Device for selectively collecting target compound, method for manufacturing said device, and method for selectively collecting target compound from biological or environmental sample

Abstract

A device for selectively collecting a target compound is disclosed. This device includes a cellulose substrate on which a detection reagent is supported. The detection reagent includes a constituent compound having at least one functional group selected from the group consisting of amino groups, carboxyl groups, sulfone groups, and phenolic hydroxyl groups, and a target compound is at least one type of substance selected from the group consisting of biological metabolites and environmental compounds.

Description

Device for selectively collecting target compounds, method for manufacturing said device, and method for selectively collecting target compounds from biological or environmental samples

[0001] The present invention relates to a device for selectively collecting a target compound, a method for producing the device, and a method for selectively collecting a target compound from a biological or environmental sample.

[0002] "Paper points" are filter paper formed into a twisted paper shape. For example, during oral examinations and treatments, they are used to absorb exudates and excess medicinal solutions from root canals, dry them, stop bleeding, apply medicinal solutions, and fill them with MTA cement.

[0003] The standard for paper points is defined by the ISO, and "paper points" of the required size are used in diagnostic and treatment settings. Paper points are also used to collect gingival crevicular fluid (GCF) that flows out from lesions, for example, in periodontal disease tests. They are particularly useful for collecting minute amounts of metabolites when analyzing them in GCF.

[0004] However, even if biometabolites can be collected using paper samples as described above, their analysis currently requires many steps. Specifically, biometabolites in gingival crevicular fluid collected using paper samples are extracted from the samples using a specific solvent, dried under reduced pressure, and then derivatized using a specific method. The resulting derivatives are then analyzed using known techniques such as gas chromatography-mass spectrometry (GCMS).

[0005] The process of analyzing biological metabolites using paper points is complicated, making it difficult to complete the analysis in a short time. Furthermore, a large amount of solvent is used and discarded during the process from extraction to drying under reduced pressure, making it undesirable from an environmental perspective.

[0006] The present invention is intended to solve the above-mentioned problems, and an object of the present invention is to provide a device for selectively collecting a target compound, which allows for easy separation of the target compound from various samples such as biological samples and environmental samples, a method for manufacturing the device, and a method for selectively collecting a target compound from a biological or environmental sample.

[0007] The present invention is a device for selectively collecting a target compound, comprising a cellulose substrate carrying a detection reagent, the detection reagent including a constituent compound having at least one functional group selected from the group consisting of an amino group, a carboxyl group, a sulfonic acid group, and a phenolic hydroxyl group, and the target compound is at least one selected from the group consisting of biological metabolites and environmental compounds.

[0008] In one embodiment, the detection reagent comprises at least one selected from the group consisting of amino acids, peptides, organic acids and salts thereof, polyethyleneimine, polydopamine, tannic acid, and lignin.

[0009] In one embodiment, the cellulose substrate is a paper point or a cellulose monolith.

[0010] The present invention also provides a method for manufacturing a device for selectively collecting a target compound, the method comprising the step of supporting a detection reagent on a cellulose substrate, the detection reagent comprising a constituent compound having at least one functional group selected from the group consisting of an amino group, a carboxyl group, a sulfonic acid group, and a phenolic hydroxyl group, and the target compound is at least one selected from the group consisting of biological metabolites and environmental compounds.

[0011] The present invention also provides a method for selectively collecting a target compound from a biological or environmental sample, the method comprising the step of contacting the biological or environmental sample with the device described above.

[0012] In one embodiment, the biological or environmental sample is saliva, blood, urine, gingival crevicular fluid, fecal suspension, exhaled breath condensate, exudate from an inflammatory focus, oral swab, nasal swab, condensate from the atmosphere, seawater, river water, lake water, fruit juice, juice squeezed from a plant, or an extract from a food.

[0013] The present invention also relates to a detection reagent for selectively collecting a target compound from a biological or environmental sample via a supported cellulose substrate, the detection reagent comprising a constituent compound having at least one functional group selected from the group consisting of an amino group, a carboxyl group, a sulfonic acid group, and a phenolic hydroxyl group, and the target compound is at least one selected from the group consisting of biological metabolites and environmental compounds.

