阅读说明：本技术 一种快速检测过氧化氢含量的方法 (Method for rapidly detecting content of hydrogen peroxide ) 是由 李剑锋 温宝英 于 2019-11-26 设计创作，主要内容包括：本发明公开了一种用于快速检测过氧化氢含量的SERS芯片,所述SERS芯片包括基底以及滴加于所述基底上的增强纳米粒子,所述基底上修饰有苯硼酸类分子,所述增强纳米粒子包括金纳米粒子以及包裹于所述金纳米粒子表面的惰性壳层。本发明还提供了一种利用所述SERS芯片快速检测过氧化氢含量的方法,包括将含有过氧化氢代谢产物的溶液滴在所述SERS芯片上,进行拉曼光谱测试,得到过氧化氢与苯硼酸类分子反应后的拉曼图谱,与标准曲线对照,从而计算出过氧化氢的含量。本发明的检测方法具有前处理简单、快速、高效、特异性强的优点。(The invention discloses a SERS chip for rapidly detecting hydrogen peroxide content, which comprises a substrate and enhanced nanoparticles dripped on the substrate, wherein phenylboronic acid molecules are modified on the substrate, and the enhanced nanoparticles comprise gold nanoparticles and an inert shell layer wrapped on the surfaces of the gold nanoparticles. The invention also provides a method for rapidly detecting the content of the hydrogen peroxide by utilizing the SERS chip, which comprises the steps of dripping a solution containing a hydrogen peroxide metabolite on the SERS chip, carrying out Raman spectrum test to obtain a Raman spectrum after the reaction of the hydrogen peroxide and phenylboronic acid molecules, and comparing the Raman spectrum with a standard curve, thereby calculating the content of the hydrogen peroxide. The detection method has the advantages of simple pretreatment, rapidness, high efficiency and strong specificity.)

1. The SERS chip for rapidly detecting the content of hydrogen peroxide is characterized by comprising a substrate and enhanced nanoparticles dropwise added on the substrate, wherein phenylboronic acid molecules are modified on the substrate, and the enhanced nanoparticles comprise gold nanoparticles and an inert shell layer wrapped on the surfaces of the gold nanoparticles.

2. The SERS chip according to claim 1, wherein the substrate is a silicon-based gold film or a silver film.

3. The SERS chip for rapidly detecting hydrogen peroxide content according to claim 1, wherein the phenylboronic acid molecule is 4-mercaptophenylboronic acid or 4-cyanophenylboronic acid.

4. The SERS chip for rapidly detecting hydrogen peroxide content according to claim 1, wherein the gold nanoparticles have a particle size of 40-60 nm.

5. The SERS chip for rapidly detecting the content of hydrogen peroxide according to claim 1, wherein the inert shell layer is a silicon dioxide shell layer, and the thickness of the inert shell layer is 1-2 nm.

6. A method for rapidly detecting the content of hydrogen peroxide is characterized by comprising the following steps:

(1) providing a SERS chip according to any of claims 1-5;

(2) and dripping a solution containing a hydrogen peroxide metabolite on the SERS chip, performing Raman spectrum test to obtain a Raman spectrum after the reaction of the hydrogen peroxide and the phenylboronic acid molecules, and comparing the Raman spectrum with a standard curve to calculate the content of the hydrogen peroxide.

7. The method for rapidly detecting the content of hydrogen peroxide according to claim 6, wherein the standard curve is established by the following steps:

(1) providing a series of standard sample solutions of hydrogen peroxide at different concentrations;

(2) and dripping the hydrogen peroxide standard sample solutions with different concentrations on an SERS chip, detecting by using a Raman spectrum tester, recording a detection result, and establishing a standard curve between the Raman detection signal intensity and the concentration of the hydrogen peroxide standard sample solution.

8. The method of claim 7, wherein the standard curve is established at 1023cm ^-1Peak of characteristic peakArea sum 1078cm ^-1And (4) drawing a standard curve by the ratio of the peak area of the characteristic peak to the concentration of the hydrogen peroxide.

Technical Field

The invention relates to the technical field of Raman spectrum rapid detection, in particular to a method for rapidly detecting the content of a cell metabolite, namely hydrogen peroxide by utilizing a surface-enhanced Raman spectrum technology.

