Analysis of biological samples obtained from in vivo experiments can be often challenging. In general it is not possible to apply the commonly used matrices that are necessary for the experiments to the desired analysis systems without further conditioning or sample purification steps. Besides possible adverse effects for instruments, interference between analytes and matrices can affect the correct measurement of analytes. Different methods of sample preparation can be used to convert biological samples into samples suitable for analysis; SPE and HS-SPME are two well established methods. Research of in vivo metabolism of triethyleneglycoledimethacrylate (TEGDMA), one of the most frequently contained comonomer in dental restorative materials, demands sample preparation methods that offer separation of TEGDMA and its related metabolites from biological matrices. In the presented study two methods for sample preparation were developed in order to analyze TEGDMA as well as its metabolites triethyleneglycole (TEG), 2,3-epoxymethacrylicacid methylester (2,3-EMME), and methacrylacid methylester (MAME) in Krebs-Henseleit buffer samples to facilitate a subsequent analysis via GC-MS. An easy and time-saving separation protocol was developed. Recovery rates of TEGDMA and TEG after SPE were 21 +/- 3% and 105 +/- 12%, respectively, recovery rate after headspace extraction of 2,3-EMME and MAME was higher at 48 degrees C compared with 20 degrees C extraction temperature. The tested range for 2,3-EMME and MAME concentration after HS-SPME extraction was 0.1-100 mg/L and both analytes showed a good linearity.