Iron-nickel supported hydroxysodalite fluorescent probe and its manufacturing method and application

Abstract

The present invention provides an iron-nickel-loaded hydroxysodalite fluorescent probe and its manufacturing method and use, which belong to the technical field of fluorescence detection. The iron-nickel-loaded hydroxysodalite fluorescent probe includes hydroxysodalite, elemental iron, elemental nickel, and ethylenediamine loaded on the surface of the hydroxysodalite, and fluorescein isothiocyanate chemically linked to the ethylenediamine. The N atom in the ethylenediamine on the surface of the hydroxysodalite is linked to Fe and Ni through a metal coordination bond. The iron-nickel-loaded hydroxysodalite fluorescent probe was used to detect the presence of L-arginine (L-Arg), Fe, and fluorescein isothiocyanate in a pure water phase. 3+ and Hg 2+ , PO4 3- and Fe in the pure water phase. 3+ and PO4 3- , Hg 2+ and PO4 3- It is possible to realize continuous detection.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority from a Chinese patent application filed with the China Patent Office on July 24, 2023, bearing application number 202310915594.8 and entitled "Iron-nickel supported hydroxysodalite fluorescent probe and its manufacturing method and application," the entire contents of which are incorporated herein by reference. [Technical Field]

[0002] The present invention relates to the technical field of fluorescence detection, in particular to an iron-nickel supported hydroxysodalite fluorescent probe and its preparation method and application. [Background technology]

[0003] The detection of L-arginine (L-Arg) is very important for the quality control of fermented foods, food additives, and beverages. Iron plays an important role in cellular metabolism and enzyme catalysis and is a trace element necessary for animals and plants. Iron deficiency can lead to anemia, liver and kidney damage, diabetes, and heart disease. Trace amounts of mercury can damage the kidneys, liver, and other organs. Phosphate plays an important role in bioenergy conversion and transmission and bone mineralization. However, abnormal phosphate levels can lead to tissue calcification, muscle weakness, white blood cell damage, rickets, or osteomalacia. Therefore, the detection of L-Arg, Fe 3+ , Hg 2+ and PO4 3- The detection of has great practical implications. In recent years, people have been using turbidity, ionization mass spectrometry, pulsed ampelography, capillary electrophoresis, high-performance liquid chromatography (HPLC), electrospray tandem mass spectrometry (HILC-ETMS), reversed-phase high-performance liquid chromatography, etc. to detect L-Arg and Fe. 3+ , Hg 2+ and PO4 3- However, most of these methods have the disadvantages of complicated sample preparation procedures and difficult operations. Summary of the Invention [Problem to be solved by the invention]

[0004] In view of this, the present invention aims to provide an iron-nickel-loaded hydroxysodalite fluorescent probe, its manufacturing method and application use. The fluorescent probe provided by the present invention is capable of detecting L-Arg, Fe in the aqueous phase. 3+ , H g2+ and PO4 3- can be detected easily, quickly and accurately. [Means for solving the problem]

[0005] In order to achieve the above-mentioned objectives of the invention, the present invention provides the following technical solutions: The present invention provides an iron-nickel-loaded hydroxysodalite fluorescent probe, which comprises hydroxysodalite, elemental iron, elemental nickel, and ethylenediamine loaded on the surface of the hydroxysodalite, and fluorescein isothiocyanate chemically linked to the ethylenediamine.

[0006] Preferably, the molar ratio of iron to nickel is 1:1.

[0007] Preferably, the particle size of the iron-nickel-supported hydroxysodalite fluorescent probe is 0.1 to 2 μm.

[0008] The present invention provides a step of mixing a soluble trivalent iron source, a soluble nickel source, a surfactant, hydroxysodalite, and water, and adjusting the pH value of the resulting mixture to 3 to 4 to obtain a mixture; mixing the mixture with a reducing agent to carry out a reduction reaction, thereby obtaining an iron-nickel-supported sodalite dispersion; mixing the iron-nickel-supported sodalite dispersion with ethylenediamine and carrying out an amination reaction to obtain aminated iron-nickel-supported sodalite; and mixing the aminated iron-nickel-loaded sodalite with a fluorescein isothiocyanate solution to carry out an addition reaction to obtain an iron-nickel-loaded hydroxysodalite fluorescent probe.

[0009] Preferably, the method for producing hydroxysodalite comprises: The method includes the steps of mixing NaAlO2, Na2SiO3, sodium hydroxide and water, and sequentially stirring, leaving the mixture to stand and microwave decomposition to obtain hydroxysodalite.

[0010] Preferably, the soluble trivalent iron source and the soluble nickel source have a molar ratio of Fe / Ni of 1:1; the mass ratio of the soluble iron (III) source to the hydroxysodalite is 1 to 3:1; the mass ratio of the soluble nickel source to the hydroxysodalite is 0.88 to 2.64:1; The mass ratio of the ethylenediamine to the hydroxysodalite is 1 to 2:1.

[0011] Preferably, the surfactant is polyethylene glycol, and the mass ratio of the surfactant to hydroxysodalite is 1 to 3:1.

[0012] Preferably, the reducing agent comprises ascorbic acid and NaI; the mass ratio of ascorbic acid to hydroxysodalite is 15 to 20:1; the mass ratio of NaI to hydroxysodalite is 0.10 to 0.25:1; The reduction reaction time is 1 to 3 hours.

[0013] Preferably, the mass ratio of the fluorescein isothiocyanate to the aminated iron-nickel-supported sodalite is 1:1 to 5; The addition reaction is carried out under a light-shielded condition, and the addition reaction time is 5 to 8 hours.

