Tyrosinase activity detection method based on recyclable SERS sensor

A technology for tyrosinase and activity detection, applied in the field of tyrosinase activity detection, can solve the problems of difficulty in the analysis of surface-enhanced Raman spectroscopy, rare surface-enhanced Raman spectroscopy research, and direct detection selectivity limitation. , to achieve the effect of convenient evaluation, low cost and overcoming technical deficiencies

Active Publication Date: 2019-02-15
EAST CHINA UNIV OF SCI & TECH
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Problems solved by technology

However, due to the complexity of protein molecules, the analysis of surface-enhanced Raman spectra is difficult, which limits the selectivity of direct detection.
In addition, tyrosinase in living organisms

Method used

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  • Tyrosinase activity detection method based on recyclable SERS sensor
  • Tyrosinase activity detection method based on recyclable SERS sensor
  • Tyrosinase activity detection method based on recyclable SERS sensor

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preparation example Construction

[0029] There are many methods for preparing the surface-enhanced Raman scattering sensor, as long as the structure of the surface-enhanced Raman scattering sensor can be formed. A preparation method of the surface-enhanced Raman scattering sensor is exemplified below, which includes the following steps:

[0030] First, a nanoparticle gold colloid substrate with surface-enhanced Raman activity was prepared. The specific process is as follows:

[0031] (1) Prepare a chloroauric acid solution with a mass fraction of 0.05% to 0.1%, and heat to boiling.

[0032] The boiling means that the solution being heated is boiling in the natural environment, and since the boiling temperature of the solution is different at different altitudes, it is not limited herein.

[0033] (2) Slowly add sodium citrate with a mass fraction of 1% to 2% into the chloroauric acid solution dropwise, and keep boiling for more than 15 minutes.

[0034] The reaction time of the reducing agent solution and the...

Embodiment 1

[0059] 1. Prepare a surface-enhanced Raman scattering sensor, specifically including the following steps:

[0060] (1) Preparation of nanoparticle gold colloid substrate with surface-enhanced Raman activity.

[0061] Prepare 100 mL of 0.05% tetrachloroauric acid trihydrate solution, place in a clean beaker and heat to 100°C. Take 0.1g of sodium citrate in 10mL of ultrapure water and dissolve it fully, then slowly add it dropwise to the above solution, continue to boil for 15 minutes, then let it cool down to room temperature, and obtain a red nanoparticle gold colloid with surface-enhanced Raman activity . figure 2 is the TEM image of red nanoparticle gold colloid with surface-enhanced Raman activity, from figure 2 It can be seen that the nanoparticle gold colloid with surface-enhanced Raman activity has a uniform particle size and is a better surface-enhanced Raman substrate.

[0062] (2) Preparation of a surface-enhanced Raman scattering sensor for the detection of tyro...

Embodiment 2

[0073] 1. Prepare a surface-enhanced Raman scattering sensor, specifically including the following steps:

[0074] (1) Preparation of nanoparticle gold colloid substrate with surface-enhanced Raman activity.

[0075] Prepare 100 mL of 0.05% tetrachloroauric acid trihydrate solution, place in a clean beaker and heat to 100°C. Take 0.1g of sodium citrate in 10mL of ultrapure water and dissolve it fully, then slowly add it dropwise to the above solution, continue to boil for 15 minutes, then let it cool down to room temperature, and obtain red nanoparticle gold colloid with surface-enhanced Raman activity .

[0076] (2) Preparation of a surface-enhanced Raman scattering sensor for the detection of tyrosinase activity.

[0077] Take a clean ITO electrode and clean it ultrasonically with deionized water and acetone for 30 minutes, then dry it with nitrogen. Subsequently, the ITO electrode was functionalized by immersing it in an acetone solution containing 1.0 mM 3-mercaptopropy...

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Abstract

The invention provides a tyrosinase activity detection method based on a recyclable SERS sensor. The method comprises the following steps of (a) providing the surface enhanced Raman scattering (SERS)) sensor, wherein the sensor is a self-assembled electrode, the electrode is provided with a nanogold particle membrane layer with SERS activity, and Raman signal molecules exist on the nanogold particle membrane layer; (b) establishing a tyrosinase activity detection fingerprint spectrum and a standard curve; (c) acquiring a surface enhanced Raman spectrum of the SERS sensor after reaction with tyrosinase in a sample solution; (d) comparing the surface enhanced Raman spectrum of the sample solution obtained in the step (c) with the fingerprint spectrum, and carrying out qualitative and quantitative analysis; and (e) enabling the sensor after detection to be recyclable by utilizing electrochemical regulation and control. The detection method has the advantages that the technical defects ofan existing detection method are overcome; the activity of the tyrosinase can be rapidly and sensitively detected; and the sensor is recyclable and is expected to realize automatic detection of the tyrosinase.

Description

technical field [0001] The invention relates to the field of biochemical analysis, in particular to a method for detecting tyrosinase activity using a recyclable SERS sensor based on the surface-enhanced Raman scattering (SERS) effect. Background technique [0002] As an important oxidoreductase, tyrosinase plays a vital role in the synthesis of melanin in the human body. In the human body, abnormal levels of tyrosinase can cause diseases such as vitiligo, freckles and Parkinson's syndrome, and tyrosinase is one of the markers of melanoma. Therefore, the development of effective methods for detecting tyrosinase activity will significantly improve the early diagnosis of tyrosinase-related diseases and promote an in-depth understanding of the physiological functions of tyrosinase. [0003] At present, the methods used to detect tyrosinase mainly include: colorimetric method, electrochemical method, high performance liquid chromatography and fluorescence method, etc. However, ...

Claims

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Application Information

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IPC IPC(8): G01N21/65
CPCG01N21/65
Inventor 李大伟汪璐甘振飞江蕾常帅詹德生李家宾
Owner EAST CHINA UNIV OF SCI & TECH
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