Off-line microextraction by packed sorbent combined with on solid support derivatization and GC-MS: Application for the analysis of five pyrethroid metabolites in urine samples.

A novel, fast and eco-friendly analytical method using microextraction by packed sorbent coupled to large volume injection-gas chromatography-mass spectrometry (MEPS-LVI-GC-MS) was developed for the determination of five pyrethroid metabolites (cis-2,2-dimethyl-3-(2-chloro-3,3,3-trifluoro-1-propenyl)-cyclopropanecarboxylic acid, cis/trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carboxylic acids, cis-(2,2-dibromovinyl)-2,2-dimethylcyclopropane-1-carboxylic acid and 3-phenoxybenzoic acid) in human urine. MEPS was performed off-line using a manually-operated semiautomatic syringe (eVol), and several parameters including the sample pH, extraction sorbent, washing solvent, volume and type of elution solvent and number of draw-eject cycles were optimized. Analytes were extracted from enzymatically hydrolyzed urine using a C18 solid phase with subsequent simultaneous derivatization and elution with a mixture of 1,1,1,3,3,3-hexafluoroisopropanol and diisopropylcarbodiimide in n-hexane (on-line derivatization). The optimized method was validated, with linearity established from 0.05 to 25ngmL-1 and R values > 0.99. Obtained quantification limits were in the range of 0.06-0.08ngmL-1 , and the precision expressed as relative standard deviation (RSD) was below 14% for all of the analytes. The method was cross-validated with a reference approach based on liquid-liquid extraction-gas chromatography-mass spectrometry (LLE-GC-MS) by analyzing 21 urine samples.