[0014] The present invention relates to a method for producing L-arginine, Fe 3+ , Hg 2+ , PO4 3- The present invention provides the use of the iron-nickel-loaded hydroxysodalite fluorescent probe in one or more of the following: [Effects of the Invention]

[0015] The present invention provides an iron-nickel-loaded hydroxysodalite fluorescent probe (functionalized SOD@Fe / Ni, abbreviated as Probe L) that includes hydroxysodalite, elemental iron, elemental nickel, and ethylenediamine loaded on the surface of the hydroxysodalite, and fluorescein isothiocyanate chemically linked to the ethylenediamine. In the present invention, the N atom in the ethylenediamine on the surface of the hydroxysodalite is linked to Fe and Ni via a metal coordination bond. Because iron-nickel clusters easily aggregate, the hydroxysodalite primarily serves as a carrier to prevent aggregation of the iron-nickel clusters. On the one hand, the iron-nickel clusters can be functionalized with ethylenediamine via a coordination bond to facilitate subsequent addition of fluorescein isothiocyanate, and on the other hand, they have a selective adsorption effect on the analyte. The present invention employs the iron-nickel-loaded hydroxysodalite fluorescent probe to detect L-arginine (L-Arg), Fe, and Ni in a pure water phase. 3+ and Hg 2+ , PO4 3- Simultaneous fluorescence detection of Fe was achieved in the pure water phase. 3+ and PO4 3- , Hg 2+ and PO4 3- The results of the examples show that the probes provided by the present invention can distinguish Arg, Fe, and 3+ and Hg 2+ , PO4 3- The detection limits for 10 and 20 are 7.7347×10, respectively. -8 mol / L, 2.4912 x 10 -8 mol / L, 4.7335 x 10 -8 mol / L and 4.8175 x 10-8 mol / L, and the linear range is 4.0 × 10 -6 mol / L ~ 3.2 × 10 -5 mol / L, 5.0 × 10 -7 mol / L ~ 3.5 × 10 -6 mol / L, 1.5 x 10 -6 mol / L ~ 4.0 × 10 -6 mol / L and 1.0×10 -6 mol / L ~ 1.4 × 10 -5 mol / L. Probe L + Fe 3+ is 2.0 x 10 -5 mol / L ~ 5.0 × 10 -5 mol / L range, PO4 3- The detection limit for -7 mol / L. Probe L + Hg 2+ is 3.0 x 10 -5 mol / L~9.0×10 -5 mol / L range, PO4 3- The detection limit for -7 The detection results showed good interference resistance against coexisting analytes, and the detection was rapid, with a response time of approximately 30 seconds.

[0016] The present invention provides a method for preparing the above iron-nickel supported hydroxysodalite fluorescent probe, which is simple in operation, low in cost, and easy to achieve industrialized batch production. [Brief explanation of the drawings]

[0017] [Figure 1] 1 is a flowchart showing the production of an iron-nickel-supported hydroxysodalite fluorescent probe. [Figure 2] These are the detection results for solid samples S1 to S5. [Figure 3] This shows the change in fluorescence spectrum caused by the action of the probe and various amino acids in H2O. [Figure 4] Fluorescence intensity of the probe when Arg and other amino acids coexist in H2O. [Figure 5] Fluorescence titration spectra of the probe and Arg in H2O. [Figure 6] The linear relationship between the fluorescence intensity of the probe and Arg. [Figure 7] This shows the change in fluorescence spectrum caused by the interaction of the probe with various cations in H2O. [Figure 8] This shows the change in fluorescence spectrum when the probe distinguishes between Fe3+ and Hg2+ in H2O. [Figure 9] Fluorescence intensity of the probe when Fe3+ and other cations coexist in H2O. [Figure 10] Fluorescence titration spectra of the probe and Fe3+ in H2O. [Figure 11] The fluorescence intensity of the probe is linearly related to Fe3+. [Figure 12] The interference of other cations on the discrimination of Hg2+ by the probe. [Figure 13] Fluorescence titration spectra of the probe and Hg2+ in H2O. [Figure 14] 1 is a linear relationship diagram between the fluorescence intensity of the probe at 514 nm in H2O and the concentration of Hg2+. [Figure 15] This shows the change in fluorescence spectrum caused by the interaction of the probe with various anions in H2O. [Figure 16] Fluorescence intensity of the probe in the presence of PO4 3- and other anions in H2O. [Figure 17] When the PO4 3- concentration is in the range of 0 to 1.4 × 10-5 mol / L, the fluorescence intensity of the probe changes accordingly. [Figure 18] When the PO4 3- concentration was within the range of 1.0 × 10-6 mol / L to 1.4 × 10-5 mol / L, a linear relationship was obtained between the two. [Figure 19] This is a spectrogram showing the identification of PO4 3- with the probe +Fe3+ in H2O. [Figure 20]The interference of other anions on the discrimination of PO4 3- by the probe +Fe3+. [Figure 21] Fluorescence titration spectra of the probes Fe3+ and PO4 3-. [Figure 22] This is a linear relationship diagram between probe Fe3+ and PO4 3- concentrations. [Figure 23] This is a spectrum that distinguishes PO4 3- from the probe +Hg2+. [Figure 24] The interference of other anions in the discrimination of PO4 3- by the probe +Hg2+. [Figure 25] The effect of PO4 3- on the fluorescence intensity of the probe + Hg2+. [Figure 26] The linear relationship curve of PO4 3- to probe Hg2+ fluorescence intensity was observed when the PO4 3- concentration was in the range of 3.0×10-5 mol / L to 9.0×10-5 mol / L. DETAILED DESCRIPTION OF THE INVENTION

[0018] The present invention provides an iron-nickel-loaded hydroxysodalite fluorescent probe, which comprises hydroxysodalite, elemental iron, elemental nickel, and ethylenediamine loaded on the surface of the hydroxysodalite, and fluorescein isothiocyanate chemically linked to the ethylenediamine.

