CN103487416A - Fluorescent sensor for detecting zinc ions based on asymmetrical porphyrin fluorescence ratio - Google Patents

Abstract

The invention discloses a fluorescent sensor for detecting zinc ions based on the asymmetric porphyrin fluorescence ratio. PVC), diisooctyl sebacate and freshly distilled tetrahydrofuran (THF) were added dropwise on a hydrophobized quartz glass slide and dried in the air, and the fluorescence sensor was installed on a polymer On the top of the solution flow cell in the tetrafluoroethylene flow cell, the sample to be tested is input into the solution flow cell at a speed of 1.5ml/min, and the fluorescence intensity is measured at the maximum excitation wavelength and two emission wavelengths of the fluorescent carrier in water, and measured according to the calibration equation The content of zinc ions has a simple detection process, high sensitivity, accurate detection results, strong practicability, and strong promotion and application value.

Description

A Fluorescent Sensor for Zinc Ion Detection Based on Asymmetric Porphyrin Fluorescence Ratio

technical field

The invention belongs to the technical field of zinc ion detection, in particular to a fluorescence sensor for detecting zinc ions based on the asymmetric porphyrin fluorescence ratio.

Background technique

Zinc ions are important substances for maintaining normal growth and metabolism of the body. However, many experiments and epidemiological investigations have confirmed that if the content of zinc in the human body is too high, it will inhibit the activity and bactericidal power of phagocytes, thereby reducing the immune function of the human body and increasing the susceptibility to diseases. How much can detect certain diseases, such as nervous system, prostate and so on.

At present, the methods for determining zinc ions at home and abroad mainly include ion chromatography, high-efficiency capillary electrophoresis, stripping voltammetry, oscillographic polarography, fluorescence analysis, and spectrophotometry. The detection process of these methods is complicated, time-consuming and laborious. The sensitivity is low and the detection result is not accurate.

Contents of the invention

The present invention provides a fluorescent sensor for detecting zinc ions based on the asymmetric porphyrin fluorescence ratio, aiming to solve the problems of complex detection process, time-consuming and labor-intensive, low sensitivity and inaccurate detection results in the method for detecting zinc ions provided by the prior art .

The object of the present invention is to provide a fluorescent sensor for detecting zinc ions based on the asymmetric porphyrin fluorescence ratio. A mixed solution composed of ethylene (PVC), diisooctyl sebacate and freshly steamed tetrahydrofuran (THF) is added dropwise on a hydrophobized quartz glass slide and dried in air.

Further, the specific steps of the preparation method of the fluorescent sensor are:

Mix 3.0 mg of 4-aminophenyl-10,15,20-diphenylporphyrin (ATPP), 50 mg of polyvinyl chloride (PVC), 100 mg of diisooctyl sebacate and 2 ml of freshly distilled tetrahydrofuran ( THF) and fully dissolve and mix under the action of ultrasonic waves;

Place the hydrophobized quartz glass slide in a saturated tetrahydrofuran (THF) atmosphere, and rotate it at a frequency of 600 revolutions per minute in a rotary film-making device. At the same time, drop 0.2ml of the above solution onto the quartz glass slide and rotate for 20s. Finally, take out the quartz glass slide and dry it in the air for several minutes to obtain a light yellow sensitive film with a thickness of 4 μm attached to the quartz glass slide, and store it in a dark and dry place.

Further, the preparation method of 4-aminophenyl-10,15,20-diphenylporphyrin (ATPP) is:

(1) 5-(4-nitrophenyl)-10,15,20-triphenylporphyrin is directly nitrated by tetraphenylporphyrin

2.0g tetraphenylporphyrin is dissolved in the chloroform of 300ml, drips fuming nitric acid 3.4g with constant pressure dropping funnel, and keeps reaction temperature at 0-5 ℃, reacts 2h, and reaction product is constantly monitored by TLC to guarantee by TLC The Rf=0.88 of the reactant to the product Rf=0.78;

Wash with 5 × 300ml water to obtain a black-green solution, then dry with magnesium sulfate and sodium carbonate, concentrate to 70ml, pass through a silica gel column, and then wash with chloroform to collect only the mononitrotetraphenylporphyrin substance to obtain 5-(4 -nitrophenyl)-10,15,20-triphenylporphyrin, the yield is 55%, M ⁺ =659;

