High-performance liquid-phase separation of glycosides. III. Determination of total glucosinolates in cabbage and rapeseed by capillary electrophoresis via the enzymatically released glucose.

A selective and sensitive method for the determination of total glucosinolates (GSs) in plant extracts by capillary electrophoresis-laser-induced fluorescence (LIF) detection was developed. It was based on the enzymatically released glucose from glucosinolates in the presence of the hydrolyzing enzyme myrosinase. The released glucose was converted to gluconic acid (GA) by the action of glucose oxidase. The resulting GA was then labeled selectively with the fluorescent tag 7-aminonaphthalene-1, 3-disulfonic acid (ANDSA). The peak area resulting from the GA-ANDSA derived from free and bound glucose was subtracted from the peak area of the GA-ANDSA resulting from the free glucose in the sample. This gave the total glucosinolates in the sample. The peak areas were normalized to the internal standard, N-acetylneuraminic acid derivatized with ANDSA. The method was validated using four different plant extracts, white cabbage leaves, rapeseed leaves, rapeseed roots, and rapeseed seeds. Furthermore, a capillary electrophoresis-UV detection method for profiling GS in plant extracts was developed. In addition to providing a fingerprint of the glucosinolates in plant extracts, the method allowed the experimenter to rapidly check the various steps involved in the extraction and sample cleanup.