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A kind of preparation method and application of uricase mimic enzyme
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A technology that simulates enzymes and uricase, applied in the field of chemical analysis and detection, can solve problems that have not been reported, and achieve the effect of wide linear range, high speed and low cost
Active Publication Date: 2022-08-02
KUNMING UNIV OF SCI & TECH
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The use of nanozymes in uric acid detection is mostly a two-step method. First, uricase oxidizes uric acid into allantoin and hydrogen peroxide, and then acts on the nanozyme and hydrogen peroxide to detect uric acid; about the copper-iron-based nanozyme synthesized by the present invention Related research has not been reported
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Embodiment 1
[0020] Example 1: Determination of uric acid in serum
[0021] 1. Preparation of Cu, Fe-N-C nanozyme: 0.27g sodium copper chlorophyll, 0.24g iron phthalocyanine, 15mL methanol and 70g potassium chloride were added to a round bottom flask, sealed, stirred at room temperature for 10h, and placed in a vacuum drying oven After drying at 80°C for 24h to remove methanol, the obtained solid particles were placed in a tube furnace, calcined at 750°C for 2h under nitrogen protection, and the product was treated with 0.5mol / L H 2 SO 4 The solution was soaked for 24 hours and washed with pure water by centrifugation for 7 times to obtain the copper-iron-based nanozyme Cu, Fe-N-C;
[0022] 2. Preparation of uric acid working curve: add 1mg / mL copper-iron-based nanozyme (Cu, Fe-N-C) 100µL, 20mmol / L diphenylamine (DPA) 100µL and 20mmol / L 4-antipyridine to a 5mL colorimetric tube with stopper Lin (4-AP) 100µL, uric acid standard solution, diluted with pH 7.4 citric acid-disodium hydrogen p...
Embodiment 2
[0031] Example 2: Determination of uric acid in plasma
[0032] 1. Preparation of Cu, Fe-N-C nanozyme: add 0.3g sodium copper chlorophyll, 0.25g iron phthalocyanine, 20mL methanol and 95g potassium chloride to a round-bottomed flask, seal it, and stir at room temperature for 8h at 75°C. The methanol was removed by vacuum drying for 48 h, and the obtained solid particles were placed in a tube furnace and calcined at 800 °C for 2 h under nitrogen protection. The product was treated with 0.7 mol / L H 2 SO 4 After soaking in the solution for 24 hours, centrifuging and washing with pure water several times to obtain the copper-iron-based nanozyme Cu, Fe-N-C;
[0033] 2. Production of uric acid working curve: the same as in Example 1;
[0034] 3. Determination of uric acid in plasma: blood samples were obtained from the laboratory staff, and three 0.5 mL portions of plasma were taken, each was added with 1.0 mL of 50 g / L trichloroacetic acid, vortexed and mixed for 30 s, put into a...
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Abstract
The invention discloses a preparation method of a uricase mimetic enzyme and its application in detecting uric acid. The method is based on the fact that an iron-copper-based nanozyme (Cu, Fe-N-C) has similar electron and Geometric structure, by adjusting the ratio of copper and iron, Cu, Fe-N-C exhibits uricase-mimicking activity, which can rapidly oxidize the substrates uric acid, diphenylamine and 4-antipyrine to generate blue oxides , thereby establishing a rapid detection method for uric acid; this method was applied to the detection and analysis of uric acid in human serum or plasma, and the result was coupled with uricase to catalyze the color development of 4-antipyrine phenol with horseradish peroxidase (HRP). This method is consistent with the detection method of uric acid in human serum by uric acid reduction phosphotungstic acid method; this method does not require natural uricase and HRP, and the whole reaction only takes 10 minutes, and has the characteristics of rapidity, strong specificity, simple operation and low cost.
Description
technical field [0001] The invention belongs to the technical field of chemical analysis and detection, in particular to the preparation of a uricase mimic enzyme and a method for detecting uric acid. Background technique [0002] Uric acid is the final product of purine metabolism, which has the functions of scavenging oxygen free radicals and anti-oxidation. Gout, kidney disease, hyperuricemia, etc. can cause abnormal uric acid. Recent studies have shown that the concentration of uric acid in serum is also positively correlated with the concentration of triglyceride and creatinine, and is associated with cardiac damage. There is a significant positive correlation between the level of serum uric acid in patients with coronary heart disease and carotid atherosclerosis. Therefore, the detection and analysis of uric acid has important reference value for clinical diagnosis and understanding of disease progression. At present, the determination methods of serum uric acid main...
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