New means to approach the detection of anabolic steroids in meat

In recent years, the analysis of veterinary drugs and growth-promoting agents has shifted from target-oriented procedures, mainly based on liquid chromatography coupled to triple-quadrupole mass spectrometry (LC-QqQ-MS), towards an accurate mass full scan MS type of approach (such as time of flight (ToF) or Fourier Transform (FT) MS). The applicability of an HR-Orbitrap-MS for confirmatory analysis for growth-promoting agents in meat was compared, in a previous study, to that of a QqQ-MS but proved somewhat inferior with respect to the sensitivity. Therefore our goal was to evaluate the sensitivity of a TripleTOF™ (QqTOF) instrument for anabolic steroids in meat, using accurate mass MS data (> 25,000 FWHM) in combination with MS/MS acquisition at high acquisition speeds. For the development of a sensitive detection method, the fast Information Dependent Acquisition (IDA) combining a TOF-MS survey scan and dependent TOF-MS/MS scans was compared to a scheduled and a more time consuming dedicated MRM algorithm. As a final attempt to increase the sensitivity, the analytical flow UHPLC was switched to a state-of-the-art microflow UHPLC, operating at a flow rate of 25 μL/min. Additionally, a full fractional statistical design was performed to simplify as well as optimize the current extraction procedure for anabolic steroids in meat. A review of the literature resulted in a selection of a subset of relevant parameters including e.g. variables related to the homogenisation of the meat (e.g. ultraturrax), solvents (e.g. solvent type and volume), sample clean-up (e.g. solid phase extraction) for optimisation. Afterwards the optimised method was validated according to CD 2002/657/EC. In this study the benefits of the TripleTOF™ instrument are compared to a QqQ-MS system including it’s ability to mine the data retrospectively for identifying non-targeted and emerging designer hormones, which serves as an additional advantage in the light of the national control plans. Compared to the single stage Orbitrap the TripleTOF™, operates at a lower resolution, however due to the acquired MS/MS data it is expected to provide an advantage especially for those compounds (i.e. some estrogens) for which the threshold of 20% intra-laboratory reproducibility, correlated to an inferior precision, was exceeded with the single stage Orbitrap. The microLC might also provide an additional sensitivity increase