A kind of production of fluorescent sensor and application with sulfonium ion, cupric ion and pH value Multiple detection function
Technical field
The present invention relates to the technical fields such as analytical chemistry, life science and environmental science, specifically, relate to fluorescent optical sensor with sulfonium ion, cupric ion and pH value Multiple detection function and preparation method thereof.
Background technology
In the negatively charged ion existed in numerous environmental and biological materials, sulfonium ion (S
2-) be a class to organism, the particularly virulent material of human body.Research finds, the S of high density
2-human body will be caused to occur various malaise symptoms, comprise stimulating mucosal, unconscious and respiratory standstill.And more seriously, in acid condition, S
2-hS is become by protonated further
-, thus become more toxic and corrodibility.Recent research shows, the HS after protonated
-the various physiological processes of human body can be affected, comprise central nervous system, respiratory system, Digestive tract and endocrine system; And have dependency to a certain extent with symptoms such as liver cirrhosis, diabetes, Down's syndromes.
Copper is the required trace element of human life activity, is distributed widely in biological tissue.In biosystem many relate to oxygen electron transmission and redox reaction be all enzymatic by cupric, these enzymes are all vital to vital process; Meanwhile, in body, the content of copper all has a certain impact for immunologic function, lipid and carbohydrate metabolism and hormone secretion etc., when in body during copper too high levels, can cause Upper abdominal pain, Nausea and vomiting, even suffer a shock, go into a coma or dead.Therefore, to the direct rapid detection of cupric ion be all the focus of scientific research all the time.
On the other hand, intracellular pH value plays considerable effect to whole Living system, and it all plays vital effect, such as ion transportation and cell proliferation etc. in cell, enzyme and tissue activity etc.The subtle change of intracellular ph value just can cause cell dysfunction, adverse consequences is caused to organism, itself and some common diseases also have very large dependency, as cancer, senile dementia etc., therefore, accurately measure intracellular pH value rapidly and can provide very important information for whole Living system.
At present, the method for the detection sulfonium ion, cupric ion and the pH value that have grown up is a lot, as electrochemical method, and atomic absorption spectrometry, atomic emission spectrometry etc.But most detection method cost drops into higher, and testing process is too complicated, these factors all seriously constrain the utilization of some method in reality detects.Therefore, a kind of simple, low cost of invention and efficiently detection technique there is considerable realistic meaning and application prospect.In recent years, fluorescent optical sensor, because of the optical property of its excellence, receives increasing concern, shows extremely wide application prospect in research fields such as chemistry, medical science and environmental sciences.For other detection means, fluorescent optical sensor has the remarkable characteristics such as do not destroy sample, designability is strong, highly sensitive, selectivity is good, and becomes the focus of multiple fields intermediate ion detection and indentification research.
Summary of the invention
The object of the present invention is to provide a kind of production of fluorescent sensor and the application with sulfonium ion, cupric ion and pH value Multiple detection function, a kind of water soluble fluorescence sensor with sulfonium ion, cupric ion and pH value Multiple detection function that this fluorescent optical sensor is is Material synthesis with rhodamine B, quadrol and fluorescein isothiocyanate.Further applied research shows, this fluorescent optical sensor can realize, to sulfonium ion and the highly sensitive of cupric ion, the rapid detection of highly selective, accurately can also detecting pH value.
The object of the invention is to be realized by following manner:
There is a production of fluorescent sensor for sulfonium ion, cupric ion and pH value Multiple detection function, comprise the following steps:
(1) a certain amount of rhodamine B is dissolved in anhydrous methanol, is placed in N
2under the condition of protection and lucifuge, after 20 ~ 50min, quadrol is added fast when being warming up to 40 DEG C, continue to be warming up to 85 DEG C of stirring and refluxing 20 ~ 40 hours, reaction terminates rear filtration, then removes major part (80 ~ 95%) anhydrous methanol, add a certain amount of distilled water precipitation again, suction filtration obtains required product, vacuum-drying, obtains quadrol lactamize rhodamine B;
(2) rhodamine B and the fluorescein isothiocyanate of getting the quadrol lactamize that step (1) is synthesized are dissolved in the DMF dewatered, and mixing solutions are placed in lucifuge and N
2stirring at room temperature 20 ~ 40h under the condition of protection; reaction terminates rear removing major part (85 ~ 95%) N; dinethylformamide; cross post purified product; vacuum-drying; obtain required fluorescent optical sensor, i.e. a kind of fluorescent optical sensor with sulfonium ion, cupric ion and pH value Multiple detection function, its structural formula is as follows:
The above-mentioned production of fluorescent sensor with sulfonium ion, cupric ion and pH value Multiple detection function, in described step (1), the mol ratio of rhodamine B and quadrol is 1:10 ~ 30; The concentration of rhodamine B in anhydrous methanol is 0.01 ~ 0.04mol/L; The concentration of quadrol in anhydrous methanol is 0.30 ~ 0.60mol/L.
