CN111879836A - Electrochemical method for detecting uric acid and urate oxidase and application thereof - Google Patents
Electrochemical method for detecting uric acid and urate oxidase and application thereof Download PDFInfo
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Abstract
本发明公开了基于银离子和谷胱甘肽的电化学传感器的制备方法及其应用，具体步骤如下：首先将氧化石墨烯分散液电化学方法镀到干净的裸玻碳电极上，利用GSH和Ag(I)之间的相互作用合成GSH‑Ag(I)复合物，使用前将该复合物溶液与100μL 0.02％wt Nafion溶液混合均匀，静置80min，滴涂到石墨烯修饰电极表面，制备GSH‑Ag(I)/GO/GCE，该电极对H2O2具有较强的电催化响应。利用UOx催化UA生成H2O2，分别固定UOx或UA浓度，通过复合物对H2O2的响应作为信号输出，实现对UA或UOx的高灵敏检测。优点是特异性好、灵敏度高、检测速度快、结果准确可靠、成本低。
The invention discloses a preparation method and application of an electrochemical sensor based on silver ions and glutathione. The specific steps are as follows: first, a graphene oxide dispersion liquid is electrochemically plated on a clean bare glassy carbon electrode, and GSH and glutathione are used for electrochemical plating. The GSH-Ag(I) complex was synthesized by the interaction between Ag(I), the complex solution was mixed with 100 μL of 0.02% wt Nafion solution before use, left for 80 min, and drop-coated on the surface of graphene modified electrode to prepare GSH‑Ag(I)/GO/GCE, the electrode exhibits strong electrocatalytic response to H 2 O 2 . UOx is used to catalyze UA to generate H 2 O 2 , the concentration of UOx or UA is fixed respectively, and the response of the complex to H 2 O 2 is used as a signal output to achieve highly sensitive detection of UA or UOx. The advantages are good specificity, high sensitivity, fast detection speed, accurate and reliable results, and low cost.
Description
技术领域technical field
本发明涉及一种电化学方法及其应用，尤其是涉及基于生物金属复合物的电化学传感器制备及其在尿酸及尿酸氧化酶分析传感中的应用，属于功能生物材料和生物传感技术领域。The invention relates to an electrochemical method and application thereof, in particular to the preparation of an electrochemical sensor based on a biological metal complex and its application in the analysis and sensing of uric acid and uric acid oxidase, belonging to the technical field of functional biological materials and biological sensing .
背景技术Background technique
尿酸(UA，2，4，6-三羟基嘌呤)主要由细胞代谢分解的核酸和其他嘌呤类化合物以及食物中的嘌呤经酶的作用分解而来，是嘌呤衍生物在人类新陈代谢中的终产物，血清中UA正常水平为240～520μM，尿排泄中UA正常水平为1.4～4.4mM。研究发现人体内尿酸含量的变化与临床中许多心血管疾病和代谢类疾病有着密切的关系，例如痛风、肥胖、高尿酸血症和Lesch-Nyhan综合征，因此尿酸可作为有价值的临床诊断指标并引起科学家广泛关注，检测方法有光谱法、荧光法、液相色谱法以及酶方法等。尽管这些检测方法有较高的灵敏度和选择性，但是存在耗时、材料昂贵和预处理复杂等缺点，而电化学方法以操作简便、灵敏度高、响应快等优点成为科学家关注的焦点。与尿酸紧密联系的生物酶——尿酸氧化酶(UOx)能使尿酸迅速氧化变成尿囊酸，不再被肾小管吸收而排泄，因此人体内这种酶的活性对尿酸相关疾病诊断和治疗具有较大影响。开发一种新的电化学方法实现尿酸及尿酸氧化酶灵敏检测对临床诊断和药物开发有着十分重要的意义。Uric acid (UA, 2,4,6-trihydroxypurine) is mainly decomposed by nucleic acids and other purine compounds decomposed by cell metabolism and purines in food by the action of enzymes, and is the end product of purine derivatives in human metabolism , the normal level of UA in serum is 240-520μM, and the normal level of UA in urinary excretion is 1.4-4.4mM. Studies have found that the changes of uric acid content in the human body are closely related to many cardiovascular and metabolic diseases in clinic, such as gout, obesity, hyperuricemia and Lesch-Nyhan syndrome, so uric acid can be used as a valuable clinical diagnostic indicator. And caused widespread concern of scientists, detection methods include spectroscopy, fluorescence, liquid chromatography and enzymatic methods. Although these detection methods have high sensitivity and selectivity, they have disadvantages such as time-consuming, expensive materials, and complicated pretreatment. Electrochemical methods have become the focus of scientists due to their advantages of simple operation, high sensitivity, and fast response. Uric acid oxidase (UOx), a biological enzyme closely related to uric acid, can rapidly oxidize uric acid into allantoic acid, which is no longer absorbed and excreted by renal tubules. Therefore, the activity of this enzyme in the human body is useful for the diagnosis and treatment of uric acid-related diseases. have a greater impact. The development of a new electrochemical method for sensitive detection of uric acid and urate oxidase is of great significance for clinical diagnosis and drug development.
