Analytical Sciences

Application of Capillary Electrophoresis to the Simultaneous Determination of Betaines in Plants

Naoki NISHIMURA,  Jinghua ZHANG, Mitsuru ABO, Akira OKUBO, and Sunao YAMAZAKI

Department of Applied Biological Chemistry, The University of Tokyo, Bunkyo, Tokyo 113-8657, Japan

The simultaneous determination of betaines, the key compounds for osmotic regulation in plants, was established by capillary electrophoresis (CE). After four betaines, glycine betaine (GB), beta-alanine betaine (AB), proline betaine (PB), and 2-hydroxyproline betaine (HPB), were esterified with p-bromophenacyl bromide, the esters were electrophoresed in 100 mM sodium phosphate at pH 3.0. A low-pH condition in CE and esterification gave an advantage of resolving each of the ester peaks as well as those of the unreacted reagent and other components. Furthermore, the addition of 4% polyethylene glycol (PEG) gave a better resolution of 4 peaks, resulting in the separation of the overlapped peaks of PB and AB. It was found from the standard addition method being applied to barley leaves that the GB content in plants could be evaluated by using a calibration curve of the GB standard solution. The extraction of GB from plant leaves was also improved by adopting water as the extraction solvent instead of a mixture of organic solvents. The present method was suitably applied to actual plant specimens collected from a saline area of China.

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