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

Journal article
Authors Rong Meng
Wanlin Xia
Mats Sandberg
Robert Stephens
Stephen G Weber
Published in Journal of chromatography. A
Volume 1071
Issue 1-2
Pages 179-84
ISSN 0021-9673
Publication year 2005
Published at Institute of Physiology and Pharmacology, Dept of Medical Biophysics
Pages 179-84
Language en
Keywords Chromatography, High Pressure Liquid, methods, Sensitivity and Specificity, Sodium Dodecyl Sulfate, chemistry, Thyrotropin-Releasing Hormone, analysis, chemistry
Subject categories Medical and Health Sciences


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 microl 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 microl.

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