[0014] According to the present invention, target compounds contained in various samples can be easily collected. This allows the target compounds contained in the samples to be analyzed in a short time. Furthermore, the number of work steps involved can be reduced.

[0015] 1 is a graph showing the results of GCMS analysis of glycine in a sample solution using the paper points (E1) to (E4) prepared in Examples 1 to 4. 2 is a graph showing the results of GCMS analysis of 2-isopropylmalic acid in a sample solution using the paper points (E1) to (E4) prepared in Examples 1 to 4. 3 is a graph (calibration curve) showing the results of GCMS analysis conducted in Example 9, in which glycine solutions of various concentrations were contacted with the chitosan-modified paper point (E3) prepared in Example 3. 4 is a graph (calibration curve) showing the results of GCMS analysis conducted in Example 10, in which 2-isopropylmalic acid solutions of various concentrations were contacted with the chitosan-modified paper point (E3) prepared in Example 3. 5 is a graph (chromatogram) showing the results of GCMS analysis conducted in Example 11, in which a saliva sample was contacted with the chitosan-modified paper point (E3) prepared in Example 3.

[0016] The present invention will be described in detail below.

[0017] (Device for Selectively Collecting a Target Compound) The device of the present invention comprises a cellulose substrate.

[0018] The cellulose substrate is any shaped article containing cellulose as a main raw material, and may take the form of a film, sheet, nonwoven fabric, woven fabric, slices thereof, or a combination thereof. The size and thickness of the cellulose substrate are not particularly limited, as long as it is large enough to absorb and collect the target compound described below.

[0019] Specific examples of the cellulose substrate include paper points, cellulose monoliths (porous cellulose bodies), filter paper, and tissue paper. Because of their easy availability, the cellulose substrate is preferably a paper point or a cellulose monolith.

[0020] In the present invention, a detection reagent is carried on the cellulose substrate.

[0021] The detection reagent contains a constituent compound having at least one functional group capable of reacting with a target compound contained in a sample, the target compound being sought to be collected. Examples of such functional groups in the constituent compound include an amino group, a carboxyl group, a sulfonic acid group, a phenolic hydroxyl group, and combinations thereof.

[0022] Examples of constituent compounds having an amino group in the present invention include chitosan, chitin, polyethyleneimine, amino acids (e.g., glycine, alanine, arginine, lysine, proline, aspartic acid, and glutamic acid), peptides (e.g., dipeptides, tripeptides, and other polypeptides preferably composed of 2 to 20 amino acids); amino group-containing low molecular weight compounds (e.g., aniline, ethylenediamine, and glycidyltrimethylammonium) and derivatives thereof; and salts thereof (e.g., sodium salts, potassium salts, magnesium salts, calcium salts, and carbonate salts); and combinations thereof.

[0023] Examples of constituent compounds having a carboxyl group in the present invention include organic acids (e.g., citric acid, maleic acid, malic acid, succinic acid, alginic acid, and adipic acid) and anhydrides thereof (e.g., succinic anhydride and maleic anhydride); cellulose oxides with TEMPO (2,2,6,6-tetramethylpiperidine 1-oxyl); cellulose oxides with sodium hypochlorite or sodium periodate; amino acids (e.g., glycine, alanine, arginine, lysine, proline, aspartic acid, and glutamic acid); peptides (e.g., dipeptides, tripeptides, and other polypeptides preferably composed of 2 to 20 amino acids); carboxyl group-containing low molecular weight compounds (e.g., citric acid, maleic acid, malic acid, succinic acid, alginic acid, adipic acid, benzoic acid, and monochloroacetic acid) and derivatives thereof; and salts thereof (e.g., sodium salts, potassium salts, magnesium salts, calcium salts, and carbonate salts); and combinations thereof.

[0024] Examples of the constituent compound having a sulfonic group in the present invention include alkylsulfonic acid, perfluorosulfonic acid, hydroxybenzenesulfonic acid, benzenesulfonic acid, toluenesulfonic acid, styrenesulfonic acid, 10-camphorsulfonic acid, magic acid, taurine, polystyrenesulfonic acid, sulfonated polyethersulfone, perfluorocarbonsulfonic acid (such as Nafion), and alkylbenzenesulfonic acid; derivatives thereof; salts thereof (e.g., sodium salt, potassium salt, magnesium salt, calcium salt, and carbonate); and combinations thereof.

