Analysis of non-steroidal anti-inflammatory drugs in milk using QuEChERS and liquid chromatography coupled to mass spectrometry: triple quadrupole versus Q-Orbitrap mass analyzers

We developed a Quick, Easy, Cheap, Effective, Rugged, and Safe (QuEChERS) method for the high throughput determination of 10 non-steroidal anti-inflammatory drugs (NSAIDs) in milk samples using high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) with a triple quadrupole (QqQ) instrument and an electrospray ionization (ESI) source. The new extraction procedure is highly efficient, and we obtained absolute recoveries in the range 78.1-97.1 % for the extraction and clean-up steps. Chromatographic separation is performed in the gradient mode with a biphenyl column and acidic mobile phases consisting of water and acetonitrile containing formic acid. The chromatographic run time was about 12 min, and NSAID peaks showed a good symmetry factor. For MS/MS detection, we used multiple reaction monitoring (MRM) mode, using ESI in both positive and negative modes. Our method has been validated in compliance with the European Commission Decision 657/2002/EC, and we obtained very satisfactory results in inter-laboratory testing. Furthermore, we explored the use of a hybrid high resolution mass spectrometer, combining a quadrupole and an Orbitrap mass analyzer, for high resolution (HR) MS/MS detection of NSAIDs. We achieved lower NSAID quantification limits with Q-Orbitrap high resolution mass spectrometry (HRMS/MS) detection than those achieved with the QqQ instrument; however, its main feature is its very high selectivity, which makes HRMS/MS particularly suitable for confirmatory analysis

Analysis of corticosteroids in samples of animal origin using QuEChERS and ultrahigh-performance liquid chromatography coupled to high-resolution mass spectrometry.

A rapid and sensitive method for the confirmatory analysis of 8 synthetic corticosteroids (betamethasone, dexamethasone, prednisolone, 6 methyl prednisolone, triamcinolone, flumethasone, beclomethasone, fluocinolone acetonide) is proposed. The method is useful for detecting illegal treatments in different animal species. It consists of an extraction and clean-up using the Quick Easy Cheap Effective Rugged Safe (QuEChERS) strategy. Quantitative determination is achieved by ultra-high performance liquid chromatography coupled to high-resolution mass spectrometry with heated electrospray ionization in negative mode. Quantification is performed using surrogate matrix matched standard calibration curve with dexamethasone-D4 as the internal standard. The method was validated for analyzing liver samples according to the criteria established by Decision 2002/657/EC. Linearity was assessed in the 1-10 μg kg-1 range and linear correlation coefficients were over 0.99 for all the analytes. CCα ranged from 0.04 to 0.16 μg kg-1 for substances without maximum residue limit. The method allows confident quantification and confirmation of corticosteroids in liver samples, and its simplicity makes it suitable for analyzing large numbers of samples

Noves propostes per a l’anàlisi de medicaments d’ús veterinari i de toxines marines en aliments destinats al consum humà

