Europium metal organic gel as well as preparation method and application thereof

Abstract

The invention specifically discloses europium metal organic gel as well as a preparation method and application thereof, and relates to the technical field of functional materials and biomedicine. The invention provides europium metal organic gel, and the europium metal organic gel comprises a europium-containing compound and 3, 5-dicarboxyphenylboronic acid. The europium metal organic gel provided by the invention can be used for specifically detecting indoxyl sulfate in a complex sample, and is efficient and economical.

Description

Technical Field

[0001] The present invention relates to the technical fields of functional materials and biomedical technologies, and particularly relates to a europium metal-organic gel and a preparation method and application thereof. Background Art

[0002] 1. Biological Significance of IS

[0003] Indoxyl sulfate (IS) is a precursor of uremic toxin produced by the metabolism of tryptophan by intestinal microorganisms. Under physiological conditions, the content of indoxyl sulfate in serum is relatively low and can be excreted through the kidneys. However, in patients with chronic kidney disease (CKD), the accumulation of IS caused by impaired renal function leads to vascular calcification, inflammatory reactions, and renal cell damage. Its serum concentration is positively correlated with the progression of CKD and is a key marker for early diagnosis.

[0004] 2. Bottlenecks of Existing Detection Technologies

[0005] (1) High-performance liquid chromatography (HPLC): It requires complex pretreatment, expensive equipment, and is difficult to popularize;

[0006] (2) Liquid chromatography-mass spectrometry (LC-MS / MS): It has high sensitivity but requires professional operation and high maintenance costs;

[0007] (3) Enzyme-linked immunosorbent assay (ELISA): It is easily interfered by cross-reactions and has insufficient accuracy;

[0008] (4) Common Defects: None of them can achieve real-time visual detection, rely on large-scale instruments, and are difficult to meet the requirements of point-of-care testing (POCT).

[0009] 3. Technical Gap and Innovation Direction

[0010] There is an urgent need to develop a detection method based on novel functional materials, which has the characteristics of high sensitivity, anti-interference, visual output, and low cost, and is suitable for the rapid detection of IS in complex media such as serum and urine. Summary of the Invention

[0011] (I) Technical Problems to be Solved

[0012] In view of this, one of the main purposes of the present invention is to provide a europium metal-organic gel, and the europium metal-organic gel includes a europium-containing compound and 3,5-dicarboxylphenylboronic acid. The europium metal-organic gel provided by the present invention can specifically detect indoxyl sulfate in complex samples and is efficient and economical.

[0013] (II) Technical Solutions

[0014] To achieve the above object, the present invention provides a europium metal-organic gel, and the europium metal-organic gel includes a europium-containing compound and 3,5-dicarboxylphenylboronic acid.

[0015] In one embodiment, the europium-containing compound includes one or a combination of halides, chalcogenides, oxygen-containing salts, or organoeuropium compounds.

[0016] In one embodiment, the europium-containing compound is an oxygen-containing salt.

[0017] In one embodiment, the oxygen-containing salt includes one or a combination of europium sulfate, europium nitrate, europium phosphate, or europium carbonate.

[0018] In one embodiment, the oxygen-containing salt is europium nitrate.

[0019] In one embodiment, the molar ratio of the europium nitrate (Eu(NO3)3·6H2O) to 3,5-dicarboxylphenylboronic acid is (0.1 - 10):1.

[0020] In one embodiment, the molar ratio of the europium nitrate to 3,5-dicarboxylphenylboronic acid is 6:5.

[0021] On the other hand, the present invention provides a method for preparing the above-mentioned europium metal-organic gel, and the preparation method includes:

[0022] S1: Dissolve the europium-containing compound in solvent A to prepare solution A, and dissolve 3,5-dicarboxylphenylboronic acid in solvent B to prepare solution B;

[0023] S2: Mix solution A and solution B to react to obtain the europium metal-organic gel.

[0024] In one embodiment, the solvent A is DMF.

[0025] In one embodiment, the dosage of the europium nitrate is 0.2787 g.

[0026] In one embodiment, the dosage of the DMF is 1 mL.

[0027] In one embodiment, the solvent B is DMF.

[0028] In one embodiment, the dosage of the 3,5-dicarboxylphenylboronic acid is 0.1049 g.

[0029] In one embodiment, the dosage of the DMF is 1 mL.

