Iron-nickel-loaded hydroxysodalite fluorescent probe, and preparation method therefor and use thereof

Abstract

The present invention belongs to the technical field of fluorescence detection. Provided are an iron-nickel-loaded hydroxysodalite fluorescent probe, and a preparation method therefor and the use thereof. The iron-nickel-loaded hydroxysodalite fluorescent probe comprises: hydroxysodalite; an elementary substance of iron, an elementary substance of nickel, and ethylenediamine, which are loaded on the surface of the hydroxysodalite; and fluorescein isothiocyanate, which is chemically bonded to the ethylenediamine. N atoms in the ethylenediamine on the surface of the hydroxysodalite are bonded to Fe and Ni via metal coordination bonds. By using the iron-nickel-loaded hydroxysodalite fluorescent probe, the simultaneous fluorescence identification of L-arginine (L-Arg), Fe3+, Hg2+ and PO4 3- in a pure water phase is achieved; and Fe3+ and PO4 3-, and Hg2+ and PO4 3- can be continuously identified in the pure water phase.

Description

A kind of iron-nickel loaded hydroxysodalite fluorescent probe and its preparation method and application

[0001] This application claims priority to the Chinese patent application filed with the Patent Office of China on July 24, 2023, with application number 202310915594.8, and invention name “A kind of iron-nickel loaded hydroxysodalite fluorescent probe and its preparation method and application”, the entire contents of which are incorporated by reference into this application. Technical Field

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

[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 cell metabolism and enzyme catalysis and is an essential trace element 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 a key role in energy transduction and bone mineralization in organisms. However, abnormal phosphate levels can lead to tissue calcification, muscle weakness, white blood cell damage, rickets or osteomalacia, etc. Therefore, for L-Arg, Fe 3+ 、Hg 2+ and PO4 3- The detection has important practical significance.

[0004] In recent years, people have developed turbidimetry, ionization mass spectrometry, pulsed amperometry, capillary electrophoresis, high performance liquid chromatography (HPLC), electrospray tandem mass spectrometry (HILC-ETMS), reversed-phase high performance liquid chromatography, etc. 3+ 、Hg 2+ and PO4 3- However, most of these methods have the disadvantages of complex sample preparation procedures and difficult operations.

[0005] Summary of the Invention

[0006] In view of this, the present invention aims to provide a fluorescent probe of hydroxysodalite loaded with iron and nickel and its preparation method and application. The fluorescent probe provided by the present invention can detect L-Arg, Fe and Cr in aqueous phase in a simple, fast and accurate manner. 3+ 、Hg 2+ and PO4 3- .

[0007] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0008] The invention provides an iron-nickel loaded hydroxysodalite fluorescent probe, comprising hydroxysodalite, iron element, nickel element and ethylenediamine loaded on the surface of the hydroxysodalite, and fluorescein isothiocyanate chemically connected to the ethylenediamine.

[0009] Preferably, the molar ratio of the iron element to the nickel element is 1:1.

[0010] Preferably, the particle size of the iron-nickel loaded hydroxysodalite fluorescent probe is 0.1-2 μm.

[0011] The present invention provides a method for preparing the above-mentioned iron-nickel loaded hydroxysodalite fluorescent probe, comprising the following steps:

[0012] Mixing a soluble ferric iron source, a soluble nickel source, a surfactant, hydroxysodalite and water, and adjusting the pH value of the resulting mixed solution to 3-4 to obtain a mixed solution;

[0013] mixing the mixed solution with a reducing agent to carry out a reduction reaction to obtain an iron-nickel loaded sodalite dispersion;

[0014] mixing the iron-nickel loaded sodalite dispersion with ethylenediamine to carry out an amination reaction to obtain the amination-loaded iron-nickel loaded sodalite;

[0015] The aminated iron-nickel loaded sodalite is mixed with a fluorescein isothiocyanate solution and subjected to an addition reaction to obtain an iron-nickel loaded hydroxysodalite fluorescent probe.

[0016] Preferably, the preparation method of hydroxysodalite comprises the following steps:

[0017] NaAlO2, Na2SiO3, sodium hydroxide and water are mixed, and the mixture is stirred, allowed to stand and microwave digested in sequence to obtain hydroxysodalite.

[0018] Preferably, the Fe / Ni molar ratio of the soluble ferric iron source and the soluble nickel source is 1:1;

[0019] The mass ratio of the soluble ferric iron source to hydroxysodalite is 1 to 3:1;

[0020] The mass ratio of the soluble nickel source to hydroxysodalite is 0.88 to 2.64:1;

[0021] The mass ratio of the ethylenediamine to hydroxysodalite is 1 to 2:1.

[0022] Preferably, the surfactant is polyethylene glycol, and the mass ratio of the surfactant to hydroxysodalite is 1 to 3:1.

[0023] Preferably, the reducing agent comprises ascorbic acid and NaI;

[0024] The mass ratio of ascorbic acid to hydroxysodalite is 15 to 20:1;

[0025] The mass ratio of NaI to hydroxysodalite is 0.10-0.25:1;

[0026] The reduction reaction time is 1 to 3 hours.

[0027] Preferably, the mass ratio of the fluorescein isothiocyanate to the amino-loaded iron-nickel sodalite is 1:1-5;

[0028] The addition reaction is carried out in a light-proof condition, and the time of the addition reaction is 5 to 8 hours.

[0029] The present invention provides the above-mentioned iron-nickel loaded hydroxysodalite fluorescent probe for detecting L-arginine, Fe 3+ 、Hg 2+ PO4 3- One or more applications.