[0019] In the present invention, the molar ratio of the iron element to the nickel element is preferably 1:1.

[0020] In the present invention, the iron and nickel form iron-nickel clusters on the surface of hydroxysodalite, and the particle size of the iron-nickel clusters is preferably 10 to 100 nm, more preferably 20 to 80 nm, and even more preferably 50 nm.

[0021] In the present invention, the N atom in the ethylenediamine on the surface of the hydroxysodalite is linked to Fe and Ni via a metal coordinate bond.

[0022] In the present invention, the particle size of the iron-nickel-supported hydroxysodalite fluorescent probe is preferably 0.1 to 2 μm, more preferably 0.5 to 1.5 μm.

[0023] The present invention provides a step of mixing a soluble trivalent iron source, a soluble nickel source, a surfactant, hydroxysodalite, and water, and adjusting the pH value of the resulting mixture to 3 to 4 to obtain a mixture; mixing the mixture with a reducing agent to carry out a reduction reaction, thereby obtaining an iron-nickel-supported sodalite dispersion; mixing the iron-nickel-supported sodalite dispersion with ethylenediamine and carrying out an amination reaction to obtain aminated iron-nickel-supported sodalite; mixing the aminated iron-nickel-loaded sodalite with a fluorescein isothiocyanate solution to carry out an addition reaction to obtain an iron-nickel-loaded hydroxysodalite fluorescent probe.

[0024] In the present invention, the method for producing hydroxysodalite includes: Preferably, the method includes a step of mixing NaAlO2, Na2SiO3, sodium hydroxide, and water, and sequentially carrying out stirring, standing, and microwave decomposition to obtain hydroxysodalite.

[0025] In the present invention, the Na2SiO3 is preferably Na2SiO3·9H2O. In the present invention, the mass ratio of NaOH, NaAlO2, Na2SiO3·9H2O, and water is preferably 1.95-21.5:1:8.67:100-200, more preferably 10-21.5:1:8.67:120-180.

[0026] In the present invention, the mixing method is preferably as follows: first, sodium hydroxide and water are mixed to obtain a sodium hydroxide solution; then the sodium hydroxide solution is divided into two parts, and NaAlO2 and Na2SiO3 are dissolved in them, respectively; and then the NaOH solution of NaAlO2 is slowly added to the NaOH solution of Na2SiO3.

[0027] In the present invention, the stirring time is preferably 1 to 3 hours, more preferably 2 hours, the standing time is preferably 0.5 to 1 hour, more preferably 0.6 to 0.8 hours, the microwave decomposition temperature is preferably 140 to 160°C, more preferably 150°C, and the time is preferably 1 to 3 hours, more preferably 2 hours.

[0028] In the present invention, after the microwave decomposition, the resulting liquid is preferably centrifuged for washing, and the centrifugal speed is preferably 6000 to 9000 r / min, more preferably 7000 to 8000 r / min. In the present invention, the washing is preferably performed with deionized water, and the solid obtained after centrifugation is preferably washed using deionized water until it becomes neutral, thereby obtaining hydroxysodalite.

[0029] In the present invention, a soluble trivalent iron source, a soluble nickel source, a surfactant, hydroxysodalite, and water are mixed together, and the pH value of the resulting mixture is adjusted to 3 to 4 to obtain a mixed solution. In the present invention, the soluble trivalent iron source is preferably FeCl3, more preferably FeCl3·6H2O, and the soluble nickel source is preferably NiCl2.

[0030] In the present invention, the surfactant is preferably polyethylene glycol, and the molecular weight of the polyethylene glycol is preferably 4,000.

[0031] In the present invention, the mixing method is preferably to first mix a soluble trivalent iron source with water, add a soluble nickel source, and obtain an Fe / Ni mixed solution; A surfactant and hydroxysodalite are added to the Fe / Ni mixed solution in sequence.

[0032] In the present invention, the pH adjusting agent used to adjust the pH is preferably HCl.

[0033] After adjusting the pH value, the present invention preferably subjects the resulting mixture to ultrasonic treatment, wherein the output power for the ultrasonic treatment is preferably 50 to 100 W, more preferably 60 to 80 W, and the duration is preferably 15 minutes. After adjusting the pH value, the soluble trivalent iron source and the soluble nickel source form Fe / Ni nanoclusters on the surface of the hydroxysodalite.

[0034] In the present invention, the Fe / Ni molar ratio in the soluble trivalent iron source and the soluble nickel source is preferably 1:1, the mass ratio of the soluble trivalent iron source to hydroxysodalite is preferably 1 to 3:1, more preferably 2:1, and the mass ratio of the soluble nickel source to hydroxysodalite is preferably 0.88 to 2.64:1, more preferably 1 to 2:1. In the present invention, the mass ratio of the surfactant to hydroxysodalite is preferably 1 to 3:1, more preferably 2:1.

[0035] In the present invention, the mixed solution is mixed with a reducing agent to carry out a reduction reaction, thereby obtaining an iron-nickel-supported sodalite dispersion. In the present invention, the reducing agent preferably contains ascorbic acid and NaI, and the mass ratio of the ascorbic acid to hydroxysodalite is preferably 15 to 20:1, more preferably 16 to 18:1, and the mass ratio of the NaI to hydroxysodalite is preferably 0.10 to 0.25:1, more preferably 0.15 to 0.16:1. In the present invention, the reducing agent is preferably added as an aqueous solution of the reducing agent, and the aqueous solution of the reducing agent further preferably contains 2 to 3 drops of hydrochloric acid.

