TW201702578A - Preparation method of microfluidic channel type SERS detection substrate, preparation method of probe type SERS detection substrate, preparation method of planar SERS detection substrate, and detection method of organic pollutants performing comparative analysis and identifying types, concentrations and the like of the organic pollutants - Google Patents

Abstract

This invention provides a preparation method of a microfluidic channel type or probe type surface enhanced Raman scattering (SERS) detection substrate, which comprises the following steps: forming at least one microchannel pattern on a glass substrate by using a photomask via a photolithography method to prepare a microfluidic channel structure master mold; coating a polydimethylsiloxane (PDMS) solution on the previously-described microfluidic channel structure master mold, heating and curing as well as stripping to obtain a PDMS curing structure with a microfluidic channel; and injecting a silver mirror reaction reagent into a microchannel of the PDMS curing structure and then injecting a reducing agent to produce a silver mirror reaction to form silver nanoparticles by reduction, and rinsing with deionized water to obtain the microfluidic channel type SERS detection substrate. Furthermore, this invention provides a detection method capable of rapidly detecting organic pollutants. The method comprises the following steps: making the organic pollutants in a to-be-detected object be adsorbed to the microfluidic channel type or probe type SERS detection substrate, and sensing via a three-dimensional nano Raman fluorescence microscopy system to obtain a Raman spectrogram; and performing comparative analysis and identifying types, concentrations and the like of the organic pollutants.

Description

Method for preparing microfluidic channel type SERS detection substrate, preparation method of probe type SERS detection substrate, preparation method of planar SERS detection substrate, and detection method of organic pollutant

The invention relates to a preparation method of a microfluidic channel type SERS detecting substrate, a preparation method of a probe type SERS detecting substrate, and a method for detecting organic pollutants; in particular, a microfluid channel type or probe A method for preparing a substrate for needle surface enhanced Raman scattering (SERS), comprising: forming a micro-channel pattern comprising at least one microchannel pattern on a glass substrate by a photolithography method using a photomask a flow channel structure master mold; a solution of polydimethylsiloxane (PDMS) is applied to the aforementioned microchannel structure master mold, heat curing and stripping to obtain a PDMS solidified structure having a microfluidic channel; The reagent is injected into the microchannel of the PDMS solidified structure, and then a reducing agent is injected to cause silver mirror reaction to be reduced to form silver nanoparticles, and rinsed with deionized water to obtain a microfluidic channel type SERS detecting substrate.

For a long time, comprehensive indicators of organic pollution such as COD and BOD have been used at home and abroad as parameters for evaluating environmental quality. However, they only reflect certain general content data, and cannot provide effective information on the characteristics of organic pollutants themselves.

For example, some compounds such as aromatic compounds have a large π bond-stabilized structure on the benzene ring, which is difficult to oxidize. Toxic compounds inhibit the microbial activity from being completely degraded. Even if TOC and TOD completely oxidize organic matter, the specific toxic substances cannot be qualitatively and quantitatively determined. Especially the triad organic substances often appear at low concentration levels in the environment, which is enough for human health and ecological environment. Hazard, so the above-mentioned conventional analytical methods such as COD and BODs cannot be used as effective control methods for organic pollutants.

Modern gas chromatography (GC), high performance liquid chromatography (HPLC), mass spectrometry (including color-mass spectrometry) (NMR) and other technologies have emerged, and have been widely used in the field of analytical chemistry, environmental monitoring. And has achieved many important results, opening up broad prospects for detecting organic pollutants in the environment. Although Europe, the United States, Japan, and the United States have successively classified chromatography (including color-mass spectrometry) as environmental monitoring analysis methods and standard analytical methods, they have been widely used in the atmosphere, water quality, soil, biology, and food in recent decades. Such environmental monitoring is used to analyze trace, complex, multi-component organic pollutants; however, chromatography still has considerable limitations on the analytical methods of complex systems.

For example, it is found in water and sediment, water, industrial water, domestic wastewater, industrial wastewater, activated sludge, sediment in farmland or ditch, or other water or sludge, especially in water bodies such as rivers, lakes and oceans. The organic matter is not only a wide variety, but also the concentration is very large, and the low concentration can be as low as several thousandths of micro ppm, so the analysis and identification are difficult.

In particular, the detection of phthalates (PAs) by conventional methods, for example, in the detection of dimethyl phthalate (DMP), diethyl phthalate (DEP), dibutyl phthalate (DBP), butyl benzyl phthalate (BBP), n-octyl phthalate (DNOP), di(2-ethylhexyl) phthalate (DEHP), before passing through the sample Processing, extraction, concentration, purification, and instrumental analysis often take about three days to know the results.

In addition, in the conventional method of detecting organic pollutants in sediments, the analytical techniques used in the past always require extremely complicated processing procedures, such as organic pollutants such as PBDEs, PAEs, PAHs, PCBs, etc., often undergo long-time extraction. And complicated purification procedures. For example, in terms of extraction method, Soxhlet extraction takes 12-24 hours, microwave extraction takes about 1 hour, and super-boundary fluid extraction takes 1 hour; accelerated solvent extraction takes about 0.5 hour. Can be processed. Not only is it time consuming, cost and manpower, but the analytical data obtained is also unsatisfactory in terms of reliability, and further breakthroughs are yet to be made.

Further, for example, polybrominated diphenyl ethers (PBDEs) containing a bromine atom are commonly used as flame retardants in the industry, and have a total of 209 homologues. PBDEs are highly soluble, are not easily decomposed and interfere with the thyroid endocrine of the organism, and thus pose an increasing threat to the environment and humans. Among the PBDEs, the most harmful pollutants to the environment are the most used decabromodiphenyl ether (BDE-209), accounting for 85% to 95% of the total PBDEs. However, since the current detection of the BDE-209 processing procedure requires long time extraction and complicated purification procedures, which is time consuming, costly and labor intensive, it is necessary to develop a rapid and sensitive method for detecting BDE-209.

Therefore, it is expected to develop an effective method for detecting organic pollutants that does not require complicated processing procedures, can shorten the lead time, is easy to operate, saves cost, analyzes fast, and has high reliability.

In view of the above problems of the conventional organic pollutant detection technology, the present inventors have devoted themselves to researching and applying the characteristics of Surface Enhanced Raman Scattering (SERS), thereby developing problems that can improve the conventional detection technology. The novel technology thus completes the present invention.

That is, according to the first aspect of the present invention, a method of preparing a substrate for a microfluidic channel type SERS detection, which is a method of preparing a surface enhanced Raman scattering (SERS) wafer having a microfluidic channel, includes: Forming a microchannel structure master mold by photolithography using a photomask to form at least one microchannel pattern on a glass substrate; applying a polydimethylsiloxane (PDMS) solution to the solution The aforementioned microchannel structure master mold is heat-cured and stripped to obtain a PDMS solidified structure having a microfluidic channel; a silver mirror reagent is injected into the microchannel of the PDMS solidified structure, and then a reducing agent is injected to generate a silver mirror reaction. The silver nanoparticles were reduced to form and washed with deionized water to obtain a microfluidic channel type SERS substrate.

