KR102208108B1 - Apparatus and Method for On-line Monitoring of Metabolic Products from Bioconversion via Surface Enhanced Raman Spectroscopy using Electrostatic Interaction of Metallic Nanostructure - Google Patents

Abstract

A bioconversion metabolite detection device comprising a metabolite enrichment and detection unit including a metal nanostructure, and detecting a bioconversion metabolite attached to the metal nanostructure provided in the metabolite enrichment and detection unit as a surface-enhanced Raman scattering signal. , A biotransformation metabolite detection device and a method for effectively concentrating and ultrafast detection of biotransformation metabolites using the same are provided.

Description

Apparatus and Method for On-line Monitoring of Metabolic Products from Bioconversion via Surface Enhanced Raman Spectroscopy using Electrostatic Interaction of Metallic Nanostructure}

The present specification relates to a method and apparatus for detecting biotransformation metabolites based on a surface-enhanced Raman scattering method using the electrostatic attraction of a metal nanostructure.

Bioconversion refers to a technology that converts raw materials into useful materials or energy sources by utilizing the functions of microorganisms or enzymes. Since biotransformation reactions use microorganisms or enzymes, unlike chemical reactions, the reaction proceeds under safe conditions that do not require high temperature or high pressure. In addition, since it does not use toxic substances such as organic solvents, it is possible to develop eco-friendly processes. In addition, the substrate specificity of the enzyme can increase the yield of production of specific metabolites. For this reason, bioconversion reactions in the production of pharmaceuticals, health functional foods, cosmetics, and energy raw materials are in the spotlight as a substitute for existing chemical reactions.

In order to replace the existing chemical process with the bioconversion process, it is necessary to solve the problem that the bioconversion process generally has a lower yield than the chemical process. To this end, in order to increase the production efficiency of metabolites through bioconversion reactions, microorganisms or enzymes with high metabolic efficiency are being developed through genetic manipulation, and the reaction conditions (pH, temperature, medium, etc.) of the bioconversion process are optimized. Research is also actively progressing. Therefore, in order to optimize the bioconversion process and increase the yield of metabolites, a technology capable of monitoring the concentration of metabolites in the bioreactor in real time is required.

Previously, to measure the concentration of metabolites in the reactor, high-performance liquid chromatography (HPLC) was relied on in combination with a mass spectrometer or photo-diode array detector. . However, the detection method using chromatography is not suitable for real-time monitoring because the response time takes more than 20 minutes, and the reactor is exposed to the risk of contamination due to contact with the external environment during the offline sampling process that collects a part of the aqueous solution inside the reactor. .

In order to improve this problem, a method of analyzing the molecules of a solution in a reactor using Raman spectroscopy has been proposed. (Patent Document 1, Non-Patent Document 1) Raman spectroscopy is an optical detection method that measures the frequency of molecular bonds by using Raman scattering generated by chemical bonds of molecules when irradiated with a laser. Since Raman spectroscopy provides information on the chemical bonds of molecules, it has the advantage of selectively distinguishing each molecule and non-destructively real-time detection. However, Raman spectroscopy has a problem that it is difficult to be used in a bioconversion reactor because of its low sensitivity.

Registered Patent No. 10-1849511

Gray, S. R., Peretti, S. W., & Lamb, H. H. Real-time monitoring of high-gravity corn mash fermentation using in situ raman spectroscopy. Biotechnol. Bioeng. 110, 1654-1662 (2013).

In one aspect of the present invention, to provide a surface-enhanced Raman scattering method-based molecular detection method and apparatus capable of monitoring metabolites in a bioconversion reactor online.

In another aspect of the present invention, it is intended to provide a method and apparatus for molecular detection based on a surface-enhanced Raman scattering method capable of detecting metabolites in a bioconversion reactor, which was difficult to detect with conventional chromatography techniques.