Dissolve 2.50g of 5-(4-nitrophenyl)-10,15,20-triphenylporphyrin in 80ml of concentrated hydrochloric acid, add 2.6g of tin dichloride, heat to 65°C, and pour it into into 300ml cold water, adjust the pH to 8.0 with saturated ammonium hydroxide, extract the aqueous phase with 6×300ml chloroform, dry with magnesium sulfate, concentrate the organic phase to 100ml with a rotary evaporator, purify it through a silica gel column, and wash with dichloromethane solution, the eluate was 1.75 g of 5-(4-aminophenyl)-10,15,20-triphenylporphyrin, the yield was 75%, and M ⁺ =629.

Further, the rotary film-making device is composed of an engine and a rotary pool, the quartz glass slide is placed in the rotary pool, and the rotary pool is connected with the power output shaft of the engine.

Further, the realization method for the hydrophobization treatment of the quartz glass slide is:

Dip the quartz glass slide with a diameter of 13mm in chromic acid washing solution, 3% HF solution, 10% H2O2 solution, and double distilled water;

The buffer solution of 0.2mlTSPM, 2ml0.2moll-1HAc-NaAc and 8ml double-distilled water are mixed to prepare TSPM solution, and the quartz slide is soaked in this solution for 3h and then rinsed with water, dried at room temperature for subsequent use, and the buffer solution of 1-1HAc-NaAc The pH is 3.6.

Another object of the present invention is to provide a polytetrafluoroethylene flow cell for detecting zinc ions. The polytetrafluoroethylene flow cell includes: a polypropylene cell body, a solution flow cell, fastening bolts, a fluorescent sensor, a solution inlet channel, solution outflow channel, dual-arm optical fiber;

The solution inlet channel and the solution outflow channel are in communication with the solution flow cell, the solution flow cell is arranged inside the polypropylene cell, the fluorescent sensor is arranged on the upper part of the solution flow cell, and the fluorescent sensor The sensor is fixed on the lower part of the polypropylene pool body through the fastening bolt, and one end of the double-arm optical fiber of the fluorescence sensor is connected to the fluorometer, and the other end is inserted into the lower part of the polypropylene pool body and tightened. Affix the fluorescent sensor.

Further, the diameter of the double-armed optical fiber is 8mm, and the length is 1m. The excitation light source in the fluorometer is a 150W xenon lamp, and the detector is an R928F infrared-sensitive photomultiplier tube.

Further, the implementation method of zinc ion detection is:

The sample to be tested is input into the solution flow cell at a speed of 1.5ml/min. At the maximum excitation wavelength and emission wavelength of the fluorescent sensor molecule, the fluorometer measures the fluorescence intensity of the sample to be tested, and the content of zinc ions is measured according to the calibration equation.

Further, when the concentration of Zn ²⁺ is in the range of 1.0×10 ^-6 ~1.0×10 ^-4 moll ^-1 , the correction equation is: F ₀ /F=1.1826+203504[Zn ²⁺ ], where r=0.9983, F ₀ and F are the fluorescence intensity at 651nm when the optode film is in contact with blank solution and Zn ²⁺ solution with different concentrations, respectively.

Further, if considering the phenomenon that the fluorescence intensity at 446nm decreases with the increase of the concentration of Zn ²⁺ , within the same concentration range, the relationship between the fluorescence intensity and the concentration of Zn ²⁺ is considered by using the change of the fluorescence intensity at 446nm and 651nm, The correction equation is: F ₀ -F ₀ '/F-F'=1.0719+390432[Zn ²⁺ ], where r=0.9992, F ₀ ' and F' are respectively the optode film and the blank solution, and different concentrations of Zn ²⁺ Fluorescence intensity at 446 nm upon solution contact.

