Modification of the analytical procedures for the determination of herbage and faecal n-alkanes used in the estimation of herbage intake

Abstract

SUMMARYThe evaluation of n-alkane concentrations in herbage and faeces is the basis of a methodology that yields precise estimates of herbage intake. Established chemical analysis procedures for the determination of n-alkane concentrations in herbage and faecal samples involve an elaborate sequence of steps utilizing n-hexane as solvent for both the liquid-liquid extraction of n-alkanes from the sample and the solid-phase separation of n-alkanes in the extract. Composites of herbage and faecal samples from studies with sheep provided the experimental material used to evaluate a series of modifications which would simplify and reduce the workload involved in the chemical analysis. The results show that a less hazardous chemical, n-heptane, can replace n-hexane at all stages of the analytical procedure. They also show that evaporation of the sample extract and redissolving it prior to the separation of the n-alkanes, using a silica gel column, is unnecessary and that the volume of solvent used can be reduced. Procedures for saponification of samples prior to extraction can also be simplified as the process can be run overnight involving a slow build-up of temperature to 90 °C. The gain in precision from processing samples in duplicate was negligible and it would be more appropriate to invest extra efforts on an increased number of experimental animals. The results snowed that the workload and cost of using the n-alkane technique to estimate herbage intake can be reduced substantially.