In another aspect of the present invention, it is intended to provide a method and apparatus for detecting a molecule based on a surface-enhanced Raman scattering method that can improve detection sensitivity by utilizing a heterogeneous metal nanostructure having strong electromagnetic field amplification and electrostatic attraction.

Exemplary embodiments of the present invention are a bioconversion metabolite detection device including a metabolite enrichment and detection unit including a metal nanostructure, and a bioconversion metabolite with a metal nanostructure provided in the metabolite enrichment and detection unit It provides a bioconversion metabolite detection device for detecting a surface-enhanced Raman scattering signal.

Exemplary embodiments of the present invention provide a method for concentrating and detecting a biotransformation metabolite, comprising: attaching the biotransformation metabolite to a metal nanostructure and concentrating; It provides a method for detecting a bioconversion metabolite comprising; detecting a bioconversion metabolite from the surface-enhanced Raman scattering signal emitted by irradiating the metal nanostructure with a light source.

According to exemplary embodiments of the present invention, it is possible to detect biotransformation metabolites at ultra-high speed while having high reliability and specificity. In exemplary embodiments of the present invention, since bioconversion metabolites in the reactor are selectively concentrated on the surface of metal nanostructures through chemical adsorption, it is possible to sensitively detect bioconversion metabolites present in a very low concentration in a solution. There is an advantage.

In addition, according to exemplary embodiments of the present invention, unlike off-line sampling technology in which an aqueous solution in the reactor is collected and analyzed outside, a bioconversion metabolite detection device can be directly employed inside the reactor, thereby reducing the risk of reactor contamination. There is an advantage that there is.

In addition, according to exemplary embodiments of the present invention, the apparatus and method for detecting bioconversion metabolites may be widely used for real-time monitoring of feeds and products of various bioconversion reactors by introducing various metal nanostructures.

1 is an exemplary embodiment of the present invention, by introducing an optical sensor in which a metal nanostructure and a laser are combined in a bioconversion reactor to concentrate metabolites around the metal nanostructure through electrostatic attraction, and surface-enhanced Raman scattering thereof. A schematic diagram of real-time monitoring by law is shown.
Figure 2 is, in the embodiment of the present invention, a single layer of gold nanoparticles in the form of a rod was formed at the interface between the water-organic solvent, and the organic solvent was evaporated and then transferred to a solid substrate and observed with a scanning electron microscope. The results are shown.
3 is a surface enhancement after forming a single layer of silver nanoparticles in the form of a rod at the interface between the water-organic solvent, evaporating the organic solvent, and transferring it to a transparent polymer substrate (PDMS, polydimethylsiloxane) in an embodiment of the present invention. It shows the result of detecting acetate (acetate) through the Raman scattering method.
Figure 4 is, in an embodiment of the present invention, a single layer of silver nanoparticles in the form of a rod is formed at the interface between the water-organic solvent, the organic solvent is evaporated, and then transferred to a transparent polymer substrate, through a surface-enhanced Raman scattering method. The result of detecting butyrate is shown.
5 shows a single layer of gold nanoparticles in the form of a rod at the water-organic solvent interface, evaporating the organic solvent, and introducing it into a microfluidic system, and changing the concentration of acetate through a surface-enhanced Raman scattering method. Real-time observation results are shown.

Hereinafter, exemplary embodiments of the present invention will be described in detail.

In the present specification, "biotransformation metabolite" refers to a substance produced in the process of metabolism of microorganisms.

In the present specification, "nano" means 1000 nm or less.

Bioconversion metabolite detection device

An exemplary embodiment of the present invention is a bioconversion metabolite detection device including a metabolite concentration and detection unit including a metal nanostructure, and a bioconversion metabolite to which a metal nanostructure provided in the metabolite concentration and detection unit is attached. It provides a bioconversion metabolite detection device that detects with a surface-enhanced Raman scattering signal.

In an exemplary embodiment, the bioconversion metabolite detection device can be directly introduced into the bioconversion reactor, so unlike the offline sampling technology in which an aqueous solution in the reactor is collected and analyzed outside, the bioconversion metabolite detection device is directly used as a reactor. Since it can be employed inside, the risk of reactor contamination can be reduced.