[0030] The present invention provides a hydroxysodalite fluorescent probe loaded with iron and nickel (functionalized SOD@Fe / Ni, referred to as probe L), comprising hydroxysodalite, iron element, nickel element and ethylenediamine loaded on the surface of the hydroxysodalite, and fluorescein isothiocyanate chemically connected to the ethylenediamine. In the present invention, the N atom in the ethylenediamine on the surface of the hydroxysodalite is connected to Fe and Ni through metal coordination bonds. In the present invention, since iron-nickel clusters are easy to aggregate, hydroxysodalite mainly acts as a carrier to avoid the aggregation of iron-nickel clusters. On the one hand, the iron-nickel clusters can be functionalized with ethylenediamine through coordination bonds to facilitate subsequent addition with fluorescein isothiocyanate; on the other hand, they have a selective adsorption effect on the analyte. The present invention uses the hydroxysodalite fluorescent probe loaded with iron and nickel to achieve simultaneous fluorescent identification of L-arginine (L-Arg), Fe 3+ and Hg 2+ PO4 3- , and can realize continuous identification of Fe in pure water phase 3+ and PO4 3- 、Hg 2+ and PO4 3- The results of the examples show that the probe provided by the present invention is sensitive to Arg, Fe 3+ and Hg 2+ PO4 3- The detection limits were 7.7347×10 -8 mol / L、2.4912×10 -8 mol / L、4.7335×10 -8 mol / L and 4.8175×10 -8mol / L, and the linear range was 4.0×10 -6 mol / L~3.2×10 -5 mol / L、5.0×10 -7 mol / L~3.5×10 -6 mol / L、1.5×10 -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+ In 2.0×10 -5 mol / L~5.0×10 -5 In the range of mol / L, for PO4 3- The detection limit was 2.7807×10 -7 mol / L. Probe L+Hg 2+ In 3.0×10 -5 mol / L~9.0×10 -5 In the range of mol / L, for PO4 3- The detection limit was 3.195210 -7 mol / L. The above tests all showed good anti-interference performance for coexisting analytes and were rapid, with response times of around 30 seconds.

[0031] The present invention provides a method for preparing the iron-nickel loaded hydroxysodalite fluorescent probe. The method is simple to operate, low in cost, and easy to realize industrialized batch production. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] FIG1 is a flow chart of the preparation of an iron-nickel loaded hydroxysodalite fluorescent probe;

[0033] Figure 2 shows the test results of solid samples S1 to S5;

[0034] Figure 3 shows the fluorescence spectrum changes of the probe in H2O when it interacts with various amino acids;

[0035] Figure 4 shows the fluorescence intensity of the probe in H2O when Arg and other amino acids coexist;

[0036] Figure 5 shows the fluorescence titration spectra of the probe and Arg in H2O;

[0037] Figure 6 shows the linear relationship between the probe fluorescence intensity and Arg;

[0038] Figure 7 shows the fluorescence spectrum changes of the probe in H2O when it interacts with various cations;

[0039] Figure 8 shows the probe reaction of Fe in H2O 3+ and Hg 2+The fluorescence spectrum changes under different identifications;

[0040] Figure 9 shows the probe in Fe 3+ Fluorescence intensity when coexisting with other cations;

[0041] Figure 10 shows the relationship between the probe and Fe in H2O. 3+ Fluorescence titration spectrum of

[0042] Figure 11 shows the relationship between the fluorescence intensity of the probe and Fe 3+ Linear relationship;

[0043] Figure 12 shows the recognition of Hg by other cation pair probes 2+ interference;

[0044] Figure 13 shows the relationship between the probe and Hg in H2O. 2+ Fluorescence titration spectrum of

[0045] Figure 14 shows the fluorescence intensity of the probe in H2O at 514 nm and the fluorescence intensity of Hg 2+ Linear relationship graph of concentration;

[0046] Figure 15 shows the fluorescence spectrum changes of the probe interacting with various anions in H2O;

[0047] Figure 16 shows the probe in H2O and PO4 3- Fluorescence intensity when coexisting with other anions;

[0048] Figure 17 shows the 3- The concentration is 0~1.4×10 -5 In the mol / L range, the fluorescence intensity of the probe changes with;

[0049] Figure 18 shows the 3- The concentration is 1.0×10 -6 mol / L~1.4×10 -5 In the range of mol / L, a linear relationship was obtained between the two;

[0050] Figure 19: Probe + Fe in H2O 3+ Identify PO4 3- Spectrum graph;

[0051] Figure 20 shows other anion pairs probe + Fe 3+ Identify PO4 3- interference;

[0052] Figure 21 is probe+Fe 3+ With PO4 3- Fluorescence titration spectrum of

[0053] Figure 22 is probe+Fe 3+ With PO4 3-Linear relationship graph of concentration;

[0054] Figure 23 is probe + Hg 2+ Identify PO4 3- spectrum;

[0055] Figure 24 shows other anion pairs probe + Hg 2+ Identify PO4 3- interference;

[0056] Figure 25 shows PO4 3- Probe+Hg 2+ Effect of fluorescence intensity;

[0057] Figure 26 shows the 3- The concentration is 3.0×10 -5 mol / L~9.0×10 -5 PO4 in the mol / L range 3- Probe+Hg 2+ Linear relationship curve of fluorescence intensity. DETAILED DESCRIPTION

[0058] The invention provides an iron-nickel loaded hydroxysodalite fluorescent probe, comprising hydroxysodalite, iron element, nickel element and ethylenediamine loaded on the surface of the hydroxysodalite, and fluorescein isothiocyanate chemically connected to the ethylenediamine.