[0036] In the present invention, the mixture of the mixed solution and the reducing agent is preferably carried out under nitrogen gas protection. In the present invention, the reduction reaction time is 1 to 3 hours, more preferably 2 hours. In the present invention, the reduction reaction is preferably carried out at room temperature.

[0037] In the present invention, the iron-nickel-supported sodalite dispersion is mixed with ethylenediamine and subjected to an amination reaction to obtain aminated iron-nickel-supported sodalite. In the present invention, the mass ratio of ethylenediamine to hydroxysodalite is preferably 1 to 2:1, more preferably 1.5:1.

[0038] In the present invention, the ethylenediamine is preferably added under magnetic stirring conditions. In the present invention, the time for the amination reaction is preferably 1 to 3 hours, more preferably 2 hours.

[0039] After the amination reaction, the present invention preferably obtains aminated iron-nickel-supported sodalite (aminated SOD@Fe / Ni), which is the supernatant of the resulting amination reaction solution.

[0040] In the present invention, the aminated iron-nickel-supported sodalite and a fluorescein isothiocyanate solution are mixed and subjected to an addition reaction to obtain an iron-nickel-supported hydroxysodalite fluorescent probe. In the present invention, the method for producing the fluorescein isothiocyanate solution preferably includes the following steps: The method includes the steps of mixing fluorescein isothiocyanate with N,N-dimethylformamide, and adding Na2CO3 and NaHCO3 buffer solutions to obtain a fluorescein isothiocyanate solution.

[0041] In the present invention, the ratio of the mass of the fluorescein isothiocyanate to the volume of the N,N-dimethylformamide is preferably 100 mg:20 mL, the pH value of the Na2CO3 and NaHCO3 buffer solution is preferably 9.2, and the volume ratio of the N,N-dimethylformamide to the Na2CO3 and NaHCO3 buffer solution is preferably 2:5.

[0042] In the present invention, the mass ratio of the fluorescein isothiocyanate to the aminated iron-nickel-supported sodalite is preferably 1:1-5, more preferably 1:2-4.

[0043] In the present invention, the addition reaction is preferably carried out under light-shielded conditions, the addition reaction time is preferably 5 hours, and the temperature is preferably room temperature. During the addition reaction, the amino groups on the surface of the aminated iron-nickel-supported sodalite undergo an addition reaction with the thiocyanate group of fluorescein isothiocyanate.

[0044] In the present invention, after the addition reaction, the addition reaction solution is preferably centrifuged and the resulting supernatant is diluted to obtain an iron-nickel-supported hydroxysodalite fluorescent probe. In the present invention, the centrifugation speed is preferably 6000 r / min, the dilution is preferably with deionized water, and the dilution ratio is preferably 2500 times.

[0045] In the present invention, a flowchart for producing the iron-nickel-supported hydroxysodalite fluorescent probe is shown in FIG.

[0046] The present invention relates to a method for producing L-arginine, Fe 3+ , Hg 2+ , PO4 3- The present invention provides the application of the iron-nickel-loaded hydroxysodalite fluorescent probe in one or more of the following: 3+ and Hg 2+ , PO4 3- and Fe in the pure water phase. 3+ and PO4 3- , Hg 2+ and PO4 3- It is possible to realize continuous detection.

[0047] Specifically, Fe 3+ and Hg 2+ If continuous detection of Fe is required, 3+ After detecting Fe, triethanolamine was added. 3+ This includes concealing the

[0048] The iron-nickel-loaded hydroxysodalite fluorescent probe provided by the present invention, its preparation method, and applications will be described in detail below with reference to examples, but these examples should not be construed as limitations on the scope of protection of the present invention.

[0049] Example 1 Preparation of Hydroxysodalite (SOD) 1.6g, 5.6g, 9.6g, 13.6g, and 17.6g of NaOH solid were dissolved in 85.95mL of water. The five NaOH solutions were divided into two portions, each containing 0.8197g of NaAlO2 and 7.1055g of Na2SiO3·9H2O. The NaAlO2-NaOH solution was then slowly added to the Na2SiO3-NaOH solution and stirred for 1 hour to obtain a homogeneous supernatant solution. After allowing to stand for 0.5 hours, the samples were placed in a microwave digester and heated at 140°C for 1 hour. The samples were numbered S1 to S5, from lowest to highest, based on the amount of NaOH added. After the reaction was complete, the samples were centrifuged at 6000 rpm and the solids were washed with deionized water until neutral.

[0050] The results of the detection of solid samples S1 to S5 are shown in Figure 2. The peaks of sample S5 appear at 14°, 24.5°, 32°, 34.5°, and 42.5°, which meet the standard XRD chromatogram of SOD. Sample S2 exhibits weak peaks and has low crystallinity. S1, S3, and S4 have many impurity peaks. Therefore, sample S5 was selected for subsequent experiments.

[0051] Example 2 Preparation of iron-nickel-supported hydroxysodalite fluorescent probe (probe L) 0.54 g of FeCl3·6H2O was dissolved in 50 mL of deionized water, followed by 0.47538 g of NiCl2, and the mixture was thoroughly dissolved to obtain a 1:1 Fe / Ni molar ratio solution. 0.54 g of polyethylene glycol-4000 (PEG-4000) was added to the mixture. After the PEG was dissolved, 0.2 g of SOD was added. The pH of the solution was adjusted to between 3 and 4 with HCl, and the mixture was sonicated for 15 min to obtain solution A. Separately, 3 g of Vc was dissolved in 50 mL of deionized water, to which 0.03 g of NaI and 2-3 drops of hydrochloric acid were added, followed by thorough stirring to obtain solution B. Solution B was slowly added dropwise to solution A under nitrogen gas protection, and the reaction was continued for 1 hour after the addition was complete. After multiple centrifugations, the supernatant was removed to obtain SOD@Fe / Ni. 0.3 mL of ethylenediamine was added to the solution under magnetic stirring, and the mixture was stirred for 2 hours. The centrifuged supernatant was then used as aminated SOD@Fe / Ni.

