Effect of sample pre-treatment on the determination of steroid esters in hair of bovine calves

The effect of three sample pre-treatment steps, washing, cutting and grinding on the determination of steroid esters in hair is studied. The study is performed by using hair samples obtained after pour-on application of steroid esters to bovine calves. After sample pre-treatment the hair is treated with a mild reducing agent [tris(2-carboxyethyl)phosphine hydrochloride] to extract the steroid esters. After a solid-phase extraction clean-up step the extracts are analysed by using liquid chromatography combined with triple–quadrupole mass spectrometric detection. For the washing step the use of non-organic washing solvents like (warm) water and a solution of 0.1% sodium dodecyl phosphate and organic solutions containing different percentages of methanol are tested. By using the non-organic solvents and the organic solvents with a percentage of methanol <20% the recovery results are as good as the results obtained without washing the hair. Cutting the hair samples increases the analyte recoveries of incurred steroid esters by 20% compared to the non-cut hair. The analyte recoveries of cut hair samples are about 60–80% that of ground hair samples. The obtained surface expansion of hair samples by grinding proves to be necessary in order to achieve the highest possible analyte yields. Finally the use of pressurised liquid extraction (PLE) for the extraction of steroid esters from plain (no washing, cutting or grinding) hair is investigated. The first results show lower (up to 40%) extraction recoveries in comparison with the classical solvent extraction procedures. If the limit of detection requirement is met, PLE may be an alternative for extracting large numbers of hair samples due to the short sample treatment procedure involved.