[0030] In one embodiment, the reaction condition is ultrasonic wave, and the reaction time is 30 min.

[0031] In one embodiment, the preparation method specifically includes:

[0032] S1: Dissolve europium nitrate in DMF to prepare solution A, and dissolve 3,5-dicarboxylphenylboronic acid in DMF to prepare solution B;

[0033] S2: Mix solution A and solution B and react to obtain the europium metal-organic gel.

[0034] In one embodiment, the preparation method specifically includes:

[0035] S1: Dissolve 0.2787 g of europium nitrate in 1 mL of DMF to prepare solution A, and dissolve 0.1049 g of 3,5-dicarboxylphenylboronic acid in 1 mL of DMF to prepare solution B;

[0036] S2: Mix solution A and solution B, and react for 30 min under ultrasonic conditions to obtain the europium metal-organic gel.

[0037] In one embodiment, triethylamine is further added in S2.

[0038] In one embodiment, the dosage of triethylamine is 230 μL.

[0039] In one embodiment, the preparation method specifically includes:

[0040] S1: Dissolve europium nitrate in DMF to prepare solution A, and dissolve 3,5-dicarboxylphenylboronic acid in DMF to prepare solution B;

[0041] S2: Mix solution A and solution B, add triethylamine and react to obtain the europium metal-organic gel.

[0042] In one embodiment, the preparation method specifically includes:

[0043] S1: Dissolve 0.2787 g of europium nitrate in 1 mL of DMF to prepare solution A, and dissolve 0.1049 g of 3,5-dicarboxylphenylboronic acid in 1 mL of DMF to prepare solution B;

[0044] S2: Mix solution A and solution B, react for X min under ultrasonic conditions, add 230 μL of triethylamine and react for 30 min under ultrasonic conditions to obtain the europium metal-organic gel.

[0045] On the other hand, the present invention also provides a europium metal-organic gel, which is obtained by the above preparation method.

[0046] On the other hand, the present invention also provides a capsule, which includes the above europium metal-organic gel.

[0047] On the other hand, the present invention also provides a fluorescence sensing material, which includes the above europium metal-organic gel.

[0048] The present invention also provides, in another aspect, a fluorescent test strip, which contains the above-mentioned europium metal-organic gel or fluorescent sensing material.

[0049] The present invention also provides the use of the above-mentioned europium metal-organic gel, capsule, fluorescent sensing material or fluorescent test strip in detecting indoxyl sulfate.

[0050] In one embodiment, the detection only involves detection for non-diagnostic purposes.

[0051] In one embodiment, the detection specifically includes contacting the above-mentioned europium metal-organic gel, capsule, fluorescent sensing material or fluorescent test strip with a sample and observing the change in fluorescence color.

[0052] In one embodiment, the sample contains indoxyl sulfate.

[0053] In one embodiment, the sample does not contain indoxyl sulfate.

[0054] The present invention also provides the use of the above-mentioned europium metal-organic gel, capsule, fluorescent sensing material or fluorescent test strip in preparing a reagent for detecting indoxyl sulfate.

[0055] Advantages

[0056] The present invention provides a europium metal-organic gel, a preparation method thereof and applications. Compared with the prior art, the following advantages are achieved:

[0057] 1. Rapid synthesis at room temperature: Without high temperature and high pressure, the process is simple and controllable;

[0058] 2. Instant visual detection: It can be judged with the naked eye or a portable ultraviolet lamp, without the need for complex instruments;

[0059] 3. Strong clinical applicability: It can directly detect serum / urine samples without pretreatment;

[0060] 4. Significant cost-effectiveness: The cost of material synthesis and detection reagent is reduced by more than 90% compared with LC-MS / MS. Description of the Drawings

[0061] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for use in the description of the embodiments or the prior art. Obviously, the following drawings are only some embodiments of the present invention. For those of ordinary skill in the art, other drawings can be obtained based on these drawings without creative efforts.

[0062] Figure 1: Schematic diagram of Eu-MOG synthesis at room temperature (including reaction conditions and gel formation process); (a) Eu-MOG that failed to form before adding triethylamine; (b) Inverted Eu-MOG; (c) Prepared Eu-MOG capsules; (d) Eu-MOG capsules under ultraviolet light.