[cat] El sector de la producció ramadera i de l’aqüicultura són pilars fonamentals de la indústria alimentària, que té plantejats grans reptes, com el de poder posar a l’abast de la població quantitat suficient d’aliments i que aquests siguin saludables als consumidors. La indústria alimentària ha anat optimitzant els seus mecanismes de producció d’aliments, i a dia d’avui són molt esteses les pràctiques de producció intensiva, que es caracteritzen per una productivitat molt elevada amb un cost econòmic relativament baix. Aquests tipus d’enfocament utilitza medicaments veterinaris per tractar els animals malalts, però també amb finalitats profilàctiques, per minimitzar el risc de contagi. L’ús de medicaments pot comportar la presència de residus dels fàrmacs en els productes alimentaris destinats a consum humà, i el risc associat és un tema que preocupa la societat. Amb l’objectiu d’oferir a la població l’accés a aliments segurs i saludables, s’han establert límits màxims de residu (LMR) per a medicaments d’us veterinari o bé prohibició per d’altres, atenent a la seva toxicitat. No és aquest l’únic àmbit on es regula la presència als aliments de compostos que poden suposar un risc per als consumidors; podem citar els pesticides, els contaminants ambientals, les toxines d’origen natural, etc. El repte pels laboratoris de control és el de poder disposar de mètodes d’anàlisi prou sensibles i selectius com per poder donar compliment a la legislació. A la vegada aquests mètodes convé que siguin suficientment transversals com per poder ser aplicats a diferents matrius alimentàries, les quals poden ser de naturalesa molt diversa pel que fa a la seva composició. Els mètodes han de ser d’elevada fiabilitat, però també ràpids i senzills. En aquesta tesi, s’han posat a punt mètodes per l’anàlisi de residus de 3 famílies de medicaments veterinaris (avermectines, nitroimidazols i antiinflamatoris no estereoideus); moltes de les substàncies considerades tenen establerts LMR, però algunes són prohibides. Per altra banda, s’ha posat a punt un mètode d’anàlisi de toxines marines lipofíliques en mol·luscs bivalves. Les toxines estudiades tenen nivells màxims admissibles legislats.[eng] Livestock plays an important role in food industry. In the last decades, the mechanisms for food production have been optimized, based mainly in an intensive livestock farming and, as a consequence an increasing use of veterinary drugs to raise the animals. As a result of the use of veterinary drugs in livestock raising, residues can be present in foodstuff for human consumption. Nowadays, this issue is currently becoming more and more important. Also some natural processes occurring in oceans and seas (red tides) may produce natural toxins that contaminate foodstuff, such as the bivalves. Food safety laboratories need to control the presence of such residues in food of animal origin. Therefore, reliable, easy and fast analytical methods should be used in order to control an increasing amount of samples. In this thesis, simple analytical methods for the analysis of avermectins, nitroimidazoles, non- steroidal anti-inflammatory drugs (NSAIDs) and lipophilic marine biotoxins (MBTXs) have been set up and validated. These methods are base in the QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) strategy, a method that was first published for pesticide residue analysis and nowadays becoming widely used. Some of the compounds have a published MRL (Maximum Residue Limit) in some matrices of animal origin, but because of their mutagenic and carcinogenic properties, some nitroimidazoles are included in the list of prohibited substances of Regulation (EU) No 37/2010, namely dimetridazole, metronidazole and ronidazole. The instrumentation involved in the present analytical methods is UHPLC-MS/MS, (triple quadrupole, QqQ) using an electrospray ionization (ESI) interface in Multiple Reaction Monitoring (MRM) mode. The use of high resolution mass spectrometry (HRMS), with Q-Orbitrap mass analyzer, has been tested as well. The use of HRMS has permitted to resolve coeluting isobaric interferences which did not permit confirmation of analytical results by QqQ. Also due to its excellent selectivity, very low regulated limits could be achieved. This is the case of diclofenac, a NSAIDs with a very low MRL in milk (0.1 µg·kg-1). Different working modes of the Q-Orbitrap mass analyzer have been tested, among them, t-SIM-ddpp (target-sim-data-dependent) and t-MS/MS (target- MS/MS). Validation of the methods included in this thesis has been performed following European legislation rules included in the 657/2002/CE Decision with excellent results. CCα (decision limit) and CCβ (detection capability) have been calculated as well. Limits of quantification (LQ) of the analytical methods for veterinary drugs residues range between 0.5- 2.5 µg·kg-1. For MBTX, LQ has been set up at 25 µg·kg-1. The validated matrices are muscle, milk and mussel, but all methods have been tested in alternatives matrices with successful results. Avermectin analysis has been tested in liver and milk; nitroimidazole analysis in kidney and liver, NSAIDs analysis in kidney and MBTXs analysis in canned and processed bivalves. The methods have been implemented in the Laboratori de l’Agència de Salut Pública de Barcelona for control analysis and are used for routine analysi

Analysis of non-steroidal anti-inflammatory drugs in milk using QuEChERS and liquid chromatography coupled to mass spectrometry: triple quadrupole versus Q-Orbitrap mass analyzers

We developed a Quick, Easy, Cheap, Effective, Rugged, and Safe (QuEChERS) method for the high throughput determination of 10 non-steroidal anti-inflammatory drugs (NSAIDs) in milk samples using high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) with a triple quadrupole (QqQ) instrument and an electrospray ionization (ESI) source. The new extraction procedure is highly efficient, and we obtained absolute recoveries in the range 78.1-97.1 % for the extraction and clean-up steps. Chromatographic separation is performed in the gradient mode with a biphenyl column and acidic mobile phases consisting of water and acetonitrile containing formic acid. The chromatographic run time was about 12 min, and NSAID peaks showed a good symmetry factor. For MS/MS detection, we used multiple reaction monitoring (MRM) mode, using ESI in both positive and negative modes. Our method has been validated in compliance with the European Commission Decision 657/2002/EC, and we obtained very satisfactory results in inter-laboratory testing. Furthermore, we explored the use of a hybrid high resolution mass spectrometer, combining a quadrupole and an Orbitrap mass analyzer, for high resolution (HR) MS/MS detection of NSAIDs. We achieved lower NSAID quantification limits with Q-Orbitrap high resolution mass spectrometry (HRMS/MS) detection than those achieved with the QqQ instrument; however, its main feature is its very high selectivity, which makes HRMS/MS particularly suitable for confirmatory analysis

Analysis of corticosteroids in samples of animal origin using QuEChERS and ultrahigh-performance liquid chromatography coupled to high-resolution mass spectrometry.

A rapid and sensitive method for the confirmatory analysis of 8 synthetic corticosteroids (betamethasone, dexamethasone, prednisolone, 6 methyl prednisolone, triamcinolone, flumethasone, beclomethasone, fluocinolone acetonide) is proposed. The method is useful for detecting illegal treatments in different animal species. It consists of an extraction and clean-up using the Quick Easy Cheap Effective Rugged Safe (QuEChERS) strategy. Quantitative determination is achieved by ultra-high performance liquid chromatography coupled to high-resolution mass spectrometry with heated electrospray ionization in negative mode. Quantification is performed using surrogate matrix matched standard calibration curve with dexamethasone-D4 as the internal standard. The method was validated for analyzing liver samples according to the criteria established by Decision 2002/657/EC. Linearity was assessed in the 1-10 μg kg-1 range and linear correlation coefficients were over 0.99 for all the analytes. CCα ranged from 0.04 to 0.16 μg kg-1 for substances without maximum residue limit. The method allows confident quantification and confirmation of corticosteroids in liver samples, and its simplicity makes it suitable for analyzing large numbers of samples