The above-mentioned production of fluorescent sensor with sulfonium ion, cupric ion and pH value Multiple detection function, in described step (2), the rhodamine B of quadrol lactamize and the mol ratio of fluorescein isothiocyanate are 1:1.0 ~ 1.6; Quadrol lactamize rhodamine is 0.01 ~ 0.04mol/L in the concentration of DMF; The concentration of fluorescein isothiocyanate in DMF is 0.01 ~ 0.06mol/L.
The above-mentioned production of fluorescent sensor with sulfonium ion, cupric ion and pH value Multiple detection function, the removing method of described anhydrous methanol and DMF is preferably rotary evaporation.
According to fluorescent optical sensor prepared by above-mentioned preparation method, its concrete reaction process is as follows:
A kind of fluorescent optical sensor prepared according to above-mentioned preparation method.
According to the application of fluorescent optical sensor in sulfonium ion, cupric ion and pH value Multiple detection prepared by above-mentioned preparation method.
The present invention adopts and quadrol lactamize rhodamine B and fluorescein isothiocyanate reaction is prepared required fluorescent optical sensor, the aqueous solution of this fluorescent optical sensor, can along with the increase appearance Enhancement of Fluorescence gradually of sulfonium ion concentration when there being sulfonium ion to exist; And when there being cupric ion to exist, the fluorescence intensity of this fluorescent optical sensor can decline gradually along with the increase of copper ion concentration, and the detection of this fluorescent optical sensor to these two kinds of ions has obvious highly selective; Meanwhile, this fluorescent optical sensor also has responsiveness well to pH value, can reach the effect of high-sensitivity detection.Compared to more existing detection techniques, less input for fluorescence chemical sensor cost in the present invention, synthetic route is simple, convenient post-treatment, directly can realize specific recognition to the cupric ion in water and sulfonium ion, carries out Accurate Measurement, be applicable to amplifying production and practical application to pH value.
Accompanying drawing explanation
Fig. 1 is the mass spectrum of fluorescent optical sensor of the present invention.
Fig. 2 is fluorescent optical sensor of the present invention to the identification schematic diagram of cupric ion and sulfonium ion.
When Fig. 3 is different sulfonium ion concentration, the fluorescence emission spectrum variation diagram (λ of fluorescent optical sensor prepared by the embodiment of the present invention 1
ex=490nm), [S
2-]=0 (a), 1.0 × 10
-6mol/l (b), 5.0 × 10
-6mol/l (c), 1.0 × 10
-5mol/l (d), 1.5 × 10
-5mol/l (e), 2.0 × 10
-5mol/l (f), 2.5 × 10
-5mol/l (g), 3.0 × 10
-5mol/l (h), 3.5 × 10
-5mol/l (i), 4.0 × 10
-5mol/l (j), 4.5 × 10
-5mol/l (k), 5.0 × 10
-5mol/l (l), 5.5 × 10
-5mol/l (m), 6.0 × 10
-5mol/l (n), 6.5 × 10
-5mol/l (o), 7.0 × 10
-5mol/l (p), (when measuring the fluorescence intensity of sample in Fig. 3, slit used is wide is 3+5+High).
Fig. 4 is the fluorescent optical sensor prepared of the embodiment of the present invention 1 with matched curve corresponding to the fluorescence intensity change value of sulfonium ion change in concentration and functional arrangement (F corresponding to this Curves
blankwith F be sulfonium ion add before and after fluorescent optical sensor taking 490nm as excitation wavelength, 530nm is the fluorescence intensity change value of transmitted wave strong point).