In an exemplary embodiment, the metal nanostructure may include a substrate and a metal nanoparticle layer formed on the substrate.

In an exemplary embodiment, the metal nanoparticle layer may include a plurality of metal nanoparticles, and the metal nanoparticles may selectively concentrate metabolites in the reactor on the surface of the metal nanostructure through electrostatic attraction.

In an exemplary embodiment, the metal nanoparticle layer includes one or more of a positive charge region and a negative charge region, the positive charge region includes first metal nanoparticles having a positive charge, and the negative charge region is a negative charge region. It may contain 2 metal nanoparticles.

Referring to FIG. 1, the metal nanoparticles may be positively charged first metal nanoparticles or negatively charged second metal nanoparticles, and depending on the electric charge of the metal nanoparticles, selectively biological Conversion metabolites can be attached.

For example, a part of the metal nanostructure may be a positively charged region, and in the positively charged region, an electrostatic attraction may be formed between a biotransformation metabolite carrying a negative charge, and thus, a negatively charged region in the positively charged region. Biotransformation metabolites can be attached. In addition, for example, a part of the metal nanostructure may be a negatively charged region, and in the negatively charged region, an electrostatic attraction may be formed between a bioconversion metabolite having a positive charge, and thus, a positive charge may be obtained in the negatively charged region. Biotransformation metabolites can be attached.

As a result, negatively charged biotransformation metabolites can be attached to some areas of the metal nanostructure by electrostatic attraction, and positively charged bioconversion metabolites are attached to other areas of the metal nanostructure by electrostatic attraction. Can be.

Here, the bioconversion metabolite may be a material that is difficult to detect due to its small molecule size and Raman cross-sectional area. However, the bioconversion metabolite detection device according to the embodiment of the present invention is a bioconversion metabolite in the reactor through electrostatic attraction. Since is selectively concentrated on the surface of the metal nanostructure, bioconversion metabolites present at a very low concentration in the aqueous solution can be sensitively detected through the surface-enhanced Raman scattering method.

In an exemplary embodiment, the bioconversion metabolite may be a metabolite produced in a metabolic process of a microorganism, and is not particularly limited as long as it is produced in a metabolic process of a microorganism. As a non-limiting example, it may include one or more selected from the group consisting of acetate, butyrate, 3-propionate, 2,3-butanediol, ethanol, and methanol.

For example, acetate is a type of metabolite that uses syngas as a raw material and is produced through the metabolism of microorganisms, and is one of the substances that is difficult to detect due to its small molecule size and Raman cross-section. Further, the acetate molecule may have a negative charge in the aqueous solution.

In an exemplary embodiment, the substrate is not particularly limited as long as it is capable of transferring the metal nanoparticle layer. As a non-limiting example, the substrate is a transparent substrate, the transparent substrate is one or more selected from the group consisting of polymer, glass, and ITO, and the polymer may be one or more selected from the group consisting of PDMS, PMMA, and hydrogel have.

In an exemplary embodiment, the metal nanoparticles may have one or more shapes selected from the group consisting of a sphere, a rod, an ellipsoid, a dendrimer, a tetrahedron, a hexahedron, an octahedron, a two-dimensional square, and a two-dimensional triangular shape.

In an exemplary embodiment, the metal nanostructure includes one or more metals selected from the group consisting of Au, Ag, Pd, Pt, Al, Cu, Co, Cr, Mn, Ni, and Fe, or an alloy of two or more metals. can do. For example, acetate or butyrate may be attached to the surface of silver by electrostatic attraction, and by using this, these molecules may be concentrated on the surface of the metal nanostructure. For example, silver (Ag) may have a positively charged surface. Accordingly, it is possible to detect negatively charged acetate using silver nanoparticles. In addition, for example, a molecule of butyrate, which is one of the bioconversion metabolites, is also negatively charged in an aqueous solution, so it can be detected using silver nanoparticles. Here, both acetate and butyrate are one of representative metabolites produced in the metabolic process of microorganisms, and are one of the substances that are difficult to detect due to the small size of the molecule and the small Raman cross-sectional area.

