 曲秀莉; 汤立军; 励建荣; 侯淑华; 徐永霞; 边延江; 周雪; 张悦
Original assignee: Bohai University
Current assignee: Bohai University
Priority date: 2014-11-28
Filing date: 2014-11-28
Publication date: 2015-05-06
Anticipated expiration: 2034-11-28
Also published as: CN104592983B

Abstract

The invention discloses fluorescent probes based on pyrene, and a preparation method and application thereof. The concrete preparation method comprises taking a mixture of THF and H2O with the ratio of 4:1 as a solvent, controlling the molar ratio of raw materials 1,8-diethynylpyrene to 2-azido-N-(quinolin-8-yl)acetamide to be 1:2-1:3, and stirring at 50-80 DEG C for a night; adding ammonia water for extraction, performing reduced-pressure distillation to remove the solvent, purifying the obtained solid by using silica gel chromatography, and using dichloromethane as an eluant for processing, so as to obtain a light yellow solid 1, and employing a mixture of ethyl acetate and dichloromethane according to the volume ratio of 1:15 as an eluant for processing, so as to obtain a yellow solid compound 2. Two fluorescent probes are obtained through the one-step reaction, tailed detection on copper ion and pyrophosphate ion in water environment can be realized, and the fluorescent probes have relatively high sensitivity and interference resistance.

Description

A kind of fluorescent probe based on pyrene and preparation method thereof and application

Technical field

The invention belongs to technical field of chemistry, relate to a kind of fluorescent probe based on pyrene and preparation method thereof and application, specifically, relate to a kind of single step reaction and synthesize two kinds of methods based on the fluorescent probe of pyrene and application.

Background technology

Copper is institute's necessarily a kind of trace heavy metal element and nutrient substance in organism, and the content in cell is only second to zinc and iron, plays an important role in various organic basic physiological process.It participates in the katalysis of more than 20 kind of enzyme (as Terminal oxidase, superoxide dismutase, tyrosine oxidase, dopamine β-hydroxylase, lysyloxidase etc.) as metalloenzyme.Cupric ion content is in vivo little, but shortage can cause metabolism and growth disorderly, and content is crossed and can be produced huge toxic action to organism at most.Cupric ion metabolic balance in body is damaged and can causes the generation of nerve degenerative diseases, such as the remote gram disease such as this syndromes, Wilson syndrome.Therefore, the detection of cupric ion has great importance in biological study and medical diagnosis.

At present, about the fluorescent probe based on pyrene in the document reported, mainly based on the derivative of monosubstituted pyrene, and the fluorescent probe of the derivative of two replacement pyrene is seldom reported; And synthesize two kinds of methods based on the fluorescent probe of pyrene by single step reaction also seldom to report; The fluorescent probe being connected quinoline and pyrene derivatives by triazole is not reported yet; Do not report based on the fluorescent probe relay identification cupric ion of pyrene and the fluorescent probe of pyrophosphate ion yet.Therefore, design and synthesis single step reaction obtain two kinds of derivatives replacing pyrenes and to connect the fluorescent probe of quinoline by triazole very important.

Summary of the invention

The object of the invention is to the defect overcoming existence of the prior art, a kind of fluorescent probe based on pyrene and preparation method thereof and application are provided.

Its concrete technical scheme is:

Based on a fluorescent probe for pyrene, structural formula is as follows:

A preparation method for fluorescent probe based on pyrene of the present invention, its reaction formula is:

Comprise the following steps:

With THF: H ₂o=4: 1 is solvent, raw material 1, the mol ratio of 8-diacetylene pyrene and 2-azido--N-(quinoline-8-yl) ethanamide is 1: 2 ~ 1: 3, stir at 50 ~ 80 DEG C and spend the night, add ammoniacal liquor extraction, decompression steams solvent, the solid silica gel chromatography obtained carries out purifying, with methylene dichloride as eluent, obtain light yellow solid Compound 1, by ethyl acetate: methylene chloride volume than 1: 15 for eluent obtains yellow solid compound 2.

