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Electrochemical detection method for sarcosine based on polyamide acid and sarcosine oxidase

A technology of sarcosine oxidase and polyamic acid, which is applied in the field of biomedical detection, can solve problems such as the inability to realize sarcosine detection, and achieve significant clinical application value, excellent analytical performance, and great clinical application value

Active Publication Date: 2018-12-11
THE SECOND HOSPITAL OF NANJING
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The invention selects pyridine ruthenium derivatives as luminescent reagents, and preferably uses bipyridyl ruthenium with high sensitivity and high stability. Through simple instruments, it realizes efficient and rapid detection of sarcosine, and this method is better than chromatographic - Mass spectrometry is relatively cheap, but the detectable sample solution of this invention is limited to diluted sarcosine solution, which cannot detect sarcosine in complex samples

Method used

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  • Electrochemical detection method for sarcosine based on polyamide acid and sarcosine oxidase
  • Electrochemical detection method for sarcosine based on polyamide acid and sarcosine oxidase
  • Electrochemical detection method for sarcosine based on polyamide acid and sarcosine oxidase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Immobilization on polyamic acid (PAA) and sarcosine oxidase (SOX) glassy carbon electrode (GCE)

[0038] The glassy carbon electrode (GCE, diameter 3mm) was pretreated, first using methylformamide (DMF) solution as a solvent to prepare a polyamic acid (PAA) solution with a concentration of 4 mg / mL, and the prepared polyamic acid ( The PAA) solution was sonicated for 30 min to ensure complete dissolution of the PAA, and then 5 μL of the sonicated polyamic acid (PAA) solution was deposited on the GCE surface and dried overnight at room temperature. Subsequently, 5 μL of a uniformly dispersed 2.5% glutaraldehyde solution was dropped onto the surface of the modified GCE and dried at 25 °C for 2 h to form a film. Finally, 5 μL of sarcosine oxidase (SOX) solution (1 mg / mL) was dropped on the modified GCE surface and left overnight at 4 °C to form a SOX layer.

Embodiment 2

[0039] Embodiment 2 electrochemical detection

[0040]Chronoamperometry was carried out in 5mL 0.5M PBS (pH7.2) containing different concentrations of sarcosine, the initial potential was -0.4V, and the sample drop interval was 150s;

[0041] Wherein, the amino acid concentration is (a) 1, (b) 2.5, (c) 5, (d) 10, (e) 25, (f) 50, (g) 75, (h) 100 and (i) 125nM;

[0042] Electrochemical Impedance Spectroscopy (EIS for short) was used in 5.0mM [Fe(CN) containing 0.1M KCl 6 ] 3- / 4- In solution, the frequency is 10-10 4 Hz.

experiment example 1

[0043] Modification process of experiment example 1 electrode

[0044] In this experimental example, the surface morphology of the modified electrode was studied by scanning electron microscopy (SEM). attached figure 2 a shows the topography of the bare GCE; figure 2 b shows PAA / GCE with a uniform PAA layer added dropwise, and the results show that the polymer has covered the electrode surface with good adhesion; figure 2 c shows the SEM image of glutaraldehyde / PAA / GCE with visible thick layer; figure 2 d shows the SEM image of SOX / glutaraldehyde / PAA / GCE, which clearly shows that SOX has been immobilized on the modified electrode surface.

[0045] In order to further verify the modification process of the electrode, this experimental example uses electrochemical impedance spectroscopy (EIS) to verify the modification process of the electrode. as attached image 3 As shown, the EIS of the bare electrode (curve a) is similar to a straight line; after PAA is immobilized,...

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Abstract

The invention establishes an electrochemical detection method for sarcosine based on polyamide acid (PAA) and sarcosine oxidase (SOX), so as to realize the detection of sarcosine in a complex sample.The method comprises the following steps: firstly, pretreating a glassy carbon electrode (GCE); taking a dimethyl formamide (DMF) solution as a solvent for preparing a PAA solution and ultrasonicallytreating; depositing the solution on GCE surface, drying under room temperature and staying overnight; dropping the uniformly dispersed glutaraldehyde solution onto the GCE surface and drying for 2 hours at 25 DEG C, thereby forming a film; dropping the SOX solution onto the modified GCE, thereby forming a SOX layer used for detecting sarcosine. PAA is a polymer with a cation complexing property.Under the action of carboxyl group of PAA, the PAA can be used as a reducing agent for improving the sensitivity of the electrochemical method, so as to realize the sensitive detection for sarcosine.

Description

technical field [0001] The invention belongs to the technical field of biomedical detection, and in particular relates to an electrochemical detection method for sarcosine, more specifically, an electrochemical detection method for sarcosine based on polyamic acid and sarcosine oxidase. Background technique [0002] Prostate cancer (PCa) is a common cancer in men with increasing incidence and high risk of metastasis and recurrence. Prostate cancer refers to epithelial malignant tumors that occur in the prostate. According to the 2004 WHO "Pathology and Genetics of Tumors of the Urinary System and Male Reproductive Organs", the pathological types of prostate cancer include adenocarcinoma (acinar adenocarcinoma), ductal adenocarcinoma, urothelial carcinoma, squamous cell carcinoma, and adenosquamous carcinoma. . Among them, prostate adenocarcinoma accounts for more than 95%. Therefore, what we usually call prostate cancer refers to prostate adenocarcinoma. In 2012, the inci...

Claims

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Application Information

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IPC IPC(8): G01N27/26G01N27/327
CPCG01N27/26G01N27/3272
Inventor 李金龙张永臣时瑞滕晓燕胡琴琴
Owner THE SECOND HOSPITAL OF NANJING
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