Secondly, according to a second aspect of the present invention, a method of preparing a probe type SERS detecting substrate, which is a method of preparing a surface enhanced Raman scattering (SERS) wafer having a needle tip, comprising: providing a stainless steel The wire is used as a core material, and a PDMS substrate for probe type SERS detection is formed by coating a polydimethylsiloxane (PDMS) solution on the surface of the outer layer of the core material; The probe type SERS detection PDMS substrate was immersed in a silver mirror reaction reagent, and then formaldehyde was injected to cause a uniform silver mirror reaction on the surface to generate silver particles.

Furthermore, according to a third aspect of the present invention, a method of preparing a probe type SERS detecting substrate can be provided, which is a method of preparing a surface enhanced Raman scattering (SERS) wafer having a needle tip, comprising: providing a a stainless steel wire is used as a core material, and a PMMA substrate for probe type SERS detection is formed by coating a polymethyl methacrylate (PMMA) solution on the surface of the outer layer of the core material; The probe type SERS detection was immersed in a silver mirror reaction reagent with a PMMA substrate, and then formaldehyde was injected to cause a uniform silver mirror reaction on the surface to generate silver particles.

Further, according to a fourth aspect of the present invention, there can be provided a method for producing a substrate for planar SERS detection, which is a method for preparing a wafer having a flat surface enhanced Raman scattering (SERS), which comprises: washing the crucible The crystal substrate is immersed in the surface modifier, and after a period of time, it is cleaned with deionized water to obtain a surface-modified and modified crystal substrate; the surface modified and modified crystal substrate, nano The gold modifier was placed in a centrifuge, and the film was formed by gravity centrifugation, and then taken out and naturally dried to obtain a substrate for planar SERS detection.

Further, according to a fifth aspect of the present invention, there can be provided a method for producing a substrate for planar SERS detection, which is a method for preparing a wafer having a flat surface enhanced Raman scattering (SERS), which comprises: washing the crucible The crystal substrate is immersed in the surface modifier, and after a period of time, it is cleaned with deionized water to obtain a surface-modified and modified crystal substrate; the surface modified and modified crystal substrate, nano The silver modifier was placed in a centrifuge, and formed into a film by gravity centrifugation, and then taken out and naturally dried to obtain a substrate for planar SERS detection.

Further, according to a fifth aspect of the present invention, a method for detecting an organic contaminant, which is a method for rapidly detecting an organic contaminant, comprising: injecting an organic contaminant in an object to be detected into the above-described preparation The microfluidic channel type SERS detection substrate obtained by the method is in the microfluidic channel; and then, for the microfluidic channel type SERS detection substrate or the probe type SERS detection substrate, by the three-dimensional nanometer The Raman fluorescence microscope system performs detection to obtain a Raman spectrum indicating a change of the Raman signal; the Raman spectrum is compared and analyzed to identify the type and concentration of the organic pollutant.

Further, according to a sixth aspect of the present invention, a method for detecting an organic contaminant, which is a method for rapidly detecting an organic contaminant, comprising: a probe type SERS prepared by the above-described preparation method, can be provided. a substrate for detection or a substrate for planar SERS detection, placed in an object to be detected, so that organic contaminants are adsorbed to the probe and left for a predetermined period of time; and then, for the microfluidic channel type SERS detection described above Using a substrate or probe type SERS detection substrate, a Raman spectrum representing a change in Raman signal is obtained by a three-dimensional nano-Raman fluoroscopy system; the Raman spectrum is analyzed and compared Identify the type and concentration of organic pollutants. [Effect of invention]

According to various novel ideas and research results of the present invention, the microfluidic channel type, probe type surface enhanced Raman scattering (SERS) wafer and the substrate developed by the present invention are used for detecting organic contamination by the SERS Raman spectroscopy of the present invention. The method can solve the problem that the organic gas detection such as the traditional gas chromatograph (GC) or the high-performance liquid chromatograph (HPLC) requires a large number of samples, complicated processing operations, long analysis time, and large labor and material resources. And can identify structurally similar organic matter or strains, and simultaneously detect qualitative and quantitative determination of a variety of different pollutants.

Moreover, since the ERS Raman spectroscopy detection organic contamination detecting method of the present invention is a method which is very simple, very rapid, and has high analytical precision and high reliability, and can be used for the SERS detecting substrate and the organic pollution detecting method of the present invention. And it becomes a portable detection system or device, so it can be widely used in various scientific applications and technical fields that need to detect organic pollution, thereby detecting substances and quantification, including in high molecular polymers, nano materials, electrochemistry, semiconductors. , film, mineralogy, biology, medical drugs, carbides, online process monitoring, quantitative control, criminal identification detection: glass materials, oxides, paints and pigments, hydroxides, polymers, sulfides, explosives, carbonic acid Salt, fiber, sulfate, chemical residues, phosphates, particulate inclusions, anesthetics and controllable substances...etc.

In the following, various embodiments of the present invention will be described with reference to the drawings, but the invention is not limited to the embodiments and examples. It will be understood by those skilled in the art that the embodiments and the embodiments may be modified or changed as long as they do not depart from the principles and spirit of the present invention, and such modifications or changes may fall into The scope of applicability of the invention and its equivalents are considered to be within the scope of the invention.

First, the characteristics of Surface Enhanced Raman Scattering (SERS) are described.

Raman spectroscopy is an important modern spectroscopy technique that has been widely used to detect molecular structures and interactions between atoms, and techniques for studying molecular vibration modes. It has become a study of molecular structure and configuration, and determination of crystal structure symmetry. A powerful tool for studying the defects and impurities in solids, environmental pollutants, biomolecules, and the microstructure of industrial materials.

Fleishmann et al. [3] discovered Raman spectra of pyridine molecules adsorbed on silver electrodes in electrochemical cells. Surface Enhanced Raman Scattering (SERS) was discovered in 1974. Significantly enhanced, Raman signal enhancement can reach the order of 10 ⁴ ~ 10 ⁶ . Although it is known that SERS can be efficiently produced on, for example, Ag, Au, Pt, Cu, Al, and alkali metal surfaces, SERS cannot occur on any surface; even on the limited metal surfaces mentioned above, even SERS can be found. Its intensity is also very weak, so it can not be arbitrarily applied to the detection of trace substances, especially the detection of organic pollutants.

In order to solve the problems and difficulties of the conventional techniques, the present inventors have developed a microfluidic channel type SERS capable of adsorbing or bonding a chemical substance in a test object well and having a surface modified or specially treated. A substrate for detection and a substrate for probe type SERS detection; and an effective and rapid method for detecting organic contaminants by Raman scattering spectroscopy by the SERS substrate of the present invention.

For example, according to a first aspect of the present invention, a method of preparing a substrate for a microfluidic channel type SERS can be provided, which is a method of preparing a surface enhanced Raman scattering (SERS) wafer having a microfluidic channel, including: Photolithography method, using a photomask, forming at least one microchannel pattern on a glass substrate to form a microchannel structure master; applying a polydimethylsiloxane (PDMS) solution to the foregoing The microchannel structure master mold is heat-cured and stripped to obtain a PDMS cured structure having a microfluidic channel; a silver mirror reagent is injected into the microchannel of the PDMS solidified structure, and then a reducing agent is injected to generate a silver mirror reaction. The silver nanoparticles were reduced to form and washed with deionized water to obtain a microfluidic channel type SERS substrate.