[0059] In the present invention, the molar ratio of the iron element to the nickel element is preferably 1:1.

[0060] 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 more preferably 50 nm.

[0061] In the present invention, the N atoms in the ethylenediamine on the surface of the hydroxysodalite are connected to Fe and Ni through metal coordination bonds.

[0062] In the present invention, the particle size of the iron-nickel loaded hydroxysodalite fluorescent probe is preferably 0.1 to 2 μm, more preferably 0.5 to 1.5 μm.

[0063] The present invention provides a method for preparing the above-mentioned iron-nickel loaded hydroxysodalite fluorescent probe, comprising the following steps:

[0064] Mixing a soluble ferric iron source, a soluble nickel source, a surfactant, hydroxysodalite and water, and adjusting the pH value of the resulting mixed solution to 3-4 to obtain a mixed solution;

[0065] mixing the mixed solution with a reducing agent to carry out a reduction reaction to obtain an iron-nickel loaded sodalite dispersion;

[0066] mixing the iron-nickel loaded sodalite dispersion with ethylenediamine to carry out an amination reaction to obtain the amination-loaded iron-nickel loaded sodalite;

[0067] The aminated iron-nickel loaded sodalite is mixed with a fluorescein isothiocyanate solution and subjected to an addition reaction to obtain an iron-nickel loaded hydroxysodalite fluorescent probe.

[0068] In the present invention, the preparation method of hydroxysodalite preferably comprises the following steps:

[0069] NaAlO2, Na2SiO3, sodium hydroxide and water are mixed, and the mixture is stirred, allowed to stand and microwave digested in sequence to obtain hydroxysodalite.

[0070] In the present invention, the Na2SiO3 is preferably Na2SiO3·9H2O. In the present invention, the mass ratio of the 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.

[0071] In the present invention, the mixing method is preferably: first mix sodium hydroxide and water to obtain a sodium hydroxide solution, divide the sodium hydroxide solution into two parts, dissolve NaAlO2 and Na2SiO3 respectively, and then slowly add the NaAlO2 NaOH solution to the Na2SiO3 NaOH solution.

[0072] 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. In the present invention, the microwave digestion temperature is preferably 140 to 160°C, more preferably 150°C, and the time is preferably 1 to 3 hours, more preferably 2 hours.

[0073] In the present invention, after the microwave digestion, the liquid obtained is preferably centrifuged and washed. The centrifugation rate is preferably 6000-9000 r / min, more preferably 7000-8000 r / min. In the present invention, the washing is preferably done with deionized water. In the present invention, deionized water is preferably used to wash the solid obtained after centrifugation until it is neutral to obtain hydroxysodalite.

[0074] The present invention comprises mixing a soluble ferric iron source, a soluble nickel source, a surfactant, hydroxysodalite, and water, and adjusting the pH of the resulting mixed solution to 3 to 4 to obtain a mixed solution. In the present invention, the soluble ferric iron source is preferably FeCl3, more preferably FeCl3·6H2O; and the soluble nickel source is preferably NiCl2.

[0075] In the present invention, the surfactant is preferably polyethylene glycol, and the molecular weight of the polyethylene glycol is preferably 4000.

[0076] In the present invention, the mixing method is preferably: firstly mixing a soluble ferric iron source with water, and then adding a soluble nickel source to obtain an Fe / Ni mixed solution;

[0077] A surfactant and hydroxysodalite are sequentially added to the Fe / Ni mixed solution.

[0078] In the present invention, the pH adjuster used to adjust the pH value is preferably HCl.

[0079] After adjusting the pH value, the present invention preferably subjects the resulting mixed solution to ultrasonic treatment, wherein the ultrasonic treatment power is preferably 50 to 100 W, more preferably 60 to 80 W, and the duration is preferably 15 minutes. After the pH value is adjusted, the soluble ferric iron source and the soluble nickel source form Fe / Ni nanoclusters on the surface of the hydroxysodalite.

[0080] In the present invention, the Fe / Ni molar ratio of the soluble ferric iron source and the soluble nickel source is preferably 1:1; the mass ratio of the soluble ferric iron source to hydroxysodalite is preferably 1-3:1, more preferably 2:1; the mass ratio of the soluble nickel source to hydroxysodalite is preferably 0.88-2.64:1, more preferably 1-2:1. In the present invention, the mass ratio of the surfactant to hydroxysodalite is preferably 1-3:1, more preferably 2:1.

[0081] The present invention mixes the mixed solution with a reducing agent and performs a reduction reaction to obtain a sodalite dispersion loaded with iron and nickel. In the present invention, the reducing agent preferably comprises ascorbic acid and NaI; the mass ratio of the ascorbic acid to hydroxysodalite is preferably 15 to 20:1, more preferably 16 to 18:1; 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 in the form of an aqueous reducing agent solution, which preferably also includes 2 to 3 drops of hydrochloric acid.

[0082] In the present invention, the mixing of the mixed solution and the reducing agent is preferably carried out under nitrogen 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.

[0083] The present invention mixes the iron-nickel loaded sodalite dispersion with ethylenediamine and performs an amination reaction to obtain the amination-loaded iron-nickel loaded sodalite. In the present invention, the mass ratio of the ethylenediamine to the hydroxysodalite is preferably 1 to 2:1, more preferably 1.5:1.

[0084] In the present invention, the ethylenediamine is preferably added under magnetic stirring. In the present invention, the amination reaction time is preferably 1 to 3 hours, more preferably 2 hours.

[0085] After the amination reaction, the present invention preferably takes the supernatant of the amination reaction solution, which is the amination-loaded iron-nickel sodalite (amination-loaded SOD@Fe / Ni).