Fig. 5 is the selectivity correlation data figure of various negatively charged ion to fluorescent optical sensor fluorescence intensity prepared by the embodiment of the present invention 1, and the concentration of the negatively charged ion after adding is 5.0 × 10
-6mol/l, F
blankwith F be each negatively charged ion add before and after fluorescent optical sensor taking 490nm as excitation wavelength, 530nm is the fluorescence intensity change value of transmitted wave strong point.
When Fig. 6 is different Cu ionic concn, the fluorescence emission spectrum variation diagram (λ of fluorescent optical sensor prepared by the embodiment of the present invention 1
ex=490nm), [Cu
2+]=0 (a), 1.0 × 10
-6mol/l (b), 2.0 × 10
-6mol/l (c), 4.0 × 10
-6mol/l (d), 8.0 × 10
-6mol/l (e), 1.5 × 10
-5mol/l (f), 2.5 × 10
-5mol/l (g), 4.0 × 10
-5mol/l (h), 5.0 × 10
-5mol/l (i) (when measuring the fluorescence intensity of sample in Fig. 6, slit used is wide is 5+10+Low).
Fig. 7 is matched curve corresponding to fluorescence intensity change value that fluorescent optical sensor prepared by the embodiment of the present invention 1 change with the copper ion concentration functional arrangement corresponding with this Curves, F
blankwith F be cupric ion add before and after fluorescent optical sensor taking 490nm as excitation wavelength, 530nm is the fluorescence intensity change value of transmitted wave strong point).
Fig. 8 is the selectivity correlation data figure of various positively charged ion to the fluorescence intensity of fluorescent optical sensor prepared by the embodiment of the present invention 1, and the various cationic concentration after adding is 5.0 × 10
-6mol/l, F
blankwith F be each positively charged ion add before and after fluorescent optical sensor taking 490nm as excitation wavelength, 530nm is the fluorescence intensity change value of transmitted wave strong point.
Fig. 9 is the fluorescence emission spectrogram that the fluorescent optical sensor of the embodiment of the present invention 1 preparation changes with pH value, and having surveyed pH value in experimentation is respectively 4,4.5,5,5.5,6,6.5,7,7.5,8,8.5, the fluorescence intensity change value (when measuring the fluorescence intensity of sample in Fig. 9, slit used is wide is 5+5+Low) of 9.
Figure 10 is matched curve corresponding to fluorescence intensity change value that fluorescent optical sensor prepared by the embodiment of the present invention 1 change with the pH value functional arrangement (F corresponding with this Curves
pH=4with F to be pH value be respectively 4 and pH value be 4.5,5,5.5,6,6.5,7, the fluorescence intensity change value of 7.5,8,8.5,9,490nm is excitation wavelength, and 530nm is emission wavelength).
Embodiment
Below in conjunction with drawings and the specific embodiments, the present invention is described in further detail.
Embodiment 1
There is a production of fluorescent sensor for sulfonium ion, cupric ion and pH value Multiple detection function, comprise the following steps:
(1) quadrol lactamize rhodamine B.
Rhodamine B (3mmol) and quadrol (90mmol) are dissolved in anhydrous methanol (250ml), are placed in N
2under the condition of protection and lucifuge; be warming up to 85 DEG C of stirring and refluxing 24h; reaction terminates rear filtration; rotary evaporation removes most of anhydrous methanol again; be about 10ml to volume, then use suitable quantity of water (200ml) to precipitate, suction filtration obtains required product; vacuum-drying, with synthesis of ethylenediamine lactamize rhodamine.
(2) quadrol lactamize rhodamine B and fluorescein isothiocyanate synthesis fluorescent optical sensor is utilized.
The rhodamine B (0.25mmol) and fluorescein isothiocyanate (0.25mmol) of getting the quadrol lactamize that step (1) is synthesized are dissolved in the N dewatered, dinethylformamide (10ml), is placed in lucifuge and N by mixing solutions
2stirring at room temperature 24h under the condition of protection, reaction terminates rear rotary evaporation and removes most of DMF; cross post purified product; vacuum-drying, to synthesize required fluorescent optical sensor, i.e. a kind of fluorescent optical sensor with sulfide, cupric ion and pH value Multiple recognition function.
Embodiment 2
There is a production of fluorescent sensor for sulfonium ion, cupric ion and pH value Multiple detection function, comprise the following steps:
(1) quadrol lactamize rhodamine B.