Secondly, according to a second aspect of the present invention, a method for preparing a probe type SERS detecting substrate, which is a method of preparing a surface enhanced Raman scattering (SERS) wafer having a needle tip, comprising: a probe The substrate is immersed in a silver ammonium solution, and then injected with formaldehyde to cause a silver mirror reaction to be reduced to form silver nanoparticles, and rinsed with deionized water to obtain a needle-shaped SERS substrate.

According to an embodiment of the present invention, the kind of the substrate to which the present invention is applied is particularly limited; as long as a substance capable of forming a fluid passage on the surface or inside thereof can be used. For example, it may be at least one selected from the group consisting of polydimethyl methoxy oxane (PDMS) and polymethyl methacrylate (PMMA).

Further, in the method for producing a microfluidic channel type SERS detecting substrate or a probe type SERS detecting substrate according to the first to third aspects of the present invention, the polydimethyloxane (PDMS) solution It is not particularly limited and may be composed of a PDMS monomer having a vinyl group at the end and a PDMS monomer having a functional group containing Si—H.

Further, in the method for producing a microfluidic channel type SERS detecting substrate or a probe type SERS detecting substrate according to the first to third aspects of the invention, the silver mirror reaction reagent is not particularly limited, however Usually a silver ammonium solution.

Further, in the method for producing a microfluidic channel type SERS detecting substrate or a probe type SERS detecting substrate according to the first to third aspects of the invention described above, the silver ammonium solution is obtained from a silver nitrate solution (AgNO) ₃ , 2%), add a few drops of sodium hydroxide (NaOH, 2.5M) solution to produce brown silver oxide, and quickly add concentrated ammonia water to the solution until the brown precipitate just dissolves to obtain the silver ammonia solution. .

Further, the constituent components of the silver ammonium solution applicable to one embodiment of the present invention are particularly limited; for example, it may include silver nitrate (AgNO ₃ , 2%), ammonia water (NH 4 OH, 2 M), hydrogen. Sodium oxide (NaOH, 2.5 M) is used, and the concentration of the silver ammonium solution is 0.1% by weight to 5% by weight.

Further, in the method of producing a microfluidic channel type SERS detecting substrate or a probe type SERS detecting substrate according to the first to third aspects of the present invention, the reducing agent is not particularly limited and may be Formaldehyde, acetaldehyde, propionaldehyde, other aldehydes, formic acid, glucose, glucose ester, maltose, fructose, and mixtures thereof constitute at least one selected from the group. However, preferred is formaldehyde, acetaldehyde, propionaldehyde, formic acid, glucose, glucose ester, maltose, or fructose; more preferably formaldehyde, acetaldehyde, formic acid, glucose, or fructose; particularly preferred is formaldehyde, acetaldehyde , glucose, or fructose; or it may be a mixture of them.

Further, the mixing ratio of the reducing agent which is suitable for use in an embodiment of the present invention is particularly limited; for example, the concentration of the formaldehyde may be 95% by volume to 99.99% by volume.

Furthermore, the size of the microfluidic channel suitable for use in an embodiment of the present invention is particularly limited; for example, the size of the microfluidic channel, for example, the width may be in the range of 0.01 to 0.5 cm, and the depth may be It is in the range of 0.01~0.5 cm nanometer.

Further, according to a fourth aspect of the present invention, a method of producing a substrate for planar SERS detection, which is a method for preparing a wafer having a flat surface-enhanced Raman scattering (SERS), comprising: a cleaned crucible The crystal substrate is immersed in the surface modifier, and after a period of time, it is cleaned with deionized water to obtain a surface-modified and modified crystal substrate; the surface modified and modified crystal substrate, nano The gold modifier was placed in a centrifuge, and the film was formed by gravity centrifugation, and then taken out and naturally dried to obtain a substrate for planar SERS detection.

Further, the constituent ratio of the surface modifier which is applicable to one embodiment of the fourth aspect of the present invention is particularly limited; for example, the surface modifier may be 3-aminopropyltriethyl a solution of oxydecane (APTMS) and ethanol; the nanogold modifier is a solution A prepared by dissolving tannic acid, sodium citrate or potassium carbonate in deionized water, and tetrabasic acid is dissolved in deionized The solution B formed by water is mixed and thoroughly stirred to obtain a nano gold colloid; followed by 11-MUA (11-Mercaptooundecanoic acid), 6-MHA (6-mercaptohexanoic acid), or 3-MPA (3-Mercaptopropionic acid) a nano gold modifier prepared by modifying the obtained nano gold colloid.

Further, according to a fifth aspect of the present invention, a method of producing a substrate for planar SERS detection, which is a method for preparing a wafer having a flat surface enhanced Raman scattering (SERS), comprising: a cleaned crucible The crystal substrate is immersed in the surface modifier, and after a period of time, it is cleaned with deionized water to obtain a surface-modified and modified crystal substrate; the surface modified and modified crystal substrate, nano The silver modifier is placed in a centrifuge, and is formed into a film by gravity centrifugation, and then taken out and naturally dried to obtain a substrate for planar SERS detection.

Further, the constituent ratio of the surface modifier which is applicable to one embodiment of the fifth aspect of the present invention is particularly limited; for example, the surface modifier may be 3-aminopropyltriethyl a solution of (3-Aminopropyl) trimethoxysilane (APTMS) and ethanol; the nano silver modifier is obtained by mixing sodium citrate, deionized water, silver nitrate, sodium borohydride and heating and stirring. Nano silver colloid; next, the nano silver colloid obtained by modifying the obtained nano silver colloid with 11-MUA, 6-MHA, or 3-MPA.

Furthermore, according to a sixth aspect of the present invention, a method for detecting an organic contaminant can be provided, which is a method for rapidly detecting an organic contaminant, comprising: injecting an organic contaminant in an object to be inspected to be as described above The microfluidic channel type SERS detection substrate obtained by the preparation method is followed by the microfluidic channel; and the microfluidic channel type SERS detection substrate or the probe type SERS detection substrate is further described by the three-dimensional The nano-Raman fluorescence microscope system detects the Raman spectrum representing the change of the Raman signal; the Raman spectrum is compared and analyzed to identify the type and concentration of the organic pollutant.

Further, according to a seventh aspect of the present invention, a method for detecting an organic contaminant, which is a method for rapidly detecting an organic contaminant, comprising: detecting a probe type SERS prepared by the above preparation method, can be provided. The substrate for substrate or planar SERS detection is placed in the object to be detected, so that the organic pollutants are adsorbed to the probe and left for a predetermined period of time; and then, for the above-mentioned microfluidic channel type SERS detection Substrate or probe type SERS detection substrate, which is detected by a three-dimensional nano-Raman fluorescence microscope system to obtain a Raman spectrum representing a change of Raman signal; the Raman spectrum is compared and analyzed by the Raman spectrum The type and concentration of organic pollutants.