Preferably, described 1,8-diacetylene pyrene and 2-azido--N-(quinoline-8-yl) ethanamide mol ratio are 1: 3.

Fluorescent probe based on pyrene of the present invention detects the cupric ion in water surrounding and the application in pyrophosphate ion process in relay.

Compared with prior art, beneficial effect of the present invention is:

(1) by single step reaction, two kinds of fluorescent probes can be obtained.

(2) fluorescent probe itself comprises pyrene this to have fluorescence lifetime long, and quantum yield is high, stablizes light, to the fluorophore of the features such as micro-environmental variation is quick on the draw.

(3) fluorescent probe identification Cu ²⁺after, with the title complex of its formation, also can continue to monitor P ₂o ₇ ^4-.

(4) fluorescent probe can detect the Cu in water surrounding ²⁺and P ₂o ₇ ^4-.

(5) fluorescent probe has good sensitivity and stronger immunity from interference.

Accompanying drawing explanation

Fig. 1 is fluorescent probe 1 of the present invention ¹h NMR spectrogram;

Fig. 2 is fluorescent probe 1 of the present invention ¹³c NMR spectrogram;

Fig. 3 is the mass spectrogram of fluorescent probe 1 of the present invention;

Fig. 4 is fluorescent probe 2 of the present invention ¹h NMR spectrogram;

Fig. 5 is fluorescent probe 2 of the present invention ¹³c NMR spectrogram;

Fig. 6 is the mass spectrogram of fluorescent probe 2 of the present invention;

Fig. 7 is the fluorescence emission spectrogram of fluorescent probe 1 of the present invention;

Fig. 8 is fluorescent probe 1 of the present invention and the fluorescence emission spectrogram adding cupric ion;

Fig. 9 is fluorescent probe 1 of the present invention and Ni ²⁺, Hg ²⁺, Ca ²⁺, Ba ²⁺, Mg ²⁺, K ⁺, Al ³⁺, Mn ²⁺, Pb ²⁺, Na ⁺, Sr ²⁺, Co ²⁺, Cr ³⁺, Ag ⁺, Fe ²⁺, Fe ³⁺, Cu ²⁺, Zn ²⁺, Cd ²⁺fluorescence emission spectrogram before and after effect;

Figure 10 is fluorescent probe of the present invention 1 couple of Cu ²⁺the fluoroscopic examination figure of other metal ion disturbances is resisted during identification;

Figure 11 is the Cu of fluorescent probe 1 of the present invention and 0 μm of ol/L ~ 100 μm ol/L ²⁺fluorescence emission spectrum variation diagram before and after effect;

Figure 12 is fluorescent probe 1-Cu of the present invention ²⁺with the fluorescence emission spectrogram before and after other negatively charged ion effects;

Figure 13 is fluorescent probe 1-Cu of the present invention ²⁺with the P of 0 μm of ol/L ~ 150 μm ol/L ₂o ₇ ^4-fluorescence emission spectrum variation diagram before and after effect;

Figure 14 is the fluorescence emission spectrogram of fluorescent probe 2 of the present invention;

Figure 15 is fluorescent probe 2 of the present invention and the fluorescence emission spectrogram adding cupric ion;

Figure 16 is fluorescent probe 2 of the present invention and Ni ²⁺, Hg ²⁺, Ca ²⁺, Ba ²⁺, Mg ²⁺, K ⁺, Al ³⁺, Mn ²⁺, Pb ²⁺, Na ⁺, Sr ²⁺, Co ²⁺, Cr ³⁺, Ag ⁺, Fe ²⁺, Fe ³⁺, Cu ²⁺, Zn ²⁺, Cd ²⁺fluorescence emission spectrogram before and after effect;

Figure 17 is fluorescent probe of the present invention 2 couples of Cu ²⁺the fluoroscopic examination figure of other metal ion disturbances is resisted during identification;

Figure 18 is the Cu of fluorescent probe 2 of the present invention and 0 μm of ol/L ~ 20 μm ol/L ²⁺fluorescence emission spectrum variation diagram before and after effect;

Figure 19 is fluorescent probe 2-Cu of the present invention ²⁺with the fluorescence emission spectrogram before and after other negatively charged ion effects;

Figure 20 is fluorescent probe 2-Cu of the present invention ²⁺with the P of 0 μm of ol/L ~ 40 μm ol/L ₂o ₇ ^4-fluorescence emission spectrum variation diagram before and after effect.