[0025] Examples of constituent compounds having a phenolic hydroxyl group in the present invention include dopamine (e.g., polydopamine), tannic acid, lignin, polyphenols (e.g., catechin and curcumin), amino acids (e.g., phenylalanine, tyrosine, and tryptophan), peptides (e.g., dipeptides, tripeptides, and other polypeptides preferably composed of 2 to 20 amino acids), oxidized polymers of other derivatives (e.g., polydopamine and tannic acid-metal ion complexes), and derivatives thereof; salts thereof (e.g., sodium salts, potassium salts, magnesium salts, calcium salts, and carbonates); and combinations thereof.

[0026] In the present invention, the detection reagent may contain at least one of the above-mentioned constituent compounds having an amino group, constituent compounds having a carboxyl group, constituent compounds having a sulfone group, and constituent compounds having a phenolic hydroxyl group.

[0027] The detection reagent preferably contains at least one constituent compound selected from the group consisting of amino acids, peptides, organic acids and their salts, polyethyleneimine, polydopamine, tannic acid, and lignin, because it is easily available and allows for stable selective collection of target compounds onto a cellulose substrate.

[0028] The detection reagent may be supported on the cellulose substrate by chemical bonding (chemisorption), physical adsorption, or a combination thereof.

[0029] The amount of detection reagent supported on the cellulose substrate is not particularly limited, but is preferably 0.01 to 100 parts by mass, more preferably 0.1 to 10 parts by mass, per 100 parts by mass of the cellulose substrate. If the amount of detection reagent supported on the cellulose substrate is less than 0.01 parts by mass, the amount of detection reagent supported may be too small, making it difficult to efficiently collect the target compound contained in the sample. On the other hand, it is technically difficult to support the detection reagent on the cellulose substrate in an amount exceeding 10 parts by mass, making it difficult to provide a device of consistent quality.

[0030] The target compounds that can be selectively collected by the device of the present invention are biological metabolites and / or environmental compounds. Examples of biological metabolites include amino acids (e.g., glycine, alanine, aspartic acid, asparagine, glutamic acid, serine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, valine, arginine, cysteine, glutamine, proline, tyrosine, γ-aminobutyric acid (GABA), β-alanine, cystine, hydroxyproline, hydroxylysine, thyroxine, O-phosphoserine, desmosine, sarcosine, ornithine, citrulline, opine, and theanine) and salts thereof;Fatty acids (e.g., acetic acid, propionic acid, butanoic acid, pentanoic acid, lauric acid, myristic acid, palmitic acid, palmitoleic acid, margaric acid, stearic acid, oleic acid, vaccenic acid, linoleic acid, linolenic acid, arachidic acid, mead acid, arachidonic acid, behenic acid, lignoceric acid, nervonic acid, cerotic acid, montanic acid, melissic acid, docosahexaenoic acid, eicosapentaenoic acid, citric acid, lactic acid, 2-hydroxybutyric acid, malic acid, succinic acid, 2-oxoglutaric acid, aconitic acid, tartaric acid, butyric acid, fumaric acid, malic acid, succinic acid, 2-oxoglutaric acid, aconitic acid, tartaric acid, butyric acid, fumaric acid, Leic acid, pyruvic acid, citramalic acid, 5-hydroxymethyl-2-furan acid, 3-oxoglutaric acid, furan-2,5-dicarboxylic acid, furancarbonylglycine, carboxilic acid, tricarballylic acid, 2-hydroxyphenylacetic acid, 4-hydroxyphenylacetic acid, 4-hydroxybenzoic acid, 4-hydroxyhippuric acid, hippuric acid, 3-indoleacetic acid, 4-cresol, 3,4-dihydroxyphenylpropionic acid, glyceric acid, glycolic acid, oxalic acid, succinic acid, 2-oxoglutaric acid, 3-(3-hydroxyphenyl)-2-methyl-2-propanol (hydroxyphenyl)-3-hydroxypropionic acid (HPHPA), homovanillic acid, vanillylmandelic acid, 5-hydroxyindoleacetic acid, quinolinic acid, kynurenic acid, uracil, thymine, 3-hydroxybutyric acid, acetoacetic acid, 4-hydroxybutyric acid, ethylmalonic acid, methylsuccinic acid, adipic acid, suberic acid, sebacic acid, methylmalonic acid, pyridoxic acid, pantothenic acid, glutaric acid, ascorbic acid, 3-hydroxy-3-methylglutaric acid, N-acetylcysteine, methylcitric acid, pyroglutamic acid, orochi lactic acid, 2-hydroxyhippuric acid, 2-hydroxyisovaleric acid, 2-oxoisovaleric acid, 3-methyl-2-oxovaleric acid, 2-hydroxyisocaproic acid, 2-oxoisocaproic acid, 2-oxo-4-metholbutyric acid, mandelic acid, phenyllactic acid, phenylpyruvic acid, homogentisic acid, 4-hydroxyphenyllactic acid, N-acetylaspartic acid, malonic acid, and 3-methylglutaric acid) and their salts; and sugars (e.g., arabinose, glucose, galactose, and fructose) and their derivatives;Compounds containing a polar functional group or a functional group having an aromatic ring are preferred because they are compounds that interact with the constituent compounds in the detection reagent.