[0086] The present invention mixes the amino-loaded iron-nickel sodalite with a fluorescein isothiocyanate solution to perform an addition reaction to obtain an iron-nickel-loaded hydroxysodalite fluorescent probe. In the present invention, the preparation method of the fluorescein isothiocyanate solution preferably includes the following steps:

[0087] Fluorescein isothiocyanate was mixed with N,N-dimethylformamide, and Na2CO3 and NaHCO3 buffer were added to obtain a fluorescein isothiocyanate solution.

[0088] In the present invention, the mass ratio of fluorescein isothiocyanate to N,N-dimethylformamide is preferably 100 mg:20 mL. In the present invention, the pH value of the Na2CO3 and NaHCO3 buffer is preferably 9.2; and the volume ratio of N,N-dimethylformamide to Na2CO3 and NaHCO3 buffer is preferably 2:5.

[0089] In the present invention, the mass ratio of fluorescein isothiocyanate to the amino-supported iron-nickel sodalite is preferably 1:1-5, more preferably 1:2-4.

[0090] In the present invention, the addition reaction is preferably carried out under light-proof 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-loaded sodalite react with the isothiocyanate groups of fluorescein isothiocyanate.

[0091] In the present invention, after the addition reaction, the present invention preferably centrifuges the resulting addition reaction solution, and dilutes the supernatant obtained by centrifugation to obtain an iron-nickel-loaded hydroxysodalite fluorescent probe. In the present invention, the centrifugation rate is preferably 6000 r / min, the dilution is preferably diluted with deionized water, and the dilution multiple is preferably 2500 times.

[0092] In the present invention, the preparation flow chart of the iron-nickel loaded hydroxysodalite fluorescent probe is shown in FIG1 .

[0093] The present invention provides the above-mentioned iron-nickel loaded hydroxysodalite fluorescent probe for detecting L-arginine, Fe 3+ 、Hg 2+ PO4 3-The present invention uses a hydroxysodalite fluorescent probe loaded with iron and nickel to realize the simultaneous fluorescent identification of L-arginine (L-Arg), Fe 3+ and Hg 2+ PO4 3 -, and can realize continuous identification of Fe in pure water phase 3+ and PO4 3- 、Hg 2+ and PO4 3- .

[0094] Specifically, when continuous identification of Fe 3+ and Hg 2+ After Fe 3+ After that, triethanolamine was added to Fe 3+ Perform masking.

[0095] The iron-nickel loaded hydroxysodalite fluorescent probe provided by the present invention, its preparation method and application are described in detail below with reference to the examples, but they should not be construed as limiting the scope of protection of the present invention.

[0096] Example 1 Preparation of Hydroxysodite (SOD)

[0097] Dissolve 1.6g, 5.6g, 9.6g, 13.6g, and 17.6g of NaOH solid in 85.95mL of water. Divide each of the five NaOH solutions into two portions, dissolving 0.8197g of NaAlO2 and 7.1055g of Na2SiO3·9H2O, respectively. Slowly add the NaAlO2-NaOH solution to the Na2SiO3-NaOH solution, stirring continuously for 1 hour, to obtain a homogeneous, clear solution. After standing for 0.5 hours, place the solution in a microwave digester and treat at 140°C for 1 hour. Samples are labeled S1 to S5, from least to most NaOH, according to the amount of NaOH used. After the reaction, centrifuge at 6000 rpm and wash the solid with deionized water until neutral.

[0098] The test results for solid samples S1 to S5 are shown in Figure 2. Sample S5 exhibits peaks at 14°, 24.5°, 32°, 34.5°, and 42.5°, consistent with the standard XRD pattern of SOD. Sample S2 exhibits a weak, broad peak, indicating low crystallinity. Samples S1, S3, and S4 exhibit numerous miscellaneous peaks. Therefore, sample S5 was selected for subsequent experiments.

[0099] Example 2 Preparation of Iron-Nickel-Loaded Hydroxysodalite Fluorescent Probe (Probe L)

[0100] Dissolve 0.54 g of FeCl₃·6H₂O in 50 mL of deionized water, then add 0.47538 g of NiCl₂ and dissolve thoroughly to obtain a mixture with an Fe / Ni molar ratio of 1:1. Add 0.54 g of polyethylene glycol-4000 (PEG-4000) to the mixture. Once the PEG has dissolved, add 0.2 g of SOD. Adjust the pH of the solution to between 3 and 4 with HCl, and sonicate for 15 minutes to obtain Solution A.

[0101] Dissolve 3g of Vc in 50mL of deionized water, add 0.03g of NaI, and 2-3 drops of hydrochloric acid, stirring thoroughly to obtain Solution B. Under nitrogen, slowly add Solution B dropwise to Solution A. Once the addition is complete, continue the reaction for 1 hour. Centrifuge the mixture several times, and collect the supernatant to obtain SOD@Fe / Ni. Add 0.3mL of ethylenediamine to this solution under magnetic stirring. Stir continuously for 2 hours. After centrifugation, the supernatant is the amination-modified SOD@Fe / Ni.

[0102] Dissolve 100 mg of fluorescein isothiocyanate (FITC) in 20 mL of N,N-dimethylformamide (DMF). Add 50 mL of a pH 9.2 NaCO and NaHCO buffer solution, mix thoroughly, and then slowly add the solution to the amino-treated SOD@Fe / Ni solution. Incubate at room temperature in the dark for 5 hours. Centrifuge the reaction solution several times at 6000 rpm, and dilute the supernatant 2500-fold with deionized water to obtain probe L.