Embodiment

Below in conjunction with specific embodiment, technical scheme of the present invention is described in more detail.

Embodiment 1

(1) single step reaction synthesizes two kinds of reaction formula based on the fluorescent probe of pyrene:

(2) single step reaction synthesizes two kinds of concrete steps based on the fluorescent probe of pyrene:

Take 1, the 8-diacetylene pyrene of 100mg and 2-azido--N-(quinoline-8-yl) ethanamide of 273mg, be dissolved in 10ml THF: H ₂in O=4: 1, stir at 60 DEG C and spend the night.Add ammoniacal liquor extraction, decompression steams solvent, the solid silica gel chromatography obtained carries out purifying, with absolute dichloromethane as eluent, obtain 73mg light yellow solid Compound 1, productive rate 38%, adopts ethyl acetate: methylene dichloride=1: 15 (volume ratios) obtain 43mg yellow solid compound 2, productive rate 15% for eluent.

Embodiment 2

Take 1, the 8-diacetylene pyrene of 200mg and 2-azido--N-(quinoline-8-yl) ethanamide of 545mg, be dissolved in 10ml THF: H ₂in O=4: 1, stir at 65 DEG C and spend the night.Add ammoniacal liquor extraction, decompression steams solvent, the solid silica gel chromatography obtained carries out purifying, with absolute dichloromethane as eluent, obtain 156mg light yellow solid Compound 1, productive rate 41%, adopts ethyl acetate: methylene dichloride=1: 16 (volume ratios) obtain 101mg yellow solid compound 2, productive rate 18% for eluent.

Embodiment 3

Take 1, the 8-diacetylene pyrene of 250mg and 2-azido--N-(quinoline-8-yl) ethanamide of 680mg, be dissolved in 10ml THF: H ₂in O=4: 1, stir at 66 DEG C and spend the night.Add ammoniacal liquor extraction, decompression steams solvent, the solid silica gel chromatography obtained carries out purifying, with absolute dichloromethane as eluent, obtain 215mg light yellow solid Compound 1, productive rate 45%, adopts ethyl acetate: methylene dichloride=1: 13 (volume ratios) obtain 155mg yellow solid compound 2, productive rate 22% for eluent.

The master data of the fluorescent probe 1,2 of embodiment 1 ~ embodiment 3:

Fluorescent probe 1:

¹h NMR (400MHz, DMSO-d ₆) δ 10.822 (s, 1H), 9.001-8.956 (dd, J=14.1,6.5Hz, 2H), 8.875 (s, 1H), 8.615 (d, J=7.6Hz, 1H), 8.533 (d, J=9.5Hz, 1H), 8.451-8.38 (m, 3H), 8.314-8.197 (m, 4H), 7.715 (d, J=8.4Hz, 1H), (7.669-7.637 m, 1H), 7.588 (t, J=8.0Hz, 1H), 5.830 (s, 2H), 4.794 (s, 1H). (as Fig. 1)

¹³c NMR (101MHz, DMSO-d ₆) δ 165.50,149.53,146.25,138.76,137.13,134.50,131.83,131.69,131.11,130.69,128.94,128.38,128.00,127.70,127.40,126.94,126.79,126.66,126.58,125.77,125.63,124.31,124.11,123.17,122.74,117.80,116.63,109.99,87.29,82.54,53.35. (as Fig. 2)

Fluorescent probe 2:

¹h NMR (400MHz, DMSO-d ₆) δ 10.81-10.77 (m, 2H), 8.96-8.93 (m, 2H), 8.97-8.93 (m, 2H), 8.92-8.86 (m, 4H), 8.92-8.85 (m, 4H), 8.62-8.59 (m, 2H), 8.46-8.35 (m, 7H), 8.44-8.37 (m, 6H), 8.28-8.24 (m, 2H), 8.27-8.25 (m, 2H), 7.72-7.67 (m, 3H), 7.71-7.68 (m, 2H), 7.66-7.62 (m, 2H), 7.67-7.60 (m, 3H), 7.61-7.54 (m, 3H), 7.59-7.54 (m, 2H), 5.84-5.79 (m, 4H). (as Fig. 4)

¹³c NMR (101MHz, DMSO-d ₆) δ 165.52 (s), 149.52 (s), 146.28 (s), 138.73 (s), 137.11 (s), 134.47 (s), 131.35 (s), 128.28 (d, J=16.2Hz), 127.67 (s), 127.38 (s), 126.84 (s), 126.19 (s), 125.84 (s), 124.91 (s), 123.15 (s), 122.73 (s), 117.78 (s), 53.34 (s). (as Fig. 5)

High resolution mass spectrum (electron spray(ES), holotype): the calculated value C of fluorescent probe 1 ₃₁h ₁₉n ₅o [M+H] ⁺, 478.1590; Measured value 478.1658. (as Fig. 3); The calculated value C of fluorescent probe 2 ₄₂h ₂₈n ₁₀o ₂[M+H] ⁺, 705.2397; Measured value 705.2462. (as Fig. 6).

Fluorescent probe 1 couple of Cu ²⁺optionally detect:

Acetonitrile/the Tris of the fluorescent probe 1 of 10 μm of ol/L cushions (V/V=1: 1, pH=7.4) solution, and the fluorescence emission spectrum of this fluorescent probe as shown in Figure 7, adds the metal ion (Ni of 10 times of equivalents respectively ²⁺, Hg ²⁺, Ba ²⁺, Mg ²⁺, K ⁺, Al ³⁺, Mn ²⁺, Pb ²⁺, Na ⁺, Sr ²⁺, Co ²⁺, Cr ³⁺, Ag ⁺, Fe ²⁺, Fe ³⁺, Cu ²⁺, Zn ²⁺, Cd ²⁺), detect the fluorescence emission spectrum change of solution, as Fig. 8, shown in 9.From Fig. 8, can find out in 9, when adding Cu ²⁺time, the emission peak at 410nm and 430nm place reduces, and when adding other ions, there is no obvious phenomenon, and fluorescent probe 1 couple of Cu is described ²⁺there is good selectivity.

The anti-interference detection of fluorescent probe 1:

Acetonitrile/the Ttis of the fluorescent probe 1 of 10 μm of ol/L cushions (V/V=1: 1, pH=7.4) solution, adds the metal ion (Ni of 10 times of equivalents respectively ²⁺, Hg ²⁺, Ba ²⁺, Mg ²⁺, K ⁺, Al ³⁺, Mn ²⁺, Pb ²⁺, Na ⁺, Sr ²⁺, Co ²⁺, Cr ³⁺, Ag ⁺, Fe ²⁺, Fe ³⁺, Cu ²⁺, Zn ²⁺, Cd ²⁺), and then add the Cu of 10 times of equivalents respectively ²⁺, detect the fluorescence emission spectrum of solution, get the value mapping corresponding to maximum emission wavelength, as shown in Figure 10.As shown in Figure 10, when there being other metal ions to exist, fluorescent probe is substantially only to Cu ²⁺there is combination, to Cu when illustrating that other metal ions exist ²⁺detection do not disturb, only have and work as Ni ²⁺when existing, there is weak interference.

