CLAM POISON: III. PAPER ELECTROPHORESIS OF CLAM POISON

In paper strip electrophoretograms on Whatman 3 MM paper, clam poison migrated toward the cathode when formic acid (1 N and 0.1 N), acetic acid (1 N), sodium acetate-acetic acid (0.1 M), monosodium dihydrogen phosphate-disodium monohydrogen phosphate (0.1 M), and sodium tetraborate (0.1 M) were used as electrolytes. Formic acid (1 N) proved the most suitable of these for examination of the quantitative electrophoretic behavior of clam poison. A method of pretreatment for the paper was developed to minimize introduction of extractable impurities into material recovered after electrophoresis by aqueous elution. Pure clam poison, on electrophoresis for 1.75 hours at 900 v on an 8 × 18-in. sheet of pretreated paper, migrated as a single, positively charged, Weber-positive band with apparent mobility 18.1 cm, which exhibited no fluorescence. Recovery was quantitative on both a weight and toxicity basis, and the toxicity, specific rotation, and infrared spectrum of the recovered toxin were identical with those of the starting material, thus providing an additional check on the homogeneity of the sample. Quantitative electrophoresis of toxin from clam poison tailings which had been purified by paper chromatography removed impurities which the latter method was incapable of eliminating, with resultant improvement in both the specific rotation and toxicity. Two such treatments provided toxin possessing a toxicity and specific rotation within the accepted limits for pure clam poison.