Substrate for spectroscopic analysis including a polycrystalline structure and method for manufacturing the same

Abstract

The present invention relates to a substrate for spectroscopic analysis including a polycrystalline structure and a method for manufacturing the same. More specifically, the present invention relates to a substrate for spectroscopic analysis of a universal material substrate including a polycrystalline structure with improved signal intensity and signal uniformity, and a method for manufacturing a substrate for spectroscopic analysis including a polycrystalline structure whose shape is controlled by a simple process on the surfaces of various materials.

Description

Technical Field

[0001] The present invention relates to a substrate for spectroscopic analysis including a polycrystalline structure and a method for manufacturing the same. More specifically, the present invention relates to a substrate for spectroscopic analysis of a universal material substrate including a polycrystalline structure with improved signal intensity and signal uniformity, and a method for manufacturing a substrate for spectroscopic analysis including a polycrystalline structure with its shape controlled by a simple process on the surfaces of various materials.

Background Art

[0002] Raman scattering is inelastic scattering in which the energy of the incident light changes. When light is applied to a specific molecular entity, it refers to a phenomenon in which light with a slightly different wavelength from the irradiated light is generated due to the vibration transition inherent in the molecular entity.

[0003] In fact, almost all organic molecules have a unique Raman shift. According to Raman spectroscopy using Raman scattering, signals can be obtained even in the case of non-polar molecules with a change in the induced polarizability of the molecule. In addition, since Raman spectroscopy is not affected by the interference of water molecules, it is more suitable for the detection of biomolecules such as proteins and genes.

[0004] Since the wavelength of the Raman emission spectrum indicates the chemical composition and structural characteristics of the light-absorbing molecules in the sample, analyzing such a Raman signal can directly analyze the analyte.

[0005] As the background art of the present application, Korean Patent No. 10-1867670 describes a method for manufacturing a paper-based surface-enhanced Raman scattering substrate using a continuous chemical reaction method. However, in the said patent, not only is the process complex for manufacturing metal nanoparticles having a diameter of 1 to 100 nm and using a first metal precursor solution, a second metal precursor solution, a first reducing agent solution, and a second reducing agent solution, but also there are problems in that process control is not easy, such as repeating several times processes such as a precursor adsorption step, a precursor washing step, a loading step into a reducing agent solution, and a reducing agent solution washing step.

Summary of the Invention

Problems to be Solved by the Invention

[0006] An object of the present invention is to provide a substrate for spectroscopic analysis that is excellent in sensitivity and signal uniformity when used for spectroscopic analysis, in which noble metal-containing nanoparticles are directly grown on a base member.

[0007] Another object of the present invention is to provide a substrate for spectroscopic analysis that is excellent in flexibility and easy to apply to various three-dimensional surfaces.

[0008] Another object of the present invention is to provide a composition for manufacturing a substrate for spectroscopic analysis that can easily adjust the shape of noble metal nanocrystalline particles formed regardless of the material of the base member and can easily form polycrystalline particles, thereby enhancing the sensitivity and signal uniformity of surface-enhanced Raman spectroscopy (SERS) signals.

[0009] Another object of the present invention is to provide a Raman spectroscopic apparatus capable of on-site diagnosis such as cancer.

[0010] Another object of the present invention is to provide a method for manufacturing a substrate for spectroscopic analysis that can efficiently manufacture a substrate for spectroscopic analysis having high sensitivity and signal uniformity of SERS signals by a simple process regardless of the material of the base member.

[0011] The object of the present invention is not limited to the objects mentioned above, and other objects not mentioned can be clearly understood from the description of the detailed description.

Means for Solving the Problem

[0012] According to one aspect, there is provided a substrate for spectroscopic analysis, which is composed of a base member; and an aggregate of a plurality of nanoparticles formed on the base member, and has a polycrystalline structure having a plurality of grain boundaries, and is formed by a solution process.

[0013] According to one embodiment, the base member may be composed of one or more of paper, polymer, well plate, wafer, and protein.

[0014] According to one embodiment, the nanoparticles are connected by a branch structure, and the polycrystalline structure may have an average particle size of 1 μm to 100 μm.

[0015] According to one embodiment, the substrate for spectroscopic analysis of the present application has an electrochemically active surface area (EASA) of 0.1 cm 2 ~10.0 cm 2 and may be.

[0016] According to one embodiment, the nanoparticles may be composed of one or more of Au, Ag, and Pt.

[0017] According to one embodiment, the substrate for spectroscopic analysis may be applicable to a Raman spectroscopy system using a wavelength of 500 nm or more or a near-IR FT Raman spectroscopy system using a wavelength of 1064 nm.

[0018] According to one embodiment, the substrate for spectroscopic analysis may be composed of a three-dimensional swap stick, a wearable substrate, or a plasmonic well plate.

[0019] According to another aspect, there is provided a composition for manufacturing a substrate for spectroscopic analysis, which contains a noble metal precursor and a reducing agent, and the ratio of the reducing agent to the noble metal precursor is 1:0.5 to 1:15.

[0020] According to one embodiment, in the composition for manufacturing a substrate for spectroscopic analysis of the present application, as the ratio of the reducing agent to the noble metal precursor increases, the size of the polycrystalline structure formed on the substrate for spectroscopic analysis decreases, and the density of the polycrystalline structure can increase.

[0021] According to one embodiment, the noble metal precursor may be one or more of HAuCl4 and NaAuCl4.

[0022] According to one embodiment, the reducing agent may be one or more of hydroxylamine, ascorbic acid, FeSO4, and hydroxyquinone.

[0023] According to still another aspect, there is provided a Raman spectroscopy apparatus including a light source; a substrate for spectroscopic analysis described in the present application; and a detector for detecting Raman spectroscopy.