Fluorescent probe 1 couple of Cu ²⁺titration experiments:

Acetonitrile/the Ttis of the fluorescent probe 1 of 10 μm of ol/L cushions (V/V=1: 1, pH=7.4) solution, adds the Cu of 0 μm of ol/L ~ 100 μm ol/L respectively ²⁺, detect the fluorescence emission spectrum change of solution as shown in figure 11.As can be seen from Figure 11, along with Cu ²⁺constantly add, reduce gradually at the emission peak at 410nm place, when joining the Cu of 100 μm of ol/L ²⁺time, no longer reduce at the emission peak at 410nm and 430nm place, illustrate reach saturated.

Fluorescent probe 1-Cu ²⁺to P ₂o ₇ ^4-optionally detect:

The fluorescent probe 1-Cu of 10 μm of ol/L ²⁺acetonitrile/Ttis cushion (V/V=1: 1, pH=7.4) solution, add (F after the negatively charged ion of 15 times amount ^-, Cl ^-, Br ^-, I ^-, SCN ^-, PO ₄ ^3-, H ₂pO ₄ ^-, HPO ₄ ^2-, NO ₂ ^-, NO ₃ ^-, ClO ₄ ^-, SO ₄ ^2-, P ₂o ₇ ^4-, HSO ₄ ^-, CO ₃ ^2-, HCO ₃ ^-), detect the fluorescence emission spectrum change of solution as shown in figure 12.As can be seen from Figure 12, only P is added ₂o ₇ ^4-,time, fluorescence just can be made to be returned to the state (or seeing Fig. 8) of acceptor self, to illustrate that this system is to P ₂o ₇ ^4-selectivity and specificity better.

Fluorescent probe 1-Cu ²⁺to P ₂o ₇ ^4-titration experiments:

The fluorescent probe 1-Cu of 10 μm of ol/L ²⁺acetonitrile/Ttis cushion (V/V=1: 1, pH=7.4) solution, add the P of 0 μm of ol/L ~ 150 μm ol/L respectively ₂o ₇ ^4-, detect the fluorescence emission spectrum change of solution as shown in figure 13.As can be seen from Figure 13, along with P ₂o ₇ ^4-constantly add, raise gradually at the emission peak at 410nm and 430nm place, when joining the P of 150 μm of ol/L ₂o ₇ ^4-time, no longer change in the emission peak intensity at 410nm and 430nm place, illustrate reach saturated.

Fluorescent probe 2 couples of Cu ²⁺optionally detect:

The THF/HEPES of the fluorescent probe 2 of 10 μm of ol/L cushions (V/V=1: 1, pH=7.4) solution, and the fluorescence emission spectrum of this fluorescent probe as shown in figure 14, adds the metal ion (Ni of 2 times of equivalents respectively ²⁺, Hg ²⁺, Ba ²⁺, Mg ²⁺, K ⁺, Al ³⁺, Mn ²⁺, Pb ²⁺, Na ⁺, Sr ²⁺, Co ²⁺, Cr ³⁺, Ag ⁺, Fe ²⁺, Fe ³⁺, Cu ²⁺, Zn ²⁺, Cd ²⁺), detect the fluorescence emission spectrum change of solution, as Figure 15, shown in 16.From Figure 15, can find out in 16, when adding Cu ²⁺time, the emission peak at 410nm and 430nm place reduces, and has basket to move, and the peak basket at 410nm place moves to 400nm place, and the peak basket at 430nm place moves to 420nm, and when adding other ions, there is no obvious phenomenon, and fluorescent probe 2 couples of Cu are described ²⁺there is good selectivity.

