Further characterization of tunicate and tunichrome electrochemiluminescence

Electrophoresis, size exclusion chromatography, fluorescence, and electrochemiluminescence (ECL) data obtained from the cell-rich perivisceral fluid ('blood') of the tunicate, Molgula occidentalis, suggests that a yellowish protein or protein subunit of < 6.5 kDa is probably responsible for the low level intrinsic ECL reported previously. Variable potential ECL scans and spectrofluorometric analyses directly indicated that chlorophylls from ingested phytoplankton were not significant contaminants of M. occidentalis blood samples. Chlorophylls were also examined indirectly from monocultures of dinoflagellate and diatom species to determine their ECL levels. While ECL was observed in these cultures, high concentrations (10(6) organisms/mL) of dinoflagellates or diatoms were needed to produce ECL levels comparable to those observed for M. occidentalis blood. Additional work with two synthetic tunichrome isomers indicated 10-fold increases in ECL when the '2,3,4-tunichrome' was reacted with Tl+ in 1:1 and 1:2 metal ion to ligand molar ratios. The '3,4,5-tunichrome' isomer exhibited a titration curve with Hg2+ suggestive of the existence of at least two Hg2+ binding sites and a 9- to 10-fold increase in ECL output.