At present, the number of patients with hyperuricemia and gout in China is increasing year by year, and the demand for uric acid detection is increasing. In the current situation, a uric acid detection chip was designed based on the redox reaction between potassium ferricyanide and uric acid and the principle of electrochemical sensor, and the uric acid content was measured by the electrochemical detection chip and the software PSTrace5.8, and the error of the result was within ±10%. The method is low cost and high accuracy, which can provide a new technical means for uric acid detection

Wang, Haowei

English

Advanced Emergency Medicine (E-Journal)

Volume 12 | Issue 3 -115-Application of Electrochemical Sensor Based on the Reaction ofPotassium Ferricyanide and Uric Acid in Uric Acid DetectionHaowei WangHunan University of Technology, Zhuzhou 412000, China.Abstract: At present, the number of patients with hyperuricemia and gout in China is increasing year by year, and thedemand for uric acid detection is increasing. In the current situation, a uric acid detection chip was designed based on theredox reaction between potassium ferricyanide and uric acid and the principle of electrochemical sensor, and the uric acidcontent was measured by the electrochemical detection chip and the software PSTrace5.8, and the error of the result waswithin ±10%. The method is low cost and high accuracy, which can provide a new technical means for uric acid detection.Keywords: Electrochemical Sensor; Uric Acid Detection; Potassium Ferricyanide1. Introduction Introduction1.1 Current situation of hyperuricemia and gout in ChinaUric acid (2,4,6 trihydroxypurine, uric acid. UA) is the end product of purine metabolism in the human body.Hyperuricemia is a metabolic abnormality syndrome caused by the disorder of purine metabolism, and it is calledhyperuricemia when the blood uric acid level exceeds 420μmol/L on two different days in both men and women. Blood uricacid exceeding its saturation level in blood or tissue fluid can form and deposit sodium urate crystals locally in the joints,inducing local inflammatory reactions and tissue destruction, known as gout; it can be deposited in the kidneys, causingacute nephropathy, chronic interstitial nephritis or kidney stones, known as uric acid nephropathy. There is much evidencethat hyperuricemia and gout are independent risk factors for chronic kidney disease, hypertension, cardiovascular diseaseand diabetes, and are independent predictors of premature death. Hyperuricemia and gout are systemic diseases withmultisystemic involvement and have received a lot of attention from multiple disciplines, and their management alsorequires multidisciplinary involvement. According to relevant data, the prevalence of hyperuricemia ranges from 2.6% to36% in different races, and the prevalence of gout ranges from 0.03% to 15.3%, with a significant increase and youthfultrend in recent years. It has become another common metabolic disease after diabetes mellitus. According to the relatedsurvey, by 2022, the number of diagnosed hyperuricemia in China will be as high as 170 million, and the diagnosis rate willbe more than 13.3% in some areas with heavy diets, so it is important to develop an accurate method to measure uric acidcontent for the health of the nation.2. Main methods of measuring uric acidCurrent methods commonly used for uric acid testing include phosphotungstic acid reduction, high performance liquidchromatography, spectrophotometric, enzymatic and uric acid sensor assays and microfluidic paper chip methods.Phosphotungstic acid reduction method is more mature, but it has the disadvantages of poor linearity, precipitation of uricacid and proteins, and susceptibility to interference by reducing substances: high performance liquid chromatographymethod has high precision, fast analysis, and low sample usage, but it has the disadvantages of pre-treatment of samples,finding suitable chromatographic conditions, and dependence on high precision instruments: spectrophotometric method is-116- Advanced Emergency Medicineeasy and rapid to operate. However, the sensitivity is poor and it is difficult to measure the uric acid content in plasmadirectly: the advantages of enzymatic method are high specificity and mild action conditions, but its disadvantages of noteasy to preserve and high acquisition cost also limit the application of the method; and the microfluidic paper chip methodhas good development prospects, has the advantages of convenient, fast, green and cheap, but at this stage of developmenttechnology is