[0103] Performance Testing

[0104] (1) Probe L fluorescently recognizes Arg in aqueous phase

[0105] Take 2 mL of deionized water in a cuvette, add 20 μL of probe solution, mix thoroughly, and then add 4 μL of Arg (1.0×10 -2 mol / L) solution, mix thoroughly, and detect the fluorescence spectra before and after the addition of Arg. Using the same operation method, detect the fluorescence spectra of the probe + other amino acids (Ile, Asn, Thr, Ser, Pro, Leu, Arg, Val, Met, Glu, Ala, Gly, Phe, Cys).

[0106] The fluorescence spectrum changes of the probe in H2O when it interacts with various amino acids are shown in FIG3 . The inset in FIG3 shows the color of the solution without and with Arg added.

[0107] As shown in Figure 3, the λem of the probe in H2O is 514 nm. -5After adding 1 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 little or no effect on its fluorescence intensity. Therefore, the probe has excellent recognition of Arg in H₂O. Irradiating the probe solution with 365nm light from a portable UV lamp reveals a clear color difference between the probe solution and the probe+Arg solution (Figure 3, inset), indicating that the solution fluorescence changes from weak to light green.

[0108] Take 2 mL of deionized water in a cuvette, add 20 μL of probe solution, 4 μL of Cys solution, and 4 μL of Arg solution, and stir thoroughly to obtain a probe + Cys + Arg solution. Use the same operation method to obtain probe + other amino acids + Arg solutions for fluorescence spectrum detection.

[0109] Figure 4 shows the fluorescence intensity of the probe in the presence of Arg and other amino acids in H₂O. Comparing the fluorescence intensity at 514 nm for the probe + other amino acids + Arg and the probe + Arg, Figure 4 shows that, with the exception of Glu and Asn, which significantly decrease the fluorescence intensity, the fluorescence intensity of the solution increases or decreases slightly when the other amino acids coexist, indicating that the presence of other amino acids has little effect on the interaction between the probe and Arg. Therefore, in H₂O, the interaction between the probe and Arg exhibits good anti-interference properties.

[0110] Take 2 mL of deionized water in a cuvette, add 20 μL of probe solution, and use the Arg cumulative increase method to make the cumulative injection volume of Arg solution 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, stir thoroughly and mix well before performing fluorescence spectrum detection to keep the fluorescence intensity of the solution stable.

[0111] The fluorescence titration spectra of the probe and Arg in H2O are shown in Figure 5. As shown in Figure 5, with the increase of Arg concentration, the fluorescence intensity of the solution at 514nm continues to increase. Further analysis and research, with Arg concentration as the horizontal axis and the fluorescence intensity of the solution at 514nm as the vertical axis, at 4.0×10 -6 mol / L~3.2×10 -5 mol / L range, the linear relationship between the two is shown in Figure 6, where R 2The value is 0.9964, indicating that the linear fit of the detection limit is good and the reliability is high. According to the detection limit calculation formula: LOD = 3δ / k (δ is the standard deviation of the fluorescence intensity of the blank solution scanned three times at 514nm; k is the absolute value of the slope of the linear fitting equation), the LOD of the probe for Arg in H2O is obtained as 7.7347×10 -8 mol / L.

[0112] In the above experiments, the recognition, interference, and detection limit experiments of the probe on amino acids in H2O were all carried out under the detection conditions of an excitation wavelength of 350 nm and a slit width of 5 nm.

[0113] (2) Identification of Fe in H2O by fluorescence spectroscopy 3+ 、Hg 2+

[0114] Take 2 mL of deionized water in a cuvette, add 20 μL of probe solution, stir thoroughly, and perform fluorescence spectrum detection to obtain the fluorescence spectrum of the probe in H2O. Then add 2 μL of Fe 3+ The solution was stirred and mixed thoroughly to allow the reaction to complete and then the fluorescence spectrum was detected again to obtain the probe + Fe 3+ The same operation method was used to detect the fluorescence spectra of the probe + other cations in turn.

[0115] The fluorescence spectra changes of the probe in H2O when it interacts with various cations are shown in Figure 7. The inset in Figure 7 shows the fluorescence spectra of the probe when it is not added and when it is added with Fe. 3+ and Hg 2+ Solution color.

[0116] As shown in Figure 7, the λem of the probe in H2O is 514 nm. -5 mol / L Fe 3+ After adding 1×10 -5 mol / L Hg 2+ After ionization, the fluorescence intensity of the solution decreased by 3.0 times, and the fluorescence of the solution changed from weak fluorescence to extremely weak fluorescence. + 、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+ 、Zr4+ ) has little or no effect on its fluorescence intensity. 3+ 、Hg 2+ It has better identification function.

[0117] Take 2mL of deionized water in a cuvette, add 20μL of probe solution, 2μL of Fe 3+ solution, 1.6 μL TEA (triethanolamine) solution (diluted 100 times), 2 μL Hg 2+ After adding each solution, the mixture was stirred thoroughly and fluorescence spectrum detection was performed.

[0118] Probe Fe in H2O 3+ and Hg 2+ The fluorescence spectrum changes under different identification are shown in Figure 8. As shown in Figure 8, when Fe 3+ After that, the fluorescence intensity of the solution decreased significantly, and TEA was then added to mask Fe 3+ , the fluorescence intensity of the solution returned to its initial position, and finally Hg 2+ , the fluorescence intensity of the solution decreases again. Therefore, the probe can recognize Fe 3+ After the masking operation, Hg was identified again. 2+ , further achieving the goal of optimizing probe selectivity.