Rhodamine B (3mmol) and quadrol (45mmol) are dissolved in anhydrous methanol (100ml), are placed in N
2under the condition of protection and lucifuge; be warming up to 85 DEG C of stirring and refluxing 24h; reaction terminates rear filtration; rotary evaporation removes most of anhydrous methanol again; be about 15ml to volume, then use suitable quantity of water (250ml) to precipitate, suction filtration obtains required product; vacuum-drying, with synthesis of ethylenediamine lactamize rhodamine.
(2) quadrol lactamize rhodamine B and fluorescein isothiocyanate synthesis fluorescent optical sensor is utilized.
The rhodamine B (0.25mmol) and fluorescein isothiocyanate (0.30mmol) of getting the quadrol lactamize that step (1) is synthesized are dissolved in the N dewatered, dinethylformamide (20ml), is placed in lucifuge and N by mixing solutions
2stirring at room temperature 24h under the condition of protection, reaction terminates rear rotary evaporation and removes most of DMF; cross post purified product; vacuum-drying, to synthesize required fluorescent optical sensor, i.e. a kind of fluorescent optical sensor with sulfide, cupric ion and pH value Multiple recognition function.
Embodiment 3
There is a production of fluorescent sensor for sulfonium ion, cupric ion and pH value Multiple detection function, comprise the following steps:
(1) quadrol lactamize rhodamine B.
Rhodamine B (3mmol) and quadrol (60mmol) are dissolved in anhydrous methanol (200ml), are placed in N
2under the condition of protection and lucifuge; be warming up to 85 DEG C of stirring and refluxing 24h; reaction terminates rear filtration; rotary evaporation removes most of anhydrous methanol again; be about 15ml to volume, then use suitable quantity of water (250ml) to precipitate, suction filtration obtains required product; vacuum-drying, with synthesis of ethylenediamine lactamize rhodamine.
(2) quadrol lactamize rhodamine B and fluorescein isothiocyanate synthesis fluorescent optical sensor is utilized.
The rhodamine B (0.25mmol) and fluorescein isothiocyanate (0.40mmol) of getting the quadrol lactamize that step (1) is synthesized are dissolved in the N dewatered, dinethylformamide (15ml), is placed in lucifuge and N by mixing solutions
2stirring at room temperature 24h under the condition of protection, reaction terminates rear rotary evaporation and removes most of DMF; cross post purified product; vacuum-drying, to synthesize required fluorescent optical sensor, i.e. a kind of fluorescent optical sensor with sulfide, cupric ion and pH value Multiple recognition function.
Embodiment 4
The test experience of sulfonium ion.
(1) getting 16 5ml sample bottles, add the fluorescent optical sensor solution 3ml of gained in embodiment 1 respectively, is then [S by concentration respectively
2-]=0 (a), 1.0 × 10
-6mol/l (b), 5.0 × 10
-6mol/l (c), 1.0 × 10
-5mol/l (d), 1.5 × 10
-5mol/l (e), 2.0 × 10
-5mol/l (f), 2.5 × 10
-5mol/l (g), 3.0 × 10
-5mol/l (h), 3.5 × 10
-5mol/l (i), 4.0 × 10
-5mol/l (j), 4.5 × 10
-5mol/l (k), 5.0 × 10
-5mol/l (l), 5.5 × 10
-5mol/l (m), 6.0 × 10
-5mol/l (n), 6.5 × 10
-5mol/l (o), 7.0 × 10
-5the 3 μ l sodium sulfide solutions of mol/l (p) add in 16 sample bottles, stir under normal temperature after 3 minutes, take 490nm as excitation wavelength, measure the fluorescence intensity of these samples respectively, obtain the fluorescence emission spectrum variation diagram of 16 samples, see Fig. 3 (when measuring the fluorescence intensity of sample in Fig. 3, slit used is wide is 3+5+High).
Measurement result shows: the fluorescence intensity of this fluorescent optical sensor progressively rises along with the increase gradually of sulfonium ion concentration, and during sulfonium ion concentration >1 μm of ol/L, the fluorescence intensity of fluorescent optical sensor obviously rises.The more satisfactory function curve diagram after corresponding matching functional arrangement (y=p1*x/ (p2+x), p1=17.37285, p2=96.10114, the R corresponding with this Curves can be made according to the fluorescence intensity change value of Fig. 3
2=0.9984), Fig. 4 is seen.
