# US 2021/0171561 A1 · Method for analysis of chondroitin sulfate > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2021/0171561 A1 - Document ID: US20210171561A1 - Status: Published application - Kind: A1 - Application number: 17/181,295 - Filed: 2021-02-22 - Published: 2021-06-10 - Earliest claimed: 2018-08-21 - Assignees: JCR Pharmaceuticals Co., Ltd. - Applicants: JCR Pharmaceuticals Co., Ltd. - Inventors: Noboru TANAKA; Sachiho KIDA - Web page: https://patentgrep.com/patent/US20210171561A1 ## CPC - G01N 33/50: Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing (measuring or testing processes involving enzymes or microorganisms, compositions or test papers therefor; processes for forming such compositions, condition responsive control in microbiological or enzymological processes C12Q) - G01N 33/6893: {related to diseases not provided for elsewhere} - C07H 5/06: Aminosugars - G01N 2030/027: {Liquid chromatography} - G01N 2030/8836: {involving saccharides} - G01N 2400/40: Glycosaminoglycans, i.e. GAG or mucopolysaccharides, e.g. chondroitin sulfate, dermatan sulfate, hyaluronic acid, heparin, heparan sulfate, and related sulfated polysaccharides - G01N 2800/52: Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) - C07H: SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS (derivatives of aldonic or saccharic acids C07C, C07D; aldonic acids, saccharic acids C07C59/105, C07C59/285; cyanohydrins C07C255/16; glycals C07D; compounds of unknown constitution C07G; polysaccharides, derivatives thereof C08B; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; sugar industry C13) ## Abstract Disclosed is a method for decomposing chondroitin sulfate contained in a sample into disaccharide. In particular disclosed is a method for decomposing chondroitin sulfate contained in a sample into disaccharide by heating the chondroitin sulfate in HCl-methanol containing 2,2-dimethoxypropane at a temperature of 60° C. to 90° C. for 50 minutes to 180 minutes, optionally in the method, the sample is selected from body fluid, a cell, a tissue, an organ, a cell culture solution, a tissue culture solution, a food, and a feed, or a derived therefrom. ## Claims ### Claim 1 A method for decomposing chondroitin sulfate contained in a sample into disaccharide represented by formula (IV) below: wherein the chondroitin sulfate is decomposed by heating in HCl-methanol containing 2,2-dimethoxypropane at a temperature of 60° C. to 90° C. for 50 minutes to 180 minutes. ### Claim 2 (depends on 1) The method according to claim 1, wherein the heating for decomposing the chondroitin sulfate is performed at a temperature of 65° C. to 75° C. for 70 minutes to 110 minutes. ### Claim 3 (depends on 1) The method according to claim 1, wherein the sample is selected from a body fluid, a cell, a tissue, an organ, a cell culture medium, a tissue culture medium, a food, and a feed, or a derivative thereof. ### Claim 4 (depends on 1) The method according to claim 1, wherein the sample is selected from the group consisting of a body fluid, a cell, a tissue, an organ, blood, a blood serum, a blood plasma, urine, a bone marrow fluid, a cerebral spinal fluid, and a derivative thereof obtained from a mammal. ### Claim 5 (depends on 4) The method according to claim 4, wherein the mammal is selected from the group consisting of a human, a monkey, a mouse, a rat, a guinea pig, a hamster, a rabbit, a horse, a cow, a pig, a dog, and a cat. ### Claim 6 (depends on 4) The method according to claim 4, wherein the mammal is a human, and the human is a patient with a disorder in which chondroitin sulfate is accumulated in the body. ### Claim 7 (depends on 6) The method according to claim 6, wherein the disorder is selected from the group consisting of Maroteaux-Lamy syndrome, Morquio syndrome type A, Morquio syndrome type B, and Sly syndrome. ### Claim 8 (depends on 6) The method according to claim 6, wherein the patient has been treated to reduce the chondroitin sulfate present in the body. ### Claim 9 (depends on 1) A method for decomposing chondroitin sulfate and heparan sulfate contained in a sample, wherein the chondroitin sulfate is decomposed into disaccharide represented by formula (IV) below, and the heparan sulfate is decomposed into disaccharide represented by formula (XIV) below, and wherein the heparan sulfate is decomposed by heating in HCl-methanol containing 2,2-dimethoxypropane at a temperature of 65° C. to 85° C. for 80 minutes to 180 minutes, and the chondroitin sulfate is decomposed by the method according to claim 1. ### Claim 10 (depends on 9) The method according to claim 9, wherein the heating for decomposing the heparan sulfate is performed at a temperature of 78° C. to 82° C. for 110 minutes to 130 minutes. ### Claim 11 (depends on 9) A method for decomposing chondroitin sulfate, heparan sulfate, and dermatan sulfate contained in a sample, wherein represented by formula (IV) below, the heparan sulfate is decomposed into disaccharide represented by formula (XIV) below, and the dermatan sulfate is decomposed into disaccharide represented by