According to an embodiment of the present invention, the potential difference range of the electrode plate is particularly limited; for example, preferably, it may be in the range of 1.0 to 110 volts; more preferably, it may be in the range of 10 to 110 volts. The range; particularly good can be in the range of 30 to 110 volts.

According to an embodiment of the present invention, the method for detecting an organic pollutant is particularly limited in terms of the object to be detected; for example, water and sediment in a water body such as a river, a lake, and the like, tap water, industrial water, and a family. Waste water, industrial wastewater, activated sludge, sediment in farmland or ditch, or other water or sludge; or food, beverages, cosmetics, health products, medical equipment, toys, packaging materials, children or elderly care products The invention can be used for detection.

According to an embodiment of the present invention, the detection of the organic contaminant of the object is particularly limited; for example, it may include polybrominated diphenyl ethers (PBDEs), PAEs, Polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), dioxins, chlorinated furans, chlorinated benzenes, monochlorobenzene, Chlorobenzene, hexachlorobenzene, phthalates, DMP, DEP, DBP, BBP, DNOP, DEHP, aldrin, chlordane, dichlorodiphenyl trichloride Ethane (4,4'-dichlorodiphenyltrichloroethane, DDT) and its derivatives, dieldrin, endrin, heptachlor, toxaphene, endosulfan Carbaryl, thymine, BDE-15, BDE-47, BDE-209, decabromodiphenyl ether or mixtures thereof can be tested using the present invention. (Example)

Hereinafter, the present invention will be further illustrated by the examples, but the present invention is of course not limited to the examples. Further, the evaluation of the characteristics in the examples was carried out in the following manner. (Preparation Example 1) "Preparation of Au-11-MUA substrate for nano-modified SERS detection" "Preparation and modification of nano-gold colloid"

First, 0.005 g of tannic acid (manufactured by Sigma-Aldrich, USA, 99% of the reagent grade), 0.4 g of sodium citrate (manufactured by JT Backer Co., Ltd., 99% of the test grade), and 0.0017 g of potassium carbonate were prepared. (Taiwan United Chemical Co., a pharmaceutical grade of 99%) was dissolved in a solution A formed by deionized water to a concentration of 200 mL. Next, a solution B prepared by dissolving 0.10 g of Hydrogen tetrachloro aurate (III) trihydrate (manufactured by Alfa Co., Ltd., reagent grade / 99%) in deionized water to 200 mL was prepared. The two solutions prepared above were mixed and baked at a temperature of 60 ° C for 4 hours to form a nano gold colloid. The gel was then placed to cool to room temperature and stored at 4 degrees Celsius to prevent agglomeration precipitation. Then, the above solution A and solution B were mixed and thoroughly stirred with a magnet to obtain a nano gold colloidal solution having a particle diameter of about 12 nm.

Next, the prepared nano gold colloid solution was put into a modification solution of 11-Mercaptoundecanoic acid (11-MUA) (manufactured by Sigma Aldrich Co., Ltd., 99%), and the modified solution was applied to the nano gold colloid. The ratio of the input volume of the solution was 10:1 (V/V), and the pH was adjusted to 10.5 with deionized water to thereby surface-modify the nano-gold colloid. "Modification of twinned substrate"

After cleaning and cleaning the tantalum wafer (made by Taiwan, Honghui Optoelectronics Co., Ltd.), the surface of the HS solution prepared by mixing 30% of hydrogen peroxide and sulfuric acid in a ratio of 3:7 (w/w) is used for preliminary surface modification. The hydrophilic surface of the Si-OH sterol group is improved; then, the surface of the substrate is rinsed with deionized water, and then placed in 3-aminopropyltriethoxydecane (APTMS) / ethanol solution, soaked in 8- After 12 hours, the surface was modified and modified to obtain a crystal substrate having a modified surface.

Next, the above-mentioned surface modified crystal substrate, nano gold colloid, and modified solution were placed in a centrifuge, and centrifuged to form a film by gravity centrifugation 643.2 ́g (Rcf). After the film formation, the substrate was taken out, naturally dried, and blown dry with a nitrogen gas gun to obtain a Au-11-MUA substrate for nano-modified SERS.

The magnified 100,000-fold SEM photograph of Fig. 1 was obtained by magnifying 100,000 times with a scanning electron microscope SEM. Fig. 1 is a SEM image showing a surface enlargement of 100,000 times of the surface of the Au-MUA substrate for nano-modified SERS of Preparation Example 3 of the present invention. As shown in the SEM image of Au-11-MUA in Fig. 1, it was confirmed that silver nanoparticles were uniformly distributed on the surface of the Au-11-MUA substrate for nano-modified SERS detection and uniformly stacked in Au-11- On the surface of the MUA substrate, the gap between the two silver particles is about 8-10 nm .

In addition, the results of image analysis by Image J are shown in FIG. 2. From the silver particle size distribution obtained by Image J analysis in Fig. 2, it was confirmed that the average particle diameter of the nano silver particles was 34.5 nm.

Then, using the three-dimensional nano-Raman fluorescence microscope system of ZTE University's Nanotechnology Center (laser wavelength is 632.8 nm and 488 nm, maximum power is 3.6 mW), and Rhodamine B (R6G) is used as the indicator molecule. In the case where R6G is present on the Au-11-MUA substrate for nanoscale modified SERS detection, and in the case of R6G and T-dimer silver, the incident detection at 633 nm is measured. The signal was used to calculate the SERS enhancement effect using Equation 1, and it was confirmed that the detection limit concentration of the T-shape dimer can be measured by the Au-11-MUA substrate using the nano-modified planar SERS detection. The minimum is 10 ^-8 M. (Formula 1) (Preparation Example 2) "Preparation of Au-6-MHA substrate for nano-modified SERS detection""Preparation and modification of nano-gold colloid"

First, the obtained nano gold colloid was operated in the same manner as in Production Example 1. Next, the prepared nano gold colloid was put into a modification of 6-mercaptohexanoic acid (6-MHA) (manufactured by Sigma Aldrich Co., Ltd., 99%) manufactured by 10:1 (V/V, colloid: modified solution). In the solution, the pH was adjusted to 10.5 with deionized water to thereby surface-modify the nano-gold colloid to obtain an Au-6-MHA-modified nano gold colloid.

Next, the surface-modified modified crystal substrate obtained in Preparation Example 1, the above-mentioned Au-6-MHA modified nano gold colloid, and the modified solution were placed in a centrifuge, in the same manner as the preparation example. The sample was centrifuged using gravity centrifugation 643.2 ́g (Rcf). After the film formation, the substrate was taken out, naturally dried, and blown dry with a nitrogen gas gun to obtain a Au-6-MHA substrate for nano-modified SERS.

Further, in the same manner as in Preparation Example 1, the magnified 100,000-fold SEM photograph of FIG. 3 was observed by magnifying 100,000 times with a scanning electron microscope SEM. Fig. 5 is a SEM image showing a surface enlargement of 100,000 times of the surface of the Au-6-MHA substrate for nano-modified SERS of Preparation Example 2 of the present invention. As shown in the SEM image of Au-6-MHA of Fig. 3, on the surface of the Au-6-MHA substrate for the nanogold-modified SERS detection, there were uniformly distributed and uniformly stacked silver particles.