生物金属复合物是一种利用金属离子和生物分子的分子内及分子间相互作用连接而形成的配位聚合物材料，其中造币金属(如：Au、Ag、Cu)和巯基生物分子的配位聚合物引起了相当大的关注。一般说这种复合物是基于生物分子的巯基和金属离子的相互作用(巯基-金属/金属-金属)所形成的超分子聚合物材料。谷胱甘肽(GSH)是一种具有特殊生物学功能的氨基酸衍生物，有三个可解离质子和十个可参与配位的原子，属于含有巯基的小分子肽类物质，是一种形成复合物较理想的生物分子模型。到目前为止，已经有很多关于GSH及金属离子相互作用的报导，但是将GSH和金属离子形成的生物金属复合物用于尿酸及尿酸氧化酶的分析检测尚未见报导，具有极大的潜力。Biometal complexes are coordination polymer materials formed by intramolecular and intermolecular interactions between metal ions and biomolecules. Bit polymers have attracted considerable attention. Generally speaking, this complex is a supramolecular polymer material formed based on the interaction of thiol groups of biomolecules and metal ions (thiol-metal/metal-metal). Glutathione (GSH) is an amino acid derivative with special biological functions. It has three dissociable protons and ten atoms that can participate in coordination. It is a small molecule peptide substance containing sulfhydryl groups. The complex is an ideal biomolecular model. So far, there have been many reports on the interaction between GSH and metal ions, but the use of biometal complexes formed by GSH and metal ions for the analysis and detection of uric acid and uric acid oxidase has not been reported yet, which has great potential.
本发明设计了一种检测尿酸及尿酸氧化酶的电化学方法，该方法采用GSH作为生物有机配体，银离子(Ag(I))作为金属节点，通过GSH中巯基与Ag(I)和Ag(I)与Ag(I)相互作用简单绿色合成GSH-Ag(I)复合物。基于金属离子的氧化还原活性，本发明发现该复合物对双氧水(H2O2)具有较好的电催化作用。此外，UOx能使UA迅速氧化变成尿囊酸和H2O2，因此通过复合物修饰电极及其对H2O2催化还原，本发明构建了一种新的电化学方法以期实现尿酸及尿酸氧化酶的分析监测，为尿酸相关临床诊断及药物开发提供了一种新思路。The present invention designs an electrochemical method for detecting uric acid and uric acid oxidase. The method adopts GSH as a biological organic ligand and silver ions (Ag(I)) as a metal node. (I) Simple green synthesis of GSH-Ag(I) complexes by interacting with Ag(I). Based on the redox activity of metal ions, the present invention finds that the composite has a better electrocatalytic effect on hydrogen peroxide (H 2 O 2 ). In addition, UOx can rapidly oxidize UA into allantoic acid and H 2 O 2 , so by modifying the electrode with the composite and its catalytic reduction of H 2 O 2 , a new electrochemical method is constructed in the present invention to realize the reduction of uric acid and H 2 O 2 . The analysis and monitoring of uric acid oxidase provides a new idea for uric acid-related clinical diagnosis and drug development.