[0024] According to one embodiment, the Raman spectroscopy apparatus of the present application is cylindrical with the first end open, the first end includes a fastening portion fastened to the Raman spectroscopy apparatus, the second end includes an adapter having a through hole formed therein for guiding light from the light source; and a spoon shape composed of a head portion and a linear body portion extending from the head portion, the head portion further includes a hemispherical seating portion on which the coupling portion of the second end of the adapter is seated, and a seating groove extending longitudinally to the hemispherical seating portion at the center of the body portion and on which a linear spectroscopic analysis strip substrate is seated, and can further include a measuring member.

[0025] According to one embodiment, the seating groove may be formed in a T shape, and it can be characterized in that the end of the spectroscopic analysis strip substrate does not come off from the hemispherical seating portion.

[0026] According to one embodiment, the target substances of the detector may be selected from one or more of cells, metabolites, proteins, nucleic acids, DNA, RNA, enzymes, organic molecules, viruses, extracellular vesicles, microvesicles, exosomes, and fats in urine, saliva, sweat, and tears.

[0027] According to one embodiment, the Raman spectrometer may be capable of on-site diagnosis of cancer.

[0028] According to still another aspect, there is provided a method for manufacturing a substrate for spectroscopic analysis described in the present application, including: a) a step of preparing a base member; and b) a step of supporting the base member on a composition for manufacturing a substrate for spectroscopic analysis containing a noble metal precursor and a reducing agent solution, and forming a polycrystalline structure composed of a cluster of a plurality of nanoparticles and having a plurality of grain boundaries on the base member, wherein in step b), the ratio of the reducing agent to the noble metal precursor solution is 1:0.5 to 1:15, and the method includes a shape adjustment step of adjusting the shape of the polycrystalline structure.

[0029] According to one embodiment, in step a) of the method for manufacturing a substrate for spectroscopic analysis of the present application, the method may further include a surface modification step of modifying the surface of the base member before the step of forming the polycrystalline structure.

[0030] According to one embodiment, in the method for manufacturing a substrate for spectroscopic analysis of the present application, the surface modification step includes treating the surface of the base member with a surface modification composition containing one or more selected from 1 to 3% of a base and 1 to 3% of an acid and containing alcohol or water as a solvent. 1-5 of C

Advantages of the Invention

[0031] According to one embodiment, the substrate for spectroscopic analysis of the present application can include a polycrystalline structure having a microparticle size, in which noble metal-containing nanoparticles are directly grown and connected to a branch structure, and is excellent in sensitivity and signal uniformity when used for spectroscopic analysis.

[0032] According to one embodiment, the base member of the substrate for spectroscopic analysis of the present application is composed of various materials, is excellent in flexibility, and can be easily applied to various three-dimensional surfaces.

[0033] According to one embodiment, the composition for manufacturing the substrate for spectroscopic analysis of the present application can adjust the ratio of the noble metal precursor and the reducing agent, easily adjust the shape of the noble metal nanocrystalline particles formed regardless of the material of the base member, easily form polycrystalline particles, and enhance the sensitivity and signal uniformity of the SERS signal.

[0034] According to one embodiment, the Raman spectroscopic apparatus of the present application can confirm the type of non-labeled analyte on-site and perform quantitative analysis.

[0035] According to one embodiment, the Raman spectroscopic apparatus of the present application can perform on-site diagnosis of cancer and the like using a small amount of biological samples such as urine.

[0036] According to one embodiment, the manufacturing method of the substrate for spectroscopic analysis of the present application can directly grow a noble metal-containing polycrystalline structure on the base member in a simple process of one solution process regardless of the material of the base member, and efficiently manufacture a substrate for spectroscopic analysis with high SERS signal intensity and signal uniformity. According to one embodiment, the manufacturing method of the substrate for spectroscopic analysis of the present application can efficiently manufacture a substrate for spectroscopic analysis having a high particle density and improved signal uniformity through surface modification.

Brief Description of the Drawings

[0037]

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[0038]

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Mode for Carrying Out the Invention

[0039] The objects, specific merits, and novel features of the present disclosure will become more apparent from the following detailed description and examples related to the attached drawings.

[0040] Prior to this, terms or words used in this specification and claims should not be construed in a normal and dictionary sense, but should be construed in a meaning and concept consistent with the technical idea of the present disclosure based on the principle that the inventor can appropriately define the concept of the terms in order to explain his own invention in the best way.

[0041] As used herein, when a component such as a layer, a portion, or a substrate is described as being "on," "connected to," or "coupled to" another component, this may mean directly "on," "connected to," or "coupled to" the other component, or there may be one or more other components intervening between the two components. In contrast, when a component is described as being "directly on," "directly connected to," or "directly coupled to" another component, there can be no other component intervening between the two components.

[0042] The terms used herein are for the purpose of describing particular embodiments only and are not intended to limit the present disclosure. Singular expressions include plural expressions unless the context clearly dictates otherwise.

[0043] As used herein, terms such as "comprising" or "having" are intended to specify the presence of the features, numbers, steps, operations, components, parts, or combinations thereof described in the specification, and should be understood not to preclude the possibility of the presence or addition of one or more other features, such as numbers, steps, operations, components, parts, or combinations thereof.

[0044] As used herein, when a portion is said to "comprise" a certain component, this means that, unless otherwise specifically stated to the contrary, it does not exclude other components and can further comprise other components. Throughout the specification, "on" means located above or below the target portion, and does not necessarily mean located on the upper side with reference to the direction of gravity.