In particular, bioconversion metabolites, such as acetate or butyrate, that are present in a solution in trace amounts are concentrated and attached using a metal nanostructure including metal nanoparticles, and the bioconversion metabolites are added to the metal nanoparticles. By attaching, strong electromagnetic field amplification can occur between metal nanoparticles on the surface.

Biotransformation metabolites generally show a weak Raman signal, but the Raman signal of the biotransformation metabolite, which is a weak signal, can be amplified by the method described above, and the position of the amplified Raman signal is analyzed to determine the biotransformation metabolite. Can be detected. Accordingly, it is possible to detect biotransformation metabolites with high reliability, specificity, and sensitivity at high speed.

In an exemplary embodiment, the diameter of the metal nanoparticles may be 5 to 200 nm.

In an exemplary embodiment, the apparatus for detecting bioconversion metabolites may further include a light irradiation unit, and the light irradiation unit may be to irradiate a light source onto the metal nanostructure. The light source may detect bioconversion metabolites by irradiating the optical sensor metal nanostructure with a light source such as a laser.

In an exemplary embodiment, the metal nanoparticle layer may be in contact with a bioconversion metabolite to detect a surface-enhanced Raman scattering signal through the substrate.

In an exemplary embodiment, the detection device may further include a microfluidic channel configured to provide a metal nanoparticle to which a bioconversion metabolite is attached to the detection unit. With the use of microfluidic channels, large volumes of samples can be processed quickly. In a non-limiting example, a microfluidic channel can be implemented, for example as shown in FIG. 5.

1 is an exemplary embodiment of the present invention, by introducing an optical sensor in which a metal nanostructure and a laser are combined in a bioconversion reactor to concentrate metabolites around the metal nanostructure through electrostatic attraction, and surface-enhanced Raman scattering thereof. It is a schematic diagram of real-time monitoring by law. Since metabolites are concentrated by electrostatic attraction around metal nanostructures where Raman signals are amplified, real-time monitoring of metabolites present at very low concentrations is possible.

As shown in FIG. 1, a solution containing a bioconversion metabolite is provided to the metal nanostructure to attach the bioconversion metabolite to the metal nanoparticle. Subsequently, a light source, such as a laser, may be irradiated to the bioconversion metabolite concentration and detection unit to which the bioconversion metabolite is attached.

The metal nanoparticles included in the bioconversion metabolite concentration and detection unit may effectively concentrate the bioconversion metabolite.

By irradiating a laser to the bioconversion metabolite concentration and detection unit, the bioconversion metabolite concentrated in the metal nanoparticles may be detected as a surface-enhanced Raman scattering signal.

In an exemplary embodiment, the metal nanoparticle layer may contact a bioconversion metabolite and detect a surface-enhanced Raman scattering signal through the transparent substrate.

In an exemplary embodiment, the detection unit may include quantifying the type and/or concentration of the bioconversion metabolite according to the position at which the bioconversion metabolite is attached to the metal nanostructure.

Biotransformation metabolite detection method

An exemplary embodiment of the present invention is a method of concentrating and detecting a biotransformation metabolite, comprising: attaching the bioconversion metabolite to a metal nanostructure and concentrating; A method for detecting a bioconversion metabolite comprising; detecting a bioconversion metabolite from the surface-enhanced Raman scattering signal emitted by irradiating the metal nanostructure with a light source is provided.

In an exemplary embodiment, the detection method may detect a bioconversion metabolite by measuring a Raman signal of a molecule surrounding a metal nanostructure.

In an exemplary embodiment, the concentrating step may be selectively concentrating a bioconversion metabolite to at least a portion of the metal nanostructure through electrostatic attraction.