The fluorescent sensor for detecting zinc ions based on the asymmetric porphyrin fluorescence ratio provided by the present invention is a combination of 4-aminophenyl-10,15,20-diphenylporphyrin (ATPP), polyvinyl chloride (PVC) , diisooctyl sebacate and freshly steamed tetrahydrofuran (THF) were added dropwise on a hydrophobized quartz glass slide and dried in the air, and the fluorescence sensor was installed on a polytetrafluoroethylene The top of the solution flow cell in the ethylene flow cell, the sample to be tested is input into the solution flow cell at a speed of 1.5ml/min, the fluorescence intensity is measured at the maximum excitation wavelength and two emission wavelengths of the fluorescent carrier in water, and the zinc ion is measured according to the calibration equation The content is simple, the detection process is simple, the sensitivity is high, the detection result is accurate, the practicability is strong, and it has strong promotion and application value.

Description of drawings

Figure 1 is the mass spectrum of 5-(4-aminophenyl)-10,15,20-triphenylporphyrin (ATPP) provided by the example of the present invention;

Fig. 2 is a schematic structural view of a rotary film forming device provided by an embodiment of the present invention;

Fig. 3 is a schematic structural view of a polytetrafluoroethylene flow cell provided by an embodiment of the present invention;

Fig. 4 is a fixed excitation wavelength of 390nm provided by the embodiment of the present invention, the fluorescence spectrum measured at the maximum emission wavelength of 448nm and 651nm, wherein the ordinate is the relative fluorescence intensity value, and the abscissa is the wavelength;

Fig. 5 is a schematic diagram of the influence of the solution acidity provided by the embodiment of the present invention on the ATPP optode film;

Fig. 6 is the calibration curve of zinc ions obtained by the fluorescence ratio method and the calibration curve of zinc ions measured by the fluorescence method alone provided by the embodiment of the present invention.

In the figure: 21, engine; 22, rotating pool; 23, quartz glass slide; 31, polypropylene pool body; 32, solution flow pool; 33, fastening bolt; 34, fluorescence sensor; 35, solution entering channel; 36, Solution outflow channel; 37. Double-arm optical fiber.

Detailed ways

In order to make the purpose, technical solution and advantages of the present invention more clear, the present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the invention.

The purpose of the present invention is to provide a fluorescent sensor 34 for detecting zinc ions based on the asymmetric porphyrin fluorescence ratio. A mixed solution composed of polyvinyl chloride (PVC), diisooctyl sebacate and freshly distilled tetrahydrofuran (THF) is dripped onto a hydrophobized quartz glass slide 23 and dried in the air.

In the embodiment of the present invention, the specific steps of the preparation method of the fluorescent sensor 34 are:

Mix 3.0 mg of 4-aminophenyl-10,15,20-diphenylporphyrin (ATPP), 50 mg of polyvinyl chloride (PVC), 100 mg of diisooctyl sebacate and 2 ml of freshly distilled tetrahydrofuran ( THF) and fully dissolve and mix under the action of ultrasonic waves;

Place the hydrophobized quartz glass slide 23 in a saturated tetrahydrofuran (THF) atmosphere, and rotate it at a frequency of 600 revolutions per minute in a rotary film forming device, and at the same time, drop 0.2ml of the above solution onto the quartz glass slide 23 After rotating for 20 s, the quartz glass slide 23 was taken out and dried in the air for several minutes to obtain a light yellow sensitive film with a thickness of 4 μm attached to the quartz glass slide 23, which was stored in a dark and dry place.

In the embodiment of the present invention, the preparation method of 4-aminophenyl-10,15,20-diphenylporphyrin (ATPP) is as follows:

(1) 5-(4-nitrophenyl)-10,15,20-triphenylporphyrin is directly nitrated by tetraphenylporphyrin

2.0g tetraphenylporphyrin is dissolved in the chloroform of 300ml, drips fuming nitric acid 3.4g with constant pressure dropping funnel, and keeps reaction temperature at 0-5 ℃, reacts 2h, and reaction product is constantly monitored by TLC to guarantee by TLC The Rf=0.88 of the reactant to the product Rf=0.78;

Wash with 5 × 300ml water to obtain a black-green solution, then dry with magnesium sulfate and sodium carbonate, concentrate to 70ml, pass through a silica gel column, and then wash with chloroform to collect only the mononitrotetraphenylporphyrin substance to obtain 5-(4 -nitrophenyl)-10,15,20-triphenylporphyrin, the yield is 55%, M ⁺ =659;