[0045] Since the present disclosure can be subjected to various conversions and can have various embodiments, specific embodiments will be illustrated in the drawings and described in detail in the detailed description. However, this is not intended to limit the present disclosure to specific embodiments, and it should be understood to include all conversions, equivalents, and alternatives included in the spirit and technical scope of the present disclosure. In explaining the present disclosure, if a specific explanation of related known technologies is determined to obscure the gist of the present disclosure, the detailed explanation thereof will be omitted.

[0046] Hereinafter, embodiments of the present disclosure will be described in detail with reference to the accompanying drawings. When describing with reference to the accompanying drawings, the same or corresponding components will be given the same drawing numbers, and duplicate descriptions thereof will be omitted.

[0047] Figure 1(a) shows a paper-based substrate for spectroscopic analysis manufactured using cellulose acetate as a base member according to an embodiment of the present invention. Figures 1(b) to 1(e) show SEM photographs (Figure 1(b)), polycrystalline verification results (Figure 1(c)), particle formation mechanisms (Figure 1(d)), and particle distribution graphs (Figure 1(e)) of the substrate for spectroscopic analysis of Figure 1(a). Figure 2 is an SEM photograph showing the shapes of particles grown using nanocellulose (NC), mixed cellulose ester (MCE), chromatography paper, and printing paper as base members according to embodiments of the present invention.

[0048] Figures 3(a) to 3(c) are drawings showing the results of measuring the electrochemical surface area according to the ratio of the reducing agent contained in the composition for manufacturing a substrate for spectroscopic analysis according to embodiments of the present invention.

[0049] The substrate for spectroscopic analysis of the present application is composed of a base member; and a plurality of clusters of nanoparticles formed on the base member, and includes a polycrystalline structure having a plurality of grain boundaries, and is a substrate formed in a solution process.

[0050] Referring to FIGS. 1(a) to 1(e) and FIG. 2, polycrystals formed by the aggregation of a plurality of nanoparticles formed on cellulose acetate (see FIGS. 1(a) to 1(e)) and various other base members can be confirmed (see FIG. 2). Referring to FIG. 1(d), although not limited thereto, among noble metals, it is shown that an Au precursor forms nanoparticles and aggregates. In particular, these grow directly on the base member using a solution process. Specifically, the aggregated nanoparticles form small-sized polygonal-structured grains (grains) by coalescence, and then the grains gradually grow larger by oriented attachment. Therefore, a polycrystalline structure having a plurality of grain boundaries is formed. Although not limited thereto, noble metal coral-shaped particles, which are polycrystalline structures, can grow to an average micro size (see FIG. 1(e)). As described above, a polycrystalline structure having a plurality of grain boundaries has increased scattering at the plurality of grain boundaries, and when used as a substrate for spectroscopic analysis, the signal intensity and signal uniformity can be remarkably improved.

[0051] The base member applicable to the present application is not particularly limited, and various materials can be utilized. Without being limited thereto, the base member may be composed of one or more of paper, polymer, well plate, wafer, and protein. Without being limited thereto, the paper may be cellulose acetate, nano cellulose (NC), mixed cellulose ester (MCE), chromatography paper, and the same paper as commonly used printing paper. Further, the substrate for analytical spectroscopy may be a protein film composed of one or more proteins such as porous polymer, plasmonic well plate, wafer, and silk, collagen, chitin, elastin, keratin. Referring to FIG. 2, it is possible to confirm the form of polycrystalline structures of similar structures grown on the base member of nano cellulose (NC) (FIG. 2(a)), mixed cellulose ester (MCE) (FIG. 2(b)), chromatography paper (FIG. 2(c)), and commonly used printing paper (FIG. 2(d)). Without being limited thereto, depending on the field of application, a base member having porosity and / or flexibility can be suitable, and paper, polymer, or protein film can be suitable. Further, the plasmonic well plate can be suitable when detecting a plurality of samples simultaneously.

[0052] Without being limited thereto, the nanoparticles are connected in a branch structure, and the average particle size of the polycrystalline structure may be 1 μm to 100 μm. The substrate for spectroscopic analysis of the present application grows into a polycrystalline structure by a solution process, is connected in a branch structure, and the average particle size may be 1 μm to 100 μm. Without being limited thereto, the average particle size of the polycrystalline structure can be suitable for improving signal intensity and signal uniformity within the above range.

[0053] Without being limited thereto, the substrate for spectroscopic analysis of the present application has an electrochemically active surface area (EASA) of 0.1 cm 2 ~10.0 cm 2It is possible. The substrate for spectroscopic analysis of the present application can grow into a polycrystalline structure by a solution process, and the surface area can be increased. Further, referring to FIGS. 3(a) to 3(c), as the ratio of the reducing agent to the metal ion precursor contained in the composition for manufacturing a substrate for spectroscopic analysis according to the present application increases, it can be confirmed that the electrochemically active surface area (EASA, cm 2 ) has increased. On the other hand, when the ratio of the reducing agent to the metal ion precursor is less than 1:0.5, the density of the noble metal polycrystalline structure may be low and current may not flow.

[0054] Although not limited thereto, the nanoparticles can be composed of one or more of Au, Ag, and Pt, and Au is most suitable in terms of signal intensity and signal uniformity.

[0055] Although not limited thereto, the substrate for spectroscopic analysis of the present application may be applicable to a Raman spectroscopy system using a wavelength of 500 nm or more or a near-IR FT Raman spectroscopy system using a wavelength of 1064 nm.

[0056] Referring to FIG. 6(b), when the substrate for spectroscopic analysis according to the present application is used, it shows a wide absorption region at a wavelength of 500 nm or more, so it can be confirmed that Raman spectroscopy systems using various currently commercial wavelengths can be used. Although not limited thereto, it is more suitable at 633 nm or 785 nm.

[0057] FIGS. 4(a) to 4(d) are photographs showing various forms of substrates for spectroscopic analysis according to embodiments of the present invention.

