The nature of bound residues derived from triforine in barley plants

The shoots of barley plants root-treated with ³H]-triforine fungicide (uniformly labelled in the piperazine ring) were analysed 30 days after treatment. Methanol extraction left a solid plant residue which contained 33% of the total ³H which had been incorporated into the shoots. Methanol acidified with hydrochloric acid extracted a further 18% of the triforine-derived bound residues as N-2,2,2-trichloro-1-(piperazin-1-yl)ethyl]formamide (8%), and piperazine (10%). In the plants, these compounds had thus been complexed to plant constituents. Hot dimethyl sulphoxide (DMSO) extracted a further 13% of the total ³H, leaving a solid residue (mainly cellulose) which contained 2% of ³H, perhaps incorporated into the cellulose. Evaporation of the solvent from the DMSO extract gave a solid, the radioactivity of which (13%) could not be extracted by methanol. A part (7%) of this radioactivity could be released by successive hydrolysis with amyloglucosidase and β-glucosidase, which generated a complex mixture of polar and water soluble unknown radioactive compounds not including piperazine. These latter compounds would be the products of extensive metabolism of triforine (and its metabolite piperazine) bound to, or incorporated into starch. Most (11%) of the radioactivity of this solid could be released by hydrochloric acid hydrolysis, which also generated a complex mixture of polar and water soluble unknown radioactive compounds not including piperazine; a part (4%) of them could have been associated with lignin in the plant.