# US 2021/0239656 A1 · Analysis system > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2021/0239656 A1 - Document ID: US20210239656A1 - Status: Published application - Kind: A1 - Application number: 17/049,249 - Filed: 2019-01-25 - Published: 2021-08-05 - Earliest claimed: 2018-04-27 - Assignees: Shimadzu Corporation - Applicants: Shimadzu Corporation - Inventors: Yanli GUO; Naoki HAMADA; Yuki HASHI - Web page: https://patentgrep.com/patent/US20210239656A1 ## CPC - G01N 30/06: Preparation - G01N 30/34: of fluid composition, e.g. gradient (G01N30/36 takes precedence) - G01N 30/46: using more than one column {(G01N30/44 takes precedence)} - G01N 30/463: {for multidimensional chromatography} - G01N 30/88: Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 - G01N 30/89: Inverse chromatography - G01N 2030/065: {using different phases to separate parts of sample} - G01N 2030/8813: {biological materials} - 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) ## Abstract The present invention relates to an analysis system that is applied to rapid automatic analysis and detection of highly lipophilic components and a method for rapidly detecting vitamin D in an oil or fat, or a biological sample. In the present invention, online pretreatment and separation of a sample containing highly lipophilic components in a sample to be analyzed are performed by a multidimensional chromatography system. The multidimensional chromatography system includes a supercritical chromatography part and a reverse phase liquid chromatography part sequentially connected, the reverse phase liquid chromatography part including one or more reverse phase liquid chromatography columns, and the supercritical chromatography part including a supercritical mobile phase, a modifier, and a supercritical packed column. ## Claims ### Claim 1 An analysis system comprising: an autosampler; a multidimensional chromatography system including a supercritical chromatography column and one or more reverse phase liquid chromatography columns, a nonpolar substance being eluted and removed in the supercritical chromatography column, and a weakly polar substance being adsorbed to a stationary phase of the supercritical chromatography column and further transported to the one or more reverse phase liquid chromatography columns; a column oven in which a column is to be provided; a liquid pump configured to transport a modifier to the supercritical chromatography column and transport a mobile phase to the one or more reverse phase liquid chromatography columns; and a mass spectrometer. ### Claim 2 (depends on 1) The system according to claim 1, wherein the modifier is selected from alcohol, nitrile, and an aqueous solution of alcohol or nitrile. ### Claim 3 (depends on 1) The system according to claim 1, wherein a multi-way valve group connects or switches connection between the supercritical chromatography column and the one or more reverse phase liquid chromatography columns in the multidimensional chromatography system. ### Claim 4 (depends on 1) The system according to claim 1, wherein the weakly polar substance adsorbed to the supercritical chromatography column is transported to the one or more reverse phase liquid chromatography columns by only an action of the modifier to remove a supercritical mobile phase flowing out of the supercritical chromatography column in the one or more reverse phase liquid chromatography columns. ### Claim 5 (depends on 1) The system according to claim 1, wherein: the multidimensional chromatography system includes one reverse phase liquid chromatography column; and the weakly polar substance adsorbed to the supercritical chromatography column is transported to the one reverse phase liquid chromatography column by only an action of the modifier to remove the supercritical mobile phase flowing out of the supercritical chromatography column in the one reverse phase liquid chromatography column, and then a component of the weakly polar substance is separated by the one reverse phase liquid chromatography column. ### Claim 6 (depends on 1) The system according to claim 1, wherein: the multidimensional chromatography system includes two reverse phase liquid chromatography columns; the weakly polar substance adsorbed to the supercritical chromatography column is transported to a first reverse phase liquid chromatography column by only an action of the modifier to remove the supercritical mobile phase flowing out of the supercritical chromatography column in the first reverse phase liquid chromatography column; and the weakly polar substance is transported to a second reverse phase liquid chromatography column to separate a component. ### Claim 7 (depends on 1) The system according to claim 1, wherein, in the supercritical chromatography column: a stationary phase is selected from a silica gel modified with a polar group selected from a hydroxy group, an amino group, and a cyano group; and a mobile phase is supercritical carbon dioxide. ### Claim 8 (depends on 1) The system according to claim 1, wherein, in the reverse phase liquid chromatography column: a stationary phase is selected from a silica gel modified with a hydrophobic group selected from a hydrocarbon group; and the mobile phase is a polar organic solvent or an aqueous solution of a polar organic solvent. ### Claim 9 (depends on 1) The system according to claim 1, wherein