[0058] Although not limited thereto, the substrate for spectroscopic analysis may be composed of a three-dimensional swap stick, a wearable substrate, or a plasmonic well plate. The substrate for optical analysis of the present application can include various base members, can be manufactured from a flexible material, and can have various forms. Referring to FIG. 4, in addition to a planar substrate, it can be applied in various forms such as a 3D swap stick using a three-dimensional porous material, a wearable substrate form, a well plate form, a protein film form, etc.

[0059] The substrate for spectroscopic analysis in the form of the three-dimensional swap stick enables easy sample collection. The wearable substrate is composed in the form of a patch or a lens and allows sample collection for a relatively long time, and sample collection may be possible through the eye. Also, in the case of a plasmonic well plate, a large amount of samples can be analyzed simultaneously, and the analysis time can be shortened.

[0060] According to another aspect of the present application, the composition for manufacturing a substrate for spectroscopic analysis of the present application includes a noble metal precursor and a reducing agent.

[0061] FIGS. 5(a) to 5(c) are graphs showing the intensities of SERS signals at 633 nm and 785 nm when methylene blue (MB) is applied at different concentrations, where the reducing agent applied to the substrate for spectroscopic analysis according to an embodiment of the present invention is hydroxylamine (the ratio of the reducing agent to the noble metal precursor is 1:2). FIGS. 5(d) and 5(e) are graphs showing the signal uniformity of the substrate for spectroscopic analysis according to an embodiment of the present invention, and FIG. 5(f) is a photograph showing the flexibility of the substrate for spectroscopic analysis according to an embodiment of the present invention.

[0062] Referring to FIGS. 5(a) to 5(f), after measuring the intensity of the SERS signal according to the concentration of methylene blue (MB) using the spectroscopic analysis substrate of the present application manufactured with cellulose acetate as the base member, the SERS signal intensity was verified at 633 nm and 785 nm, respectively, according to the concentration of methylene blue. When using the spectroscopic analysis substrate of the present application, it can be confirmed that the uniformity of the signal is high and constant. Further, in the case of the spectroscopic analysis substrate of the present application, it is small and flexible, so it is convenient to carry and can be applied to various surfaces.

[0063] Although not limited thereto, if the ratio of the reducing agent to the noble metal precursor is less than 1:0.5, Raman spectroscopy systems of various wavelengths cannot be used, and if it exceeds 1:15, it may be difficult to obtain a spectroscopic analysis substrate that forms a film and has excellent signal intensity and signal uniformity.

[0064] Therefore, in the composition for manufacturing the spectroscopic analysis substrate of the present application, when the ratio of the reducing agent to the noble metal precursor is 1:0.5 to 1:15, it is suitable for manufacturing a spectroscopic analysis substrate with excellent signal intensity and signal uniformity, and it can be 1:0.5 to 1:12, it can be 1:0.5 to 1:10, it can be 1:1 to 1:10, it can be 1:1 to 1:9, it can be 1:1 to 1:8, it can be 1:1 to 1:7, it can be 1:1 to 1:6, it can be 1:1 to 1:5, it can be 1:1 to 1:4, it can be 1:1 to 1:3. Also, the range may vary slightly depending on the type of reducing agent.

[0065] Although not limited thereto, in the composition for manufacturing the spectroscopic analysis substrate of the present application, as the ratio of the reducing agent to the noble metal precursor increases, the size of the polycrystalline structure formed on the spectroscopic analysis substrate decreases, and the density of the polycrystalline structure can increase.

[0066] FIG. 6(a) is an SEM photograph showing the change in particle shape according to the ratio of the metal precursor to the reducing agent in the composition for manufacturing a substrate for spectroscopic analysis according to an embodiment of the present invention. FIG. 6(b) shows the change in surface plasmon resonance (SPR) of the light-absorbing surface of the spectroscopic analysis substrate according to the wavelength due to the ratio of the metal ion precursor to the reducing agent in the composition for manufacturing a substrate for spectroscopic analysis according to an embodiment of the present invention. FIG. 6(c) is a graph showing the change in SERS signal intensity of the spectroscopic analysis substrate according to the wavelength due to the ratio of the metal ion precursor to the reducing agent in the composition for manufacturing a substrate for spectroscopic analysis according to an embodiment of the present invention.

[0067] Referring to FIG. 6, it can be confirmed that at a low ratio of the reducing agent to the metal precursor, the size of the polycrystalline structure is large and the connection of the branch structures is small, but as the ratio of the reducing agent to the metal precursor increases, the size of the polycrystalline structure becomes smaller and it is connected by more branch structures. On the other hand, within the composition for manufacturing a substrate for spectroscopic analysis, depending on the optimal ratio range of the reducing agent to the metal precursor, the density of the polycrystalline structure increases, the transmittance decreases, and the absorbance increases, so that a Raman spectroscopic apparatus with higher sensitivity can be manufactured.

[0068] FIGS. 7(a) and 7(b) are an SEM photograph (FIG. 7(a)) showing the change in particle shape according to the reaction time after supporting the base member on the composition for manufacturing a substrate for spectroscopic analysis according to the present invention, and a graph (FIG. 7(b)) showing the change in light-absorbing SPR thereby.

[0069] Also, referring to FIG. 7, the shape of the particles according to the time of supporting the substrate on the composition for manufacturing a substrate for spectroscopic analysis according to the present application is shown. In particular, it can be confirmed that the absorbance increases as the supporting time increases.

[0070] Although not limited thereto, the noble metal precursor may be selected from the group consisting of HAuCl4, AuCl, AuCl2, AuCl3, Na2Au2Cl8, and NaAuCl2, and one or more of HAuCl4 and NaAuCl4 are more suitable. Although not limited thereto, the reducing agent may be one or more of hydroxylamine, ascorbic acid, FeSO4, and hydroxyquinone.