[0031] Examples of environmental compounds include PFAS (perfluoroalkylsulfonic acid); pesticides (e.g., diethyl paranitrophenyl thiophosphate (parathion), dimethyl paranitrophenyl thiophosphate (methyl parathion), dimethyl ethyl mercaptoethyl thiophosphate (methyl demeton), ethyl paranitrophenyl thionobenzene phosphonate (EPN), thiuram, simazine, thiobencarb, dichlorvos, dimethyl ethyl sulfinyl isopropyl thiophosphate, methylcarbamate-2-(1-methylpropyl)phenol, protease inhibitors, etc.) Examples of suitable antibacterial agents include thiazolinone, ...

[0032] Such target compounds have the property of actively interacting with the constituent compounds in the detection reagent of the present invention. Therefore, in the device of the present invention, a predetermined target compound contained in a sample preferentially interacts with the constituent compounds in the detection reagent supported on the cellulose substrate, thereby transferring the target compound from the sample side to the cellulose substrate side, thereby enabling the selective collection of the target compound.

[0033] Furthermore, the migration of the target compound from the sample side to the cellulose substrate side depends on the concentration of the target compound, and therefore, by measuring the amount of the target compound that has migrated to the cellulose substrate side, it is possible to quantify the concentration of the target compound contained in the sample.

[0034] (Device Manufacturing Method) The above device can be manufactured, for example, as follows.

[0035] In the production method of the present invention, first, a detection reagent is supported on a cellulose substrate.

[0036] Specifically, for example, when the detection reagent contains a constituent compound composed of an organic acid, an aqueous solution of the detection reagent prepared to a predetermined concentration is applied to the cellulose substrate by immersion, spraying, coating, or a combination thereof.

[0037] When a detection reagent consisting of two or more constituent compounds is applied to a cellulose substrate, a solution (e.g., an aqueous solution) containing the two or more constituent compounds may be prepared in advance and applied to the cellulose substrate, or an aqueous solution containing one constituent compound may be applied to the cellulose substrate, and then dried as necessary, and then an aqueous solution containing another constituent compound may be applied to the cellulose substrate.

[0038] After the detection reagent is applied to the cellulose substrate, heating may be performed. The temperature that can be used for this heating is not necessarily limited, as it varies depending on the constituent compounds contained in the detection reagent and / or the type of cellulose substrate, etc., but is preferably 40°C to 240°C, more preferably 80°C to 160°C. The heating time is also not limited, but is preferably 0.5 hours to 48 hours, more preferably 1 hour to 24 hours.

[0039] In the present invention, after a detection reagent is once loaded onto the cellulose substrate, another detection reagent may be loaded onto the cellulose substrate. In this case, an aqueous solution containing the other detection reagent may be applied to the cellulose substrate loaded with a certain detection reagent, and heating may be performed as described above, if necessary.