[0052] 100 mg of fluorescein isothiocyanate (FITC) was dissolved in 20 mL of N,N-dimethylformamide (DMF), and 50 mL of 50 mL of pH 9.2 Na2CO3 and NaHCO3 buffer was added. After thorough mixing, the solution was slowly added to the aminated SOD@Fe / Ni solution and incubated at room temperature for 5 hours in the dark. The reaction solution was centrifuged multiple times at 6000 r / min, and the resulting supernatant was diluted 2500-fold with deionized water to obtain probe L.

[0053] Performance Test (1) Fluorescence recognition of probe L to Arg in the aqueous phase 2 mL of deionized water was placed in a cuvette, 20 μL of the probe solution was added, and after mixing thoroughly, Arg (1.0 × 10 -2 The fluorescence spectra were measured before and after the addition of Arg. The same procedure was used to measure the fluorescence spectra of the probe and other amino acids (Ile, Asn, Thr, Ser, Pro, Leu, Arg, Val, Met, Glu, Ala, Gly, Phe, and Cys).

[0054] The changes in the fluorescence spectrum caused by the action of the probe and various amino acids in H2O are shown in Figure 3. The interpolated images in Figure 3 are the colors of the solutions with and without Arg added. From Figure 3, the λem of the probe in H2O is 514 nm, and 2 × 10 -5 After adding 100 mol / L Arg, the fluorescence intensity of the solution increased by 1.8-fold. The other 14 amino acids (Ile, Asn, Thr, Ser, Pro, Leu, Arg, Val, Met, Glu, Ala, Gly, Phe, and Cys) had relatively little or no effect on the fluorescence intensity. Therefore, in HO, the probe had a good discrimination effect on Arg. When the probe solution was irradiated with 365 nm light using a portable UV lamp, the colors of the probe solution and the probe + Arg solution were clearly distinguishable (interpolated images in Figure 3), i.e., the fluorescence of the solution changed from weak fluorescence to light green fluorescence.

[0055] 2 mL of deionized water was placed in a cuvette, and 20 μL of probe solution, 4 μL of Cys solution, and 4 μL of Arg solution were added. The mixture was thoroughly stirred to obtain a probe + Cys + Arg solution. The same procedure was used to obtain probe + other amino acids + Arg solutions, which were then subjected to fluorescence spectral detection.

[0056] The fluorescence intensity of the probe in the presence of Arg and other amino acids in HO is shown in Figure 4. In Figure 4, the fluorescence intensity at 514 nm of the probe + other amino acids + Arg and the probe + Arg solution was compared. It was found that, except for the significant decrease in the fluorescence intensity of the solution caused by Glu and Asn, the fluorescence intensity of the solution increased or decreased slightly in the presence of other amino acids, demonstrating that the presence of other amino acids had little effect on the interaction between the probe and Arg. Therefore, the interaction between the probe and Arg in HO exhibited good anti-interference performance.

[0057] 2 mL of deionized water was placed in a cuvette, and 20 μL of probe solution was added. The cumulative injection volume of Arg solution was 0.8 μL, 1.2 μL, 1.6 μL, 2.0 μL, 2.4 μL, 2.8 μL, 3.2 μL, 3.6 μL, 4.0 μL, 4.4 μL, 4.8 μL, 5.6 μL, and 6.4 μL, respectively. After each addition of Arg solution, the solution was thoroughly stirred to ensure uniform mixing before fluorescence spectral detection, maintaining a stable change in the fluorescence intensity of the solution.

[0058] The fluorescence titration spectrum of the probe and Arg in H2O showed that the fluorescence intensity of the solution at 514 nm continued to increase with increasing Arg concentration, as shown in Figure 5. Further analysis and study revealed that the fluorescence intensity of the solution at 514 nm increased by 4.0 x 10 -6 mol / L ~ 3.2 × 10 -5 A linear relationship between the two was obtained within the mol / L range, as shown in Figure 6. 2 The LOD of the probe for Arg in HO was 7.7347 × 10, which was determined by the formula: LOD = 3δ / k (δ is the standard deviation of the fluorescence intensity at 514 nm obtained by scanning the blank solution three times, and k is the absolute value of the slope of the linear fitting equation). -8 mol / L was obtained. In the above experiments, the experiments on the discrimination, interference, and detection limit of the probe for amino acids in H2O were all carried out under detection conditions of an excitation wavelength of 350 nm and a slit width of 5 nm.

[0059] (2) Fe in H2O by fluorescence spectroscopy 3+ , Hg 2+ Identification of 2 mL of deionized water was placed in a cuvette, 20 μL of the probe solution was added, and the mixture was thoroughly mixed to obtain a fluorescence spectrum. 3+Add 2 μL of the solution, stir thoroughly to mix evenly, and after the reaction is complete, perform further fluorescence spectrum detection. 3+ The fluorescence spectrum of the probe and other cations was obtained in the same manner.