[0071] Figures 8(a) to 12 show the optimal ratio of the reducing agent to the noble metal precursor through the comparison of the SERS signal intensity when using hydroxylamine, ascorbic acid, FeSO4, and hydroxykinone reducing agents.

[0072] Figure 8(a) is a photograph showing the hue change of the substrate for spectroscopic analysis according to the ratio of the metal ion precursor and the hydroxylamine reducing agent in the composition for manufacturing the substrate for spectroscopic analysis according to an embodiment of the present invention. Figure 8(b) is a graph showing the absorption SPR change of the substrate for spectroscopic analysis in Figure 8(a). Figure 8(c) is a graph showing the change in the SERS signal intensity measured at 633 nm of the substrate for spectroscopic analysis in Figure 8(a). Figure 8(d) is a graph showing the change in the SERS signal intensity measured at 785 nm of the substrate for spectroscopic analysis in Figure 8(a). Figure 8(e) is an SEM photograph showing the particle shape of the substrate for spectroscopic analysis according to the ratio of the metal ion precursor and the hydroxylamine reducing agent in the composition for manufacturing the substrate for spectroscopic analysis according to an embodiment of the present invention.

[0073] Referring to Figure 8, in the case of the hydroxylamine reducing agent, the particle density began to increase from the 1:1 ratio, and the SERS intensity increased as the ratio of the reducing agent to the noble metal precursor increased. The 1:2 ratio at which the SERS signal was observed to be the strongest can be set as the optimized ratio, and a range of 1:1 or more and less than 1:5 can be set as the optimized ratio range.

[0074] Figure 9(a) is a photograph showing the hue change of a substrate for spectroscopic analysis according to the ratio of a metal ion precursor and an ascorbic acid reducing agent in a composition for manufacturing a substrate for spectroscopic analysis according to an embodiment of the present invention. Figure 9(b) is a graph showing the absorbance SPR change of the substrate for spectroscopic analysis of Figure 9(a). Figure 9(c) is a graph showing the change in SERS signal intensity measured at 633 nm of the substrate for spectroscopic analysis of Figure 9(a). Figure 9(d) is a graph showing the change in SERS signal intensity measured at 785 nm of the substrate for spectroscopic analysis of Figure 9(a). Figure 9(e) is an SEM photograph showing the particle shape of a substrate for spectroscopic analysis according to the ratio of a metal ion precursor and an ascorbic acid reducing agent in a composition for manufacturing a substrate for spectroscopic analysis according to an embodiment of the present invention.

[0075] Referring to Figure 9, in the case of the ascorbic acid reducing agent, at 785 nm, the intensity of the SERS signal increased as the ratio of the reducing agent to the noble metal precursor increased. The ratio of 1:10 at which the SERS signal was observed to be the strongest can be set as the optimized ratio, and a range of 1:0.5 to 1:15 can be set as the optimized ratio range. However, in the case of 633 nm, the ratio of 1:1 at which the SERS signal was observed to be the strongest can be set as the optimized ratio, and a range of 1:0.5 to 1:15 can be set as the optimized ratio range.

[0076] Figure 10(a) is a photograph showing the hue change of a substrate for spectroscopic analysis according to the ratio of a metal ion precursor and an FeSO4 reducing agent in a composition for manufacturing a substrate for spectroscopic analysis according to an embodiment of the present invention. Figure 10(b) is a graph showing the absorbance SPR change of the substrate for spectroscopic analysis of Figure 10(a). Figure 10(c) is a graph showing the change in SERS signal intensity measured at 633 nm of the substrate for spectroscopic analysis of Figure 10(a). Figure 10(d) is a graph showing the change in SERS signal intensity measured at 785 nm of the substrate for spectroscopic analysis of Figure 10(a). Figure 10(e) is an SEM photograph showing the particle shape of a substrate for spectroscopic analysis according to the ratio of a metal ion precursor and an FeSO4 reducing agent in a composition for manufacturing a substrate for spectroscopic analysis according to an embodiment of the present invention.

[0077] Referring to Fig. 10, in the case of the FeSO4 reducing agent, the SERS intensity increased as the ratio of the reducing agent to the noble metal precursor increased. The ratio of 1:5 at which the SERS signal was observed to be the strongest can be set as the optimized ratio, and a range of more than 1:1 and less than or equal to 1:15 can be set as the optimized ratio range.

[0078] Fig. 11(a) is a photograph showing the hue change of the substrate for spectroscopic analysis according to the ratio of the metal ion precursor and the hydroxyquinone reducing agent in the composition for manufacturing the substrate for spectroscopic analysis according to an embodiment of the present invention. Fig. 11(b) is a graph showing the absorption SPR change of the substrate for spectroscopic analysis in Fig. 11(a). Fig. 11(c) is a graph showing the change in the SERS signal intensity measured at 633 nm of the substrate for spectroscopic analysis in Fig. 11(a). Fig. 11(d) is a graph showing the change in the SERS signal intensity measured at 785 nm of the substrate for spectroscopic analysis in Fig. 11(a). Fig. 11(e) is an SEM photograph showing the particle shape of the substrate for spectroscopic analysis according to the ratio of the metal ion precursor and the hydroxyquinone reducing agent in the composition for manufacturing the substrate for spectroscopic analysis according to an embodiment of the present invention.

[0079] Referring to Fig. 11, in the case of the hydroxyquinone reducing agent, the ratio of 1:2 at which the SERS signal was observed to be the strongest can be set as the optimized ratio, and a range of 1:0.5 to less than 1:15 can be set as the optimized ratio range.

