Rapid estimation of corynetoxins in bacterial galls from annual ryegrass (Lolium rigidum Gaudin) by high-performance liquid chromatography

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Abstract
A rapid and sensitive method is described for the extraction, detection and measurement of corynetoxins in ryegrass seed galls colonized by Corynebucterium rathayi. Excised galls are extracted with aqueous methanol and the extract analysed, without further purification, by reversed-phase high-performance liquid chromatography with detection by uItraviolet absorption. The method is applicable down to the level of a single gall or part of a gall with a minimum level of detection of about 0.25 ¦g using the equipment described. The corynetoxin content of the individual galls examined varied from below the detection limit to 10.6 ¦g. The highest level of corynetoxin in a single gall was 1.15% of gall weight. The corynetoxin composition of galls collected during outbreaks of annual ryegrass toxicity in South Australia, Western Australia and South Africa during a 19-year period were found to be similar.