In an exemplary embodiment, the step of forming a metal nanostructure further includes, wherein the forming of the metal nanostructure includes: forming a metal nanoparticle layer at an interface between the first material and the second material; And transferring the formed metal nanoparticle layer to a substrate.

In an exemplary embodiment, the forming of the metal nanoparticle layer may include forming a metal nanoparticle layer through self-assembly of the metal nanoparticles at a liquid-liquid interface of the first material and the second material; It may include a step of forming a liquid-phase interface by changing the phase of the second material.

On the other hand, metal nanoparticles may be self-assembled. Here, self-arrangement means that the corresponding components can be arranged in a specific baton or can form a structure without an external force applied separately to perform the arrangement.

In one embodiment, the metal nanoparticles at the liquid-liquid interface of the first material and the second material may form a single layer of metal nanoparticles through self-assembly. By forming a single layer, it is possible to have excellent detection performance.

Meanwhile, alcohol may be added to the liquid-liquid interface for self-assembly of the metal nanoparticles. For example, ethanol can be added to the liquid-liquid interface.

For example, the nucleic acid may be added to form an interface with an aqueous solution. Metal nanoparticles are generally stable to exist at the interface due to the energy caused by the surface tension, but when only nucleic acids are added, the electrostatic repulsion between the metal nanoparticles is stronger and self-assembly does not occur. Since ethanol weakens the charge of molecules surrounding the nanoparticle surface, it can induce self-assembly by reducing electrostatic repulsion.

Then, the second material may be phase-changed. For example, the phase change can be made from liquid to gaseous. The means used for the phase change are not limited within the purpose of the present invention. Through this phase change process, the first material and the second material may form a metal nanoparticle layer on the liquid-vapor interface.

In one embodiment, the formed metal nanoparticle layer may be transferred to the substrate, and specifically, the first material and the second material may be transferred from the liquid-vapor interface to the substrate through the vapor-phase interface.

For example, in the case of a metal nanoparticle layer formed at a liquid-liquid interface, the liquid phase present in the upper layer (eg, a second material) is evaporated, and then the metal nanoparticle layer present at the formed liquid-gas interface can be transferred. have. In an embodiment, after forming a metal nanoparticle layer on the water-nucleic acid interface and evaporating the nucleic acid, the formed metal nanoparticle layer may be transferred to a substrate.

In addition, in one embodiment, in the transfer step, the metal nanoparticle layer may be transferred to the substrate by contacting the interface and the substrate so that the formed interface and the surface of the substrate, such as the transparent substrate, are parallel to each other and then separated.

In an exemplary embodiment, when the first material is water (liquid), the second material may be an organic solvent to form an interface with the first material. For example, the organic solvent is benzene, toluene, It may be an organic solvent such as a fatty acid series such as chloroform, a nucleic acid, or oleic acid, or an aliphatic amine series such as oleylamine.

In an exemplary embodiment, the liquid phase interface between the first material and the second material can be directly formed, and in this case, the second material includes air and the first material includes an organic solvent such as benzene, toluene, nucleic acid, chloroform, etc. can do.

In an exemplary embodiment, it may be to provide a metal nanostructure to a solution containing a bioconversion metabolite in which the metal nanostructure and electrostatic attraction act.

In an exemplary embodiment, the step of detecting the biotransformation metabolite may include quantifying the concentration of the detected biotransformation metabolite. For example, it may include normalizing the result of detecting the biotransformation metabolite from the surface-enhanced Raman scattering signal to a certain standard.

In an exemplary embodiment, detecting the biotransformation metabolite may include quantifying the concentration of the biotransformation metabolite according to the position attached to the metal nanostructure.

Hereinafter, specific embodiments according to exemplary embodiments of the present invention will be described in more detail. However, the present invention is not limited to the following examples, and various types of examples may be implemented within the scope of the appended claims, and only the following examples are common in the art to complete the disclosure of the present invention. It will be understood that it is intended to facilitate the implementation of the invention to those of skill.