[0040] Through such heating, the detection reagent is carried on the cellulose substrate, and the device of the present invention is produced.

[0041] (Method for selectively collecting target compounds from biological or environmental samples)

[0042] Next, a method for selectively collecting a target compound from a biological or environmental sample (hereinafter sometimes referred to as the collection method of the present invention) will be described.

[0043] In the collection method of the present invention, a biological or environmental sample is contacted with the device.

[0044] Biological or environmental samples include biological samples collected from animals, including humans, and / or environmental samples collected from the environment, including plants, air, soil, and water. Biological samples include, but are not limited to, saliva, blood, urine, gingival crevicular fluid, fecal suspension, exhaled breath condensate, exudate from inflammatory foci, oral swabs, and nasal swabs. Environmental samples include, but are not limited to, condensate from air, seawater, river water, lake water, fruit juice, squeezed juice from plants, and food extracts.

[0045] The biological or environmental sample can be brought into contact with the device by absorbing the biological or environmental sample itself or a solution containing the biological or environmental sample (e.g., an aqueous solution) into the device of the present invention. The device after absorbing the biological or environmental sample may be dried as needed.

[0046] In this way, target compounds in biological or environmental samples can be selectively collected through the device of the present invention.

[0047] The target compound collected on the device side can be quantified using a method known in the art.

[0048] For example, a device that has absorbed a biological or environmental sample is washed to remove excess biological or environmental sample. The device that has absorbed the biological or environmental sample is then placed in a predetermined solvent to extract the target compound present in the device. The extract of the target compound can then be analyzed using means known in the art to quantify the target compound.

[0049] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples.

[0050] Example 1: Preparation of citric acid-modified paper point (E1) 0.2 g of paper point (Zippeler) was immersed in 10 mL of 40% by mass aqueous citric acid solution for 30 minutes. The paper point was then removed and heated on a wire mesh at 130°C for 12 hours to cause a reaction. The paper point was then washed with deionized water and dried to obtain citric acid-modified paper point (E1).

[0051] (Example 2: Preparation of polydopamine-modified paper point (E2)) 60 mg of dopamine hydrochloride was dissolved in 10 mL of Tris-HCl buffer (pH 8.5) to prepare a Tris solution, and 0.2 g of paper point (Zippeler) was immersed in this solution and allowed to react for 4 hours under atmospheric conditions. After that, the paper point was washed with deionized water and dried to obtain polydopamine-modified paper point (E2).

[0052] Example 3: Preparation of chitosan-modified paper point (E3) A 1.0% by mass chitosan-acetic acid aqueous solution (acetic acid concentration: 1.0% by mass) containing chitosan (Chitosan 100 manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) and acetic acid was prepared. 0.2 g of paper point (manufactured by Zipperer) was immersed overnight in 10 mL of this chitosan-acetic acid aqueous solution. The paper point was then removed and heated on a wire mesh at 130°C for 6 hours to obtain chitosan-modified paper point (E3).

[0053] (Example 4: Preparation of chitosan-modified citric acid-modified paper point (E4)) The citric acid-modified paper point (E1) prepared in Example 1 was immersed overnight in the 1.0% by mass chitosan acetic acid aqueous solution prepared in Example 3. The paper point was then removed and heated on a wire mesh at 130°C for 6 hours to obtain chitosan-modified citric acid-modified paper point (E4).

[0054] Example 5: Adsorption of target compounds onto citric acid-modified paper points (E1) and analysis thereof The citric acid-modified paper points (E1) prepared in Example 1 were cut into pieces of approximately 1 milligram size, washed with 100 μL of ultrapure water, then with 100 μL of acetonitrile, and then washed twice with 90% by volume acetonitrile / water (100 μL) for initialization. The cut pieces were then placed in a 0.5 mL Eppendorf tube.

[0055] Glycine and 2-isopropylmalic acid were selected as target compounds. Specifically, an aqueous glycine solution prepared at an arbitrary concentration and an aqueous 2-isopropylmalic acid solution prepared at an arbitrary concentration were mixed, and this mixture was used to prepare a sample solution consisting of 90% by volume of acetonitrile / water solution. 10 μL of this sample solution was added to the Eppendorf tube containing the initialized paper point (E1) obtained above and allowed to stand for an appropriate period of time.