(2) getting 11 5ml sample bottles, be respectively charged into the fluorescent optical sensor solution 3ml of gained in embodiment 1, is then 5.0 × 10 by concentration respectively
-3the Cl of mol/l
-, NO
3 -, SO
4 2-, HCO
3 -, CO
3 2-, S
2-, Br
-, F
-, S
2o
3 2-, ClO
4 -and HPO
4 -respectively getting 3 μ l adds in 11 sample bottles, and then measure 11 samples respectively and excite at 490nm wavelength, the fluorescent emission intensity at 530nm wavelength emission place, the results are shown in Figure 5.Measurement result shows: except sulfonium ion, and the fluorescence intensity of other above-mentioned various negatively charged ion to prepared fluorescent optical sensor does not have a significant effect, and this illustrates that this fluorescent optical sensor has very strong specific recognition capability to sulfonium ion.
Embodiment 5
The test experience of cupric ion.
(1) getting 9 5ml sample bottles, add the fluorescent optical sensor solution 3ml of gained in embodiment 1 respectively, is then [Cu by concentration respectively
2+]=0 (a), 1.0 × 10
-6mol/l (b), 2.0 × 10
-6mol/l (c), 4.0 × 10
-6mol/l (d), 8.0 × 10
-6mol/l (e), 1.5 × 10
-5mol/l (f), 2.5 × 10
-5mol/l (g), 4.0 × 10
-5mol/l (h), 5.0 × 10
-5the 3 μ l copper-baths of mol/l (i) add in 9 sample bottles, stir under normal temperature after 3 minutes, take 490nm as excitation wavelength, measure these fluorescent high lights respectively, obtain the fluorescence emission spectrum variation diagram of 9 samples, see Fig. 6 (when measuring the fluorescence intensity of sample in Fig. 6, slit used is wide is 5+10+Low).Measurement result shows: the fluorescence intensity of this fluorescent optical sensor progressively declines along with the increase gradually of copper ion concentration, and during copper ion concentration >1 μm of ol/L, the fluorescence intensity of fluorescent optical sensor obviously declines.The more satisfactory function curve diagram after corresponding matching functional arrangement (y=a*x^b, a=0.10565, b=0.75094, the R corresponding with this Curves can be made according to the fluorescence intensity change value of Fig. 6
2=0.99882), the results are shown in Figure 7.
(2) getting 9 5ml sample bottles, be respectively charged into the fluorescent optical sensor solution 3ml of gained in embodiment 1, is then 5.0 × 10 by concentration respectively
-3the Mg of mol/l
2+, Ni
2+, Mn
2+, Ca
2+, Cu
2+, Pb
2+, Zn
2+, Co
2+, and Fe
2+respectively getting 3 μ l adds in other 9 sample bottles, and then measure 9 samples respectively and excite at 490nm wavelength, the fluorescent emission intensity at 530nm wavelength emission place, the results are shown in Figure 8.Measurement result shows: except cupric ion, and the fluorescence intensity of other above-mentioned various positively charged ion to prepared fluorescent optical sensor does not have a significant effect, and this illustrates that this fluorescent optical sensor also has very strong specific recognition capability to cupric ion.
Embodiment 6
The test experience of pH value.
Get 11 5ml sample bottles, be respectively charged into the fluorescent optical sensor solution 30 μ l of gained in embodiment 1, in 13 sample bottles, then add pH value is respectively 4,4.5,5,5.5,6,6.5,7,7.5,8,8.5, the buffered soln 3ml of 9, stirs after 3 minutes under normal temperature, measures the fluorescence intensity change value of each sample, the results are shown in Figure 9 (when measuring the fluorescence intensity of sample in Fig. 9, slit used is wide is 5+5+Low).Measurement result shows: 490nm excites down, along with pH rises gradually from acidity to the fluorescence of alkaline fluorescent optical sensor.Functional arrangement (y=A2+ (A1-A2)/(1+exp ((the x-x0)/d)) corresponding with this Curves of the more satisfactory function curve diagram after corresponding matching can be made according to the fluorescence intensity change value of Fig. 9, A1=-1.83163, A2=83.75594, x0=6.90942, d=0.6307, R
2=0.99805), Figure 10 is seen.
Above-described embodiment is used for explaining and the present invention is described, instead of limits the invention, and in the protection domain of spirit of the present invention and claim, to any amendment done by the present invention and change, all falls into protection scope of the present invention.