[0060] The changes in the fluorescence spectrum resulting from the interaction of the probe with various cations in HO are shown in Figure 7. The interpolated image in Figure 7 shows the Fe 3+ and Hg 2+ Unadded and Fe 3+ and Hg 2+ is the color of the solution to which

[0061] From Figure 7, the λem of the probe in H2O is 514 nm, and -5 mol / L Fe 3+ After adding 1 × 10, the fluorescence intensity of the solution decreased by 3.3 times. -5 mol / L Hg 2+ After adding 15 other cations (Ag), the fluorescence intensity of the solution decreased by 3.0 times, and the fluorescence of the solution changed from weak to extremely weak. + , Al 3+ , Ba 2+ , Ca 2+ , Cd 2+ , Co 2+ , Cr 3+ , Cu 2+ , K. + , Mg 2+ , Mn 2+ , Ni 2+ , Pb 2+ , Zn 2+ , Zr 4+ ) has a smaller or almost no effect on its fluorescence intensity. Therefore, in HO, the probe 3+ , Hg 2+ has good discrimination effect.

[0062] 2 mL of deionized water was placed in a cuvette, and 20 μL of probe solution and Fe 3+ Solution 2 μL, TEA (triethanolamine) solution (100 times diluted) 1.6 μL, Hg2+ 2 μL of each solution was added, and after each addition, the solution was thoroughly stirred to mix uniformly, and then the fluorescence spectrum was detected.

[0063] In H2O, the probe is Fe 3+ and Hg 2+ The change in the fluorescence spectrum after identifying each is shown in Figure 8. From Figure 8, it can be seen that when Fe is added to the probe solution, 3+ After adding TEA, the fluorescence intensity of the solution decreased significantly. 3+ The fluorescence intensity of the solution recovers to its initial value. Finally, 2+ The fluorescence intensity of the solution was found to decrease again when Fe was added. 3+ After identifying Hg 2+ This enabled us to distinguish between the two and further achieve the goal of optimizing the selectivity of the probe.

[0064] 2 mL of deionized water was placed in a cuvette, and 20 μL of probe solution and Ag + 2μL of solution, Fe 3+ Add 2 μL of solution and stir thoroughly to mix evenly. + +Fe 3+ The solution was obtained by the same procedure, and the probe + other cations + Fe3 + The solutions were obtained and the fluorescence spectra were detected.

[0065] Fe in H2O 3+ The fluorescence intensity of the probe in the presence of other cations is shown in Figure 9. In Figure 9, the fluorescence intensity of the probe + other cations + Fe 3+ and probe + Fe 3+ The fluorescence intensity at 514 nm of the probe and Fe was compared. When other cations were present, the fluorescence intensity of the solution either increased or decreased slightly. 3+ Therefore, in H2O, the effect of the probe and Fe 3+ Both functions have good anti-interference performance.

[0066] Place 2 mL of deionized water in a cuvette, add 20 μL of the probe solution, and measure the Fe 3+ The cumulative increase method was adopted, and Fe 3+ The cumulative injection volumes of the solutions were set to 0.1 μL, 0.2 μL, 0.3 μL, 0.4 μL, 0.5 μL, 0.6 μL, and 0.7 μL, respectively. 3+ After adding the solutions, they were all thoroughly stirred to be uniformly mixed, and then the fluorescence spectrum was detected to maintain a stable change in the fluorescence intensity of the solution.

[0067] Probe and Fe in HO 3+ The fluorescence titration spectrum of the probe is shown in Figure 10. 3+ The linear relationship is shown in Figure 11. From Figures 10 and 11, Fe 3+ Concentration is 5.0 x 10 -7 mol / L ~ 3.5 × 10 -6 When the concentration is within the mol / L range, the linear relationship between the two is shown in Figure 11, R 2 is 0.9926, and Fe in H2O 3+ LOD of probe for = 2.4912 × 10 -8 mol / L.

[0068] (3) Hg 2+ Identification of Hg in the presence of other ions 2+ The study method for the effect on detection is Fe 3+ The same as the Hg probe 2+ The interference of other cations on the discrimination of is shown in Figure 12. Place 2 mL of deionized water in a cuvette, add 20 μL of the probe solution, and measure Hg. 2+ The cumulative increase method was adopted, and Hg 2+ The cumulative injection volumes of the solutions were set to 0.3 μL, 0.4 μL, 0.5 μL, 0.6 μL, and 0.8 μL, respectively, and Hg 2+ After adding the solution, all were thoroughly stirred to ensure uniform mixing, and then fluorescence spectra were detected to maintain a stable change in the fluorescence intensity of the solution. 2+The fluorescence titration spectrum of the probe in H2O at 514 nm and Hg 2+ The linear concentration relationship is shown in FIG.

[0069] From Figures 13 and 14, Hg 2+ Concentration is 1.5 x 10 -6 mol / L ~ 4.0 × 10 -6 mol / L range, R 2 is 0.9870, and Hg in H2O 2+ LOD of the probe for = 4.7335 × 10 -8 mol / L. Experiments on the discrimination, interference, and detection limit of the probe for cations in H2O were all carried out under detection conditions of an excitation wavelength of 350 nm and a slit width of 10 nm.

[0070] (4) PO4 in H2O by fluorescence spectroscopy 3- Identification of 2 mL of deionized water was placed in a cuvette, and 20 μL of the probe solution was added. The mixture was thoroughly stirred and mixed uniformly. The fluorescence spectrum was then detected to obtain the fluorescence spectrum of the probe in H2O. 3- (1.0×10 -2 mol / L) solution was added, and the mixture was thoroughly stirred to mix evenly. After the reaction was complete, the fluorescence spectrum was detected again. 3- The fluorescence spectrum of the probe and other anions was obtained using the same procedure.