still immature. The uric acid detector method has become a popular method in the field of uric acid detectiondue to its high sensitivity, good selectivity and practicality. Therefore, this study is based on the uric acid sensing detectormethod, and the current generated by the redox reaction between potassium ferricyanide and uric acid is measured by anelectrochemical detection chip, so that the magnitude of uric acid concentration in the test sample can be visually reflectedby the current data.2. Materials and Methods2.1 Experimental principleIn this study, a uric acid detection chip was designed based on the redox reaction between potassium ferricyanide anduric acid, with the principle of (1) K3 Fe (CN)6 + uric acid → K4 Fe (CN)6 + uric acid oxide.(2)K4 Fe(CN)6 -e- →K3 Fe(CN)6 Uric acid is oxidized by potassium ferricyanide, which loses electrons to generatepotassium ferricyanide, and then the oxidation potential is applied to the surface of the working electrode, and potassiumferricyanide loses electrons to become potassium ferricyanide again, and the oxidation current on the surface of the uric aciddetection chip is collected by the detection instrument to realize the conversion of the uric acid concentration size of thedetected sample into the value of the circuit. The uric acid concentration in the sample can be measured by the detectioncircuit. The instrument is designed based on the principle of electrochemical sensor, and the oxidation potential is applied tothe surface of the working electrode of the uric acid detection chip relative to the reference electrode, and the oxidationcurrent generated on the surface of the working electrode of the cell phone due to the oxidation of uric acid is convertedbetween the chemical reaction signal and the electrical signal by the electrochemical detection chip to obtain more accurateresults of uric acid.2.2 Experimental materials and apparatusExperimental material: uric acid quality control solution with different uric acid concentrations (Since human bloodsamples are not easily available and inconvenient to use, a uric acid quality control solution was designed to replace theblood in this experiment, as follows: 100 mL of ultrapure water was taken, to which the following substances were added inorder and the mass concentrations of each substance were made: sodium chloride 0.58% (w/w), phosphate buffer 1.82%(w/w) (pH 7.4), polyethylene glycol (molecular weight 40,000) 4 g/L, cuscuta red 0.1% (w/w), ProClin 300 0.04% (w/w).Uric acid was added to the above base solution at different concentrations (Uric acid was pre-dissolved with 0.lmol/L NaOH)to prepare a concentration gradient of 0.25 mmol/L, 0.5 mmol/L, 0.8 mmol/L, and 1.0 mmol/L of Uric acid quality controlsolution, respectively.)Uric acid detection chip (by screen printing a conductive silver film on a PET sheet as a silver conductive layerconnecting the three electrodes, and a carbon film printed on the front of the silver conductive layer as a counter electrode,working electrode, reference electrode, and suction sample judgment electrode. The reference electrode and the counterelectrode share a common electrode, and the working electrode and the reference electrode form a circuit to measure thepotential value of the research electrode. The working electrode and the counter electrode form another circuit for measuringthe current value. The aspiration judgment electrode is used to detect whether the sample fills the entire biochemical reactionzone. Modify the reaction active layer on the electrode surface: the reaction layer solution is configured with ascorbateoxidase, potassium ferricyanide, carboxymethyl cellulose and phosphate in certain ratio, where ascorbate oxidase is theanti-interference substance, potassium ferricyanide is the electron mediator for the reaction with uric acid, andVolume 12 | Issue 3 -117-carboxymethyl cellulose is the film-forming auxiliary component. 