[0063] Figure 2 : Infrared and X-ray photoelectron (XPS) spectra of Eu-MOG material; (a) Infrared spectra of Eu-MOG and 5-bop. The peak of the ligand at 1701 cm -1 corresponding to the C=O stretching vibration of the ligand disappears, indicating that the carboxyl group coordinates with Eu 3+ to form Eu-MOG. The absorption peak at 1331 cm -1 originating from the boric acid group can be observed in both 5-bop and Eu-MOG, indicating the successful synthesis of boric acid-functionalized Eu-MOG. The absorption peak at 1598 cm -1 is the stretching vibration of C=C on the benzene ring; (b) XPS spectrum of Eu-MOG; Elements Eu, O, N, C, and B can be seen in the XPS survey spectrum of Eu-MOG, which exactly proves the successful synthesis of Eu-MOG.

[0064] Figure 3 : Fluorescence spectrum of Eu-MOG; Excitation spectrum (gray), emission spectrum (red).

[0065] Figure 4 : Dissolution process of the capsule in artificial urine.

[0066] Figure 5 : Fluorescent color change of the capsule with the change of IS concentration in artificial urine.

[0067] Figure 6 : Fluorescent sensing diagram of the capsule for different analytes in artificial urine.

[0068] Figure 7 : Fluorescent diagram of the capsule in artificial urine when other ions, amino acids, and IS coexist.

[0069] Figure 8 : Linear relationship diagram of the fluorescence intensity of the capsule with the IS concentration in artificial urine

[0070] Figure 9 : Fluorescent sensing diagram of the capsule for different analytes in serum.

[0071] Figure 10 : Fluorescent diagram of the capsule in serum when other metal ions, amino acids, and IS coexist.

[0072] Figure 11 : Linear relationship diagram of the fluorescence intensity of the capsule with the IS concentration in serum.

[0073] Figure 12 : Detection mechanism diagram of the capsule for IS.

[0074] Figure 13 : Gel states at different ratios of europium-containing compounds and 3,5-dicarboxylphenylboronic acid. Detailed implementation manners

[0075] To make the objectives, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention. Apparently, the described embodiments are some but not all of the embodiments of the present invention. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0076] Terms and definitions

[0077] As used herein, the terms "sample" and "biological sample" refer to a composition obtained from or derived from a target subject, which contains cell entities and / or other molecular entities to be characterized and / or identified, for example, based on physical, biochemical, chemical, and / or physiological characteristics. The sample can be obtained from the subject's blood and other fluid samples and tissue samples of biological origin, such as biopsy tissue samples or tissue cultures or cells derived therefrom. The source of the tissue sample can be solid tissue, such as from fresh, frozen, and / or preserved organ or tissue samples, biopsy tissues, or aspirates; blood or any blood component; body fluid; cells at any time during an individual's pregnancy or development; or plasma.

[0078] In one embodiment, it includes but is not limited to whole blood, plasma, serum, red blood cell fraction, saliva, cerebrospinal fluid, semen, feces, urine, rectal swab, bile, lymph, sputum, lavage fluid, or a combination thereof.

[0079] As used herein, the term "subject" includes mammals and non-mammals.

[0080] Examples of mammals include but are not limited to any member of the class Mammalia: humans, non-human primates such as chimpanzees and other apes and monkeys; farm animals such as cows, horses, sheep, goats, pigs; domestic animals such as rabbits, dogs, and cats; laboratory animals, including rodents such as rats, mice, and guinea pigs, etc. Examples of non-mammals include but are not limited to birds, fish, or other non-mammals, etc.

[0081] As used herein, "comprising", "having", or "including" includes "containing", "consisting essentially of", "substantially consisting of", and "consisting of"; "consisting essentially of", "substantially consisting of", and "consisting of" are subordinate concepts of "comprising", "having", or "including".

[0082] The experimental methods used in the following examples are conventional methods unless otherwise specified, and the reagents, methods, and equipment used are conventional reagents, methods, and equipment in the technical field unless otherwise specified.

[0083] Example 1

[0084] Preparation of capsules at room temperature:

[0085] First, weigh 0.2787 g of Eu(NO3)3·6H2O and dissolve it in 1 mL of DMF, and ultrasonically dissolve for 3 minutes to obtain solution A; then weigh 0.1049 g of 3,5-dicarboxylphenylboronic acid in 1 mL of DMF, and continue to ultrasonically dissolve for 3 minutes to obtain solution B; mix solution A and B, ultrasonically treat for 30 minutes, add 230 μL of triethylamine dropwise, shake in one direction for 10 minutes, continue to ultrasonically treat for half an hour, and then stand for 2 h to obtain an inverted gel.

