Comprehensive two‐dimensional liquid chromatography (NPLC×RPLC) with vacuum‐evaporation interface

A comprehensive two-dimensional liquid chromatographic system incorporating a vacuum-evaporation interface was developed. Normal-phase liquid chromatography with a CN microcolumn was used as the first dimension (1(st)-D), and reversed-phase liquid chromatography with a C(18) monolithic column was used as the second dimension (2(nd)-D). An electronically controlled dual-position, ten-port valve with two identical storage loops served as the interface and the analysis time in the 2(nd)-D was 1.5 min. The solvent in the loops of the interface was evaporated at 25 degrees C under vacuum conditions, leaving the analytes on the inner wall of the loops. The mobile phase of the 2(nd)-D dissolved the analytes in the loop and injected them onto the second column, allowing an on-line solvent exchange of the fractions from the 1(st)-D to the 2(nd)-D. The chromatographic resolution of analytes on the two dimensions was evaluated. Sample loss due to evaporation in the interface was investigated with standard samples having different boiling points. The usefulness of the comprehensive 2-DLC system was demonstrated in the analysis of a traditional Chinese medicine Radix salviae miltiorrhiza bage extract.