发明内容SUMMARY OF THE INVENTION
本发明所要解决的技术问题是提供一种特异性好、灵敏度高、检测速度快、结果准确可靠、成本低的检测尿酸及尿酸氧化酶的电化学方法及其应用。The technical problem to be solved by the present invention is to provide an electrochemical method for detecting uric acid and uric acid oxidase and its application with good specificity, high sensitivity, fast detection speed, accurate and reliable results, and low cost.
本发明解决上述技术问题所采用的技术方案为：一种检测尿酸及尿酸氧化酶的电化学方法及其应用，具体步骤如下：The technical solution adopted by the present invention to solve the above-mentioned technical problems is: an electrochemical method for detecting uric acid and uric acid oxidase and its application, and the specific steps are as follows:
(1)GSH-Ag(I)复合物的制备(1) Preparation of GSH-Ag(I) complex
依次取1～10μL浓度为0.1～20mM的硝酸银水溶液、1～10μL浓度为0.1～20mM谷胱甘肽水溶液，混合均匀后加入磷酸缓冲溶液(10mM，pH 7.0，Na2HPO4/NaH2PO4)配成50～120μL的溶液，将溶液在25～40℃下缓缓震荡8～17min，即得到GSH-Ag(I)复合物，使用前将该复合物溶液与50～120μL0.02％wt Nafion溶液混合均匀，静置5～20min。Take 1-10 μL of silver nitrate aqueous solution with a concentration of 0.1-20 mM and 1-10 μL of an aqueous solution of glutathione with a concentration of 0.1-20 mM in turn, and then add phosphate buffer solution (10 mM, pH 7.0, Na 2 HPO 4 /NaH 2 PO after mixing evenly). 4 ) Make up a solution of 50-120 μL, and shake the solution slowly at 25-40° C. for 8-17 minutes to obtain a GSH-Ag(I) complex. Mix the complex solution with 50-120 μL of 0.02% before use. The wt Nafion solution is mixed evenly and left to stand for 5-20 minutes.
(2)电化学生物传感器的制备(2) Preparation of electrochemical biosensors
a.将玻碳电极(GCE，直径为3mm)在麂皮上依次用粒径0.3μm、0.05μm的三氧化二铝粉末抛光0.5～5min，抛光后将电极置于超声清洗器中用超纯水超声清洗1～5min，然后用N2吹干，记为GCE；a. Polish the glassy carbon electrode (GCE, 3mm in diameter) on the chamois with Al2O3 powder with particle size of 0.3μm and 0.05μm in turn for 0.5-5min. After polishing, place the electrode in an ultrasonic cleaner with ultrapure Ultrasonic cleaning with water for 1-5 min, and then drying with N 2 , recorded as GCE;
b.利用循环伏安法，设置电位范围为-1.2～0.5V、扫速5～20mV/s将0.5～2mg/mL石墨烯分散液(GO)电沉积到裸玻碳电极上得到GO/GCE；然后用取5～15μL(1)中溶液滴涂于GO/GCE上，室温下静置20～60min，用超纯水缓缓冲洗电极，记为GSH-Ag(I)/GO/GCE。b. Using cyclic voltammetry, set the potential range to -1.2 to 0.5 V and the scan rate to 5 to 20 mV/s to electrodeposit 0.5 to 2 mg/mL graphene dispersion (GO) on the bare glassy carbon electrode to obtain GO/GCE Then, 5-15 μL of the solution in (1) was applied dropwise on GO/GCE, left at room temperature for 20-60 min, and the electrode was slowly rinsed with ultrapure water, which was recorded as GSH-Ag(I)/GO/GCE.