[0086] Wash the inverted gel with DMF, centrifuge at 8000 r for 3 minutes, repeat centrifugation five times, and then freeze-dry at -101 °C in a freeze dryer for 12 h to obtain a freeze-dried powder. Weigh 5 mg of the powder into a capsule shell to obtain the capsule.

[0087] Figure 1 It is a physical diagram of the gel formation process and the capsule preparation process. Figure 2 It is the fluorescence spectrum of Eu-MOG solid. It can be seen from the figure that an inverted gel could not be formed before adding triethylamine. After adding triethylamine, deprotonation occurred, facilitating the self-assembly between the ligand and the metal to form an inverted gel, and obvious red light could be observed under ultraviolet light. Figure 3 It is the infrared and XPS spectra of Eu-MOG. In Eu-MOG, the peak of the ligand at 1701 cm -1 corresponding to the C=O stretching vibration of the ligand disappeared, indicating that the carboxyl group coordinated with Eu 3+ to form Eu-MOG. The absorption peak at 1331 cm -1 derived from the boric acid group was observed in both 5-bop and Eu-MOG, indicating the successful synthesis of boric acid-functionalized Eu-MOG. The absorption peak at 1598 cm -1 is the stretching vibration of C=C on the benzene ring. The elements Eu, O, N, C, and B can be seen in the XPS survey spectrum of Eu-MOG, which exactly proves the successful synthesis of Eu-MOG.

[0088] Example 2

[0089] Urine sample detection:

[0090] Place one capsule prepared in Example 1 in a cuvette and add 4 mL of artificial urine (the concentration of IS is 5×10 -3 mol / L) (Regen Biotech, CZ0350), let it stand still. After 10 minutes, observe the dissolution of the capsule and perform ultrasonic treatment for 2 - 3 minutes.

[0091] From Figure 4 It can be observed that the capsule dissolves and releases the fluorescent substance, and the fluorescent color of the mixture gradually changes from red to blue.

[0092] Place the capsule prepared in Example 1 in 10 mL of artificial urine (the concentrations of IS from left to right are 9×10 - 3 mol / L, 5×10 -4 mol / L, 9×10 -5 mol / L, 1×10 -5 mol / L and 1×10 -6 mol / L) (Regen Biotech, CZ0350), and further detect whether there is a macroscopic color change with the change of the IS concentration.

[0093] From Figure 5 It can be observed that with the decrease of the concentration, the color of the solution gradually changes from blue to red.

[0094] According to the above results, it can be found that the capsule solution can dissolve well and there are obvious color differences.

[0095] Next, add L-lysine, L-phenylalanine, L-methionine, L-threonine, L-isoleucine, L-leucine, L-valine, L-histidine, L-alanine, L-arginine, L-asparagine, L-aspartic acid, L-cysteine, L-glutamic acid, L-glutamine, L-glycine, L-proline, L-serine, L-tyrosine, indoxyl sulfate, magnesium chloride, calcium chloride, sodium chloride and glucose to 6 mL of artificial urine (Regen Biotech, CZ0350) solution respectively, so that the solution concentration is 3.32×10 -6 mol / mL, and then add one capsule prepared in Example 1 to each of them respectively.

[0096] From Figure 6 It can be seen that IS can significantly enhance the fluorescence emission peak of the capsule solution at 393 nm.

[0097] After the above operations, except for the indoxyl sulfate solution and the blank sample, add 5 mg of indoxyl sulfate to the other solutions and continue the fluorescence sensing experiment. Figure 7 It shows that the detection effect of IS will not be affected by the interference of other substances in urine.

[0098] From Figure 8 It can be seen that when the concentration of IS is 1×10 -6 -9×10 -6 mol / L, with the increase of the concentration, the fluorescence intensity at 393 nm can be seen to increase, which indicates that the fluorescence probe of the capsule has certain feasibility for the detection of IS in urine in real life. The detection limit is calculated to be 43.61 ppm.