[0119] Take 2mL of deionized water in a cuvette, add 20μL of probe solution, 2μL of Ag + solution, 2 μL Fe 3+ Solution, stir and mix thoroughly to obtain probe + Ag + +Fe 3+ Solution, using the same operation method, respectively obtain probe + other cations + Fe 3+ Fluorescence spectrum detection was performed on the solution.

[0120] The probe in H2O is Fe 3+ The fluorescence intensity when coexisting with other cations is shown in Figure 9. In Figure 9, the fluorescence intensity of the probe + other cations + Fe 3+ With probe+Fe 3+ The fluorescence intensity at 514 nm was compared, and it was found that when other cations coexisted, the fluorescence intensity of the solution increased or decreased slightly, indicating that the presence of other cations had a significant effect on the interaction between the probe and Fe. 3+ Therefore, in H2O, the probe and Fe 3+ The effect of both has good anti-interference performance.

[0121] Take 2mL of deionized water in a cuvette, add 20μL of probe solution, and use Fe 3+ Cumulative increase method, Fe3+ The cumulative injection volumes of the solution were 0.1μL, 0.2μL, 0.3μL, 0.4μL, 0.5μL, 0.6μL, and 0.7μL, respectively. 3+ After the solution is prepared, it is stirred thoroughly and then the fluorescence spectrum is detected to keep the fluorescence intensity of the solution stable.

[0122] Probe in H2O and Fe 3+ The fluorescence titration spectrum of the probe is shown in Figure 10. 3+ The linear relationship is shown in Figure 11.

[0123] As shown in Figures 10 and 11, 3+ The concentration is 5.0×10 -7 mol / L~3.5×10 -6 mol / L range, the linear relationship between the two is shown in Figure 11, where R 2 The probe in H2O has a relative affinity to Fe 3+ LOD = 2.4912 × 10 -8 mol / L.

[0124] (3) Identification of Hg 2+

[0125] Other ions coexisting on Hg 2+ The impact of the detection method is the same as Fe 3+ Other cation pair probes recognize Hg 2+ The interference is shown in Figure 12.

[0126] Take 2mL of deionized water in a cuvette, add 20μL of probe solution, and use Hg 2+ Cumulative increase method, Hg 2+ The cumulative injection volumes of the solution were 0.3 μL, 0.4 μL, 0.5 μL, 0.6 μL, and 0.8 μL, respectively. 2+ After the solution is fully stirred and mixed, the fluorescence spectrum is detected to keep the fluorescence intensity of the solution stable. 2+ The fluorescence titration spectrum of the probe is shown in Figure 13. The fluorescence intensity of the probe at 514 nm in H2O is similar to that of Hg 2+ The linear relationship graph of the concentrations is shown in FIG14 .

[0127] As shown in Figures 13 and 14, in the case of Hg 2+ The concentration is 1.5×10 -6 mol / L~4.0×10 -6 mol / L range, where R 2 The probe in H2O is 0.9870 for Hg 2+LOD = 4.7335 × 10 -8 mol / L.

[0128] The recognition, interference, and detection limit experiments of the probe for cations in H2O were all carried out under the detection conditions of an excitation wavelength of 350 nm and a slit width of 10 nm.

[0129] (4) Identification of PO4 in H2O by fluorescence spectroscopy 3-

[0130] Take 2mL of deionized water in a cuvette, add 20μL of probe solution, stir thoroughly, and perform fluorescence spectrum detection to obtain the fluorescence spectrum of the probe in H2O. Then add 4μL of PO4 3- (1.0×10 -2 mol / L) solution, stir and mix thoroughly, make it react completely and then perform fluorescence spectrum detection again to obtain probe+PO4 3- The same operation method was used to detect the fluorescence spectra of the probe + other anions in turn.

[0131] The fluorescence spectra changes of the probe in H2O when it interacts with various anions are shown in Figure 15. The inset in Figure 15 is PO4 3- As shown in Figure 15, the λem of the probe in H2O is 514 nm. -5 mol / L, the fluorescence intensity of the solution increased by 2.0 times, and the solution fluorescence changed from weak fluorescence to light green fluorescence. - 、SO3 2- 、SO4 2- 、S2O3 2- 、SCN - , HS - 、S2 - 、NO2 - , I - 、AlO2 - 、CH3COO - 、Cl - 、HCO3 - 、HPO4 2- ) has little or no effect on its fluorescence intensity. Therefore, in H2O, the probe has little or no effect on PO4 3- It has better identification function.

[0132] Take 2mL of deionized water in a cuvette, add 20μL of probe solution, 4μL of HCO3 - solution, 4 μL PO4 3- Solution, stir and mix thoroughly to obtain probe + HCO3 - +PO43- Solution, using the same operation method, respectively obtain probe + other anions + PO4 3- Fluorescence spectrum detection was performed on the solution.

[0133] The probe in H2O is in PO4 3- The fluorescence intensity when coexisting with other anions is shown in Figure 16. In Figure 16, the fluorescence intensity of the probe + other anions + PO4 3- With probe+PO4 3- The fluorescence intensity at 514 nm was compared and it was found that when other anions coexisted, the fluorescence intensity of the solution increased or decreased slightly, indicating that the presence of other anions had a significant effect on the interaction between the probe and PO4. 3- Therefore, in H2O, the probe and PO4 3- The effect of both has good anti-interference performance.

