Search

CN-121978089-A - Colorimetric sensing array based on hierarchical pore nano enzyme and construction method and application thereof

CN121978089ACN 121978089 ACN121978089 ACN 121978089ACN-121978089-A

Abstract

The invention discloses a colorimetric sensing array based on hierarchical pore nano-enzyme and a construction method and application thereof, and belongs to the technical field of analysis and detection. The array is composed of two cerium-based MOF nano enzymes of micropore UiO-66 and hierarchical pore HP-UiO-66, which are respectively combined with TMB and ABTS chromogenic substrates to form a four-way sensing system. The method comprises the steps of synthesizing hierarchical pore HP-UiO-66 through a template method, collecting absorbance of each channel under characteristic wavelength by utilizing an array to generate a specific fingerprint of an object to be detected, and combining with mode identification technologies such as linear discriminant analysis and the like to realize rapid, synchronous and quantitative detection of at least eight structurally similar antioxidants such as glycyrrhizic acid, caffeic acid and the like in the traditional Chinese medicine decoction. According to the invention, the catalysis and mass transfer efficiency is improved by regulating and controlling the pore structure of the nano enzyme, the difficulty of multi-target synchronous identification in a complex matrix is solved by utilizing the array cross response, and the method has the advantages of simplicity and convenience in operation, high flux and good accuracy.

Inventors

  • LIU JUNLEI
  • LI WENYING
  • YANG KAIQIANG

Assignees

  • 宁波大学

Dates

Publication Date
20260505
Application Date
20251225

Claims (9)

  1. 1. A colorimetric sensing array based on hierarchical pore nano enzyme is characterized by comprising a microporous cerium-based metal organic framework material UiO-66, a hierarchical pore cerium-based metal organic framework material HP-UiO-66, a chromogenic substrate 3,3', 5' -tetramethyl benzidine and a chromogenic substrate 2,2 '-azino-bis- (3-ethylbenzodihydrothiazoline-6-sulfonic acid, wherein the microporous cerium-based metal organic framework material UiO-66 and the hierarchical Kong Shi-based metal organic framework material HP-UiO-66 are respectively combined with 3,3',5 '-tetramethyl benzidine and the 2,2' -azino-bis- (3-ethylbenzodihydrothiazoline-6-sulfonic acid to form four signal output channels, and the hierarchical Kong Shi-based metal organic framework material HP-UiO-66 is synthesized through a surfactant template method and has a pore diameter larger than that of the microporous cerium-based metal organic framework material UiO-66.
  2. 2. The colorimetric sensing array of claim 1, wherein the method of synthesizing the multi-stage Kong Shi-based metal-organic framework material HP-UiO-66 comprises the steps of: a) Dissolving a template agent P123 in deionized water, and adding glacial acetic acid and 1,3, 5-trimethylbenzene to form microemulsion; b) Adding a cerium source (NH 4 ) 2 Ce(NO 3 ) 6 and an organic ligand terephthalic acid; c) The system was stirred at 50 ℃, and after the stirring reaction was completed, the product was centrifuged, washed and dried in vacuo.
  3. 3. A method of constructing a colorimetric sensing array as claimed in any one of claims 1 to 2 comprising the steps of: a) Sequentially adding a buffer solution, a chromogenic substrate, a sample to be detected and a nano enzyme solution into a reaction container to form a reaction system, wherein the chromogenic substrate is 3,3', 5' -tetramethylbenzidine or 2,2' -azino-bis- (3-ethylbenzodihydrothiazoline-6-sulfonic acid), and the nano enzyme solution is a microporous cerium-based metal organic frame material UiO-66 or multi-stage Kong Shi-based metal organic frame material HP-UiO-66 aqueous dispersion; b) Measuring absorbance of the reaction system at a specific wavelength after the reaction, wherein when the chromogenic substrate is 3,3', 5' -tetramethylbenzidine, the measuring wavelength is 652 nm, and when the chromogenic substrate is 2,2' -azino-bis- (3-ethylbenzothiazoline-6-sulfonic acid), the measuring wavelength is 425 nm; c) And combining the absorbance signals of the four signal output channels to generate a specific fingerprint of the sample to be detected.
  4. 4. The method of claim 3, wherein in step a), the buffer solution is NaAc-HAc buffer solution with ph=4.0, the chromogenic substrate is 5mM, and the nano-enzyme solution is 1:1 mg/mL.
  5. 5. A method for detecting an antioxidant of a traditional Chinese medicine, characterized in that at least one antioxidant selected from glycyrrhizic acid, caffeic acid, coumaric acid, vanillic acid, ferulic acid, gallic acid, naringenin and berberine is synchronously identified by adopting the colorimetric sensor array according to any one of claims 1-2.
  6. 6. The method according to claim 5, wherein the method is used for quantitative detection of glycyrrhizic acid, caffeic acid, ferulic acid and gallic acid; The glycyrrhizic acid concentration is in a linear relation with a linear discrimination Factor 1 within the range of 0-50 mu M, and a linear regression equation is y= 1.3689x-28.3486, and R 2 =0.994; Caffeic acid concentration is in linear relation with Factor 1 in the range of 0-25 μm, and linear regression equation is y= 6.7574x-109.0898, and r 2 =0.992; the concentration of ferulic acid is in linear relation with Factor 1 in the range of 0-50 mu M, the linear regression equation is y= 1.2407x-20.0008, and R 2 =0.996; The gallic acid concentration is in a linear relation with the Factor 1 in the range of 0-25 mu M, the linear regression equation is y= 4.3039x-63.9522, and R 2 =0.990, wherein y represents the value of the Factor 1, and x represents the antioxidant concentration.
  7. 7. The method according to claim 6, wherein the limit of detection of glycyrrhizic acid is 2.66. Mu.M, the limit of detection of caffeic acid is 0.57. Mu.M, the limit of detection of ferulic acid is 1.69. Mu.M, and the limit of detection of gallic acid is 0.78. Mu.M.
  8. 8. The method according to claim 5, further comprising a sample pretreatment step of mixing the raw materials of the Chinese medicine to be tested with water, heating and refluxing at 100 ℃, then filtering, centrifuging to obtain a supernatant, diluting, and adding the supernatant as the sample to be tested into the reaction system.
  9. 9. The method of claim 8, wherein the time of the heat reflow is 1 hour.

