SOLID-PHASE EXTRACTION AND FLUOROMETRIC DETECTION OF BENZOPHENANTHRIDINE ALKALOIDS FROM SANGUINARIA-CANADENSIS CELL-CULTURES

A solid-phase extraction (SPE) method using Sep-Pak C18 reversed-phase cartridges was developed for the prepurification of benzophenanthridine alkaloids produced by Sanguinaria canadensis cell cultures. Efficient recoveries (>90%) of chelirubine, sanguinarine and chelerythrine were obtained from synthetic solutions using the SPE method. Up to 20 mg of alkaloids were extracted from spiked biomass and culture-medium samples on a 400-mg adsorbent cartridge. This extraction method was more reproducible and as selective as a conventional liquid-liquid extraction technique. Fluorimetric detection was used for the identification and quantification of benzophenanthridine alkaloids after high-performance liquid chromatographic separation. This detection method was compared to conventional UV absorption at 280 nm. The detection limits of both detectors were in the 100-pg range for chelirubine and sanguinarine and in the 1-ng range for chelerythrine. The main advantage of fluorimetry over UV spectrometry was its improved selectivity for the benzophenanthridine alkaloids. © 1990.