[0080] Figs. 12(a) and 12(b) show a comparison of the SERS intensities at wavelengths of 633 nm (Fig. 12(a)) and 785 nm (Fig. 12(b)) of the substrate for spectroscopic analysis manufactured from the composition for manufacturing the substrate for spectroscopic analysis containing the metal ion precursor and each reducing agent at the optimal ratio according to the embodiments of the present invention.

[0081] Referring to Fig. 12, the optimal ratio was derived for each reducing agent, and the optimization conditions for all reducing agents were confirmed at wavelengths of 633 nm and 785 nm.

[0082] According to another aspect of the present application, a Raman spectroscopy apparatus is provided that includes a light source; a spectroscopic analysis substrate described in the present application; and a detector that detects Raman spectroscopy.

[0083] FIGS. 13(a) to 13(d) schematically show a method for on-site diagnosis of cancer using a portable Raman spectroscopy apparatus (FIG. 13(a)) according to an embodiment of the present invention.

[0084] Referring to FIGS. 13(a) to 13(d), as an example, urine excretions containing metabolites of cancer cells are used to derive a Raman spectrum using the analysis spectroscopic substrate (sensor) according to the present application and a portable Raman spectroscopy apparatus, and multivariate analysis is used to perform on-site analysis to diagnose prostate cancer and pancreatic cancer. A Raman spectroscopy apparatus and an analysis method are schematically shown.

[0085] FIG. 14 is a drawing showing a Raman spectroscopy apparatus according to an embodiment of the present invention. FIG. 15(a) is a perspective photograph of an adapter for a Raman spectroscopy apparatus according to an embodiment of the present invention, FIG. 15(b) is a front view of the adapter of FIG. 15(a), FIG. 15(c) is a plan view of the adapter of FIG. 15(a), FIG. 15(d) is a bottom view of the adapter of FIG. 15(a), and FIG. 15(e) is a perspective view of the adapter of FIG. 15(a).

[0086] FIG. 16(a) is a plan photograph of an analysis member for a Raman spectroscopy apparatus according to an embodiment of the present invention, FIG. 16(b) is a plan view of the analysis member of FIG. 16(a), FIG. 16(c) is a front view of the analysis member of FIG. 16(a in the A direction), FIG. 16(d) is a side view of the analysis member of FIG. 16(a in the B direction), and FIG. 16(e) is a perspective view of the analysis member of FIG. (a).

[0087] Referring to FIGS. 14 to 16, the Raman spectroscopy apparatus 200 of the present application may further include an adapter 100 fastened to the Raman spectroscopy apparatus 200 and an analysis member 10 on which the adapter 100 and the spectroscopic analysis strip substrate are seated and analyzed.

[0088] Referring to FIGS. 14 to 16, the adapter 100 includes a cylindrical body portion 110 having a hollow 112 with an open first end, and a fastening portion 112a at the first end for fastening to the Raman spectrometer 200. The fastening portion 112a can be a thread formed inside the cylindrical body portion 110. Also, the second end of the adapter 100 includes a coupling portion 114 in which a through hole 116 for guiding light from a light source is formed. The coupling portion 114 can be configured in a shape that can be stably coupled to the analysis member 10.

[0089] Referring to FIGS. 15 and 16, the analysis member 10 is spoon-shaped and includes a head portion 20 and a linear body portion 22 extending from the head portion 20. The head portion 20 includes a hemispherical seating portion 30 on which the coupling portion 114 at the second end of the adapter 100 is seated, and a seating groove 40 extending longitudinally to the hemispherical seating portion 30 at the center of the body portion 22, on which a linear spectroscopic analysis strip substrate is seated.

[0090] Although not limited thereto, the seating groove 40 includes a terminal portion 42 extending to the seating portion 30 so as to be formed in a T shape, and it can be characterized in that the accuracy of analysis can be increased by preventing the end of the spectroscopic analysis strip substrate from shifting the hemispherical seating portion 30.

[0091] FIG. 17 is a photograph schematically showing a method of proceeding with analysis by coupling an adapter and an analysis member to a Raman spectrometer according to an embodiment of the present invention.

[0092] Referring to FIG. 17, first, the adapter is screwed to the Raman spectrometer (FIGS. 17(a) and 17(b)). Next, the adapter fastened to the Raman spectrometer is coupled to the analysis member (FIG. 17(c)). After pushing the spectroscopic analysis strip substrate into the seating groove of the analysis member, the analysis is proceeded.

[0093] Without being limited thereto, the target substance of the detector may be selected from one or more of cells, metabolites, proteins, nucleic acids, DNA, RNA, enzymes, organic molecules, viruses, extracellular vesicles, microvesicles, exosomes, and fats in urine, saliva, sweat, and tears.

[0094] Without being limited thereto, the target substance is a liquid sample. Conventionally, in the case of a substrate for spectroscopic analysis, the hygroscopicity is low, and the sensitivity during measurement of the liquid sample is very low. However, the substrate for spectroscopic analysis according to the present application has hygroscopicity, and even with a liquid sample, rapid diagnosis can be performed on-site.

[0095] Without being limited thereto, the Raman spectroscopic apparatus may be capable of on-site diagnosis of cancer. Without being limited thereto, through the metabolite analysis of the target substances described above, information necessary for cancer diagnosis can be provided, and cancer can be diagnosed at an early stage by a non-invasive examination, and multiplex analysis may be possible with extremely high sensitivity.

[0096] The cancer may be one or more selected from the group consisting of lung cancer, bronchial cancer, colorectal cancer, prostate cancer, breast cancer, gastric cancer, ovarian cancer, bladder cancer, brain cancer, thyroid cancer, esophageal cancer, uterine cancer, liver cancer, kidney cancer, biliary tract cancer, and pancreatic cancer. Without being limited thereto, prostate cancer or pancreatic cancer may be more suitable.