In addition, the results of image analysis by Image J are shown in FIG. From the silver particle size distribution obtained by Image J analysis in Fig. 4, it was confirmed that the nano silver particles had a particle diameter of about 32.0 nm. (Preparation Example 3) "Preparation of Au-3-MPA substrate for nano-modified SERS detection" "Preparation and modification of nano-gold colloid"

First, the obtained nano gold colloidal solution was operated in the same manner as in Production Example 1. Next, the prepared nano gold colloid was put into a modification solution of 10:1 (V/V, colloid: modified solution) 3-Mercaptopropionic acid (3-MPA) (manufactured by Sigma Aldrich Co., Ltd., test grade 99%). In the middle, the pH was adjusted to 11.5 with deionized water to surface-modify the nano-gold colloid to obtain a Au-3-MPA-modified nano gold colloid.

Next, the surface-modified twin crystal substrate obtained in Preparation Example 1, the above-mentioned nano Au-3-MPA colloidal solution, and the ruthenium wafer were placed in a centrifuge, and gravity was used in the same manner as in Preparation Example 1. Centrifuge at 643.2 ́g (Rcf) to form a film. After film formation, the substrate was taken out, naturally dried, and blown dry with a nitrogen gas gun to obtain a Au-3-MPA substrate for nano-modified SERS.

Further, in the same manner as in Preparation Example 1, the SEM photograph was magnified 100,000 times by SEM magnification by a scanning electron microscope. Fig. 5 is a SEM image showing the surface magnified 100,000 times of the Au-3-MPA substrate for nanogold-modified SERS detection of Preparation Example 3 of the present invention. As shown in the SEM image of Au-3-MPA of Fig. 5, silver particles having uniform distribution and uniform stacking were formed on the surface of the Au-3-MPA substrate for the nanogold-modified SERS.

In addition, the results of image analysis by Image J are shown in FIG. 6. From the silver particle size distribution obtained by Image J analysis in Fig. 6, it was confirmed that the nano silver particles had a particle diameter of about 29.1 nm. (Preparation Example 4) "Preparation of Ag substrate for nano silver modified planar SERS detection"

First, 0.2 g of sodium citrate (manufactured by JT Backer Co., Ltd., 99% of the reagent grade) was dissolved in deionized water, and 0.017 g of silver nitrate (manufactured by Sigma Aldrich Co., Ltd.) was added while heating (temperature: 50 ° C). Pharmaceutical grade 99%), 0.2 g of sodium borohydride (manufactured by Panreac Co., USA, 99% of the test grade), and heating was continued for about 1 hour to remove excess sodium borohydride to form a nanoparticle containing silver particles of a predetermined particle diameter. Rice silver colloidal solution (grown twice with a particle size of 30 nm).

Next, the nano silver colloid solution prepared above is put into a modification solution of 6-mercaptohexanoic acid (6-MHA), so that the input volume ratio of the modification solution to the nano silver colloid becomes 10:1 (V/V, colloid : Modified), and adjusted to pH 10.5 with deionized water to surface-modify the nano-silver colloid. "Modification of twinned substrate"

After cleaning the enamel wafer (Taiwan, Honghui Optoelectronics), first upgrade it with an oxygen plasma machine, intermittently upgrade the oxygen for 10 seconds, stagnate for 2 minutes, repeat 3 times, and then place 30% of hydrogen peroxide. Surface pre-modification treatment with HS in a ratio of 3:7 (w/w) in a ratio of 3:7 (w/w) to make a hydrophilically enhanced surface of Si-OH sterol group; then, rinse with deionized water The surface of the substrate was placed in a 3-aminopropyltriethoxydecane (APTMS) / ethanol solution at 10% (APTMS:Et 10:90 v/v). After soaking for 12 hours, it was ultrasonically cleaned with deionized water. A crystal substrate having a surface modified and modified is obtained.

Next, the above-mentioned surface modified crystal substrate, nano silver colloid, and polydimethyloxane (PDMS) solution were placed in a centrifuge, and centrifuged to form a film by gravity centrifugation 643.2 ́g (Rcf). . After the film formation, the substrate was taken out, naturally dried, and blown dry with a nitrogen gas gun to obtain a nano substrate for nano silver-modified planar SERS detection. The magnified 100,000-fold SEM photograph of Fig. 7 was obtained by SEM observation with a scanning electron microscope.

Fig. 7 is a SEM image showing a surface enlargement of 100,000 times of the surface of the Ag-6-MHA substrate for nano silver-modified SERS of Preparation Example 4 of the present invention. As shown in the SEM image of Ag- 11-MUA in Fig. 7, it was confirmed that the surface of the Ag substrate for the nano silver-modified planar SERS detection had uniform distribution of silver particles and was uniformly stacked on the Ag substrate, and two The gap between the silver particles is about 1 nm.

In addition, the results of analysis by Image J image are shown in FIG. From the silver particle size distribution obtained by Image J analysis in Fig. 8, it was confirmed that the nano silver particles had a particle diameter of about 34.5 nm. (Preparation Example 5) "Preparation of Microfluidic Channel Type SERS Detection Substrate" "Preparation of PDMS Microfluidic Channel"

First, in a clean room (National Chung Hsing University Mengzi Biochip Center Class 100), the cleaned glass substrate was placed on a photoresist coater (SCD-6000, Yanxin Instruments, Taiwan) and coated with a negative photoresist (Japan) JSR Co., Ltd., JSR THB 121N) After the photoresist, the glass substrate was placed in an infrared double-sided alignment exposure machine (OAI-500), and optical micro-etching was performed by optical projection, and the microfluidic channel was made by the photomask. The pattern is formed on a glass substrate. After the UV exposure is completed, the glass substrate and the photoresist are immersed in a developing solution (AD-TMAH 0.5) to remove the unexposed photoresist, and then baked and annealed to remove the residual solvent to obtain a microchannel structure master.

Next, the A agent (main agent, long PDMS monomer) containing a vinyl group at the end and the B agent (hardener, short PDMS monomer) containing a functional group of Si–H in a weight ratio of A:B = 10 :1 (w / w) After mixing and stirring well until it is filled with air bubbles, put it into a vacuum drying pot and vacuum it with a vacuum pump (GVD050A, manufactured by Taiwan Cognac Technology Co., Ltd.) to remove the thick PDMS in the mixing and stirring. The bubbles generated at the time gave a PDMS solution.

Next, the obtained PDMS solution was poured into the above-described microchannel structure master, and placed on a hot plate (CO-PC600D, manufactured by Corning, USA) at 85 ° C for 3 hours. After the PDMS is cured, it is sufficiently cooled at room temperature, and then the fully cured and cooled PDMS is removed from the master to obtain a PDMS having a microchannel structure. Preparation of Silver Mirror Reaction Reagents

Silver mirror reagents, also known as Duolun reagents, toluene reagents, or Tollens' reagents, refer to aqueous solutions containing silver (I) ions ([Ag(NH ₃ ) ₂ ]+), generally It is prepared by reacting silver nitrate or other silver compounds with ammonia water. Theoretically, in addition to all substances containing an aldehyde group, formic acid, glucose, glucose ester, maltose, and fructose can also be used as a reducing agent, so that the silver mirror reaction reagent undergoes a silver mirror reaction. Generally used reducing agents such as formaldehyde, acetaldehyde and glucose.