[Production Example 1] Preparation of metal nanostructures

Forming a single layer of metal nanoparticles at the liquid-gas interface

After forming a liquid-liquid interface by carefully adding 3 ml of n-hexane to 9 ml of an aqueous solution containing rod-shaped silver nanoparticles, 4.5 ml of ethanol was added to the nanoparticle solution, and then allowed to stand at room temperature for 6 hours. -All of the hexane (n-hexane) was evaporated.

Transfer of a single layer of nanoparticles to a solid substrate

A PDMS substrate with a length of 1 cm on one side was horizontally brought into contact with the interface where a single layer of nanoparticles exists, and then removed to prepare a metal nanostructure.

FIG. 2 shows a scanning electron microscope image measured after transferring a single layer of gold nanoparticles in the form of a rod existing at a water-air interface to a PDMS substrate having a side length of 1 cm in an embodiment of the present invention.

[Example 1] Acetate detection through surface-enhanced Raman scattering method

A Raman signal was measured by dropping 30 μL of an aqueous acetate solution onto the metal nanostructure of Preparation Example 1 and then exposing a laser having a wavelength of 785 nm for 3 seconds.

FIG. 3 shows the result of detecting acetate through a surface-enhanced Raman spectroscopy method using a single layer of silver nanoparticles in the form of a rod transferred to a PDMS substrate in an exemplary embodiment of the present invention. .

From FIG. 3, it can be seen that a characteristic Raman peak appeared in the region of 928cm ^-1 appearing from the CC binding of acetate, and the lower limit of detection was measured as 10 μM.

[Example 2] Butyrate detection through surface-enhanced Raman scattering method

After 30 μL of an aqueous butyrate solution was dropped onto the metal nanostructure of Preparation Example 1, a laser having a wavelength of 785 nm was irradiated for 3 seconds to measure a Raman signal.

FIG. 4 is a result of detecting butyrate through a surface-enhanced Raman scattering method using a single layer of silver nanoparticles in the form of a rod transferred to a PDMS substrate in an exemplary embodiment of the present invention.

From FIG. 4, it can be seen that a characteristic Raman peak appeared in the region of 877cm ^-1 appearing from CO bonding of butyrate, and the lower limit of detection was measured as 1mM.

As described above, in the case of exemplary embodiments of the present invention, since metabolites in the biotransformation reactor are selectively concentrated on the surface of the metal nanostructure through electrostatic attraction, metabolism exists at a very low concentration in the aqueous solution and has a small Raman cross-sectional area. Products can be sensitively detected.

In addition, the chromatography technique used to analyze bioconversion metabolites takes more than 20 minutes to detect, but according to exemplary embodiments, the present invention is capable of exposing an optical sensor to a solution and irradiating a laser to detect it. Real time continuous detection is possible by requiring only seconds

In addition, unlike the offline sampling technology in which the aqueous solution in the reactor is collected and analyzed outside, the optical sensor in which the metal nanostructure and the laser are combined can be combined together inside the reactor, thereby reducing the risk of reactor contamination.

As a result, according to exemplary embodiments of the present invention, by introducing various metal nanostructures into the reactor, it can be widely used for real-time monitoring of feeds and products of various bioconversion reactors.

Claims (19)