[0099] Example 3

[0100] Serum sample detection:

[0101] Add L-lysine, L-phenylalanine, L-methionine, L-threonine, L-isoleucine, L-leucine, L-valine, L-histidine, L-alanine, L-arginine, L-asparagine, L-aspartic acid, L-cysteine, L-glutamic acid, L-glutamine, L-glycine, L-proline, L-serine, L-tyrosine, indoxyl sulfate, magnesium chloride, calcium chloride, sodium chloride and glucose to 6 mL of serum (Adamas life, C8142-100 mL) respectively, so that the solution concentration is 3.32×10 -6 mol / mL, and then add one capsule prepared in Example 1 to each of them respectively.

[0102] From Figure 9 It can be seen that IS can enhance the fluorescence emission peak of the capsule solution at 393 nm.

[0103] Add 5 mg of indoxyl sulfate to 3.32×10 -6 mol / mL of L-lysine, L-phenylalanine, L-methionine, L-threonine, L-isoleucine, L-leucine, L-valine, L-histidine, L-alanine, L-arginine, L-asparagine, L-aspartic acid, L-cysteine, L-glutamic acid, L-glutamine, L-glycine, L-proline, L-serine, L-tyrosine, magnesium chloride, calcium chloride, sodium chloride and glucose serum solutions respectively, and conduct fluorescence sensing experiments under the coexistence conditions of IS and the above substances.

[0104] Figure 10 It shows that the fluorescence effect of the capsule in serum is basically the same as that in urine solution.

[0105] From Figure 11It can be seen that when the concentration of IS is 1×10 -6 -9×10 -4 mol / L, with the increase of the concentration, it can be seen that the fluorescence intensity at 393 nm increases, indicating that the fluorescence probe of the capsule is feasible, convenient and sensitive for the detection of IS in serum. The detection limit is calculated to be 43.61 ppm.

[0106] Example 4

[0107] Verification of detection mechanism:

[0108] Detect the fluorescence excitation spectrum of the detection capsule and the ultraviolet absorption spectrum of IS. According to Figure 12 the spectral diagrams in it, it can be observed that there is a large area of overlap, and its detection mechanism may be related to the competitive absorption of energy.

[0109] It should be noted that in this article, relational terms such as first and second are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any actual relationship or order between these entities or operations. Moreover, the term "comprising", "including" or any other variant thereof is intended to cover non-exclusive inclusion, so that a process, method, article or device comprising a series of elements not only includes those elements, but also includes other elements not expressly listed, or also includes elements inherent to such process, method, article or device. Without further limitation, an element defined by the statement "comprising an..." does not exclude the presence of additional identical elements in the process, method, article or device comprising the element.

[0110] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that: they can still modify the technical solutions recorded in the foregoing embodiments, or perform equivalent replacements on some of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A europium metal-organic gel, characterized in that, The europium metal organic gel comprises a europium compound and 3,5-dicarboxyphenylboronic acid.

2. The europium metal-organic gel according to claim 1, wherein The molar ratio of the europium-containing compound to 3,5-dicarboxyphenylboronic acid is (0.1-10):

1.

3. The preparation method of the europium metal-organic gel according to claim 1 or 2, characterized in that, The preparation method comprises: S1: dissolving a europium-containing compound in solvent A to prepare solution A, and dissolving 3,5-dicarboxyphenylboronic acid in solvent B to prepare solution B; S2: mixing solution A and solution B to react to obtain the europium metal organic gel.

4. The preparation method according to claim 3, characterized in that The step S2 further comprises adding triethylamine.

5. A europium metal organic gel, characterized in that The europium metal organic gel is obtained by the preparation method according to claim 3 or 4.

6. A capsule, characterized in that The capsule comprises the europium metal organic gel according to claim 1 , 2 or 5 .

7. A fluorescent sensing material, characterized in that, The fluorescent sensing material comprises the europium metal organic gel according to claim 1, 2 or 5.

8. A fluorescent test paper, characterized in that The fluorescent test paper comprises the europium metal organic gel according to claim 1, 2 or 5 or the fluorescent sensing material according to claim 7.

9. Use of the europium metal organogel according to claim 1, 2 or 5, the capsule according to claim 6, the fluorescent sensing material according to claim 7 or the fluorescent test paper according to claim 8 in detecting indoxyl sulfate.

10. The application according to claim 9, characterized in that, The detection specifically comprises: contacting the europium metal organogel according to claim 1, 2 or 5, the capsule according to claim 6, the fluorescent sensing material according to claim 7 or the fluorescent test paper according to claim 8 with a sample, and observing changes in fluorescent color.

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