[0071] The changes in the fluorescence spectrum caused by the action of the probe and various anions in H2O are shown in Figure 15. The interpolated image in Figure 15 is 3- The color of the solution before and after addition was shown in Figure 15. From Figure 15, the λem of the probe in H2O was 514 nm, and 2 × 10 -5 After adding 14 other anions (HSO3 -, SO3 2- , SO42-, S2O3 2- , SCN - , H.S. - , S2 - , NO2 - , I - , AlO2 - , CH3COO - , Cl - , HCO3 - , HPO4 2- ) had little or no effect on the fluorescence intensity. 3- It had good discrimination effect against

[0072] Add 2 mL of deionized water to a cuvette, add 20 μL of probe solution, and add HCO3 - Solution 4μL, PO4 3- Add 4 μL of the solution, stir thoroughly to mix evenly, and mix the probe + HCO3 - +PO4 3- The solution was obtained by the same procedure, and the probe + other anions + PO4 3- The solutions were obtained and the fluorescence spectra were detected. PO4 in H2O 3- The fluorescence intensity of the probe in the presence of other anions is shown in Figure 16. In Figure 16, the fluorescence intensity of the probe in the presence of other anions is shown in Figure 16. 3- and probe + PO4 3- From the comparison of the fluorescence intensity at 514 nm of the probe and PO4, it was found that when other anions coexisted, the fluorescence intensity of the solution either increased or decreased slightly. 3- Therefore, in H2O, the effect of the probe and PO4 3- Both functions have good anti-interference performance.

[0073] Place 2 mL of deionized water in a cuvette, add 20 μL of the probe solution, and measure the PO4 3- Adopting the cumulative increase method, PO4 3-The cumulative injection volumes of the solutions were 0.2 μL, 0.4 μL, 0.6 μL, 0.8 μL, 1.2 μL, 1.6 μL, 2.0 μL, 2.4 μL, and 2.8 μL, respectively, and PO4 3- After adding the solutions, they were all thoroughly stirred to be uniformly mixed, and then the fluorescence spectrum was detected to maintain a stable change in the fluorescence intensity of the solution. PO4 3- Concentration is 0 to 1.4 × 10 -5 The corresponding changes in the fluorescence intensity of the probe are shown in Figure 17. 3- The concentration is 1.0×10 -6 mol / L ~ 1.4 × 10 -5 When the concentration is within the mol / L range, the linear relationship between the two is shown in Figure 18, R 2 is 0.9952, and in H2O the PO4 3- LOD of probe for = 4.8175 × 10 -8 mol / L. The experiments on the discrimination, interference, and detection limit of the probe for anions in H2O were all carried out under the detection conditions of an excitation wavelength of 350 nm and a slit width of 5 nm.

[0074] (5) Probe + Fe in H2O 3+ PO4 by fluorescence spectroscopy 3- Identification of Place 2 mL of deionized water in a cuvette, 20 μL of probe solution, and Fe 3+ Add 2 μL of each solution in order, stir thoroughly to mix evenly, and perform fluorescence spectrum detection. 3+ After that, the cuvette was filled with PO4 3- Add 20 μL of the solution, stir thoroughly to mix evenly, and after the reaction is complete, perform fluorescence spectrum detection again. 3+ +PO4 3- The same procedure was used to obtain the fluorescence spectrum of the probe + Fe. 3+ + The fluorescence spectra of the other anions were detected in turn.

[0075] Probe + Fe in H2O3+ by PO4 3- The spectrogram of the identification is shown in Figure 19. From Figure 19, the probe + Fe in HO 3+ λem is 514 nm and 1×10 -4 mol / L PO4 3- After adding 14 other anions (HSO3), the fluorescence intensity of the solution increased by 3.3 times, and the fluorescence of the solution changed from very weak fluorescence to pale green fluorescence. - , SO3 2- , SO4 2- , S2O3 2- , SCN - , H.S. - , S2-, NO2 - , I - , AlO2 - , CH3COO - , Cl - , HCO3 - , HPO4 2- ) had little or no effect on the fluorescence intensity. 3+ is PO4 3- It had good discrimination effect against

[0076] Probe + Fe 3+ by PO4 3- The interference of other anions on the discrimination of probe + Fe is shown in Figure 20. 3+ +Other anions+PO4 3- and probe + Fe 3+ +PO4 3- From the control of the fluorescence intensity at 514 nm, S2 - , AlO2 - The presence of HS significantly increased the fluorescence intensity of the solution. - The presence of other anions significantly reduces the fluorescence intensity of the solution, but the presence of other anions only slightly increases or decreases the fluorescence intensity of the solution. 3+ and PO4 3- Therefore, in H2O, the effect of the probe + Fe 3+and PO4 3- Both functions have high anti-interference performance.

[0077] Place 2 mL of deionized water in a cuvette, 20 μL of probe solution, and Fe 3+ Add 2 μL of the solution, stir to mix evenly, and add PO4 3- Adopting the cumulative increase method, PO4 3- The cumulative injection volume of the solution was set to 4 μL, 5 μL, 6 μL, 7 μL, 8 μL, 9 μL, and 10 μL, respectively, and PO4 3- After adding the solutions, they were all thoroughly stirred to be uniformly mixed, and then the fluorescence spectrum was detected to maintain a stable change in the fluorescence intensity of the solution.

[0078] Probe + Fe 3+ and PO4 3- The fluorescence titration spectrum of the probe + Fe is shown in Figure 21. 3+ and PO4 3- The linear relationship of concentration is shown in Figure 22. PO4 3- Concentration is 2.0 x 1 0-5 mol / L ~ 5.0 × 10 -5 When the concentration is within the mol / L range, the linear relationship between the two is shown in Figure 22, R 2 is 0.9863, and the PO4 in H2O 3- Probe for Fe 3+ LOD = 2.7807 x 10 -7 mol / L. Probe for anions +Fe in H2O 3+ All experiments on discrimination, interference, and detection limit were carried out under detection conditions of an excitation wavelength of 350 nm and a slit width of 10 nm.