[0097] FIG. 18 shows Raman spectra (FIGS. 18(b) and 18(c)) of prostate cancer and pancreatic cancer measured using a Raman spectroscopic apparatus (FIG. 18(a)) including a substrate for analytical spectroscopy according to an embodiment of the present invention, and multivariate analysis data (FIGS. 18(d) and 18(e)) of prostate cancer and pancreatic cancer based thereon.

[0098] According to another aspect, the cancer diagnosis method using the Raman spectroscopic apparatus according to the present invention can be performed by a known multivariate analysis method. The multivariate analysis method for the overall spectrum is a known analysis method, for example, a commonly used partial least squares discriminant analysis (PLS-DA), general discriminant analysis (GDA), principal component analysis (PCA), parallel factor analysis (PARAFAC), neural network analysis (NNA), and / or support vector machine technique (SVM), etc., but is not limited thereto.

[0099] Although not limited thereto, the cancer diagnosis method according to an embodiment can include the following steps. A step of removing the background signal for the obtained spectrum; a step of performing partial least squares discriminant analysis (PLS-DA) on the overall spectrum to obtain latent variables, a step of distinguishing two input groups based on the variables, and a step of confirming the reliability thereof.

[0100] As described above, according to the present invention, cancer can be accurately and quickly diagnosed on site. In particular, prostate cancer and pancreatic cancer showed a diagnosis accuracy of 90% or more.

[0101] According to still another aspect, the method for manufacturing a substrate for spectroscopic analysis is the method for manufacturing a substrate for spectroscopic analysis described in the present application, including: a) a step of preparing a base member; and b) a step of supporting the base member on a composition for manufacturing a substrate for spectroscopic analysis to form a polycrystalline structure. In the above, for the parts overlapping with the description of the substrate for spectroscopic analysis, detailed description is omitted.

[0102] Step a) is the step of preparing a base member. As described above, various base members can be used in the present application. Without being limited thereto, a base member composed of one or more of paper, polymer, well plate, wafer, and protein can be prepared.

[0103] Step b) is the step of supporting the base member on a composition for manufacturing a substrate for spectroscopic analysis to form a polycrystalline structure. The composition for manufacturing a substrate for spectroscopic analysis contains a noble metal precursor and a reducing agent solution, and in a solution process, a polycrystalline structure composed of a plurality of clusters of nanoparticles directly on the base member and having a plurality of grain boundaries can be formed.

[0104] In step b), the ratio of the reducing agent to the noble metal precursor solution can be set to 1:0.5 to 1:15, and a shape adjustment step of adjusting the shape of the polycrystalline structure can be included. By adjusting with the optimal ratio of the reducing agent according to the type of the reducing agent, the shape of the polycrystalline structure formed on the substrate for spectroscopic analysis can be easily adjusted. Also, by adjusting the time for supporting the base member on the composition for manufacturing a substrate for spectroscopic analysis, the shape of the polycrystalline structure formed on the substrate for spectroscopic analysis can be easily adjusted. The method for manufacturing a substrate for spectroscopic analysis of the present application can improve the signal intensity and signal uniformity simultaneously by a single process of a single loading.

[0105] Without being limited thereto, in step a) of the method for manufacturing a substrate for spectroscopic analysis of the present application, a surface modification step of surface-modifying the surface of the base member before the step of forming a polycrystalline structure can be further included. The surface modification step may, without being limited thereto, regulate particle formation through chemical surface treatment, or increase the density of particles during particle formation after surface modification.

[0106] Without being limited thereto, in the method for manufacturing a substrate for spectroscopic analysis of the present application, the surface modification step contains one or more selected from 1 to 3% of a base and 1 to 3% of an acid, and as a solvent, C 1-5It can include treating the surface of the base member with a surface modification composition containing alcohol or water. Without being limited thereto, using 1% of the base and acid and using ethanol as the solvent can be compatible from the viewpoints of improving signal intensity and signal uniformity.

[0107] Figures 19(a) and 19(b) show the results of surface modification using a surface modification composition according to an embodiment of the present invention. That is, FIG. 19(a) is an SEM photograph showing the result of an increase in the density of particles as a result of surface modification using a surface modification composition according to an embodiment of the present invention, and FIG. 19(b) is a graph showing the result of an improvement in signal uniformity thereby. Therefore, it can be confirmed that the density of particles has increased using the surface modification composition according to an embodiment of the present invention, and it can be confirmed that the signal uniformity has been improved by the increased density of the particles.

[0108] Examples 1. Production of a substrate for spectroscopic analysis Base members in the form of paper, three-dimensional swap sticks composed of porous polymers, wearable patches, and films were prepared. The ratio of the reducing agent solution to the noble metal precursor solution was set to 5:1, 2:1, 1:1 to 1:15 and mixed to produce a composition for producing a substrate for spectroscopic analysis. However, the composition must be prepared and the base member must be supported within 1 minute. However, in the case of a well plate, it is prepared by a method of supporting the composition inside each well of the well plate. The prepared base member was supported on the mixed composition for producing a substrate for spectroscopic analysis over 30 seconds to 24 hours to form a polycrystalline structure formed from a cluster of a plurality of nanoparticles on the base member. The composition for producing the substrate used HAuCl4 as the noble metal precursor solution, and hydroxylamine, ascorbic acid, FeSO4, and hydroxyquinone were used as the reducing agent. After the formation of the polycrystalline structure was completed, it was washed two or more times with water or ethanol to remove the remaining precursor and reducing agent. Then, it was dried at room temperature for 1 hour or more to complete the production of the substrate for spectroscopic analysis.