at least the reverse phase liquid chromatography column configured to perform separation of the weakly polar substance is placed in a column oven. ### Claim 10 (depends on 1) The system according to claim 1, wherein the system has a pretreatment column configured to separate a protein component in a sample to be analyzed. ### Claim 11 (depends on 1) The system according to claim 1, wherein the system is used for automatically detecting a highly lipophilic component in an oil or fat, or a biological sample. ### Claim 12 (depends on 11) The system according to claim 11, wherein the highly lipophilic component is vitamin D. ### Claim 13 A method for rapidly detecting vitamin D in an oil or fat or a biological sample, comprising: step S1 of preparing a sample to be analyzed; step S2 of separating vitamin D in the sample to be analyzed by using a multidimensional chromatography system; and step S3 of detecting the vitamin D separated in the step S2, wherein: in the step S2, the multidimensional chromatography system includes a supercritical chromatography part and a reverse phase liquid chromatography part which are sequentially connected; the reverse phase liquid chromatography part includes one or more reverse phase liquid chromatography columns; the supercritical chromatography part includes a supercritical mobile phase and a modifier; and the modifier is selected from alcohol, nitrile, and an aqueous solution of alcohol, or nitrile. ### Claim 14 (depends on 13) The method according to claim 13, wherein the supercritical chromatography part includes one supercritical chromatography column, and the reverse phase liquid chromatography part includes two reverse phase liquid chromatography columns. ### Claim 15 (depends on 13) The method according to claim 13, wherein a multi-way valve group connects the one or more reverse phase liquid chromatography columns in the multidimensional chromatography system. ### Claim 16 (depends on 13) The method according to claim 13, wherein, in the supercritical chromatography part: a stationary phase is selected from a silica gel modified with a polar group selected from a hydroxy group, an amino group, and a cyano group; and the supercritical mobile phase is supercritical carbon dioxide. ### Claim 17 (depends on 13) The method according to claim 13, wherein, in the reverse phase liquid chromatography part: a stationary phase is selected from a silica gel modified with a hydrophobic group selected from an alkyl group; and a mobile phase is a polar organic solvent or an aqueous solution of the polar organic solvent. ### Claim 18 (depends on 13) The method according to claim 13, wherein the one or more reverse phase liquid chromatography columns of the reverse phase liquid chromatography part are identical or different. ### Claim 19 (depends on 13) The method according to claim 13, wherein, in the step S2, a weakly polar substance adsorbed to a supercritical chromatography column is transported to the reverse phase liquid chromatography part by only an action of the modifier to remove a supercritical mobile phase flowing out of the supercritical chromatography part in the reverse phase liquid chromatography part. ### Claim 20 (depends on 13) The method according to claim 13, wherein, the step S2 further includes: step S21 of separating a component of the sample to be analyzed by using supercritical chromatography and transporting a weakly polar substance to the reverse phase liquid chromatography part; step S22 of removing a supercritical mobile phase flowing out of the supercritical chromatography part in the reverse phase liquid chromatography part; and step S23 of introducing a mobile phase into the reverse phase liquid chromatography part and separating a substance present in the one or more reverse phase liquid chromatography columns to obtain a vitamin D component separated, wherein transport of the weakly polar substance to the one or more reverse phase liquid chromatography columns in the step S21 may be performed at the same time as the step S22. ### Claim 21 (depends on 13) The method according to claim 13, wherein, the step S2 further includes: step S21′ of separating a component of the sample to be analyzed by using supercritical chromatography and transporting a weakly polar substance to a first reverse phase liquid chromatography column of the reverse phase liquid chromatography part; step S22′ of removing a supercritical mobile phase flowing out of the supercritical chromatography part in the first reverse phase liquid chromatography column; and step S23′ of further introducing a mobile phase into the first reverse phase liquid chromatography column, and transporting a weakly polar substance present in the first reverse phase liquid chromatography column to a second reverse phase liquid chromatography column to obtain a vitamin D component by separation in the second reverse phase liquid chromatography column, wherein transport of the weakly polar substance to the first reverse phase liquid chromatography column in the step S21′ may be performed at the same time as the step S22′. ### Claim 22 (depends on 13) The method according to claim 13, further includes, before the step S2, a step of separating a protein component in the sample to be analyzed by using a pretreatment chromatography column, wherein, in the step S3, detection is performed by using a mass spectrometer.