[0134] Take 2mL of deionized water in a cuvette, add 20μL of probe solution, and use PO4 3- Cumulative increase method, so that PO4 3- The cumulative injection volumes of the solutions were 0.2μL, 0.4μL, 0.6μL, 0.8μL, 1.2μL, 1.6μL, 2.0μL, 2.4μL, and 2.8μL, and PO4 was added each time. 3- After the solution is prepared, it is stirred thoroughly and then the fluorescence spectrum is detected to keep the fluorescence intensity of the solution stable.

[0135] At PO4 3- The concentration is 0~1.4×10 -5 mol / L range, the change of the fluorescence intensity of the probe is shown in Figure 17. 3- The concentration is 1.0×10 -6 mol / L~1.4×10 -5 In the range of mol / L, the linear relationship between the two is shown in Figure 18, where R 2 The probe in H2O is 0.9952 for PO4 3- LOD = 4.8175 × 10 -8 mol / L.

[0136] The recognition, interference, and detection limit experiments of the probe on 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.

[0137] (5) Probe + Fe 3+ Identification of PO4 in H2O by Fluorescence Spectroscopy 3-

[0138] Take 2mL of deionized water in a cuvette, add 20μL of probe solution, 2μL of Fe3+ The solution was stirred thoroughly and the fluorescence spectrum was detected to obtain the probe + Fe 3+ Fluorescence spectrum in H2O. Then 20μL PO4 was added to the cuvette. 3- The solution was stirred and mixed thoroughly to allow the reaction to complete and then the fluorescence spectrum was detected again to obtain the probe + Fe 3+ +PO4 3- The same operation method was used to detect the fluorescence spectrum of probe+Fe 3+ + Fluorescence spectra of other anions.

[0139] Probe+Fe in H2O 3+ Identify PO4 3- The spectrum is shown in Figure 19. As shown in Figure 19, the probe+Fe 3+ The λem is 514nm, adding 1×10 -4 mol / L PO4 3- After that, the fluorescence intensity of the solution increased by 3.3 times, and the fluorescence of the solution changed from extremely weak fluorescence to light green fluorescence. - 、SO3 2- 、SO4 2- 、S2O3 2- 、SCN - , HS - 、S2 - 、NO2 - , I - 、AlO2 - 、CH3COO - 、Cl - 、HCO3 - 、HPO4 2- ) has little or no effect on its fluorescence intensity. 3+ PO4 3- Has good identification function.

[0140] Other anion pair probes + Fe 3+ Identify PO4 3- The interference of is shown in Figure 20. In Figure 20, the probe + Fe 3+ + other anions + PO4 3- With probe+Fe 3+ +PO4 3- The fluorescence intensity comparison at 514nm shows that, except for S2 - 、AlO2 - The presence of HS makes the fluorescence intensity of the solution increase significantly; -The presence of Fe+Fe makes the fluorescence intensity of the solution decrease significantly. When other anions coexist, the fluorescence intensity of the solution increases or decreases slightly, indicating that the presence of other anions has a significant effect on the probe+Fe 3+ With PO4 3- Therefore, in H2O, the probe + Fe 3+ With PO4 3- The effect of both has good anti-interference performance.

[0141] Take 2mL of deionized water in a cuvette, add 20μL of probe solution, 2μL of Fe 3+ Solution, stir and mix, use PO4 3- Cumulative increase method, so that PO4 3- The cumulative injection volumes of the solutions were 4 μL, 5 μL, 6 μL, 7 μL, 8 μL, 9 μL, and 10 μL, and PO4 was added each time. 3- After the solution is prepared, it is stirred thoroughly and then the fluorescence spectrum is detected to keep the fluorescence intensity of the solution stable.

[0142] Probe+Fe 3+ With PO4 3- The fluorescence titration spectrum of the probe+Fe 3+ With PO4 3- The linear relationship graph of the concentration is shown in FIG22 .

[0143] At PO4 3- The concentration is 2.0×10 -5 mol / L~5.0×10 -5 mol / L range, the linear relationship between the two is shown in Figure 22, where R 2 0.9863, probe+Fe in H2O 3+ PO4 3- LOD = 2.7807 × 10 -7 mol / L.

[0144] Probe+Fe in H2O 3+ The identification, interference, and detection limit experiments of anions were all carried out under the detection conditions of an excitation wavelength of 350 nm and a slit width of 10 nm.

[0145] (6) Probe + Hg 2+ Identification of PO4 in H2O by Fluorescence Spectroscopy 3-

[0146] Take 2mL of deionized water in a cuvette, add 20μL of probe solution, 2μL of Hg 2+ The solution was stirred thoroughly and the fluorescence spectrum was detected to obtain the probe + Hg 2+Fluorescence spectrum in H2O. 60 μL PO4 was then added to the cuvette. 3- The solution was stirred and mixed thoroughly to allow it to react completely and then the fluorescence spectrum was detected again to obtain the probe + Hg 2+ +PO4 3- The same operation method was used to detect the probe+Hg 2+ + Fluorescence spectra of other anions.

[0147] Probe+Hg 2+ Identify PO4 3- The spectrum of other anions is shown in Figure 23. 2+ Identify PO4 3- The interference is shown in Figure 24.

[0148] In Figure 24, the probe + Hg 2+ + other anions + PO4 3- With probe+Hg 2+ +PO4 3- The fluorescence intensity comparison at 514nm shows that, except for HCO3 - 、NO2 - 、S2 - The presence of Hg+ makes the fluorescence intensity of the solution increase significantly. When other anions coexist, the fluorescence intensity of the solution increases or decreases slightly, indicating that the presence of other anions has a significant effect on the probe 2+ With PO4 3- Therefore, in H2O, the probe + Hg 2+ With PO4 3- The effect of both has good anti-interference performance.