[0109] Result Regardless of the composition and form of the base member, such as a three-dimensional swap stick, patch, and film form composed of paper and a porous sponge, as the ratio of the reducing agent solution to the noble metal precursor solution contained in the composition for manufacturing a spectroscopic analysis substrate increases, the size of the polycrystalline structures aggregated on the manufactured spectroscopic analysis substrate decreases, and the density of the polycrystalline structures increases.

[0110] As a result, the transmittance decreased and the absorbance increased. In addition, the polycrystalline structures had a large number of distributions with an average particle size between 1 μm and 100 μm, and considering the SERS sensitivity and the strength of the SERS signal depending on the type of reducing agent, the optimal ratio range of the reducing agent to the noble metal precursor was derived.

[0111] Table 1 below shows the optimal ratio range of the reducing agent to the noble metal precursor depending on the type of reducing agent.

[0112] [Table 1]

[0113] By describing specific parts of the content of the present invention in detail above, it will be apparent to those with ordinary knowledge in the art that such specific technologies are merely preferred embodiments, and thus the scope of the present invention is not limited thereby. Therefore, it can be said that the substantial scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A base member; and A spectroscopic analysis substrate, which is composed of a plurality of clusters of nanoparticles formed on the base member, includes a polycrystalline structure having a plurality of grain boundaries, and is formed by a solution process.

2. The spectroscopic analysis substrate according to claim 1, wherein the base member is composed of one or more of paper, polymer, well plate, wafer, and protein.

3. The spectroscopic analysis substrate according to claim 1, wherein the nanoparticles are connected by a branch structure, and the polycrystalline structure has an average particle size of 1 μm to 100 μm.

4. The electrochemically active surface area (EASA) is 0.1 cm ２ to 10.0 cm ２ The substrate for spectroscopic analysis according to claim 1, wherein the substrate has the electrochemically active surface area (EASA) within the range of 0.1 cm

5. The spectroscopic analysis substrate according to claim 1, wherein the nanoparticles are composed of one or more of Au, Ag, and Pt.

6. The spectroscopic analysis substrate according to claim 1, which is applicable to a Raman spectroscopy system using a wavelength of 500 nm or more or a near-IR FT Raman spectroscopy system using a wavelength of 1064 nm.

7. The spectroscopic analysis substrate according to claim 1, which is composed of a three-dimensional swap stick, a wearable substrate, or a plasmonic well plate.

8. A composition for manufacturing a spectroscopic analysis substrate, including a noble metal precursor and a reducing agent, wherein the ratio of the reducing agent to the noble metal precursor is 1:0.5 to 1:

15.

9. The composition for manufacturing a spectroscopic analysis substrate according to claim 8, wherein as the ratio of the reducing agent to the noble metal precursor increases, the size of the polycrystalline structure formed on the spectroscopic analysis substrate decreases, and the density of the polycrystalline structure increases.

10. The noble metal precursor is HAuCl ４ and NaAuCl ４ The composition for producing a substrate for spectroscopic analysis according to claim 8, which is one or more of them.

11. The reducing agent is one or more of hydroxylamine, ascorbic acid, FeSO ４ , and hydroxyquinone. The composition for manufacturing a substrate for spectroscopic analysis according to claim 8.

12. A light source; The spectroscopic analysis substrate according to claim 1; and A Raman spectroscopy apparatus including a detector for detecting Raman spectroscopy.

13. An adapter having a cylindrical shape with an open first end, the first end having a fastening portion fastened to the Raman spectroscopy apparatus, and the second end having a coupling portion formed with a through hole for guiding light from the light source; and A spoon-shaped member composed of a head portion and a linear body portion extending from the head portion, the head portion further including a hemispherical seating portion on which the coupling portion of the second end of the adapter is seated, and a seating groove extending longitudinally to the hemispherical seating portion at the center of the body portion for seating a linear spectroscopic analysis strip substrate, the Raman spectroscopy apparatus according to claim 12.

14. The Raman spectroscopic apparatus according to claim 13, wherein the seating groove is formed in a T shape so that the end of the spectroscopic analysis strip substrate does not come off from the hemispherical seating portion.

15. The Raman spectroscopic apparatus according to claim 12, wherein the target substance of the detector is selected from one or more of cells, metabolites, proteins, nucleic acids, DNA, RNA, enzymes, organic molecules, viruses, extracellular vesicles, microvesicles, exosomes, and fats in urine, saliva, sweat, and tears.

16. The Raman spectroscopic apparatus according to claim 12, wherein the Raman spectroscopic apparatus can perform in-situ diagnosis of cancer.

17. A method for manufacturing a spectroscopic analysis substrate according to claim 1, comprising: a) a step of preparing a base member; and b) supporting the base member on a composition for manufacturing a spectroscopic analysis substrate containing a noble metal precursor and a reducing agent solution, and forming a polycrystalline structure composed of a plurality of clusters of nanoparticles and having a plurality of grain boundaries on the base member; The method for manufacturing a spectroscopic analysis substrate according to claim 17, further comprising a shape adjustment step of adjusting the shape of the polycrystalline structure by setting the ratio of the reducing agent to the noble metal precursor solution to 1:0.5 to 1:15 in step b).

18. The method for manufacturing a spectroscopic analysis substrate according to claim 17, further comprising a surface modification step of modifying the surface of the base member before the step of forming the polycrystalline structure in step a).

19. The surface modification step includes one or more selected from 1 to 3% of a base and 1 to 3% of an acid, and uses, as a solvent, an alcohol or water of C １－５ The method for manufacturing a substrate for spectroscopic analysis according to claim 18, including treating the surface of the base member with a surface modification composition containing the alcohol or water of C

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