Optimization of the solid-phase microextraction method in the determination of Ixodes ricinus (L.) volatiles.

For the first time, headspace solid-phase microextraction coupled with GC-MS analysis was used to study volatile compounds emitted by the tick Ixodes ricinus (L.). Variables such as the type of SPME fibre, equilibration time and extraction time have been evaluated with the mixture of four selected standards, a so-called artificial tick (acetophenone, racemic 4-heptanolide, methyl 2-methoxybenzoate and methyl 3-chloro-4-methoxybenzoate). Optimized conditions were obtained by the use of polydimethylsiloxan 100 microm fibre at 30 min equilibration and 15 min extraction time. The method proved to have a good linearity (r2 >0.98) at a concentration range from 0.5 ng (1 ng for methyl 3-chloro-4-methoxybenzoate) to 25 ng. LODs for a compound ranged between 0.19 and 1 ng, RSD (%) ranged from 13.76 to 25.08. The determination of 1.99 ng of methyl 3-chloro-4-methoxybenzoate emitted by five engorged females proved the usefulness of the developed method to identify and quantify the volatile compounds emitted by I. ricinus ticks.