Abstract
In accordance with Commission Decision 2002/657/EC, confirmatory methods for the detection of prohibited substances should comply with specific requirements, including the criteria for confirmation. Two strategies: multiple reaction monitoring (MRM) and enhanced product ion (EPI) scanning functions were compared for confirming the anabolic compounds from synthetic stilbenes group in bovine urine samples. In the research, twenty samples fortified at the Recommended Concentration (RC) of 1 µg L−1 with diethylstilbestrol, dienestrol and hexestrol were analyzed by liquid chromatography-tandem mass spectrometry on a QTRAP 5500 instrument. The analytical procedure, validated in accordance with the Commission Decision 2002/657/EC, used in the official control of hormones in Poland was applied. The validation parameters were in agreement with 2002/657/EC performance criteria. The effectiveness of MRM and EPI scanning modes for confirmation purposes was evaluated based on the percentage of the results confirmed. In all urine samples recorded in the MRM mode, the confirmation criteria (retention time, relative intensities between transitions) have been fulfilled. The presence of stilbenes in all urine samples using EPI scan mode was confirmed too as evidenced by a good matching of stilbenes spectra in the samples to the reference spectra with critical match factor above 0.7. The results of the research show that EPI scanning function provides the same effectiveness for confirmation of banned compounds as the mostly used MRM scan mode and can be an additional tool to confirm the doubtful case results in the analysis of hormones residues, even at such low concentration levels.
Highlights
IntroductionFor effective control of illegal use of hormones and the determination of residues in samples from the area of food safety, highly specific and sensitive analytical methods are required [4]
In the European Union, the use of hormonal growth-promoting active drugs is prohibited for fattening purposes under Council Directive 96/22/EC in order to protect consumers against residues with possible toxic effects on public health [1,2,3].For effective control of illegal use of hormones and the determination of residues in samples from the area of food safety, highly specific and sensitive analytical methods are required [4]
For the compounds being the subject of research, the confirmatory criteria required for the multiple reaction monitoring (MRM) and enhanced product ion (EPI) scan modes were checked
Summary
For effective control of illegal use of hormones and the determination of residues in samples from the area of food safety, highly specific and sensitive analytical methods are required [4]. Confirmatory methods for the detection of banned, hormonally active substances in biological samples of animal origin must be approved and meet special requirements including the criteria for identification as defined in Commission Decision 2002/657/EC, the norm obligatory in all Member States of the European Union [5, 6]. A number of chromatographic methods, including gas chromatography (GC) and liquid chromatography (LC) in combination with different types of mass spectrometry (MS) have been demonstrated to determine hormones residues at low concentration levels in samples of animal origin [7,8,9,10,11,12]. LC tandem mass spectrometry (MS/MS) with triple quadrupole (QqQ) mass analyzers operating in
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