The anti-interference detection of fluorescent probe 2:

The THF/HEPES of the fluorescent probe 2 of 10 μm of ol/L cushions (V/V=1: 1, pH=7.4) solution, adds the metal ion (Ni of 2 times of equivalents respectively ²⁺, Hg ²⁺, Ba ²⁺, Mg ²⁺, K ⁺, Al ³⁺, Mn ²⁺, Pb ²⁺, Na ⁺, Sr ²⁺, Co ²⁺, Cr ³⁺, Ag ⁺, Fe ²⁺, Fe ³⁺, Cu ²⁺, Zn ²⁺, Cd ²⁺), and then add the Cu of 2 times of equivalents respectively ²⁺, detect the fluorescence emission spectrum of solution, get the value mapping corresponding to maximum emission wavelength, as shown in figure 17.As shown in Figure 17, when there being other metal ions to exist, fluorescent probe is substantially only to Cu ²⁺there is combination, to Cu when illustrating that other metal ions exist ²⁺detection do not disturb, only have and work as Ni ²⁺when existing, there is weak interference.

Fluorescent probe 2 couples of Cu ²⁺titration experiments:

The THF/HEPES of the fluorescent probe 2 of 10 μm of ol/L cushions (V/V=1: 1, pH=7.4) solution, adds the Cu of 0 μm of ol/L ~ 20 μm ol/L respectively ²⁺, detect the fluorescence emission spectrum change of solution as shown in figure 18.As can be seen from Figure 18, along with Cu ²⁺constantly add, reduce gradually at the emission peak at 410nm and 430nm place, and have basket to move, when joining the Cu of 20 μm of ol/L ²⁺time, the emission peak at 410nm and 430nm place no longer reduces and moves with basket, illustrate reach saturated.

Fluorescent probe 2-Cu ²⁺to P ₂o ₇ ^4-optionally detect:

The fluorescent probe 2-Cu of 10 μm of ol/L ²⁺tHF/HEPES cushion (V/V=1: 1, pH=7.4) solution, add (F after the negatively charged ion of 4 times amount ^-, Cl ^-, Br ^-, I ^-, SCN ^-, PO ₄ ^3-, H ₂pO ₄ ^-, HPO ₄ ^2-, NO ₂ ^-, NO ₃ ^-, ClO ₄ ^-, SO ₄ ^2-, P ₂o ₇ ^4-, HSO ₄ ^-, CO ₃ ^2-, HCO ₃ ^-), detect the fluorescence emission spectrum change of solution as shown in figure 19.As can be seen from Figure 19, only P is added ₂o ₇ ^4-time, fluorescence just can be made to be returned to the state (or seeing Figure 15) of acceptor self, to illustrate that this system is to P ₂o ₇ ^4-selectivity and specificity better.

Fluorescent probe 2-Cu ²⁺to P ₂o ₇ ^4-titration experiments:

The fluorescent probe 2-Cu of 10 μm of ol/L ²⁺tHF/HEPES cushion (V/V=1: 1, pH=7.4) solution, add the P of 0 μm of ol/L ~ 40 μm ol/L respectively ₂o ₇ ^4-, detect the fluorescence emission spectrum change of solution as shown in figure 20.As can be seen from Figure 20, along with P ₂o ₇ ^4-constantly add, raise gradually at the emission peak at 410nm and 430nm place, when joining the P of 40 μm of ol/L ₂o ₇ ^4-time, no longer change in the emission peak intensity at 410nm and 430nm place, illustrate reach saturated.

The above; be only the present invention's preferably embodiment; protection scope of the present invention is not limited thereto; anyly be familiar with those skilled in the art in the technical scope that the present invention discloses, the simple change of the technical scheme that can obtain apparently or equivalence are replaced and are all fallen within the scope of protection of the present invention.

Claims

1. based on a fluorescent probe for pyrene, it is characterized in that, structural formula is as follows:

2. described in claim 1 based on a preparation method for the fluorescent probe of pyrene, it is characterized in that, its reaction formula is:

Comprise the following steps:

3. according to claim 2 based on the preparation method of the fluorescent probe of pyrene, it is characterized in that, described 1,8-diacetylene pyrene and 2-azido--N-(quinoline-8-yl) ethanamide mol ratio are 1: 3.

4. detect the cupric ion in water surrounding and the application in pyrophosphate ion process based on the fluorescent probe of pyrene in relay described in claim 1.