# US 2024/0201145 A1 · Volatile pH Gradient Mobile Phase Kit > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2024/0201145 A1 - Document ID: US20240201145A1 - Status: Published application - Kind: A1 - Application number: 18/085,179 - Filed: 2022-12-20 - Published: 2024-06-20 - Earliest claimed: 2022-12-20 - Assignees: Dionex Corporation - Applicants: Dionex Corporation - Inventors: Jung Un Baek; Shanhua LIEHR; Yuan SUN; Christopher A POHL - Web page: https://patentgrep.com/patent/US20240201145A1 ## CPC - C07K 16/065: {Purification, fragmentation} - G01N 30/34: of fluid composition, e.g. gradient (G01N30/36 takes precedence) - B01D 15/166: {Fluid composition conditioning, e.g. gradient} - B01D 15/361: {Ion-exchange} - C07K 16/22: against growth factors {; against growth regulators} - C07K 16/241: {Tumor Necrosis Factors} - C07K 16/32: against translation products of oncogenes - G01N 30/20: using a sampling valve - G01N 30/72: Mass spectrometers {(mass spectrometers per se H01J49/00)} - G01N 30/96: using ion-exchange (G01N30/02, G01N30/90 take precedence) - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - 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) - B01D: SEPARATION (separating solids from solids by wet methods B03B, B03D; by pneumatic jigs or tables B03B; by other dry methods B07; magnetic or electrostatic separation of solid materials from solid materials or fluids, separation by high-voltage electric fields B03C; centrifuges, vortex apparatus B04; presses per se for squeezing-out liquid from liquid-containing material B30B9/02; treatment of water C02F, e.g. softening by ion-exchange C02F1/42; {arrangements of air intake cleaners in gas turbine plants F02C7/05} ; arrangements or mounting of filters in air-conditioning, air-humidification or ventilation F24F13/28) ## Abstract A pH gradient mobile phase preparation kit is described. The preparation kit includes a first component and a second component. The first component includes a first reagent separate from a second reagent. The first reagent, the second reagent, and a solvent together provide a volatile buffer solution having a low pH. The second component includes a base. The base mixed in the solvent provide a volatile base solution having a high pH. A method of separating and detecting one or more analytes comprised in a matrix of components in a sample, using the kit is also described. ## Claims ### Claim 1 A pH gradient mobile phase preparation kit comprising: a first component comprising a first reagent separate from a second reagent, wherein the first reagent, the second reagent, and a solvent together provide a volatile buffer solution having a low pH; and a second component comprising a base, wherein the base combined with the solvent provide a volatile base solution having a high pH. ### Claim 2 (depends on 1) The kit according to claim 1, wherein the first reagent, the second reagent, the solvent and the base are volatile components. ### Claim 3 (depends on 1) The kit according to claim 1, wherein the volatile buffer solution is less stable than the first reagent separate from the second reagent. ### Claim 4 (depends on 1) The kit according to claim 1, wherein combining the first component with the second component in varied proportions provides a volatile mobile phase having a pH in a range between the low pH and the high pH. ### Claim 5 (depends on 1) The kit according to claim 1, wherein the first reagent is provided in an undiluted form and the second reagent is provided in a form diluted by the solvent. ### Claim 6 (depends on 1) The kit according to claim 1, wherein the first reagent is provided in an undiluted form and the second reagent is provided in an undiluted form, and the solvent is added to form the volatile buffer solution. ### Claim 7 (depends on 1) The kit according to claim 1, wherein the base is provided in an undiluted form. ### Claim 8 (depends on 1) The kit according to claim 1, wherein the base is ammonia or an amine. ### Claim 9 (depends on 1) The kit according to claim 1, wherein the second reagent is a carboxylic acid. ### Claim 10 (depends on 1) The kit according to claim 1, wherein the first reagent includes ammonia or an amine. ### Claim 11 (depends on 10) The kit according to claim 10, wherein the first reagent is ammonium bicarbonate. ### Claim 12 (depends on 1) The kit according to claim 1 including a first container containing the first reagent, a second container containing the second reagent, and a third container containing the second component, wherein the first container, the second container, and the third container are substantially gas impermeable and gas tight. ### Claim 13 (depends on 12) The kit according to claim 12, wherein the first container has a carbon dioxide gas permeability of less than 225 cc.-mm/m2-24 hr.-Bar. ### Claim 14 (depends on 1) The kit according to claim 1, wherein the solvent includes water. ### Claim 15 A method of detecting one or more analytes comprised in a matrix of components in a sample, the method comprising: injecting the sample into an injection valve, the injection valve being in fluid communication with a first end of a chromatography column; pumping a volatile buffer solution having a low pH into the first end of the chromatography column, the volatile buffer solution comprising a first reagent and a second reagent provided from a kit; pumping a volatile base solution having a high pH into the first end of the chromatography column, the volatile base solution including a base provided in the kit; varying a proportion of the volatile buffer solution to the volatile base solution in the chromatography column by varying corresponding amounts of the volatile buffer solution and the volatile base solution pumped to the first end of the chromatography column, thereby providing a volatile mobile phase with a pH time-gradient flowing through the chromatography column; eluting the sample through the chromatography column in the volatile mobile phase thereby separating the analyte from the matrix components; injecting the sample into a mass spectrometer to provide a mass spectrum of the one or more analytes. ### Claim 16 (depends on 15) The method according to claim 15, wherein the first reagent and the second reagent from the kit are combined to form the volatile buffer solution less than 10 days prior to pumping the volatile buffer solution into the first end of the chromatography column. ### Claim 17 (depends on 15) The method according to claim 15, wherein the mass spectrometer includes an electrospray ionization source to create an aerosol of the analyte and the volatile mobile phase. ### Claim 18 (depends on 15) The method according to claim 15, wherein the first reagent includes ammonium bicarbonate, and the second reagent includes acetic acid. ### Claim 19 (depends on 15) The method according to claim 15, wherein the chromatograph column is an ion exchange column. ### Claim 20 (depends on 15) The method according to claim 15, wherein the analyte is a biomolecule having a pI between pH 4 and pH 10. ### Claim 21 (depends on 15) The method according to claim 15, wherein the pH time-gradient includes a positive gradient, wherein the pH increases with time while separating the analyte from the matrix components. ### Claim 22 (depends on 15) The method according to claim 15, wherein the first reagent and the second reagent from the kit are combined in-line to form the volatile buffer solution.