The silver mirror reaction reagent is prepared by first adding a few drops of sodium hydroxide solution to the silver nitrate solution. As shown in the following chemical formula, a very unstable silver hydroxide white precipitate is initially produced, which immediately produces a brown silver oxide precipitate. Next, concentrated ammonia water was added dropwise to the solution until the brown precipitate just dissolved, thereby obtaining a silver ammonia solution containing Ag(NH ₃ ) 2 NO ₃ (aq), which is a silver mirror reaction reagent. AgNO ₃ + NaOH → AgOH↓ + NaNO ₃ AgOH + 2 NH ₃ → [Ag(NH ₃ ) ₂ ]OH _(aq) Preparation of Microfluidic Channel Type SERS Detection Substrate

Continuing, the obtained silver ammonia solution (silver mirror reaction reagent) is injected into the inlet end of the flow path of the PDMS microchannel structure prepared by the syringe, and the pump is pumped at the other end to uniformly distribute the ammonia solution. In the micro flow channel. Then, formaldehyde is injected into the flow path, so that the silver mirror reaction occurs uniformly throughout the micro flow path, thereby generating silver particles for use as a substrate for SERS signal enhancement, thereby obtaining a microfluidic channel type SERS detection substrate.

The magnified 100,000-fold SEM photograph of Fig. 10 was obtained by SEM observation with a scanning electron microscope. Fig. 10 is a SEM image showing a 100,000-fold magnification of the channel surface of the Ag substrate for the nano silver-modified microfluidic channel type SERS detection of Preparation Example 5 of the present invention. In addition, it was confirmed by Image J image analysis that the average particle diameter of the nano silver particles was about 71.0 ± 38.2 nm. (Preparation Example 6) "Preparation of PDMS substrate for probe type SERS detection" "Preparation of PDMS probe"

First, a stainless steel was used as a core material, and a single layer of polydimethyl siloxane (PDMS) was coated on the outer layer to prepare a probe-type SERS substrate. The outer covering material is not particularly limited as long as it is capable of adsorbing contaminants; for example, it may be a hydrophobic polymer material. The shape is also not particularly limited; for example, it may be elongated.

Next, after coating the PDMS, it was further immersed in the silver ammonia solution (silver mirror reaction reagent) obtained in Preparation Example 5, followed by injecting formaldehyde to cause a uniform silver mirror reaction on the surface, thereby generating the SERS signal enhancement station. The silver particles of the substrate are required to obtain a PDMS substrate for probe type SERS detection.

A 50,000-fold SEM photograph was obtained by observation with a scanning electron microscope SEM. In addition, it was confirmed by Image J image analysis that the average particle diameter of the nano silver particles was about 71.0 ± 38.2 nm. (Preparation Example 7) "Preparation of PMMA substrate for probe type SERS detection" "Preparation of PMMA probe"

First, in the same manner as in Preparation Example 6, a stainless steel was used as a core material, and a single layer of polymethyl methacrylate (PMMA) was coated on the outer layer to prepare a probe-type SEMA substrate for SERS detection.

Next, after coating the PMMA, it was further immersed in the silver ammonia solution (silver mirror reaction reagent) obtained in Preparation Example 5, followed by injecting formaldehyde to cause a uniform silver mirror reaction on the surface, thereby generating a SERS signal enhancement. The silver particles of the substrate are required to obtain a PMMA substrate for probe type SERS detection.

The magnified 50,000-fold SEM photograph of Fig. 11 was obtained by SEM observation with a scanning electron microscope. Fig. 11 is a SEM image showing a 100,000-fold magnification of the channel surface of the Ag substrate for the nano silver-modified microfluidic channel type SERS detection of Preparation Example 5 of the present invention. In addition, it was confirmed by Image J image analysis that the average particle diameter of the nano silver particles was about 71.0 ± 38.2 nm. (Example 1) "Detecting Phthalates (PAs)"

Ag-11-MUA substrate, Ag-6-MHA substrate, Ag-3-MPA for silver-modified planar SERS detection prepared in Preparation Example 1, Preparation Example 2, Preparation Example 3, and Preparation Example 4, respectively. The substrate and the Ag substrate were cut to an appropriate size at a ratio of about 1 cm ² to detect an object of about 10 mL in volume to detect three different concentrations of 100 mg / L and 10 mg / L in methanol. 1 mg / L PAs (DMP, DEHP, DNOP, DBP and DEP) were used to establish Raman spectra of phthalate ester (PAs) organic pollutants. The results are shown in Figure 9.

Fig. 9 is a Raman spectrum diagram showing the detection of phthalic acid esters (PAs) by the substrate for SERS detection of the present invention shown in Example 1; wherein (a) represents dibutyl phthalate ( Raman spectrum of DBP); (b) shows the Raman spectrum of diethyl phthalate (DEP); (c) shows the Raman spectrum of dimethyl phthalate (DMP); ) represents a Raman spectrum of di-n-octyl phthalate (DNOP); (e) represents a Raman spectrum of di(2-ethylhexyl) phthalate (DEHP).

As shown in Fig. 9, it can be seen that their PAs are 403 cm ^-1 , 654 cm ^-1 , 1041 cm ^-1 , 1126 cm ^-1 , 1166 cm ^-1 , 1581 cm ^-1 , 1602 cm ^-1 and 1727 cm . ^-1 has a common characteristic peak, which can be used as a basis for determining the presence or absence of PAs (phthalates) in the detected object.

Then, the SERS substrate cut by the substrate for the nano- or nano-silver-modified planar SERS detection of the above Preparation Example 1, Preparation Example 2, Preparation Example 3, and Preparation Example 4 was immersed in an environmental sample. a phthalate extract, a sediment sample from Errenxi (to confirm the presence or absence of organic contamination of phthalate), and a blank sample of the extract (with n-hexane and acetone as solvent) After 12 hours, measure the SERS Raman spectrum. Each sample is measured at least 500 points and measured three times in different regions. The characteristic peaks are 635 cm ^-1 , 703 cm ^-1 , 1013 cm ^-1 , 1277 cm . ^-1 and 1607 cm ^-1 , the results are shown in Fig. 12 and Fig. 13.

Figure 12 is a surface enhanced Raman spectrum showing a sample of Errenxi bottom mud extract and blank extract. Figure 13 is a surface enhanced Raman spectrum showing a sample of Errenxi sediment. As shown in FIG. 12 and FIG. 13, it can be confirmed that the Ag-11-MUA substrate, the Ag-6-MHA substrate, the Ag-3-MPA substrate, and the Ag substrate for the silver-modified planar SERS detection can be used. Quickly measure the concentration of phthalates (PAs) in objects such as sediment. (Example 2) "Detecting decabromodiphenyl ether (BDE-209)"

Using the microfluidic channel type SERS substrate prepared in Preparation Example 5, an appropriate size was cut at a ratio of about 1 cm ² to detect an object of about 10 mL in volume to detect ethyl acetate. A sample of 0.09 ppm, 0.9 ppm, 9.0 ppm, and 90 ppm of decabromodiphenyl ether was prepared in a solvent to establish a Raman spectrum of the decabromodiphenyl ether (BDE-209) organic pollutant standard. The spectrum is shown in Figure 14.

