A method of determining buffer conditions for analyzing a biomolecule with a chromatography column using a salt gradient, the method comprising: A) performing a first, a second, and a third salt gradient chromatography runs using mobile phases having a first, a second, and a third constant pH values, respectively, to analyze the biomolecule, in which the mobile phases have the first, the second, and the third constant pH values were prepared with a first, a second, and a third predetermined proportions of a first eluent solution and a second eluent solution, respectively, wherein the first, the second, and the third predetermined proportions are different, in which the first, the second, and the third salt gradient chromatography runs each used a same salt gradient concentration range, a same gradient time, and a same salt gradient concentration change rate, in which a) the first eluent solution comprises at least four buffer salts where at least three of the four buffer salts are a monovalent buffer salt, have a net negative charge or a net neutral zwitterionic charge over a pH range of about 6 to about 10, and include a sulfonate group and an amine, where the first eluent solution has a first pH of about 6, b) the second eluent solution comprises at least four buffer salts where at least three of the four buffer salts are a monovalent buffer salt, have a net negative charge or a net neutral zwitterionic charge over a pH range of about 6 to about 10, and include a sulfonate group and an amine, where the second eluent solution has a second pH of about 10; B) measuring one or more peaks during each of the first, the second, and the third salt gradient chromatography runs; and C) performing a chromatography optimization algorithm on the measured one or more peaks of the first, the second, and the third salt gradient chromatography runs to identify at least one optimized salt gradient chromatography run from the first, the second, and the third salt gradient chromatography runs, in which the performing of the chromatography optimization algorithm identifies at least one optimized salt gradient chromatography run from the first, the second, and the third salt gradient chromatography runs, and in which the first eluent solution and the second eluent solution both have a total buffer salt concentration of greater than about 250 millimolar. A) performing a first, a second, and a third salt gradient chromatography runs using mobile phases having a first, a second, and a third constant pH values, respectively, to analyze the biomolecule, in which the mobile phases have the first, the second, and the third constant pH values were prepared with a first, a second, and a third predetermined proportions of a first eluent solution and a second eluent solution, respectively, wherein the first, the second, and the third predetermined proportions are different, in which the first, the second, and the third salt gradient chromatography runs each used a same salt gradient concentration range, a same gradient time, and a same salt gradient concentration change rate, in which a) the first eluent solution comprises at least four buffer salts where at least three of the four buffer salts are a monovalent buffer salt, have a net negative charge or a net neutral zwitterionic charge over a pH range of about 6 to about 10, and include a sulfonate group and an amine, where the first eluent solution has a first pH of about 6, b) the second eluent solution comprises at least four buffer salts where at least three of the four buffer salts are a monovalent buffer salt, have a net negative charge or a net neutral zwitterionic charge over a pH range of about 6 to about 10, and include a sulfonate group and an amine, where the second eluent solution has a second pH of about 10; a) the first eluent solution comprises at least four buffer salts where at least three of the four buffer salts are a monovalent buffer salt, have a net negative charge or a net neutral zwitterionic charge over a pH range of about 6 to about 10, and include a sulfonate group and an amine, where the first eluent solution has a first pH of about 6, b) the second eluent solution comprises at least four buffer salts where at least three of the four buffer salts are a monovalent buffer salt, have a net negative charge or a net neutral zwitterionic charge over a pH range of about 6 to about 10, and include a sulfonate group and an amine, where the second eluent solution has a second pH of about 10; B) measuring one or more peaks during each of the first, the second, and the third salt gradient chromatography runs; and C) performing a chromatography optimization algorithm on the measured one or more peaks of the first, the second, and the third salt gradient chromatography runs to identify at least one optimized salt gradient chromatography run from the first, the second, and the third salt gradient chromatography runs, in which the performing of the chromatography optimization algorithm identifies at least one optimized salt gradient chromatography run from the first, the second, and the third salt gradient chromatography runs, and in which the first eluent solution and the second eluent solution both have a total buffer salt concentration of greater than about 250 millimolar.