Methods of performing high-flow chromatography under adiabatic conditions

A method of separating a plurality of oligonucleotides, the method comprising:

• (a) loading a sample comprising the plurality of oligonucleotides, the plurality of nucleotides including a plurality of a first oligonucleotide and a plurality of a second oligonucleotide, onto a chromatography system;
• wherein the chromatography system comprises a chromatography column, a jacket surrounding the liquid chromatography column, and a vacuum chamber formed in an area between the liquid chromatography column and the jacket;
• wherein the chromatography column comprises a column body and a plurality of particles, the column body comprising an inlet region, an outlet region, an interior surface, and an exterior surface;
• (b) eluting the sample from the chromatography column by contacting the chromatography column with a mobile phase;
• wherein the chromatography column is characterized by a flow rate and a pressure differential between the inlet region and the outlet region, wherein the product of the flow rate and the pressure differential is at least 1.5 W/m;
• and
• (c) detecting the plurality of the first oligonucleotide and/or the plurality of the second oligonucleotide in the eluent, wherein the plurality of the first oligonucleotide form at least one distinct peak and wherein the plurality of the second oligonucleotide form at least one distinct peak;
• wherein the chromatography column is characterized by a radial temperature gradient of at most 0.1 K and a pressure differential of from the inlet region of the chromatography column to the outlet region of the chromatography column of at least 100 bar.