It is a bioconversion metabolite detection device comprising a; metabolite concentration and detection unit including a metal nanostructure,
The bioconversion metabolite attached to the metal nanostructure provided in the metabolite concentration and detection unit is detected as a surface-enhanced Raman scattering signal,
The metal nanostructure includes a substrate and a metal nanoparticle layer formed on the substrate,
The metal nanoparticle layer includes a plurality of metal nanoparticles, and the metal nanoparticle selectively concentrates bioconversion metabolites on the surface of the metal nanostructure through electrostatic attraction,
The metal nanoparticle layer includes at least one of a positively charged region and a negatively charged region, the positively charged region includes first metal nanoparticles having a positive charge, and the negatively charged region includes second metal nanoparticles having a negative charge And
The first metal nanoparticles concentrate a biotransformation metabolite carrying a negative charge,
The second metal nanoparticles are bioconversion metabolites detection device for concentrating biotransformation metabolites having a positive charge. delete delete delete The method of claim 1,
The biotransformation metabolite comprises at least one selected from the group consisting of acetate, butyrate, 3-propionate, 2,3-butanediol, ethanol, and methanol. The method of claim 1,
The substrate is a transparent substrate, the transparent substrate is one or more selected from the group consisting of polymers, glass, and ITO, and the polymer is one or more selected from the group consisting of PDMS, PMMA, and hydrogel, bioconversion metabolites Detection device. The method of claim 1,
The metal nanoparticles have at least one shape selected from the group consisting of a sphere, a rod, an ellipsoid, a dendrimer, a tetrahedron, a hexahedron, an octahedron, a two-dimensional square, and a two-dimensional triangular shape. The method of claim 1,
The metal nanostructure is a biotransformation metabolite comprising at least one metal selected from the group consisting of Au, Ag, Pd, Pt, Al, Cu, Co, Cr, Mn, Ni, and Fe or an alloy of two or more metals Detection device. The method of claim 1,
The diameter of the metal nanoparticles is 5-200 nm, bioconversion metabolite detection device. The method of claim 1,
The bioconversion metabolite detection device further includes a light irradiation unit,
The light irradiation unit to irradiate a light source to the metal nanostructure, bioconversion metabolite detection device. The method of claim 1,
The metal nanoparticle layer is in contact with the bioconversion metabolite and detects a surface-enhanced Raman scattering signal through the substrate. The method of claim 1,
The detection device further comprises a microfluid channel configured to provide a metal nanoparticle to which a biotransformation metabolite is attached to the detection unit.
As a method for concentrating and detecting biotransformation metabolites,
Attaching the biotransformation metabolite to the metal nanostructure and concentrating;
Including; detecting bioconversion metabolites from the surface-enhanced Raman scattering signal emitted by irradiating the metal nanostructure with a light source,
The metal nanostructure includes a substrate and a metal nanoparticle layer formed on the substrate,
The metal nanoparticle layer includes a plurality of metal nanoparticles, and the metal nanoparticle selectively concentrates bioconversion metabolites on the surface of the metal nanostructure through electrostatic attraction,
The metal nanoparticle layer includes at least one of a positively charged region and a negatively charged region, the positively charged region includes first metal nanoparticles having a positive charge, and the negatively charged region includes second metal nanoparticles having a negative charge And
The first metal nanoparticles concentrate a biotransformation metabolite carrying a negative charge,
The second metal nanoparticles are bioconversion metabolites detection method for concentrating a biotransformation metabolite bearing a positive charge. delete The method of claim 13,
Further comprising the step of forming a metal nano structure, the step of forming the metal nano structure,
Forming a metal nanoparticle layer at the interface between the first material and the second material; And
Transforming the formed metal nanoparticle layer to a substrate; containing, bioconversion metabolite detection method. The method of claim 15,
The step of forming the metal nanoparticle layer,
Forming a metal nanoparticle layer through self-assembly of the metal nanoparticles at the liquid-liquid interface of the first material and the second material;
Forming a liquid-phase interface by phase-changing the second material; comprising, bioconversion metabolite detection method. The method of claim 13,
A method for detecting a biotransformation metabolite to provide a metal nanostructure to a solution containing a bioconversion metabolite in which the metal nanostructure and electrostatic attraction act. The method of claim 13,
The step of detecting the biotransformation metabolite comprises quantifying the concentration of the detected biotransformation metabolite. The method of claim 18,
The step of detecting the biotransformation metabolite comprises quantifying the concentration of the biotransformation metabolite according to the position attached to the metal nanostructure.

Images