Online preconcentration of thyrotropin-releasing hormone (TRH) by SDS-modified reversed phase column for microbore and capillary high-performance liquid chromatography (HPLC)

Thyrotropin-releasing hormone (TRH, pGlu-His-Pro-amide) is an important tripeptide existing in biological systems at low concentrations. It is a fairly hydrophilic peptide, cationic in acidic solutions. Preconcentration online before reversed phase chromatography separation can enhance concentration detection limits of hydrophobic, but not hydrophilic species. The hydrophilic TRH can be preconcentrated using a reversed phase precolumn charged with sodium dodecyl sulfate (SDS). The separation also uses SDS. The preconcentration is effective for a microbore system, achieving detection limit of 250 pM for a sample size of 500 μl with electrochemical detection of the biuret complex formed post column. Preconcentration using an online precolumn is also effective in packed capillary high-performance liquid chromatography (HPLC) with a detection limit of 3 nM in 24 μl.