formula (XII) below, and wherein the dermatan sulfate is decomposed by heating in HCl-methanol containing 2,2-dimethoxypropane at a temperature of 60° C. to 80° C. for 20 minutes to 100 minutes, and the chondroitin sulfate and the heparan sulfate are decomposed by the method according to claim 9. ### Claim 12 (depends on 11) The method according to claim 11, wherein the heating for decomposing the dermatan sulfate is performed at a temperature of 63° C. to 67° C. for 30 minutes to 80 minutes. ### Claim 13 (depends on 9) The method according to claim 9, wherein the sample is selected from a body fluid, a cell, a tissue, an organ, a cell culture medium, a tissue culture medium, a food, a feed, and the derivative thereof. ### Claim 14 (depends on 9) The method according to claim 9, wherein the sample is selected from the group consisting of a body fluid, a cell, a tissue, an organ, a blood, a blood serum, a blood plasma, a urine, a bone marrow fluid, a cerebral spinal, and the derivative thereof obtained from a mammal. ### Claim 15 (depends on 14) The method according to claim 14, wherein the mammal is selected from the group consisting of a human, a monkey, a mouse, a rat, a guinea pig, a hamster, a rabbit, a horse, a cow, a pig, a dog, and a cat. ### Claim 16 (depends on 14) The method according to claim 14, wherein the mammal is a human, and the human is a patient with a disorder in which one or a plurality of chondroitin sulfate, heparan sulfate, and dermatan sulfate have been accumulated in the body. ### Claim 17 (depends on 16) The method according to claim 16, wherein the disorder is selected from the group consisting of Maroteaux-Lamy syndrome, Morquio syndrome type A, Morquio syndrome type B, Hunter syndrome, Hurler syndrome, Scheie syndrome, Hurler-Scheie syndrome, Sanfilippo syndrome, and Sly syndrome. ### Claim 18 (depends on 16) The method according to claim 16, wherein the patient has been treated to reduce one or a plurality of chondroitin sulfate, heparan sulfate, and dermatan sulfate present in the body. ### Claim 19 (depends on 1) A method for measuring the amount of chondroitin sulfate contained in a sample, the method including: a step of obtaining an eluate by applying disaccharide obtained by the method according to claim 1 to liquid chromatography; and a step of applying the eluate to mass spectrometry. ### Claim 20 (depends on 9) A method for measuring the amount of chondroitin sulfate and heparan sulfate contained in a sample, the method including: a step of obtaining an eluate by applying disaccharide obtained by decomposing chondroitin sulfate and heparan sulfate by the method according to claim 9 to liquid chromatography; and a step of applying the eluate to mass spectrometry. ### Claim 21 (depends on 10) A method for measuring the amount of chondroitin sulfate, heparan sulfate, and dermatan sulfate contained in a sample, the method including: a step of obtaining an eluate by applying disaccharide obtained by decomposing chondroitin sulfate, heparan sulfate, and dermatan sulfate by the method according to claim 10 to liquid chromatography; and a step of applying the eluate to mass spectrometry. ### Claim 22 (depends on 19) A method for detecting an individual with a disorder in which chondroitin sulfate is accumulated in the body from among a mammal providing a sample, the sample obtained by the method according to claim 19, on the basis of a measurement value of chondroitin sulfate contained in the sample. ### Claim 23 (depends on 20) A method for detecting an individual with a disorder in which chondroitin sulfate and/or heparan sulfate is accumulated in the body from among a mammal providing a sample, the sample obtained by the method according to claim 20, on the basis of a measurement value of chondroitin sulfate and/or heparan sulfate contained in the sample. ### Claim 24 (depends on 21) A method for detecting an individual with a disorder in which one or a plurality of chondroitin sulfate, heparan sulfate, and dermatan sulfate are accumulated in the body from a mammal providing a sample, the sample obtained by the method according to claim 21, on the basis of measurement values of chondroitin sulfate, heparan sulfate, and dermatan sulfate. ### Claim 25 (depends on 22) The method according to claim 22, wherein the disorder is selected from the group consisting of Maroteaux-Lamy syndrome, Morquio syndrome type A, Morquio syndrome type B, Hunter syndrome, Hurler syndrome, Scheie syndrome, Hurler-Scheie syndrome, Sanfilippo syndrome, and Sly syndrome. ### Claim 26 (depends on 19) A method for confirming an effect of a medical treatment, on the basis of a measurement value obtained by measuring the amount of a substance accumulated in a body and contained in samples by the method according to claim 19, wherein the samples are obtained from a patient, the patient suffering from a disorder in which one or a plurality of chondroitin sulfate are accumulated in the body and receiving a medical treatment to reduce the amount of the substance in the body, before and after receiving the treatment.