Dissolve 2.50g of 5-(4-nitrophenyl)-10,15,20-triphenylporphyrin in 80ml of concentrated hydrochloric acid, add 2.6g of tin dichloride, heat to 65°C, pour into into 300ml cold water, adjust the pH to 8.0 with saturated ammonium hydroxide, extract the aqueous phase with 6×300ml chloroform, dry with magnesium sulfate, concentrate the organic phase to 100ml with a rotary evaporator, purify it through a silica gel column, and wash it with dichloromethane solution, the eluate was 1.75 g of 5-(4-aminophenyl)-10,15,20-triphenylporphyrin, the yield was 75%, and M ⁺ =629.

Fig. 2 shows the structure of the rotary film forming device provided by the embodiment of the present invention. For ease of illustration, only the parts relevant to the present invention are shown.

The rotary film forming device is composed of an engine 21 and a rotary pool 22 , the quartz glass slide 23 is placed in the rotary pool 22 , and the rotary pool 22 is connected with the power output shaft of the engine 21 through transmission.

In the embodiment of the present invention, the implementation method for the hydrophobization treatment of the quartz glass slide 23 is as follows:

The quartz glass slide 23 with a diameter of 13 mm is immersed in chromic acid washing solution, 3% HF solution, 10% H2O2 solution, and double distilled water in sequence;

0.2mlTSPM, 2ml0.2moll-1HAc-NaAc buffer solution and 8ml twice distilled water are mixed to prepare TSPM solution, the quartz glass slide 23 is soaked in this solution for 3h, rinsed with water, dried at room temperature for later use, 1-1HAc-NaAc buffer solution The pH is 3.6.

Fig. 3 shows the structure of a polytetrafluoroethylene flow cell for detecting zinc ions provided by an embodiment of the present invention. For ease of illustration, only the parts relevant to the present invention are shown.

The polytetrafluoroethylene flow cell includes: a polypropylene cell body 31, a solution flow cell 32, fastening bolts 33, a fluorescent sensor 34, a solution inlet channel 35, a solution outflow channel 36, and a dual-arm optical fiber 37;

The solution inlet channel 35 and the solution outflow channel 36 are in communication with the solution flow cell 32, the solution flow cell 32 is arranged inside the polypropylene cell, the upper part of the solution flow cell 32 is provided with a fluorescence sensor 34, and the fluorescence sensor 34 is fixed by fastening bolts 33 At the bottom of the polypropylene pool body 31 , a fluorescence sensor 34 , one end of the double-armed optical fiber 37 is connected to the fluorometer, and the other end is inserted into the bottom of the polypropylene pool body 31 and is close to the fluorescence sensor 34 .

In the embodiment of the present invention, the double-arm optical fiber 37 has a diameter of 8mm and a length of 1m, the excitation light source in the fluorometer is a 150W xenon lamp, and the detector is an R928F infrared-sensitive photomultiplier tube.

In the embodiment of the present invention, the implementation method of zinc ion detection is:

The sample to be tested is input into the solution flow cell 32 at a speed of 1.5ml/min. At the maximum excitation wavelength and emission wavelength of the fluorescent sensor 34 molecules, the fluorescence meter measures the fluorescence intensity of the sample to be tested, and the content of zinc ions is measured according to the calibration equation .

In the embodiment of the present invention, when the concentration of Zn ²⁺ is in the range of 1.0×10 ^-6 to 1.0×10 ^-4 moll ^-1 , the correction equation is: F ₀ /F=1.1826+203504[Zn ²⁺ ], where r=0.9983, F ₀ and F are the fluorescence intensity at 651nm when the optode film is in contact with the blank solution and Zn ²⁺ solutions with different concentrations, respectively.

In the embodiment of the present invention, if considering the phenomenon that the fluorescence intensity at 446nm decreases with the increase of the concentration of Zn ²⁺ , within the same concentration range, the change of the fluorescence intensity at 446nm and 651nm is used to consider the relationship between the fluorescence intensity and the concentration of Zn ²⁺ The relationship between the concentration and the correction equation is: F ₀ -F ₀ '/F-F'=1.0719+390432[Zn ²⁺ ], where r=0.9992, F ₀ ' and F' are respectively the optode film and the blank solution, Fluorescence intensity at 446nm when Zn ²⁺ solutions with different concentrations are in contact.