[0056] The solution remaining in the Eppendorf tube was then removed using a micropipette, and the residue was washed with 90% acetonitrile / water for an appropriate number of times. After confirming that no solution remained in the Eppendorf tube, the paper point was dehydrated twice by adding 100 μL of acetonitrile.

[0057] After confirming that no solution remained in the Eppendorf tube after the dehydration operation, 5 μL of MSTFA solution (TMS reagent) was added to the Eppendorf tube and allowed to react for an appropriate period of time. Thereafter, the MSTFA solution in the Eppendorf tube was drained, and an appropriate amount of hexane was added and stirred. The entire hexane solution was then placed in a 2 mL vial.

[0058] The hexane solution in the vial was subjected to GCMS analysis using a GCMS (a gas chromatograph (7890) manufactured by Agilent Technologies Inc. and a mass spectrometer (JMS-Q1000GC) manufactured by JEOL Ltd.) under the analytical conditions shown in Table 1. The GCMS results for the detected glycine are shown in FIG. 1(a), and the GCMS results for the detected 2-isopropylmalic acid are shown in FIG. 2(a).

[0059]

[0060] (Example 6: Adsorption of target compounds onto polydopamine-modified paper points (E2) and analysis) Glycine and 2-isopropylmalic acid were adsorbed onto the paper points (E2) from a sample aqueous solution containing glycine and 2-isopropylmalic acid and analyzed by GCMS in the same manner as in Example 5, except that the polydopamine-modified paper points (E2) prepared in Example 2 were used instead of the citric acid-modified paper points (E1) prepared in Example 1. The GCMS results for the detected glycine are shown in Figure 1(b), and the GCMS results for the detected 2-isopropylmalic acid are shown in Figure 1(b).

[0061] Example 7: Adsorption of target compounds onto chitosan-modified paper points (E3) and analysis thereof Glycine and 2-isopropylmalic acid were adsorbed onto the chitosan-modified paper points (E3) from a sample aqueous solution containing glycine and 2-isopropylmalic acid, and GCMS analysis was performed in the same manner as in Example 5, except that the chitosan-modified paper points (E3) prepared in Example 3 were used instead of the citric acid-modified paper points (E1) prepared in Example 1. The GCMS results for the detected glycine are shown in Figure 1(c), and the GCMS results for the detected 2-isopropylmalic acid are shown in Figure 1(c).

[0062] Example 8: Adsorption of target compounds onto chitosan-modified, citric acid-modified paper points (E4) and analysis thereof Glycine and 2-isopropylmalic acid were adsorbed onto the paper points (E4) from a sample aqueous solution containing glycine and 2-isopropylmalic acid, and GCMS analysis was performed in the same manner as in Example 5, except that the chitosan-modified, citric acid-modified paper points (E4) prepared in Example 4 were used instead of the citric acid-modified paper points (E1) prepared in Example 1. The GCMS results for the detected glycine are shown in Figure 1(d), and the GCMS results for the detected 2-isopropylmalic acid are shown in Figure 1(d).

[0063] As shown in Figures 1 and 2, all of the paper points (E1) to (E4) prepared in Examples 1 to 4 appropriately adsorbed glycine (Figures 1(a) to (d)) and 2-isopropylmalic acid (Figures 2(a) to (d)), and it was possible to analyze this adsorbed state by GCMS.

[0064] Example 9: Preparation of a calibration curve for a glycine solution A 90% by volume aqueous solution of acetonitrile containing 50 ppm of glycine was prepared, and this was diluted 1-fold, 10-fold, 100-fold, and 1000-fold with a 90% by volume aqueous solution of acetonitrile to prepare test solutions.

[0065] GCMS analysis was carried out in the same manner as in Example 5, except that the chitosan-modified paper point (E3) prepared in Example 3 and the above test solution were used. The GCMS results for the detected glycine are shown in Figure 3.

[0066] As shown in FIG. 3, the obtained results show good linearity (R ２ = 0.996), which indicates that the glycine in the sample could be quantified using the above analytical method.