(3)尿酸及尿酸氧化酶的分析检测(3) Analysis and detection of uric acid and uric acid oxidase
总体积为2mL的反应液，包括UA(0～5mM)，UOx(0～2000U/L)，磷酸缓冲溶液(Na2HPO4/NaH2PO4，0.1M，pH 8.0)，于25～45℃下反应2～10min，随后用于GSH-Ag(I)/GO/GCE的电化学响应检测。The total volume of the reaction solution is 2mL, including UA (0～5mM), UOx (0～2000U/L), phosphate buffer solution (Na 2 HPO 4 /NaH 2 PO 4 , 0.1M, pH 8.0), at 25～45 The reaction was carried out at ℃ for 2-10 min, and then used for the electrochemical response detection of GSH-Ag(I)/GO/GCE.
基于以上反应液，通过改变UA浓度(0.001～1mM)，其他步骤同上，可实现对UA的检测；Based on the above reaction solution, the detection of UA can be realized by changing the UA concentration (0.001-1mM), and other steps are the same as above;
基于以上反应液，通过改变UOx浓度(1～2000U/L)，其他步骤同上，可实现对UOx的检测。Based on the above reaction solution, by changing the concentration of UOx (1-2000U/L), and other steps are the same as above, the detection of UOx can be realized.
利用上述检测尿酸及尿酸氧化酶的电化学方法及其应用，利用电流-时间法(Amperometric i-t Curve)，设置电位为+0.4V，利用所制备电化学传感器对H2O2的电化学响应，在含有UOx反应液的电解质溶液中获得一系列不同浓度UA(UOx)对应的电流大小，建立电流响应与UA(UOx)之间的定量关系，根据两者之间的定量关系，确定待测样品中UA(UOx)的含量。Utilize the above-mentioned electrochemical method for detecting uric acid and uric acid oxidase and its application, utilize Amperometric it Curve, set the potential to +0.4V, utilize the electrochemical response of the prepared electrochemical sensor to H 2 O 2 , Obtain a series of current magnitudes corresponding to different concentrations of UA(UOx) in the electrolyte solution containing the UOx reaction solution, establish a quantitative relationship between the current response and UA(UOx), and determine the sample to be tested according to the quantitative relationship between the two. The content of UA(UOx).
发明原理：本发明是一种检测尿酸及尿酸氧化酶的电化学方法及其应用，首先采用GSH作为生物配体，Ag(I)作为金属节点，基于巯基-金属和金属-金属之间的相互作用合成了一种能够对H2O2有电催化效应的生物金属复合物(GSH-Ag(I)复合物)，再将聚合物滴涂到GO修饰电极上，成功制备传感器。利用UOx催化UA生成H2O2，分别固定UOx或UA浓度，通过复合物对H2O2的响应作为信号输出，实现对UA或UOx的分析检测。基于此，构建了一种简单、快速、高灵敏、高选择性、免标记的UA(UOx)电化学分析方法。Principle of the invention: The present invention is an electrochemical method for detecting uric acid and uric acid oxidase and its application. First, GSH is used as a biological ligand and Ag(I) is used as a metal node. Based on the interaction between sulfhydryl-metal and metal-metal A biometallic complex (GSH-Ag(I) complex) with electrocatalytic effect on H 2 O 2 was synthesized, and then the polymer was drop-coated on the GO modified electrode, and the sensor was successfully prepared. UOx is used to catalyze UA to generate H 2 O 2 , the concentration of UOx or UA is fixed respectively, and the response of the complex to H 2 O 2 is used as a signal output to realize the analysis and detection of UA or UOx. Based on this, a simple, rapid, highly sensitive, highly selective, label-free electrochemical analysis method for UA(UOx) was constructed.