The application principle of the present invention will be further described below in conjunction with the accompanying drawings and specific embodiments.

Technical scheme of the present invention is as follows:

1. Preparation of fluorescent sensor 34

ATPP (3.0 mg), PVC (50 mg), di-isooctyl sebacate (100 mg) and 2 ml of freshly distilled THF were mixed and dissolved with the help of ultrasound. In the self-made rotary film-making device, the hydrophobized glass slide was placed in a saturated THF atmosphere, and rotated at a frequency of 600 revolutions per minute. At the same time, 0.2ml of the above solution was added dropwise to the glass slide. The slides were taken out and dried in the air for several minutes to obtain a light yellow sensitive film with a thickness of about 4 μm attached to the slides, and stored in a dark and dry place.

2. Application of fluorescence sensor 34

The above-mentioned fluorescence sensor 34 is packed into the top of the polytetrafluoroethylene flow cell, and the sample to be tested is input into the flow cell at a speed of 1.5ml/min, and the fluorescence intensity is measured at the maximum excitation wavelength and emission wavelength of the fluorescence sensor 34 molecules in water, and according to The calibration equation measures the content of zinc ions.

The preparation method of the fluorescent sensor 34 of the present invention preferably follows the steps:

(1) Sample preparation 4-aminophenyl-10,15,20-diphenylporphyrin (ATPP)

5-(4-nitrophenyl)-10,15,20-triphenylporphyrin is directly nitrated by tetraphenylporphyrin: 2.0g tetraphenylporphyrin is dissolved in 300ml of chloroform (nitrogen protection), and Add 3.4g of fuming nitric acid dropwise into the constant pressure dropping funnel, and keep the reaction temperature at 0-5°C, react for 2h, and the reaction product is constantly monitored by TLC to ensure that the Rf=0.88 of the reactant to the product Rf=0.78 (with chloroform silicone board). Wash with 5×300ml of water to obtain a black-green solution, then dry over magnesium sulfate and sodium carbonate, concentrate to 70ml, pass through a silica gel column, and then wash with chloroform. Only the mononitrotetraphenylporphyrin material was collected to obtain 5-(4-nitrophenyl)-10,15,20-triphenylporphyrin in a yield of 55%, M ⁺ =659.

Dissolve 2.50g of 5-(4-nitrophenyl)-10,15,20-triphenylporphyrin in 80ml of concentrated hydrochloric acid (nitrogen protection), add 2.6g of tin dichloride, heat to 65°C for one hour Then pour into 300ml of cold water, adjust the pH to 8.0 with saturated ammonium hydroxide, extract the aqueous phase with 6×300ml of chloroform, dry with magnesium sulfate, concentrate the organic phase to 100ml with a rotary evaporator, purify it through a silica gel column, dichloro Methane was used as the eluent, and the eluate was 1.75 g of 5-(4-aminophenyl)-10,15,20-triphenylporphyrin, with a yield of 75%, M ⁺ =629.

(2) Preparation of fluorescent sensor 34

Hydrophobic treatment of the glass slide: the quartz glass slide 23 (diameter: 13 mm) was sequentially immersed in chromic acid washing solution, 3% HF, 10% H ₂ O ₂ , and double distilled water. 0.2mlTSPM, 2ml0.2moll-1HAc-NaAcpH3.6 buffer and 8ml double distilled water were mixed to prepare TSPM solution. The slides were soaked in this solution for 3 hours, rinsed with water, and dried at room temperature for later use.

Preparation of optode film: ATPP (3.0mg), PVC (50mg), diisooctyl sebacate (100mg) and 2ml of freshly distilled tetrahydrofuran were mixed and dissolved with the help of ultrasonic waves. In the self-made rotary film-making device (Figure 2), place the hydrophobized slide in a saturated THF atmosphere, rotate at a frequency of about 600 revolutions per minute, and drop 0.2ml of the above solution onto the slide at the same time After rotating for about 20 seconds, take out the slide and let it dry in the air for several minutes to obtain a light yellow sensitive film with a thickness of about 4 μm attached to the slide. Store in a dark and dry place.

