Use of borate complexation for the separation of non-UV-absorbing calystegines by capillary electrophoresis.

A capillary zone electrophoresis method was developed for the simultaneous analysis of seven closely related polyhydroxyalkaloids called calystegines. Successful results were obtained with a fused-silica capillary, 80 mM sodium tetraborate at pH 9.2 and temperature of 50 degrees C. Detection of non-UV-absorbing calystegines was achieved through in-situ complexation with borate ions. To further improve method sensitivity, a capillary with a bubble cell was used and detection performed at low wavelength (191 nm). Effects of buffer concentration, pH and temperature on migration times and efficiency are discussed. Migration behavior of selected compounds was significantly affected by their chemical structure (i.e., number and position of hydroxy groups). Under optimized conditions, baseline separation of the selected compounds was achieved in less than 12 min. Precision was evaluated by measuring repeatability and intermediate precision of migration times and corrected peak areas. Finally, the method was applied to the qualitative analysis of calystegines in plant extracts and results were confirmed by GC-MS.