Background

Reactive Oxygen Species (ROS) play a very important role as a highly reactive chemical species in physiological and pathological processes. The Reactive Oxygen Species (ROS) is a general term for a substance that is composed of oxygen in the body or in the natural environment, contains oxygen, and is active in nature. If the body is lack of active oxygen, certain diseases (such as chronic granulomatosis, certain autoimmune diseases and the like) can be induced; excessive production of active oxygen in vivo can also induce diseases (such as cancer)Arthritis, arteriosclerosis, etc.). ROS in organisms include neutral molecules (e.g., H) ₂O ₂， ¹O ₂) Free radicals (for example: hydroxyl radical), ions (e.g.: superoxide, hypochlorous acid, nitrosyl peroxide), and the like. Hydrogen peroxide is an important component of ROS and is mainly used as an important signal molecule for cell growth, proliferation and differentiation in cells, and excessive hydrogen peroxide attacks organelles or active biomolecules in cells, such as proteins, liposomes, DNA and the like. Meanwhile, hydrogen peroxide in cells is also related to physiological and pathological processes such as aging, Alzheimer disease and cancer. The methods commonly used at present for detecting the content of hydrogen peroxide mainly include electrochemical analysis methods, chromatography, spectrophotometry and the like. For example, in patent CN106404873B, hydrogen peroxide is quantitatively detected by a method of preparing an electrochemical biosensor; in patent CN103712930B, hydrogen peroxide is reacted with a cerium sulfate standard solution, and then the hydrogen peroxide is quantitatively detected by a spectrophotometer. Although the above methods can accurately perform quantitative analysis of hydrogen peroxide, they have some disadvantages such as complicated pretreatment, expensive apparatus, long operation time, and the like. Therefore, it is important to develop a rapid, simple and cheap method for quantitatively detecting the hydrogen peroxide, which is a metabolic product in cells.

The surface enhanced Raman spectroscopy technology has the advantages of high detection speed, high detection sensitivity (capable of reaching a single molecule level), capability of providing molecular fingerprint spectroscopy and the like. Since the discovery of the technology, the technology can be rapidly and widely applied to a plurality of fields such as food, medicine, biology, chemistry and the like.

Disclosure of Invention

The invention aims to provide an SERS chip for rapidly detecting hydrogen peroxide content and a method for rapidly detecting hydrogen peroxide content, wherein the detection method has the advantages of simple pretreatment, rapidness, high efficiency and strong specificity.

In order to solve the technical problem, the invention provides a SERS chip for rapidly detecting hydrogen peroxide content, which comprises a substrate and enhanced nanoparticles dropwise added on the substrate, wherein phenylboronic acid molecules are modified on the substrate, and the enhanced nanoparticles comprise gold nanoparticles and inert shell layers wrapped on the surfaces of the gold nanoparticles.

In the invention, the inert shell layer mainly plays a role of isolating the external environment, so that the nano particles are not influenced by the environment.

Further, the substrate is a silicon-based gold film or a silver film.

Further, the phenylboronic acid molecules are 4-mercaptophenylboronic acid or 4-cyanophenylboronic acid.

Further, the particle size of the gold nanoparticles is 40-60 nm.

Further, the inert shell layer is a silicon dioxide shell layer, and the thickness of the inert shell layer is preferably 1-2 nm.

The invention also provides a method for rapidly detecting the content of hydrogen peroxide, which comprises the following steps:

(1) providing the SERS chip;

(2) and dripping a solution containing a hydrogen peroxide metabolite on the SERS chip, performing Raman spectrum test to obtain a Raman spectrum after the reaction of the hydrogen peroxide and the phenylboronic acid molecules, and comparing the Raman spectrum with a standard curve to calculate the content of the hydrogen peroxide.

Further, the standard curve is established by the following steps:

(1) providing a series of standard sample solutions of hydrogen peroxide at different concentrations;

(2) and dripping the hydrogen peroxide standard sample solutions with different concentrations on an SERS chip, detecting by using a Raman spectrum tester, recording a detection result, and establishing a standard curve between the Raman detection signal intensity and the concentration of the hydrogen peroxide standard sample solution.

Further, when a standard curve is established, 1023cm is used ^-1Peak area of characteristic peak and 1078cm ^-1And (4) drawing a standard curve by the ratio of the peak area of the characteristic peak to the concentration of the hydrogen peroxide.

The invention has the beneficial effects that:

the invention provides a surface enhanced Raman spectroscopy method for rapidly detecting the content of hydrogen peroxide, which has the advantages that: the quantitative analysis of the hydrogen peroxide can be realized rapidly, efficiently and nondestructively without pretreatment and addition of an additional internal standard substance.

Drawings

FIG. 1 is a reaction formula of 4-mercaptophenylboronic acid and hydrogen peroxide to produce 4-mercaptophenol;

FIG. 2 shows molecular formulae and Raman spectra of 4-mercaptophenylboronic acid and 4-mercaptophenol;

FIG. 3 is a Raman spectrum of hydrogen peroxide at various concentrations after reaction with 4-mercaptophenylboronic acid;

FIG. 4 is a standard curve for different concentrations of hydrogen peroxide;

FIG. 5 is a Raman spectrum of a glucose decomposition product at different concentrations after reaction with 4-mercaptophenylboronic acid;

FIG. 6 is a standard curve of glucose at different concentrations.

Detailed Description

The present invention is further described below in conjunction with the following figures and specific examples so that those skilled in the art may better understand the present invention and practice it, but the examples are not intended to limit the present invention.