(3) Application of fluorescence sensor 34

Fix the quartz glass slide 23 with the optode membrane on the top of the self-made polytetrafluoroethylene flow cell (Figure 1) by nuts, with the membrane facing down, so that the optode membrane is in contact with the sample solution. One end of a double-armed optical fiber 37 (diameter 8mm, length 1m) is connected to the fluorometer, and the other end is inserted into the flow cell and clings to the reverse side of the glass slide.

Fluorescence measurement was performed on a PerkingElmer LS55 fluorometer with a computer data processing system, the light source was a 150W xenon lamp, and the detector was an R928F infrared-sensitive photomultiplier tube.

The radiation emitted by the excitation light source is transmitted to the flow cell through one end of the double-armed optical fiber 37 to irradiate the surface of the glass slide, and excites the fluorescent substance in the photopolar film, and the emitted fluorescence is transmitted back to the detector by the other end for measurement. The sample solution is fed into the flow cell by the peristaltic pump at a speed of 1.5ml/min, and a stable fluorescence intensity value can be obtained after the optode membrane and the sample solution reach equilibrium. The fluorescence chemical fluorescence sensor 34 of the present invention can be applied to the determination of zinc ion content in water, as shown in FIG. 3 .

Example 1: The prepared fluorescent chemical sensor was loaded into a flow cell, and the peristaltic pump fed the sample into the flow cell at a speed of 1.5 ml/min, and the sample solution was a solution containing Zn ²⁺ in different concentrations. At a fixed maximum excitation wavelength of 390nm, measure the fluorescence intensity at the emission wavelength 446nm and 651nm of the optode film. When the pH value of the solution is 7.0, Zn ²⁺ can quench the fluorescence of the ATPP optode film. Fluorescence intensity of the sample solution when it reaches equilibrium, draw the fluorescence response diagram of the fluorescence intensity of the optode film with the change of Zn ²⁺ content, when the concentration of Zn ²⁺ is within the range of 1.0×10 ^-6 ~ 1.0×10 ^-4 moll ^-1 There is a certain linear relationship between them. The correction equation is: F ₀ /F=1.1826+203504[Zn ²⁺ ](r=0.9983), (here F ₀ and F are respectively at 651nm when the optode film is in contact with the blank solution and Zn ²⁺ solutions with different concentrations fluorescence intensity). If considering the phenomenon that the fluorescence intensity at 446nm decreases with the increase of Zn ²⁺ concentration, in the same concentration range, use the change of fluorescence intensity at 446nm and 651nm (fluorescence ratio) to consider the difference between the fluorescence intensity and the concentration of Zn ²⁺ relationship, the correction equation is: F ₀ -F ₀ '/F-F'=1.0719+390432[Zn ²⁺ ](r=0.9992), (here F ₀ ' and F' are respectively the optode film and the blank solution, Fluorescence intensity at 446nm when exposed to different concentrations of Zn ²⁺ solutions). It can be used as an ATPP sensor to measure the quantitative relationship of Zn ²⁺ content. By comparing the two figures together, the following conclusions can be drawn: the slope of the Zn ²⁺ calibration curve (curve 2 in Figure 6) obtained by the fluorescence ratio method is higher than that obtained by the fluorescence ratio method alone. The slope error of the Zn ²⁺ calibration curve measured by the method (curve 1 in Figure 6) is small.

Fig. 4 is a fluorescence spectrogram measured at a fixed excitation wavelength of 390nm and a maximum emission wavelength of 448nm and 651nm provided by an embodiment of the present invention, wherein the ordinate is the relative fluorescence intensity value, and the abscissa is the wavelength. Zn ²⁺ content from high to low is (moll ^-1 ): (1) 0; (2) 1.0×10 ^-5 moll ^-1 ; (3) 4.0×10 ^-5 moll ^-1 ; (4) 8.0× 10 ^-5 moll ^-1 ; (5) 1.0×10 ^-4 moll ^-1 ; (6) 2.0×10 ^-4 moll ^-1 .

