[Development of online conventional array-based two-dimensional liquid chromatographic system for proteins separation in human plasma].

Human plasma is one of the proteins-containing samples most difficult to characterize on account of the w ide dynamic concentration range of its intact proteins. Herein,w e developed a highthroughput conventional array-based tw o-dimensional liquid chromatographic system for proteins separation in human plasma in online mode. In the system,a conventional strong-anion exchange chromatographic column w as used as the first separation dimension and eight parallel conventional reversed-phase liquid chromatographic columns w ere integrated as the second separation dimension. T he fractions from the first dimension w ere sequentially transferred into the corresponding reversed-phase liquid chromatographic precolumns for retention and enrichment using a 10-port electrically actuated multi-position valve. T he second dimensional solvent flow w as directly and identically split into 8 channels. T he fractions w ere concurrently back-flushed from the precolumns into the 8 conventional R P columns and w ere separated simultaneously. An 8-channel fraction collector w as refitted to collect the reversed-phase liquid chromatographic fractions for further investigation. Bicinchoninic acid( BC A) dyeing solution w as conveniently used for high-abundance protein location. T w o separation dimensions w ere relatively independent parts, as w ell as each channel of the second dimensional array separation. T herefore,the new system could improve the separation throughput and total peak capacity. T he system w as successfully applied for the separation of human plasma intact proteins. T he results indicated the established system is an effective method for removing high abundance proteins in plasma and in-depth research in plasma proteomics.