与现有技术相比，本发明的优点在于：本发明构建了一种检测尿酸及尿酸氧化酶的电化学方法及其应用。首先，通过GSH中巯基与Ag(I)和Ag(I)与Ag(I)相互作用简单绿色合成GSH-Ag(I)复合物，该复合物对H2O2有着十分明显的催化效应。其次，利用配合物修饰电极，采用电流-时间法检测传感器对不同浓度UA(UOx)的电化学响应。显然，在浓度一定范围内，UA浓度越大，产生的H2O2越多，电流响应越明显；同理，UOx浓度越大，电流响应越明显。实验结果表明，电流的大小与UA(UOx)的浓度在一定范围内呈线性关系，实现对UA(UOx)的检测。其优点在于：Compared with the prior art, the present invention has the advantages that the present invention constructs an electrochemical method for detecting uric acid and uric acid oxidase and its application. First, the GSH-Ag(I) complex was synthesized simply and greenly through the interaction of thiol groups in GSH with Ag(I) and Ag(I) and Ag(I), which has a very obvious catalytic effect on H 2 O 2 . Secondly, the complex was used to modify the electrode, and the electrochemical response of the sensor to different concentrations of UA(UOx) was detected by the amperometric-time method. Obviously, within a certain concentration range, the greater the concentration of UA, the more H 2 O 2 produced, and the more obvious the current response; similarly, the greater the concentration of UOx, the more obvious the current response. The experimental results show that the magnitude of the current has a linear relationship with the concentration of UA(UOx) within a certain range, and the detection of UA(UOx) is realized. Its advantages are:
(1)高催化活性。利用GSH和Ag(I)制备的GSH-Ag(I)复合物对H2O2具有较高的催化活性，可用来制备新型电化学传感器。(1) High catalytic activity. The GSH-Ag(I) complexes prepared by using GSH and Ag(I) have high catalytic activity towards H 2 O 2 and can be used to prepare novel electrochemical sensors.
(2)高灵敏度。本发明基于GSH-Ag(I)复合物制备电化学传感器，利用UOx催化UA生成的H2O2，得到两条线性方程：电流响应对UA浓度线性相关方程为y＝53.47CUA+0.01，r＝0.9939，检测限为0.3μM；电流响应对UOx浓度线性相关方程为y＝18.07lgCUOx+0.05，r＝0.9980，检测限为0.5U/L；说明该传感器可对UA(UOx)实现高灵敏度检测。(2) High sensitivity. The invention prepares an electrochemical sensor based on GSH-Ag(I) complex, utilizes H 2 O 2 generated by UOx catalyzing UA, and obtains two linear equations: the linear correlation equation of current response to UA concentration is y=53.47C UA +0.01, r=0.9939, the detection limit is 0.3μM; the linear correlation equation of the current response to UOx concentration is y=18.07lgC UOx +0.05 , r=0.9980, the detection limit is 0.5U/L; it shows that the sensor can achieve high sensitivity to UA(UOx). Sensitivity detection.
(3)高特异性。对UA检测：其他对照物质如尿素(Urea)、葡萄糖(Glucose)、咖啡因(Caffeine)、抗坏血酸(AA)对体系均无干扰；对UOx检测：乙酰胆碱酯酶(AChE)、末端转移酶(TdT)、碱性磷酸酶(ALP)、核酸外切酶I(Exo I)、焦磷酸酶(PPase)和溶菌酶(LZM)对体系均无干扰；(3) High specificity. UA detection: other control substances such as urea (Urea), glucose (Glucose), caffeine (Caffeine), ascorbic acid (AA) have no interference to the system; for UOx detection: acetylcholinesterase (AChE), terminal transferase (TdT) ), alkaline phosphatase (ALP), exonuclease I (Exo I), pyrophosphatase (PPase) and lysozyme (LZM) did not interfere with the system;
(4)结果准确。回收率均在90％～110％之间。(4) The results are accurate. The recoveries were all between 90% and 110%.
(5)制备与检测方法试剂用量少、成本低。本发明只需消耗少量材料和试剂就可实现对UA(UOx)的高灵敏检测。(5) Preparation and detection method with less reagent consumption and low cost. The present invention can realize highly sensitive detection of UA(UOx) only by consuming a small amount of materials and reagents.
综上所述，本发明是构建了一种GSH-Ag(I)复合物基底的电化学传感器用于UA(UOx)的检测，具有灵敏度高、选择性好、操作简单、分析快速、易于操作等优点，可以实现较低浓度UA(UOx)的检测，具有良好的应用前景。In summary, the present invention is to construct a GSH-Ag(I) composite substrate electrochemical sensor for the detection of UA(UOx), which has the advantages of high sensitivity, good selectivity, simple operation, rapid analysis, and easy operation. and other advantages, the detection of lower concentrations of UA(UOx) can be realized, which has a good application prospect.