Description

Colorimetric sensing array based on hierarchical pore nano enzyme and construction method and application thereof Technical Field The invention relates to the field of nano analysis and detection, in particular to a colorimetric sensing array based on hierarchical pore nano enzyme, and a construction method and application thereof. Background Antioxidant therapy plays a key role in preventing and treating diseases related to oxidative stress, which has prompted increasing attention to Traditional Chinese Medicine (TCM) which is rich in various natural antioxidant ingredients. The Chinese medicinal decoction is a complex matrix, and contains phenolic acid (such as caffeic acid, ferulic acid, and gallic acid), flavonoids (such as naringenin), and alkaloids (such as berberine) with antioxidant activity. The specific components are rapidly and accurately distinguished and detected, and the method is very important for quality control of traditional Chinese medicines, elucidation of pharmacodynamic substance basis and standardized research. At present, qualitative and quantitative analysis of traditional Chinese medicine antioxidants mainly depends on chromatographic techniques (such as High Performance Liquid Chromatography (HPLC) and Gas Chromatography (GC)) and the combined technology of the traditional Chinese medicine antioxidants and mass spectrum. Although these methods are highly sensitive, they generally require complex sample pretreatment, expensive instrumentation, specialized operators, and long analysis times, making high throughput, rapid field detection difficult. More importantly, most of the methods are targeted analysis aiming at single or few target objects, and have limited synchronous recognition and distinguishing capability when facing a complex system of coexistence of multiple structural analogues in traditional Chinese medicines. To simulate the overall perception and discrimination capabilities of biological sensory systems (e.g., smell, taste) on complex mixtures, sensor array (or "electronic tongue", "electronic nose") technologies have evolved. The technology utilizes a series of sensing units with cross reactivity to respond to an object to be detected, generates a multi-dimensional signal mode (namely a fingerprint), and then realizes the differentiation and identification of different targets through a mode identification algorithm. Among them, the colorimetric sensor array based on the nano-enzyme shows unique advantages due to visual signal output (color change), simple equipment (commonly used enzyme-labeled instrument) and adjustable catalytic activity of the nano-enzyme. Various nanomaterials, including noble metal nanoparticles, metal oxides, and carbon materials, have been used as nanoenzymes to construct sensing channels. Metal-organic framework Materials (MOFs) are a class of porous crystalline materials formed by self-assembly of metal ions/clusters with organic ligands. Among them, cerium-based UiO-66 has been reported to have oxidase-like activity as a structurally stable MOF, and can catalyze color reaction of chromogenic substrates such as 3,3', 5' -Tetramethylbenzidine (TMB) or 2,2' -azino-bis- (3-ethylbenzothiazoline-6-sulfonic Acid) (ABTS), thereby having application potential in biosensing. However, in the existing research of using MOFs as nano enzymes for sensing, most of the MOFs focus on composition and macro morphology, and the influence of the key physical property of the internal pore structure (such as pore diameter and pore channel distribution) on substrate mass transfer efficiency and final catalysis and sensing performance is often neglected. In particular, how to design a sensing unit with differentiated performance by finely controlling the pore structure of MOFs, and thus construct a high-efficiency sensor array, has not been fully explored. In addition, the application of the sensor array based on MOF nano-enzyme to a traditional Chinese medicine system with extremely complex components, so as to realize synchronous identification and quantitative detection of various specific antioxidants, still faces challenges. This is mainly due to the severe interference of the traditional Chinese medicine matrix and the need for the sensing array to have sufficient discrimination capability for a plurality of targets with similar structures. Therefore, the development of the colorimetric sensing array and the colorimetric sensing method based on the novel nano enzyme material has simple and convenient construction method, can effectively overcome the interference of the traditional Chinese medicine matrix, can synchronously identify and detect various traditional Chinese medicine antioxidants, and has important scientific significance and application value. Disclosure of Invention The invention provides a colorimetric sensing array based on hierarchical pore nano-enzyme and a construction method and application thereof, aiming at overcoming the defects of c