Figure 14 is a Raman spectrum of decabromodiphenyl ether showing a PDMS probe type SERS substrate. From Fig. 14, it was confirmed that a peak having an ether bond at a wavenumber position of about 1192 cm ^-1 and 1227 cm ^-1 has a peak of a benzene ring structure at a wave number position of a wave number of about 1512 cm ^-1 . Therefore, according to the present invention, the decabromodiphenyl ether detection example can confirm that the microfluidic channel type SERS detection substrate can be used for rapid detection of whether or not decabromodiphenyl ether (BDE-209) has been contained in objects such as sediment. Pollution.

In addition, the Raman spectrum of each concentration is compared with the signal intensity of the wave number of 1620 cm ^-1 to obtain a regression equation: y = 14.452 x + 664.732 R2 = 0.992 (where y represents the Raman signal intensity; x represents the concentration (ppm) and its R ^{2 is} greater than 0.99. (Example 3) "Detecting decabromodiphenyl ether (BDE-209)"

The PDMS substrate and the PMMA substrate prepared by the probe type SERS prepared in Preparation Example 6 were cut to an appropriate size, and the concentration of decabromodiphenyl ether was 0.1 ppm and 0.2 ppm in the laboratory using ethanol as a solvent. , 0.5 ppm, 1.0 ppm, 2.0 ppm, 4.0 ppm samples, and Raman signal detection in the "3D Nano-Raman Fluorescence Microscopy System" to establish decabromodiphenyl ether (BDE-209) organic pollutants The standard Raman spectrum shows the Raman spectrum as shown in FIG.

Figure 15 is a Raman spectrum diagram showing decabromodiphenyl ether of a PMMA probe type SERS substrate. As shown in Fig. 15, it can be confirmed that the peaks at about 1808 cm ^-1 and 1974 cm ^-1 are relatively obvious, and are present on the spectra of the respective concentrations, and are in the range of about 1750 cm ^-1 to 2080 cm ^-1 . An overtone signal having a benzene ring structure at a wave number position. Therefore, it can be confirmed that the probe type SERS detecting substrate of the present invention can be used for rapid detection of whether or not an object such as a sediment has been contaminated with decabromodiphenyl ether (BDE-209) in terms of the decabromodiphenyl ether detecting example. Invention effect

According to the present invention, a large number of samples can be analyzed, and the preparation time of the fine needle type SERS substrate plus the time required for Raman signal detection can be shortened by more than 90% over the time required for the conventional method. This study confirms the possibility that SERS can be used to rapidly detect PBDEs in sediments. Based on the results of this study, SERS should be applied to the rapid detection of trace hydrophobic contaminants in environmental media.

The above description of the preferred embodiments of the present invention will be understood by those of ordinary skill in the art, There are various modifications, additions and alternatives available. In addition, such modifications, additions and substitutions should be considered as falling within the scope of the invention.

no.

Fig. 1 is a SEM photograph showing a surface enlargement of 100,000 times of the surface of the Au-11-MUA substrate for nano-modified SERS of Preparation Example 1 of the present invention. Fig. 2 is a graph showing the particle size distribution of silver obtained by analyzing the Au-11-MUA substrate for nano-modified SERS shown in Fig. 1 by Image J. Fig. 3 is a SEM photograph showing a surface magnification of 100,000 times of the surface of the Au-6-MHA substrate for nano-modified SERS of Preparation Example 2 of the present invention. Fig. 4 is a graph showing the distribution of silver particle size obtained by analyzing the Au-6-MHA substrate for nano-modified SERS shown in Fig. 3 by Image J. Fig. 5 is a SEM photograph showing the surface magnified 100,000 times of the Au-3-MPA substrate for the nanogold-modified SERS detection of Preparation Example 3 of the present invention. Fig. 6 is a graph showing the particle size distribution of silver obtained by analyzing the Au-MPA substrate for nano-modified SERS shown in Fig. 5 by Image J. Fig. 7 is a SEM photograph showing a surface magnification of 100,000 times of the surface of the Ag-6-MHA substrate for nano silver-modified SERS of Preparation Example 4 of the present invention. Fig. 8 is a graph showing the particle size distribution of silver obtained by analyzing the nano silver-modified SERS for Ag-6-MHA substrate shown in Fig. 5 by Image J. 9 is a Raman spectrum diagram showing the detection of phthalate esters (PAs) by the substrate for SERS detection of the present invention shown in Example 1; wherein (a) represents dibutyl phthalate. a Raman spectrum of the ester (DBP); (b) a Raman spectrum of diethyl phthalate (DEP); (c) a Raman spectrum of dimethyl phthalate (DMP); (d) shows a Raman spectrum of di-n-octyl phthalate (DNOP); (e) shows a Raman spectrum of di(2-ethylhexyl) phthalate (DEHP). Fig. 10 is a SEM photograph showing a 100,000-fold magnification of the channel surface of the Ag substrate for the nano silver-modified microfluidic channel type SERS detection of Preparation Example 4 of the present invention. Fig. 11 is a SEM photograph showing the surface of the probe of the Ag substrate for the silver-modified probe type SERS detection of the preparation example 5 of the present invention magnified 100,000 times. Figure 12 is a surface enhanced Raman spectrum showing a sample of Errenxi bottom mud extract and blank extract. Figure 13 is a surface enhanced Raman spectrum showing a sample of Errenxi sediment. Figure 14 is a Raman spectrum of decabromodiphenyl ether showing a PDMS probe type SERS substrate. Figure 15 is a Raman spectrum diagram showing decabromodiphenyl ether of a PMMA probe type SERS substrate.

no

Claims (9)