附图说明Description of drawings
图1为本发明传感器对H2O2的电化学响应图；Fig. 1 is the electrochemical response diagram of the sensor of the present invention to H 2 O 2 ;
图2为本发明传感器对UA(UOx)分析检测的可行性实验图；Fig. 2 is a feasibility experiment diagram of the sensor of the present invention analyzing and detecting UA (UOx);
图3为本发明传感器对不同浓度UA(UOx)的电流响应对浓度的校准曲线图；Fig. 3 is the calibration curve diagram of the current response of the sensor of the present invention to different concentrations of UA (UOx) to concentration;
图4为本发明传感器对UA(UOx)的特异性实验图。FIG. 4 is an experimental diagram of the specificity of the sensor of the present invention to UA(UOx).
具体实施方式Detailed ways
以下结合附图实施例对本发明作进一步详细描述。The present invention will be further described in detail below with reference to the embodiments of the accompanying drawings.
实施例1 GSH-Ag(I)复合物的制备Example 1 Preparation of GSH-Ag(I) complex
依次取10μL浓度为10mM的硝酸银水溶液、10μL浓度为10mM谷胱甘肽水溶液，混合均匀后加入磷酸缓冲溶液(10mM，pH 7.0，Na2HPO4/NaH2PO4)配成100μL的溶液，将溶液在30℃下缓缓震荡12min，即得到GSH-Ag(I)复合物，使用前将该复合物溶液与100μL 0.02％wtNafion溶液混合均匀，静置80min。Take 10 μL of silver nitrate aqueous solution with a concentration of 10 mM and 10 μL of 10 mM glutathione aqueous solution in turn, mix them evenly, and add phosphate buffer solution (10 mM, pH 7.0, Na 2 HPO 4 /NaH 2 PO 4 ) to make a 100 μL solution, The solution was slowly shaken at 30° C. for 12 min to obtain a GSH-Ag(I) complex. Before use, the complex solution was mixed with 100 μL of 0.02% wtNafion solution and allowed to stand for 80 min.
实施例2电化学生物传感器的制备Example 2 Preparation of electrochemical biosensors
a.首先将玻碳电极(GCE，直径为3mm)在麂皮上依次用粒径0.3μm、0.1μm、0.05μm的三氧化二铝粉末抛光2min，抛光后将电极置于超声清洗器中用超纯水中超声清洗2min，然后用N2吹干，得到裸玻碳电极，记为GCE；a. First, the glassy carbon electrode (GCE, diameter of 3mm) was polished on the chamois with Al2O3 powder with particle size of 0.3μm, 0.1μm and 0.05μm for 2min. After polishing, the electrode was placed in an ultrasonic cleaner for use Ultrapure water was ultrasonically cleaned for 2 min, and then blown dry with N 2 to obtain a bare glassy carbon electrode, denoted as GCE;
b.利用循环伏安法，设置电位范围为-1.2～0.5V、扫速10mV/s将1.2mg/mL石墨烯分散液(GO)电沉积到裸玻碳电极上得到GO/GCE；然后用取4μL实施例1中溶液滴涂于GO/GCE上，室温下静置30min，用超纯水缓缓冲洗电极，记为GSH-Ag(I)/GO/GCE。b. Using cyclic voltammetry, set the potential range from -1.2 to 0.5V and scan rate of 10mV/s to electrodeposit 1.2mg/mL graphene dispersion (GO) on the bare glassy carbon electrode to obtain GO/GCE; then use Take 4 μL of the solution in Example 1 and drop it on GO/GCE, let it stand for 30 min at room temperature, and rinse the electrode slowly with ultrapure water, which is recorded as GSH-Ag(I)/GO/GCE.
检测以上所述电极对PBS(0.1M，pH 7.0)电解质溶液的电化学响应，每100s加入0.025mM H2O2。如图1，可看出制备的传感器相比较于其他两种电极，对H2O2的电化学响应很明显。说明传感器对H2O2有良好的电催化活性。 The electrochemical response of the electrodes described above to a PBS (0.1 M, pH 7.0) electrolyte solution was tested with 0.025 mM H2O2 added every 100 s. As shown in Fig. 1, it can be seen that the prepared sensor has an obvious electrochemical response to H 2 O 2 compared with the other two electrodes. It shows that the sensor has good electrocatalytic activity for H 2 O 2 .
