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CN-121988317-A - Porous RuO for detecting creatinine2Preparation method and application of nano-enzyme

CN121988317ACN 121988317 ACN121988317 ACN 121988317ACN-121988317-A

Abstract

The application belongs to the technical field of nano-enzymes, and particularly relates to a preparation method and application of porous RuO 2 nano-enzyme for detecting creatinine. The preparation method comprises the steps of taking Ca (NO 3 ) 2 ·4H 2 O、RuCl 3 and glycine) to ultrasonically disperse in deionized water to obtain a mixed solution A, dissolving sodium alginate in the deionized water, stirring uniformly to obtain a mixed solution B, injecting the mixed solution B into the mixed solution A, slowly stirring to form bimetallic hydrogel, washing by deionized water, freeze-drying to obtain a solid A, calcining the solid A to obtain a solid B, adding the solid B into hydrochloric acid to ultrasonically disperse, stirring for a period of time, centrifugally washing the precipitate, and drying in vacuum to obtain porous RuO 2 nano-particles.

Inventors

  • SONG HUIJUN
  • JIN JUAN
  • HE QIANG
  • HE XINXIN
  • LIU YAN
  • YANG NAN

Assignees

  • 浙江省中医院、浙江中医药大学附属第一医院（浙江省东方医院）

Dates

Publication Date
20260508
Application Date
20260410

Claims (10)

  1. 1. The preparation method of the porous RuO 2 nano-enzyme for detecting creatinine is characterized by comprising the following steps: S.1, dispersing Ca (NO 3 ) 2 ·4H 2 O、RuCl 3 and glycine in deionized water by ultrasonic to obtain a mixed solution A, dissolving sodium alginate in deionized water, and uniformly stirring to obtain a mixed solution B; S.2, injecting the mixed solution B into the mixed solution A, stirring to form bimetallic hydrogel, washing with deionized water, and freeze-drying to obtain a solid A; S.3, calcining the solid A to obtain a solid B; S.4, adding the solid B into hydrochloric acid for ultrasonic dispersion, stirring, taking the precipitate for centrifugal washing, and vacuum drying to obtain loose porous RuO 2 nano particles.
  2. 2. The method for preparing the porous RuO 2 nano-enzyme for detecting creatinine according to claim 1, wherein Ca (NO 3 ) 2 ·4H 2 O、RuCl 3 , glycine and deionized water in the ratio of 3mmol:3-8mmol:4-8mmol:15-30 mL) in the step S.1.
  3. 3. The method for preparing the porous RuO 2 nano-enzyme for detecting creatinine according to claim 1, wherein the ratio of sodium alginate to deionized water in the step S.1 is 1g:70-120mL.
  4. 4. The method for preparing the porous RuO 2 nano-enzyme for detecting creatinine according to claim 1, wherein the ultrasonic time in the step S.1 is 5-15min, and the stirring time is 2-4h.
  5. 5. The method for preparing the porous RuO 2 nano-enzyme for detecting creatinine according to claim 1, wherein in the step S.2, the stirring time is 1.5-2.5h, the washing times of deionized water are 2-4 times, and the washing conditions are 9000-11000rpm and centrifuge for 9-11min.
  6. 6. The method for preparing the porous RuO 2 nano-enzyme for detecting creatinine according to claim 1, wherein the calcining treatment condition in the step S.3 is that the temperature is 300-600 ℃, the heating rate is 10 ℃ per minute, and the time is 0.5-2h.
  7. 7. The method for preparing the porous RuO 2 nano-enzyme for detecting creatinine according to claim 1, wherein the dosage ratio of the solid B to hydrochloric acid in the step S.4 is 100mg:20-40mL, the concentration of hydrochloric acid is 1M, the ultrasonic time is 1.5-2.5min, and the stirring time is 25-40min.
  8. 8. The method for preparing the porous RuO 2 nano-enzyme for detecting creatinine according to claim 1, wherein the washing condition in the step S.4 is that deionized water is centrifuged and washed for 2-4 times, the washing condition is that 9000-11000rpm is centrifuged for 9-11min, and the vacuum drying condition is that the drying temperature is 50-70 ℃ and the drying time is 7-9h.
  9. 9. The porous RuO 2 nano-enzyme for detecting creatinine obtained by the preparation method according to any one of claims 1-8.
  10. 10. The use of the porous RuO 2 nano-enzyme for detecting creatinine according to claim 9 in the detection of creatinine content.