[0149] Take 2mL of deionized water in a cuvette, add 20μL of probe solution, 2μL of Hg 2+ Solution, stir and mix, use PO4 3- Cumulative increase method, so that 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, and PO4 was added each time. 3- After the solution is prepared, it is stirred thoroughly and then the fluorescence spectrum is detected to keep the fluorescence intensity of the solution stable.

[0150] PO4 3- Probe+Hg 2+ The effect of fluorescence intensity is shown in Figure 25. 3- The concentration is 3.0×10 -5 mol / L~9.0×10 -5 mol / L range, the linear relationship between the two is shown in Figure 26, where R2 0.9839, probe+Hg in H2O 2+ PO4 3- LOD = 3.1952 × 10 -7 mol / L.

[0151] Probe in H2O+Hg 2+ The identification, interference, and detection limit experiments of anions were all carried out under the detection conditions of an excitation wavelength of 350 nm and a slit width of 10 nm.

[0152] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A hydroxysodalite fluorescent probe loaded with iron and nickel, comprising hydroxysodalite, iron element, nickel element and ethylenediamine loaded on the surface of the hydroxysodalite, and fluorescein isothiocyanate chemically connected to the ethylenediamine.

2. The iron-nickel loaded hydroxysodalite fluorescent probe according to claim 1, characterized in that: The N atom in the ethylenediamine is connected to the iron element and the nickel element via a metal coordination bond.

3. The iron-nickel loaded hydroxysodalite fluorescent probe according to claim 1, characterized in that: The molar ratio of the iron element to the nickel element is 1:

1.

4. The iron-nickel loaded hydroxysodalite fluorescent probe according to claim 1 or 3, characterized in that: The particle size of the iron-nickel loaded hydroxysodalite fluorescent probe is 0.1-2 μm.

5. The method for preparing the iron-nickel loaded hydroxysodalite fluorescent probe according to any one of claims 1 to 4, comprising the following steps: Mixing a soluble ferric iron source, a soluble nickel source, a surfactant, hydroxysodalite and water, and adjusting the pH value of the obtained mixed solution to 3-4 to obtain a mixed solution; The mixed solution is mixed with a reducing agent to perform a reduction reaction to obtain a sodalite dispersion loaded with iron and nickel; The iron-nickel loaded sodalite dispersion is mixed with ethylenediamine to carry out an amination reaction to obtain the amination iron-nickel loaded sodalite; The aminated iron-nickel loaded sodalite is mixed with a fluorescein isothiocyanate solution and subjected to an addition reaction to obtain an iron-nickel loaded hydroxysodalite fluorescent probe.

6. The preparation method according to claim 5, characterized in that: The preparation method of hydroxysodalite comprises the following steps: NaAlO2, Na2SiO3, sodium hydroxide and water are mixed, and the mixture is stirred, allowed to stand and microwave digested in sequence to obtain hydroxysodalite.

7. The preparation method according to claim 6, characterized in that: The mass ratio of NaOH, NaAlO2, Na2SiO3·9H2O and water is 1.95-21.5:1:8.67:100-200.

8. The preparation method according to claim 6, characterized in that: The stirring time is 1 to 3 hours; the standing time is 0.5 to 1 hour; the microwave digestion temperature is 140 to 160° C., and the time is 1 to 3 hours.

9. The preparation method according to claim 5, characterized in that: The molar ratio of Fe / Ni in the soluble ferric iron source and the soluble nickel source is 1:1; The mass ratio of the soluble ferric iron source to hydroxysodalite is 1 to 3:1; The mass ratio of the soluble nickel source to hydroxysodalite is 0.88-2.64:

1.

10. The preparation method according to claim 5 or 6, characterized in that: The surfactant is polyethylene glycol, and the mass ratio of the surfactant to hydroxysodalite is 1 to 3:

1.

11. The preparation method according to claim 5, characterized in that: The reducing agent includes 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 0.10-0.25:1; The reduction reaction time is 1 to 3 hours.

12. The preparation method according to claim 5, characterized in that: The mass ratio of the ethylenediamine to hydroxysodalite is 1 to 2:

1.

13. The preparation method according to claim 5 or 12, characterized in that: The amination reaction time is 1 to 3 hours.

14. The preparation method according to claim 5, characterized in that: The preparation method of the fluorescein isothiocyanate solution comprises the following steps: Fluorescein isothiocyanate was mixed with N,N-dimethylformamide, and Na2CO3 and NaHCO3 buffer were added to obtain a fluorescein isothiocyanate solution.

15. The preparation method according to claim 5 or 14, characterized in that: The mass ratio of fluorescein isothiocyanate to aminated iron-nickel loaded sodalite is 1:1-5; The addition reaction is carried out under light-proof conditions, and the time of the addition reaction is 5 to 8 hours.

16. The Fe-Ni-loaded hydroxysodalite fluorescent probe according to any one of claims 1 to 4 or the Fe-Ni-loaded hydroxysodalite fluorescent probe prepared by the preparation method according to any one of claims 5 to 15 is effective in detecting L-arginine, Fe 3+ , Hg 2+ PO4 3- One or more of the applications.

17. The use according to claim 16, characterized in that When continuous detection of Fe 3+ and Hg 2+ After Fe 3+ After that, triethanolamine was added to Fe 3+ Perform masking.

18. A method for detecting L-arginine, Fe 3+ , Hg 2+ or PO4 3- The kit comprises the iron-nickel-loaded hydroxysodalite fluorescent probe according to any one of claims 1 to 4 or the iron-nickel-loaded hydroxysodalite fluorescent probe prepared by the preparation method according to any one of claims 5 to 15.

Images