实施例3可行性实验Example 3 Feasibility Experiment
按上述实施例1和实施例2的传感器制备步骤，总体积为2mL的反应液，包括UA(0.001、0.01、0.1mM)，UOx(500U/L)，磷酸缓冲溶液(Na2HPO4/NaH2PO4，0.1M，pH 8.0)，于37℃下反应3min，随后用于GSH-Ag(I)/GO/GCE的电化学响应检测。结果如图2A，可以看出，GSH-Ag(I)/GO/GCE对不同浓度的UA有良好的电化学响应，可应用于对UA的检测。According to the sensor preparation steps of Example 1 and Example 2 above, the total volume of the reaction solution is 2mL, including UA (0.001, 0.01, 0.1mM), UOx (500U/L), phosphate buffer solution (Na 2 HPO 4 /NaH 2 PO 4 , 0.1 M, pH 8.0), reacted at 37° C. for 3 min, and then used for the electrochemical response detection of GSH-Ag(I)/GO/GCE. The results are shown in Figure 2A, it can be seen that GSH-Ag(I)/GO/GCE has good electrochemical response to different concentrations of UA, which can be applied to the detection of UA.
随后，按上述实施例1和实施例2的传感器制备步骤，总体积为2mL的反应液，包括UA(1mM)，UOx(500U/L)，磷酸缓冲溶液(Na2HPO4/NaH2PO4，0.1M，pH 8.0)，于37℃下反应3min，随后用于GSH-Ag(I)/GO/GCE的电化学响应检测。结果如图2B，UOx浓度为0时，传感器基本无响应，而UOx浓度为500U/L时，传感器响应明显。Then, according to the sensor preparation steps of Example 1 and Example 2, the total volume of the reaction solution was 2 mL, including UA (1 mM), UOx (500 U/L), phosphate buffer solution (Na 2 HPO 4 /NaH 2 PO 4 ). , 0.1 M, pH 8.0), reacted at 37 °C for 3 min, and then used for the electrochemical response detection of GSH-Ag(I)/GO/GCE. The results are shown in Figure 2B. When the UOx concentration is 0, the sensor basically has no response, while when the UOx concentration is 500 U/L, the sensor responds significantly.
以上结果证明传感器对UA和UOx有良好的电化学响应，可以应用于UA(UOx)的检测。The above results demonstrate that the sensor has a good electrochemical response to UA and UOx, and can be applied to the detection of UA(UOx).
实施例4 UA和UOx的检测Example 4 Detection of UA and UOx
按上述实施例1和实施例2的传感器制备步骤，以及实施例3对UA(UOx)的响应，通过改变UA(0.001～1mM)或UOx(1～2000U/L)的浓度，利用传感器检测对反应液的响应，结果如图3。如图3A，传感器对UA(UOx)的电流响应与浓度呈良好的线性关系，传感器的电流响应对UA浓度线性相关方程为y＝53.47CUA+0.01，r＝0.9939，线性范围为0.001～1mM，检测限为0.3μM；如图3B，电流响应对UOx浓度线性相关方程为y＝18.07lgCUOx+0.05，r＝0.9980，线性范围为1～2000U/L，检测限为0.5U/L，说明传感器对UA(UOx)实现高灵敏检测。According to the above-mentioned sensor preparation steps of Example 1 and Example 2, and the response of Example 3 to UA (UOx), by changing the concentration of UA (0.001-1mM) or UOx (1-2000U/L), the sensor is used to detect the The response of the reaction solution is shown in Figure 3. As shown in Figure 3A, the current response of the sensor to UA (UOx) has a good linear relationship with the concentration. The linear correlation equation of the current response of the sensor to the UA concentration is y=53.47C UA +0.01, r=0.9939, and the linear range is 0.001～1mM , the detection limit is 0.3 μM; as shown in Figure 3B, the linear correlation equation of the current response to UOx concentration is y=18.07lgC UOx +0.05 , r=0.9980, the linear range is 1～2000U/L, and the detection limit is 0.5U/L, indicating that The sensor realizes highly sensitive detection of UA(UOx).
