Abstract
Abstract An anion-exchange isocratic high-performance liquid chromatographic method for the separation and determination of two kainic acid analogues, namely (2S,3S,4S)-4-[l-(4′-azidobenzoy l)aminomethy lethenyl] -2-carboxy-3-pyrrolidineacetic acid and (2S.3S.4S)-4-[l-(4′-azido-phenyl)thioureylenernethy lethenyl]-2-carboxy-3-pyrro-lidineacetic acid is described. These kainic acid analogues were obtained by coupling dimethyl (2S.3S.4S)-4-(1-aminomethylethenyl)-2-carboxy-1-(9-fluorenylmethoxy-carbonyl)-3-pyrrolidineacetic acid with N-hydroxysuccinimidyl 4-azidobenzoate and 4-azidophenyl isothiocyanate respectively, followed by saponification. The thus obtained products were purified by reversed phase flash column chromatography and separated on Econosphere -NK2, using 5% (v/v) acetonitrile in 50 mM acetate buffer pH 5.5, at a flow rate of 1.5 ml/min. The effect of mobile phase pH, and of the acetonitrile concentration on the resolution between kainic acid and its analogues was also studied. Kainic acid was ...
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