Development of a subcritical water extraction approach for trace analysis of chloramphenicol, thiamphenicol, florfenicol, and florfenicol amine in poultry tissues

Subcritical water extraction was investigated as a novel and alternative technology for the separation of trace amounts of chloramphenicol, thiamphenicol, florfenicol and its major metabolite florfenicol amine from poultry tissues and its results were compared with those of conventional shaking extraction, ultrasonic extraction, and pressurized liquid extraction. Decreasing the polarity of water by successively increasing the extraction temperature from 50 °C to 200 °C at the moderate pressure enabled selective, highly effective extractions to be performed. Rapid quantification of the target compounds was carried out by ultra-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS). The critical parameters of subcritical water extraction such as solvent modifier, temperature, pressure, extraction time, and static cycles were varied with control. The optimized extraction procedures using subcritical water as extraction solvent, were carried out on a pressurized liquid extractor operated at 150 °C and 100 bar, applying two static cycles for 3 min. Average recoveries of the four analytes from fortified samples ranged between 86.8% and 101.5%, with relative standard deviations (RSDs) lower than 7.7%. The limits of detection (LODs) and quantification (LOQs) for the target compounds were in the ranges of 0.03-0.5 μg kg−1 and 0.1-2.0 μg kg−1, respectively. The proposed method is fast, sensitive, water-based thus more environmental acceptable, making it a suitable replacement for conventional organic solvent extraction in veterinary drug residue analysis.