1.5 μL of the configured solution is sucked and droppedonto the reaction working area of the prepared electrode substrate, and (Dried at 37℃ for 20 min, and after drying,double-sided adhesive and hydrophilic film were applied to produce the uric acid detection chip.)Test apparatus: open electrochemical detection chip, computer with detection software PSTrace 5.82.3 Experimental step designStep 1: Insert the uric acid test chip into the electrochemical test chip and connect the electrochemical test chip to thecomputer via the data cable.Step 2: Open the software and apply a drop of uric acid quality control solution to the surface of the uric acid test chipafter the current display is smooth.Step 3: Read the software value after 15s and compare it with the actual value.Step 4: Repeat the above steps using different concentrations of uric acid quality control solution.2.4 Analysis of experimental resultsAfter several tests, it was shown that the measurement error was within ±10%, and the uric acid detection chip designedby the modified method has a high accuracy for measuring uric acid concentration.3. Discussion of experimental resultsAt the end of the experiment, for further determination of the accuracy of the results, i.e. further testing of thefunctionality of this uric acid detection chip for uric acid sensing recognition, cyclic voltammetry and constant potentialmethods were used and tested again, as recorded below.Cyclic voltammograms of uric acid quality control solution at different concentrations. It can be seen from the figurethat a pair of redox peaks appear between -0.2 V and 0.2 V during cyclic voltammetry scan, which is the redox of Fe3+ /Fe2+ion pair with the oxidation peak potential at about 0.03 V. As the concentration of uric acid QC solution increased from 0.25mmol/L to 0.8 mmol/L, the redox peak current in cyclic voltammetry scan increased, which demonstrated the good sensingfunction of the chip for uric acid.-118- Advanced Emergency MedicineRelationship between uric acid quality control solution and response current for different uric acid concentrations atconstant potential.It can be seen from the figure that at a constant potential of 0.12 V, when the uric acid concentration in the uric acidquality control solution sample increased from 0.5 mmol/L to 0.8 mmol/L and then to 1.0 mmol/L, the response currentmagnitude increased sequentially with the increase of uric acid concentration after the addition of uric acid quality controlsolution, which further proved that the detection chip has a good function of uric acid sensing and recognition.These two measurements further validate the accuracy of the results of the uric acid test chip.4. ConclusionThe experiment showed that the electrochemical sensor based on the reaction between potassium ferricyanide and uricacid is more accurate in uric acid detection. Although the human blood experiment has not been conducted due to technicalreasons, this experiment still provides a new idea for the design of uric acid detection chip, and we hope to provide our sharefor better uric acid detection.References[1] Jiang MC, Zhao Y, Ma FF, Zhang W, Liu CY. Research progress in uric acid detection methods [J]. ChemicalScience and Technology, 2021,29 (3): 75-79.[2] Professional Committee of Endocrinology and Metabolism of Chinese Physicians Association of IntegratedTraditional and Western Medicine. Guidelines for Diagnosis and Treatment of Hyperuricemia and Gout (2021-01-20) [J].World Journal of Traditional Chinese Medicine, 2021, 16 (2): 183-189.[3] Cui M. Research on Electrochemical Sensors Based on Nanomaterials and Their Applications [D]. Beijing: BeijingUniversity of Technology, 2014:1-15.[4] Wang L, Peng A. Interpretation of Chinese Guidelines for Diagnosis and Treatment of Hyperuricemia and Gout(2019) [J]. Western Medicine, 2021,33 (9): 1254~1257.[5] Peng DD, Liu DY, Li LN, Liu ZY, Lu DQ, Pang GC. Detection of Allergens in Milk Based on ElectrochemicalNanoimmune Sensors α- Casein [J] Food Industry Technology, 2022,43 (16): 307-314.[6] Shi P, Li JP, Fang C, Li YP. Development of electrode strip for detecting uric acid and glucose [J]. Chemical Sensor,2010,30 (3): 59-64.[7] Cao R. Preparation and performance study of electrochemical uric acid biosensor enzyme membrane [D]. Beijing:Beijing University of Chemical Technology, 2017:1-17.[8] Shang S, Lu Y, Cui XY, Gu TT. Research progress on enzymatic glucose electrochemical sensors [J]. LiaoningVolume 12 | Issue 3 -119-Chemical Industry, 2022,51 (10): 1453-1457.[9] Jiang L. Application of electrochemical sensors modified with oxide nanomaterials in the detection of glucose andhydrogen peroxide [D]. Yangzhou: Yangzhou University, 2021:1-13.[10] Zhang Y, Sun YL, Shi KX, Guo SL, Ma SJ. Research progress in glucose measurement based on electrochemicalsensors [J]. Shandong Chemical Industry, 2022,51 (4): 105~107.

Application of Electrochemical Sensor Based on the Reaction of Potassium Ferricyanide and Uric Acid in Uric Acid Detection

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Application of Electrochemical Sensor Based on the Reaction of Potassium Ferricyanide and Uric Acid in Uric Acid Detection

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