A method for preparing a substrate for microfluidic channel type SERS detection, which is a method for preparing a surface enhanced Raman scattering (SERS) wafer having a microfluidic channel, comprising: using a photolithography method, using a photomask, to include At least one or more microchannel patterns are formed on the glass substrate to form a microchannel structure master mold; a polydimethylsiloxane (PDMS) solution is applied to the aforementioned microchannel structure master mold, and is heated and solidified. Stripping to obtain a PDMS cured structure having a microfluidic channel; injecting a silver mirror reaction reagent into the microchannel of the PDMS solidified structure, and then injecting a reducing agent to cause a silver mirror reaction to be reduced to form silver nanoparticles, and deionized Water-flushing to obtain a microfluidic channel type SERS detecting substrate; wherein the PDMS solution is composed of a PDMS monomer having a vinyl group at the terminal end and a functional group containing Si—H; the silver mirror reagent is silver hydroxide solution, the silver-based solution of ammonium hydroxide in a few drops of a solution of silver nitrate (AgNO _3, 2%) of (NaOH, 2.5M) solution to make a brown silver oxide to the solution and rapidly The silver ammonia solution is obtained by adding concentrated ammonia water until the brown precipitate just dissolves; the reducing agent is composed of formaldehyde, acetaldehyde, propionaldehyde, other aldehydes, formic acid, glucose, glucose ester, maltose, fructose and mixtures thereof. At least one of the groups selected. A method for preparing a probe type SERS detecting substrate, which is a method for preparing a surface enhanced Raman scattering (SERS) wafer having a needle tip, comprising: providing a stainless steel wire as a core material, in the core material The surface of the outer layer is coated with a polydimethylsiloxane (PDMS) solution and cured by heating to form a probe type SERS detection PDMS substrate; the obtained probe type SERS detection PDMS substrate is immersed in In the silver mirror reaction reagent, formaldehyde is injected into the surface to generate a uniform silver mirror reaction to form silver particles; wherein the PDMS solution is composed of a PDMS monomer having a vinyl group at the end and a functional group containing Si—H; The silver mirror reaction reagent is a silver ammonium solution, which is prepared by adding a few drops of sodium hydroxide (NaOH) solution in a silver nitrate solution (AgNO ₃ ) to produce brown silver oxide and rapidly adding concentrated ammonia water to the solution. a silver ammonia solution obtained until the brown precipitate just dissolves; the reducing agent is a group consisting of formaldehyde, acetaldehyde, propionaldehyde, other aldehydes, formic acid, glucose, glucose ester, maltose, fructose and mixtures thereof The selection of at least one. A method for preparing a probe type SERS detecting substrate, which is a method for preparing a surface enhanced Raman scattering (SERS) wafer having a needle tip, comprising: providing a stainless steel wire as a core material, in the core material The surface of the outer layer is coated with a polymethyl methacrylate (PMMA) solution and heat-cured to form a probe-type SEMA detection PMMA substrate; the obtained probe-type SERS detection PMMA substrate is immersed in a silver mirror In the reaction reagent, formaldehyde is injected to cause a uniform silver mirror reaction on the surface to form silver particles; wherein the PMMA solution is composed of a PMMA monomer having a vinyl group at the terminal end and a PMMA monomer having a functional group containing Si—H; The silver mirror reaction reagent is a silver ammonium solution, which is obtained by adding a few drops of sodium hydroxide (NaOH) solution in a silver nitrate solution (AgNO ₃ ) to produce brown silver oxide and rapidly adding concentrated ammonia water to the solution until the solution is concentrated. a silver ammonia solution obtained by dissolving a brown precipitate; the reducing agent is a group consisting of formaldehyde, acetaldehyde, propionaldehyde, other aldehydes, formic acid, glucose, glucose ester, maltose, fructose and mixtures thereof At least one of those selected. A method for preparing a planar SERS detecting substrate, which is a method for preparing a flat surface enhanced Raman scattering (SERS) wafer, comprising: immersing a washed twin substrate in a surface modifying agent, After a period of time, the surface is modified and modified by a deionized water to obtain a crystal substrate; the crystal substrate and the nano gold modifier which are modified and modified on the surface are placed in a centrifuge for use. The film is formed by gravity centrifugation, and then taken out and naturally dried to obtain a substrate for planar SERS detection; wherein the surface modifier is a solution composed of 3-aminopropyltriethoxydecane (APTMS) and ethanol; The rice gold modifier is a solution B prepared by dissolving tannic acid, sodium citrate, potassium carbonate in deionized water, and solution B in which tetrachloroauric acid is dissolved in deionized water, and mixing them sufficiently to obtain the naphtha. a gold colloid; then the nano-colloid obtained by modifying the nano-gold colloid obtained by 11-MUA (11-Mercaptooundecanoic acid), 6-MHA (6-mercaptohexanoic acid), or 3-MPA (3-Mercaptopropionic acid) Gold modifier. A method for preparing a planar SERS detecting substrate, which is a method for preparing a flat surface enhanced Raman scattering (SERS) wafer, comprising: immersing a washed twin substrate in a surface modifying agent, After a period of time, it is cleaned with deionized water to obtain a crystal-modified substrate whose surface has been modified and modified; the crystal substrate and the nano silver modifier which have been modified and modified on the surface are placed in a centrifuge and used. The film is formed by gravity centrifugation, and then taken out and naturally dried to obtain a substrate for planar SERS detection; wherein the surface modifier is a solution composed of 3-aminopropyltriethoxydecane (APTMS) and ethanol; The rice silver modifier is a nano silver colloid obtained by mixing sodium citrate, deionized water, silver nitrate, sodium borohydride and heating and stirring; followed by 11-MUA (11-Mercaptoundecanoic acid), MHA (6-mercaptohexanoic) Acid or a 3-MPA (3-Mercaptopropionic acid) modification of the nano silver colloid obtained by the nano silver colloid. A method for detecting an organic contaminant, which is a method for rapidly detecting an organic contaminant, comprising: injecting an organic contaminant in an object to be detected into a microfluidic channel prepared by the preparation method of claim 1 In the microfluidic channel of the substrate for SERS detection; next, the microfluidic channel type SERS detection substrate or the probe type SERS detection substrate is detected by a three-dimensional nano-Raman fluorescence microscope system A Raman spectrum representing a change in Raman signal is obtained by measurement; the type, concentration, and the like of the organic pollutant are analyzed and identified through the Raman spectrum comparison. A method for detecting an organic contaminant, which is a method for rapidly detecting an organic contaminant, comprising: a probe type SERS detection substrate prepared by the preparation method of claim 2 or claim 3, Or the substrate for planar SERS detection prepared by the preparation method of claim 4 or claim 5, placed in the object to be detected, so that the organic pollutant is adsorbed to the probe and placed for a predetermined period of time; For the microfluidic channel type SERS detection substrate or the probe type SERS detection substrate described above, a Raman spectrum showing a Raman signal change is obtained by detecting by a three-dimensional nano-Raman fluorescence microscope system. The type and concentration of organic pollutants are analyzed and identified through the Raman spectrogram comparison. The method for detecting organic pollutants according to claim 7 or 8, wherein the object to be detected is water and sediment in a water body including rivers, lakes, oceans, tap water, industrial water, domestic wastewater, industrial wastewater, activated sludge, farmland or The sediment in the ditch, or other water or sludge; or food, beverages, cosmetics, health supplies, medical equipment, toys, packaging materials, children or elderly care products. The method for detecting organic pollutants according to claim 7 or 8, wherein the organic pollutants are derived from decabromodiphenyl ether, PAEs, polybrominated diphenylethers (PBDEs), polychlorinated biphenyls (PCBs) ), dioxins, chlorinated furans, chlorinated benzenes, monochlorobenzene, dichlorobenzene, hexachlorobenzene, polycyclic aromatic hydrocarbons (PAHs), ortho-benzene Phthalates, DMP, DEP, DBP, BBP, DNOP, DEHP, aldrin, chlordane, dichlorodiphenyltrichloroethane (4,4'-dichlorodiphenyltrichloroethane) , DDT) and its derivatives, dieldrin, endrin, heptachlor, toxaphene, endosulfan, carbaryl, thymus Thiymine, BDE-15, BDE-47, and BDE-209, at least one selected from the group consisting of or a mixture thereof.