[0079] (6) Probe + Hg in H2O 2+ PO4 by fluorescence spectroscopy 3- Identification of Place 2 mL of deionized water in a cuvette, 20 μL of probe solution, and Hg 2+ Add 2 μL of each solution in order, stir thoroughly to mix evenly, and perform fluorescence spectrum detection.2+ After that, the cuvette was filled with PO4 3- Add 60 μL of the solution, stir thoroughly to mix evenly, and after the reaction is complete, perform fluorescence spectrum detection again. 2+ +PO4 3- The same procedure was used to obtain the fluorescence spectrum of the probe + Hg 2+ + The fluorescence spectra of the other anions were detected in turn.

[0080] Probe + Hg 2+ by PO4 3- The identification spectrum is shown in Figure 23. Probe + Hg 2+ by PO4 3- The interference of other anions on the discrimination of β-glucan is shown in FIG. In Figure 24, probe + Hg 2+ +Other anions+PO4 3- and probe + Hg 2+ +PO4 3- From the comparison of the fluorescence intensity at 514 nm, HCO3 - , NO2 - , S2 - The presence of Hg significantly increases the fluorescence intensity of the solution, but the presence of other anions only slightly increases or decreases the fluorescence intensity of the solution. 2+ and PO4 3- Therefore, in H2O, the effect of the probe + Hg 2+ and PO4 3- Both functions have high anti-interference performance. Place 2 mL of deionized water in a cuvette, 20 μL of probe solution, and Hg 2+ Add 2 μL of each solution in order, stir to mix evenly, and add PO4 3- Adopting the cumulative increase method, PO4 3- The cumulative injection volumes of the solutions were 6 μL, 8 μL, 10 μL, 12 μL, 14 μL, 16 μL, and 18 μL, respectively, and PO4 3-After adding the solutions, they were all thoroughly stirred to be uniformly mixed, and then the fluorescence spectrum was detected to maintain a stable change in the fluorescence intensity of the solution.

[0081] PO4 3- probe + Hg 2+ The effect on fluorescence intensity is shown in Figure 25. 3- The concentration is 3.0×10 -5 mol / L~9.0×1 0-5 When the concentration is within the mol / L range, the linear relationship between the two is shown in Figure 26, R 2 is 0.9839, and the PO4 3- Probe for Hg 2+ LOD = 3.1952 x 10 -7 mol / L. Probe for anions + Hg in H2O 2+ All experiments on discrimination, interference, and detection limit were carried out under detection conditions of an excitation wavelength of 350 nm and a slit width of 10 nm.

[0082] The above are only preferred embodiments of the present invention, and those skilled in the art may make further improvements and modifications without departing from the principles of the present invention, and these improvements and modifications are also considered to be within the scope of protection of the present invention.

Claims

1. An iron-nickel-loaded hydroxysodalite fluorescent probe, comprising hydroxysodalite, elemental iron loaded on the surface of the hydroxysodalite, elemental nickel and ethylenediamine, and fluorescein isothiocyanate chemically linked to the ethylenediamine. An iron-nickel-supported hydroxysodalite fluorescent probe characterized by:

2. The molar ratio of the iron element to the nickel element is 1:

1. The iron-nickel-supported hydroxysodalite fluorescent probe according to claim 1.

3. The particle size of the iron-nickel-supported hydroxysodalite fluorescent probe is 0.1 to 2 μm. The iron-nickel-supported hydroxysodalite fluorescent probe according to claim 1.

4. A method for producing the iron-nickel-supported hydroxysodalite fluorescent probe according to claim 1, comprising the steps of: mixing a soluble trivalent iron source, a soluble nickel source, a surfactant, hydroxysodalite, and water, and adjusting the pH value of the resulting mixture to 3 to 4 to obtain a mixture; mixing the mixture with a reducing agent to carry out a reduction reaction, thereby obtaining an iron-nickel-supported sodalite dispersion; mixing the iron-nickel-supported sodalite dispersion with ethylenediamine to carry out an amination reaction, thereby obtaining aminated iron-nickel-supported sodalite; mixing the aminated iron-nickel-loaded sodalite with a fluorescein isothiocyanate solution to carry out an addition reaction to obtain an iron-nickel-loaded hydroxysodalite fluorescent probe. A manufacturing method characterized by:

5. The method for producing hydroxysodalite comprises: NaAlO ２ , Na ２ SiO ３ , sodium hydroxide and water, and then stirring, standing, and microwave decomposition are carried out in this order to obtain hydroxysodalite. The method of claim 4.

6. the soluble trivalent iron source and the soluble nickel source have a molar ratio of Fe / Ni of 1:1; the mass ratio of the soluble iron(III) source to the hydroxysodalite is 1 to 3:1; the mass ratio of the soluble nickel source to the hydroxysodalite is 0.88 to 2.64:1; The mass ratio of the ethylenediamine to the hydroxysodalite is 1 to 2:

1. The method of claim 4.

7. The surfactant is polyethylene glycol, and the mass ratio of the surfactant to hydroxysodalite is 1 to 3:

1. The method of claim 4.

8. the reducing agent comprises ascorbic acid and NaI; the mass ratio of ascorbic acid to hydroxysodalite is 15 to 20:1; the mass ratio of NaI to hydroxysodalite is 0.10 to 0.25:1; The reduction reaction time is 1 to 3 hours. The method of claim 4.

9. the mass ratio of the fluorescein isothiocyanate to the aminated iron-nickel-supported sodalite is 1:1 to 5; The addition reaction is carried out under a light-shielded condition, and the addition reaction time is 5 to 8 hours. The method of claim 4.

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