实施例4特异性检测Example 4 Specific detection
为了验证该传感器的特异性，按上述实施例1、2、3的传感器制备步骤，UA的反应液中，加入其它相同浓度的干扰物，如与UA相同浓度的尿素(Urea)、葡萄糖(Glucose)、咖啡因(Caffeine)、抗坏血酸(AA)，与UOx相同浓度的乙酰胆碱酯酶(AChE)、末端转移酶(TdT)、碱性磷酸酶(ALP)、核酸外切酶I(Exo I)、焦磷酸酶(PPase)和溶菌酶(LZM)，检测传感器对UA(UOx)的特异性。结果如图4所示，说明传感器对于UA(如图4A)(UOx(如图4B))的检测有很好的特异性。In order to verify the specificity of the sensor, according to the sensor preparation steps of the above-mentioned embodiments 1, 2, and 3, other interfering substances of the same concentration are added to the reaction solution of UA, such as urea (Urea) and glucose (Glucose (Glucose) of the same concentration as UA. ), caffeine (Caffeine), ascorbic acid (AA), acetylcholinesterase (AChE), terminal transferase (TdT), alkaline phosphatase (ALP), exonuclease I (Exo I), Pyrophosphatase (PPase) and lysozyme (LZM), to detect the specificity of the sensor for UA (UOx). The results are shown in Fig. 4, indicating that the sensor has good specificity for the detection of UA (Fig. 4A) (UOx (Fig. 4B)).
当然，上述说明并非对本发明的限制，本发明也并不限于上述举例。本技术领域的普通技术人员在本发明的实质范围内做出的变化、改型、添加或替换，也应属于本发明保护范围。Of course, the above description does not limit the present invention, and the present invention is not limited to the above examples. Changes, modifications, additions or substitutions made by those skilled in the art within the essential scope of the present invention should also belong to the protection scope of the present invention.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050189240A1 (en) * | 2004-02-26 | 2005-09-01 | Tamkang University | Method and chemical sensor for determining concentrations of hydrogen peroxide and its precusor in a solution |
| CN108061750A (en) * | 2017-11-30 | 2018-05-22 | 宁波大学 | Hydrogen peroxide and glucose detection are used for based on the proteinoid nano wire structure electrochemica biological sensor with electro catalytic activity |
| CN108120761A (en) * | 2017-11-30 | 2018-06-05 | 宁波大学 | Electrochemica biological sensor based on the peptidomimetics with electro catalytic activity detects for acetylcholinesterase |
-
2020
- 2020-07-10 CN CN202010707334.8A patent/CN111879836B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050189240A1 (en) * | 2004-02-26 | 2005-09-01 | Tamkang University | Method and chemical sensor for determining concentrations of hydrogen peroxide and its precusor in a solution |
| CN108061750A (en) * | 2017-11-30 | 2018-05-22 | 宁波大学 | Hydrogen peroxide and glucose detection are used for based on the proteinoid nano wire structure electrochemica biological sensor with electro catalytic activity |
| CN108120761A (en) * | 2017-11-30 | 2018-06-05 | 宁波大学 | Electrochemica biological sensor based on the peptidomimetics with electro catalytic activity detects for acetylcholinesterase |
Non-Patent Citations (1)
| Title |
|---|
| SHILPI VERMA,ET AL.: "Uricase grafted nanoconducting matrix based electrochemical biosensor for ultrafast uric acid detection in human serum samples", 《INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113758983A (en) * | 2021-08-18 | 2021-12-07 | 齐鲁医药学院 | Construction method of glucose biosensor based on glutathione assembly |
| CN113758983B (en) * | 2021-08-18 | 2024-02-02 | 齐鲁医药学院 | Construction method of glucose biosensor based on glutathione assembly |
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