Figure 6 shows the correction equation provided by the embodiment of the present invention when the concentration of Zn ²⁺ is in the range of 1.0×10 ^-6 to 1.0×10 ^-4 moll ^-1 : F ₀ /F=1.1826+203504[Zn ²⁺ ] (r=0.9983), (where F ₀ and F are the fluorescence intensity at 651nm when the optode film is in contact with the blank solution and Zn ²⁺ solutions with different concentrations). If considering the phenomenon that the fluorescence intensity at 446nm decreases with the increase of Zn ²⁺ concentration, in the same concentration range, use the change of fluorescence intensity at 446nm and 651nm (fluorescence ratio) to consider the difference between the fluorescence intensity and the concentration of Zn ²⁺ relationship, the correction equation is: F ₀ -F ₀ '/F-F'=1.0719+390432[Zn ²⁺ ](r=0.9992), (here F ₀ ' and F' are respectively the optode film and the blank solution, Fluorescence intensity at 446nm when exposed to different concentrations of Zn ²⁺ solutions).

The embodiment of the present invention provides a fluorescent sensor 34 for detecting zinc ions based on the asymmetric porphyrin fluorescence ratio. A mixed solution composed of ethylene (PVC), diisooctyl sebacate and freshly distilled tetrahydrofuran (THF) was added dropwise on a hydrophobized quartz glass slide 23 and dried in the air, and the fluorescence The sensor 34 is installed on the top of the solution flow cell 32 in the polytetrafluoroethylene flow cell, the sample to be tested is input into the solution flow cell 32 at a speed of 1.5ml/min, and the fluorescence is measured at the maximum excitation wavelength and two emission wavelengths of the fluorescent carrier in water Intensity, the content of zinc ions is measured according to the calibration equation, the detection process is simple, the sensitivity is high, the detection result is accurate, the practicability is strong, and it has strong promotion and application value.

The above are only preferred embodiments of the present invention, and are not intended to limit the present invention. Any modifications, equivalent replacements and improvements made within the spirit and principles of the present invention should be included in the protection scope of the present invention. Inside.

Claims (10)

1. One kind based on asymmetric porphyrin fluorescence than the fluorescent optical sensor that detects zinc ion, it is characterized in that, this fluorescent optical sensor is by 4-aminophenyl-10,15, the mixed solution that 20-diphenyl porphyrin (ATPP), Polyvinylchloride (PVC), Plexol 201 and the new tetrahydrofuran (THF) steamed form drips on the quartzy slide of processing at hydrophobization, and makes at air drying.
2. 2. fluorescent optical sensor as claimed in claim 1, is characterized in that, the preparation method's of this fluorescent optical sensor concrete steps are:

   4-aminophenyl-10 by 3.0mg, 15, the new tetrahydrofuran (THF) steamed of the Polyvinylchloride (PVC) of 20-diphenyl porphyrin (ATPP), 50mg, the Plexol 201 of 100mg and 2ml mixes, and under hyperacoustic effect, carries out fully dissolving and mixing;

   The quartzy slide that hydrophobization was processed is placed in saturated tetrahydrofuran (THF) atmosphere, the frequency rotation turned with per minute 600 in the rotation film forming apparatus, simultaneously to the above-mentioned solution that drips 0.2ml on quartzy slide, after rotation 20s, quartzy slide is taken out and is placed on air drying number minute, obtain being attached to the thick faint yellow sensitive membrane of 4 μ m on quartzy slide, be placed in dark dry place and preserve.
3. 3. fluorescent optical sensor as claimed in claim 2, is characterized in that, 4-aminophenyl-10,15, and the preparation method of 20-diphenyl porphyrin (ATPP) is:

   (1) 5-(4-nitrobenzophenone)-10,15, the 20-Triphenylporphyrin is directly nitrated by tetraphenylporphyrin

   2.0g tetraphenylporphyrin is dissolved in the chloroform of 300ml, with constant pressure funnel, drips fuming nitric aicd 3.4g, and keeps temperature of reaction at 0-5 ℃, reaction 2h, reaction product by TLC constantly monitoring to guarantee that Rf=0.88 by reactant is to generation product Rf=0.78;

   Obtain blackish green solution with 5 * 300ml water washing, then use magnesium sulphate and sodium carbonate drying, be concentrated to 70ml, cross silicagel column, then with the chloroform washing, only collect single nitro tetraphenylporphyrin material, obtain the 5-(4-nitrobenzophenone)-10,15, the 20-Triphenylporphyrin, productive rate is 55%, M ⁺=659;