[0067] Furthermore, the detection limit (S / N=3, RMS) was estimated to be 0.65 pg (glycine) from the S / N ratio of the chromatogram obtained by GCMS analysis of the lowest concentration (1000-fold dilution) of the test solutions used above.

[0068] Example 10: Preparation of a calibration curve for 2-isopropylmalic acid A 90% by volume aqueous solution of acetonitrile containing 50 ppm of 2-isopropylmalic acid was prepared, and this was diluted 1-fold, 10-fold, 100-fold, and 1000-fold with a 90% by volume aqueous solution of acetonitrile to prepare test solutions.

[0069] GCMS analysis was carried out in the same manner as in Example 5, except that the chitosan-modified paper point (E3) prepared in Example 3 and the above test solution were used. The GCMS results for the detected 2-isopropylmalic acid are shown in Figure 4.

[0070] As shown in FIG. 4, the obtained results show good linearity (R ２ = 0.9991), which indicates that the quantification of 2-isopropylmalic acid in the sample was possible using the above analytical method.

[0071] Furthermore, the detection limit (S / N=3, RMS) was estimated from the S / N ratio of the chromatogram obtained by GCMS analysis of the lowest concentration (1000-fold dilution) of the test solutions used above, and was found to be 0.51 pg (2-isopropylmalic acid).

[0072] (Example 11: Analysis of saliva samples) GCMS analysis was carried out in the same manner as in Example 5, except that the chitosan-modified paper point (E3) prepared in Example 3 was used and a saliva sample collected from a human was used as the test solution. The GCMS results are shown in Figure 5.

[0073] As shown in Figure 5, in comparison with the chromatogram of the entire saliva sample (Figure 5(a)), chromatograms including fragments of the TMS-derivatized compound at m / z 73 (Figure 5(b)), 2-isopropylmalic acid at m / z 275 (Figure 5(c)), glycine at m / z 174 (Figure 5(d)), and lactic acid at m / z 147 (Figure 5(d)) were obtained. As such, it can be seen that the chitosan-modified paper point (E3) prepared in Example 3 was able to selectively collect and analyze various target compounds contained in saliva samples.

[0074] The present invention is applicable to various analyses using, for example, biological samples or environmental samples, and is useful in, for example, the fields of medicine and dentistry, pharmacy, environmental science, food chemistry, science and engineering, and the like.

Claims

1. A device for selectively collecting a target compound, comprising a cellulose substrate carrying a detection reagent, the detection reagent comprising a constituent compound having at least one functional group selected from the group consisting of an amino group, a carboxyl group, a sulfonic acid group, and a phenolic hydroxyl group, and the target compound is at least one selected from the group consisting of biological metabolites and environmental compounds.

2. The device of claim 1, wherein the detection reagent comprises at least one selected from the group consisting of amino acids, peptides, organic acids and salts thereof, polyethyleneimine, polydopamine, tannic acid, and lignin.

3. The device of claim 1, wherein the cellulose substrate is a paper point or a cellulose monolith.

4. A method for manufacturing a device for selectively collecting a target compound, comprising the step of supporting a detection reagent on a cellulose substrate, wherein the detection reagent comprises a constituent compound having at least one functional group selected from the group consisting of an amino group, a carboxyl group, a sulfonic acid group, and a phenolic hydroxyl group, and the target compound is at least one selected from the group consisting of biological metabolites and environmental compounds.

5. A method for selectively collecting a target compound from a biological or environmental sample, comprising the step of contacting the biological or environmental sample with a device according to any one of claims 1 to 3.

6. The method of claim 5, wherein the biological or environmental sample is saliva, blood, urine, gingival crevicular fluid, fecal suspension, exhaled breath condensate, exudate from an inflammatory focus, oral swab, nasal swab, condensate from the atmosphere, seawater, river water, lake water, fruit juice, or juice squeezed from a plant or animal.

7. A detection reagent for selectively collecting a target compound from a biological or environmental sample via a supported cellulose substrate, comprising a constituent compound having at least one functional group selected from the group consisting of an amino group, a carboxyl group, a sulfonic acid group, and a phenolic hydroxyl group, and the target compound is at least one selected from the group consisting of biological metabolites and environmental compounds.

Images