GC analysis of essential oils in the rumen fluid after incubation of Thuja orientalis twigs in the Rusitec system

Methods for the chemical analysis of toxic plant substances in the rumen of ruminants are of importance for the diagnosis of intoxications with poisonous plants. The present work establishes a method to estimate monoterpene components of the essential oil of thuja (Thuja orientalis, Cupressaceae) in these types of samples. alpha-Thujone, which is regarded as the toxic principle, is present at a concentration of 50-60% in the essential oil. The rumen simulation technique (Rusitec) was used to simulate natural digestion. Chopped twigs of thuja were subjected to rumen content in a closed container with an overflow device. The flow of saliva was simulated by the continuous addition of a buffer solution. Samples for analysis were taken from the overflow at 24 and 48 h. A further sample was taken from the remaining liquid fraction of the rumen content in the container at 48 h. The essential oils were extracted with hexane and concentrated. A quantitative determination was done by capillary gas chromatography. Together in the three fractions analysed this resulted in total mean recoveries of 6.8% for alpha-thujone, 5.3% for beta-thujone, 18.9% for fenchone and 27.8% for camphor. The observation that the thujones were recovered to a lesser extent than other oil components is evidence of their fast decomposition in the rumen medium. Under these circumstances the calculated detection limit is 100-200 g thuja twigs in cows with rumen volumes of 60-100 litres. The main essential oil degradation products found in the rumen fluid of all three fractions in the Rusitec system were discovered to be iso-3-thujanol, neo-3-thujanol, carvomenthol and carvomenthone. (C) 2003 Elsevier Ltd. All rights reserved.