   2.50g5-(4-nitrobenzophenone)-10,15, the 20-Triphenylporphyrin is dissolved in the concentrated hydrochloric acid of 80ml, add stannous chloride 2.6g, be heated to 65 ℃, after one hour, be poured in 300ml cold water, regulate pH to 8.0 with saturated ammonium hydroxide, 6 * 300ml chloroform extraction for water, use dried over mgso, Rotary Evaporators simmer down to 100ml for organic phase, by silicagel column, purify, methylene chloride is leacheate, and eluate is the 5-(4-aminophenyl)-10,15,20-Triphenylporphyrin 1.75g, productive rate 75%, M ⁺=629.
4. 4. fluorescent optical sensor as claimed in claim 2, is characterized in that, described rotation film forming apparatus consists of engine, rotation pond, and described quartzy slide is placed in described rotation pond, and the power output shaft of described rotation pond and described engine is in transmission connection.
5. 5. fluorescent optical sensor as claimed in claim 1, is characterized in that, the implementation method that quartzy slide hydrophobization is processed is:

   The quartzy slide of diameter 13mm is immersed in chromic acid lotion, 3% HF solution, 10% H2O2 solution, redistilled water successively;

   0.2mlTSPM, the damping fluid of 2ml0.2moll-1HAc-NaAc and 8ml redistilled water mix and make TSPM solution, quartzy slide soaks water flushing after 3h in this solution, drying for standby under room temperature, the pH of l-1HAc-NaAc damping fluid is 3.6.
6. 6. the flow cell of the teflon for detection of zinc ion, is characterized in that, this teflon flow cell comprises: polypropylene pond body, flow of solution pond, fastening bolt, fluorescent optical sensor, solution admission passage, solution flow pass, both arms optical fiber;

   Described solution admission passage and solution flow pass are connected with described flow of solution pond, described flow of solution pond is arranged on the inside in described polypropylene pond, the top in described flow of solution pond is provided with described fluorescent optical sensor, described fluorescent optical sensor is fixed on Ti bottom, described polypropylene pond by described fastening bolt, described fluorescent optical sensor, one of described both arms optical fiber terminates on luminoscope, and the other end is inserted into Ti bottom, described polypropylene pond and is close to described fluorescent optical sensor.
7. 7. teflon flow cell as claimed in claim 6, is characterized in that, the diameter of described both arms optical fiber is 8mm, and length is 1m, and the excitation source in described luminoscope is the 150W xenon lamp, and detecting device is R928F infrared-sensitive photomultiplier.
8. 8. teflon flow cell as claimed in claim 6, is characterized in that, the implementation method that zinc ion detects is:

   Testing sample is with the speed input flow of solution pond of 1.5ml/min, and in maximum excitation wavelength and the transmitted wave strong point of fluorescent optical sensor molecule, luminoscope is measured the fluorescence intensity of testing sample, and measures the content of zinc ion according to the correction equation formula.
9. 9. teflon flow cell as claimed in claim 6, is characterized in that, when the concentration of Zn2+ 1.0 * 10 ^-6～1.0 * 10 ^-4moll ^-1in scope, correction equation is: F ₀/ F=1.1826+203504[Zn ²⁺], r=0.9983 here, F ₀with F be respectively auroral poles film and blank solution, variable concentrations Zn ²⁺in solution when contact, is in the fluorescence intensity at 651nm place.
10. 10. teflon flow cell as claimed in claim 9, is characterized in that, if consider, 446nm place fluorescence intensity is with Zn ²⁺the phenomenon that reduces on the contrary of the increase of concentration, in the same concentrations scope, utilize the variation of the fluorescence intensity at 446nm and 651nm place to consider fluorescence intensity and Zn ²⁺the relation of concentration, correction equation is: F ₀-F ₀'/F-F '=1.0719+390432[Zn ²⁺], r=0.9992 here, F ₀' and F ' be respectively auroral poles film and blank solution, variable concentrations Zn ²⁺in solution when contact, is in the fluorescence intensity at 446nm place.

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