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 sarco

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
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Example Embodiment

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

[0038] The glassy carbon electrode (GCE, diameter 3mm) was pretreated. First, methylformamide (DMF) solution was used 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 minutes to ensure that PAA was completely dissolved, and then 5 μL of the sonicated polyamic acid (PAA) solution was deposited on the surface of the GCE, and dried overnight at room temperature. Subsequently, 5 μL of a 2.5% glutaraldehyde solution uniformly dispersed was added dropwise to the surface of the modified GCE, and dried at 25° C. for 2 hours to form a film. Finally, 5 μL of sarcosine oxidase (SOX) solution (1 mg / mL) was dropped on the surface of the modified GCE and left overnight at 4° C. to form a SOX layer.

Example Embodiment

[0039] Example 2 Electrochemical detection

[0040] Use chronoamperometry in 5mL 0.5M PBS (pH 7.2) containing different concentrations of sarcosine, starting potential is -0.4V, and the interval of dripping is 150s;

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

[0042] Use electrochemical impedance spectroscopy (Electrochemical Impedance Spectroscopy, referred to as EIS) in 5.0mM [Fe(CN) containing 0.1M KCl 6 ] 3- / 4- In solution, the frequency is 10-10 4 Hz.

Example Embodiment

[0043] Experimental example 1 electrode modification process

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

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

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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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