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Updated: May 11, 2025

Separation of Bioactive Small Molecules, Peptides from Natural Sources and Proteins from Microbes by Preparative Isoelectric Focusing IEF Method
Published on: June 14, 2020
Electrokinetic Chromatography-based Micro Methods for Separation and Physiochemical Characterization of Very
Robert Minkner1,2, Hermann Wätzig1
1Institute of Medicinal and Pharmaceutical Chemistry, TU Braunschweig, Braunschweig, Germany.
Abstract:
This study established an efficient separation of the highly hydrophobic analytes fenofibrate, orlistat, lumefantrine and a micelle marker using microemulsion electrokinetic capillary chromatography. A starting buffer with sodium dodecyl sulphate, heptane and butan-1-ol resulted in a partial separation of the analytes. Various experimental parameters were investigated, including voltage, sample loading time, buffer composition and temperature. An excellent separation was achieved, and compounds with hydrophobicity of up to a calculated log P of 10.2 (est. 12) could be clearly distinguished. Due to observed variabilities in retention time when changing the capillaries, it is advisable to calibrate with a standard mixture before each measurement series.
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