Description

Preparation method and application of porous RuO 2 nano-enzyme for detecting creatinine Technical Field The invention belongs to the technical field of nano-enzymes, and particularly relates to a preparation method and application of porous RuO 2 nano-enzyme for detecting creatinine. Background Creatinine is one of the end products produced by muscle metabolism, is mainly filtered by glomeruli and is discharged with urine, and the concentration of creatinine in serum and urine is closely related to the glomerular filtration function, so that creatinine is an important biochemical index for clinically evaluating the damage degree of renal function, monitoring the progress of chronic kidney disease and guiding the adjustment of drug dosage. Therefore, the creatinine detection method is constructed sensitively, accurately and simply and conveniently operated, and has important clinical application value for early screening and curative effect evaluation of kidney-related diseases. The current creatinine detection method mainly comprises a traditional Jaffe colorimetric method, an enzymatic colorimetric/electrochemical method, a high performance liquid chromatography, a mass spectrum and the like. The Jaffe method is used as a most common conventional method in clinic, and color reaction is carried out on creatinine and picric acid under alkaline conditions to realize colorimetric quantification, so that the Jaffe method has the advantages of simplicity in operation, low cost, easiness in matching with a biochemical analyzer and the like, but has poor selectivity, limited anti-interference capability and difficulty in fully ensuring repeatability and accuracy. In contrast, the enzyme method generally utilizes creatininase, creatininase and the like to construct a cascade catalytic system, realizes specific recognition and signal amplification of creatinine, and has the advantages of high substrate specificity, high sensitivity, mild reaction conditions and the like. However, the natural enzymes have high cost, complex preparation and purification processes, high sensitivity to temperature and pH, easy inactivation and poor storage stability, and severely restrict the application of the natural enzymes in large-scale, long-period and complex sample detection. In recent years, a nanomaterial (nanoenzyme) having enzyme-like catalytic activity has been considered as an ideal substitute for a natural enzyme, and can exhibit various enzyme activities such as peroxidase, oxidase, superoxide dismutase, and the like. Compared with natural enzymes, the nano-enzyme has controllable synthesis, easily controllable structure and surface, is insensitive to temperature and pH, is not easy to inactivate, and is convenient for long-term storage and application. In creatinine detection, nano enzyme with peroxidase-like activity is introduced to participate in creatinine-related enzymatic cascade reaction, H 2O2 generated in a high-efficiency catalytic system can be oxidized into a colored product by a colorless substrate (such as TMB), and amplified colorimetric response to creatinine concentration is realized, so that detection sensitivity is improved, detection limit is reduced, linear range is widened, and meanwhile, the nano enzyme has good selectivity and anti-interference capability in complex matrixes such as serum and urine. Particularly, ruO 2、Fe2O3、MoO2 and other metal oxide nano enzymes, by virtue of the excellent oxidation-reduction characteristics, the nano enzymes exhibit outstanding peroxidase-like activity in the field of biosensing. However, most metal oxide nano-enzymes (such as RuO 2) still have the problems of insufficient utilization of metal active sites, low intrinsic activity, insufficient affinity with substrate molecules and the like, and the overall catalytic efficiency is limited, so that the requirement of high-sensitivity detection is difficult to meet. Therefore, the development of the nano-enzyme with high-efficiency catalytic activity for serum creatinine detection has important scientific significance and application value. Disclosure of Invention Aiming at the problems existing in the prior art, the invention aims to design and provide a preparation method and application of porous RuO 2 nano-enzyme for detecting creatinine. The invention is realized by the following technical scheme: The first aspect of the invention provides a preparation method of porous RuO 2 nano-enzyme for detecting creatinine, which comprises the following steps: S.1, dispersing Ca (NO 3)2·4H2O、RuCl3 and glycine in deionized water by ultrasonic to obtain a mixed solution A, dissolving sodium alginate in deionized water, and stirring uniformly to obtain a mixed solution B. S.2, injecting the mixed solution B into the mixed solution A, stirring to form bimetallic hydrogel, washing with deionized water, and freeze-drying to obtain a solid A. S.3, calcining the solid A to obtain a solid B. S.4, adding the solid B into hydroch