Multiresidue method for identification and quantification of avermectins, benzimidazoles and nitroimidazoles residues in bovine muscle tissue by ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) using a QuEChERS approach (original) (raw)
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Journal of the Brazilian Chemical Society, 2016
A fast and sensitive method using high performance liquid chromatography-fluorescence detection (HPLC-FD) and ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) was developed and validated for the determination of avermectins residues in ovine muscle samples. QuEChERS (quick, easy, cheap, effective, rugged and safe) sample preparation based on acetonitrile extraction followed by partitioning with NaCl and Na 2 SO 4 and dispersive solid-phase extraction (d-SPE) clean-up with C18 was applied. Na 2 SO 4 was used instead of MgSO 4 due to lower amounts of co-extractives in the final extract. The procedure was validated according to the Commission Decision 2002/657/EC. The method showed determination coefficients (r 2) higher than 0.99, recoveries between 93.2 and 124.3% for spike levels between 0.5 and 2.0 times the maximum residues limit (MRL) values. The repeatability and intermediate precision RSD values ranged from 1 to 19%. Decision limits (CCα) and detection capabilities (CCβ) ranged from 10.7 to 59.4 μg kg-1 and 11.4 to 68.8 μg kg-1 , respectively. Method performance was successfully evaluated by analyzing real samples and proficiency test with a z-score in the range of ±1.
Meat Science, 2014
A multi-residue quantitative screening method covering 41 antibiotics from 7 different families, by ultra-highperformance-liquid-chromatography tandem mass spectrometry (UHPLC-MS/MS), is described. Sulfonamides, trimethoprim, tetracyclines, macrolides, quinolones, penicillins and chloramphenicol are simultaneously detected after a simple sample preparation of bovine muscle optimized to achieve the best recovery for all compounds. A simple sample treatment was developed consisting in an extraction with a mixture of acetonitrile and ethylenediaminetetraacetic acid (EDTA), followed by a defatting step with n-hexane. The methodology was validated, in accordance with Decision 2002/657/EC by evaluating the required parameters: decision limit (CCα), detection capability (CCβ), specificity, repeatability and reproducibility. Precision in terms of relative standard deviation was under 20% for all compounds and the recoveries between 91% and 119%. CCα and CCβ were determined according the maximum residue limit (MRL) or the minimum required performance limit (MRPL), when required.
Analytica Chimica Acta, 2010
A confirmatory method for the determination of residues of nine non-steroidal anti-inflammatory drugs and one metabolite in animal muscles has been developed. After enzymatic hydrolysis samples were extracted with acetonitrile and cleaned up using alumina and C 18 SPE cartridges. Liquid chromatography-tandem mass spectrometry was used for the separation and determination of analytes. The method was validated in bovine muscles, according to the Commission Decision 2002/657/EC criteria. Applicability of the method in the analysis of swine, horse and chicken muscles was checked by precision and recovery experiment. The influence of matrix effect on the quantification of nonsteroidal anti-inflammatory drugs residues was investigated. The method was used for the confirmation of phenylbutazone and oxyphenbutazone in horse muscle sample.
Talanta, 2012
A capillary electrophoresis (CE) method with ultraviolet detection has been developed for simultaneous detection and quantification of five nitroimidazoles including benzoylmetronidazole, dimetridazole, metronidazole, ronidazole, and secnidazole in porcine muscles. Nitroimidazoles in samples were extracted by ethyl acetate with subsequent clean-up by a strong cation exchange solid phase extraction column. The clean extracts were subjected to CE separation with optimal experimental conditions: pH 3.0 running buffer containing 25 mM sodium phosphate and 0.10 mM tetrabutylammonium bromide, 5 s hydrodynamic injection at 0.5 psi and 28 kV separation voltage. The nitroimidazoles could be monitored and detected at 320 nm within 18 min. The limits of detection were below 1.0 g/kg and limits of quantification were lower than 3.2 g/kg for all nitroimidazoles in the muscle samples. The recoveries and relative standard deviations were 85.4-96.0, 83.5-92.5, 1.3-3.9, and 1.1-4.2%, respectively for the intra-day and inter-day analyses. The proposed CE method has been successfully applied to determine nitroimidazoles in artificial porcine muscle samples with good accuracy and recovery, demonstrating that it has potential for detection and quantification of multi-nitroimidazole residue in real muscle samples.
Food Additives & Contaminants: Part A, 2011
A qualitative multiresidue method that facilitates rapid monitoring of veterinary drugs in porcine muscle is described. The method comprises the application of an innovative extraction/clean-up procedure, namely liquidliquid extraction with partition at very low temperature (LLE-FPVLT), and analysis by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). Besides the high selectivity, sensitivity and specificity, this high-throughput method proved to be quite general as 34 veterinary drugs (from six distinct classes: tetracyclines, sulfonamides, penicillins, quinolones, macrolides and benzimidazoles) could be successfully detected. The whole screening procedure was validated according to the directives from European Commission Decision 2002/657/EC and guidelines for the validation of screening methods. Acceptable values for the evaluation parameters were achieved for all analytes (except for ampicillin, clindamycin and erythromycin). Finally, these very promising results have strengthened the possibility of inclusion of such a methodology as an integral part of the National Residue Control Plan scope of the Ministry of Agriculture, Livestock and Food Supply of Brazil.
Journal of Chromatography B, 2009
A rapid LC-MS/MS method has been developed and validated for the simultaneous identification, confirmation and quantitation of 10 nitroimidazoles in plasma. The method validated in accordance with Commission Decision (CD) 2002/657/EC is capable of analysing for metronidazole (MNZ), dimetridazole (DMZ), ronidazole (RNZ), ipronidazole (IPZ) and their hydroxy metabolites MNZ-OH, HMMNI (hydroxymethyl, methyl nitroimidazole), IPZ-OH. The method is also capable of analysing carnidazole (CRZ), ornidazole (ORZ) and ternidazole (TRZ) which are rarely analysed by modern methods. MNZ, DMZ and RNZ have a recommended level (RL) of 3 ng mL −1 which this method is easily able to detect for all the nitroimidazole compounds. Plasma samples are extracted with acetonitrile, and NaCl is added to help remove matrix contaminants. The acetonitrile extract undergoes a liquid-liquid wash step with hexane; it is then evaporated and reconstituted in mobile phase. The reconstituted samples are analysed by liquid chromatography tandem mass spectrometry (LC-MS/MS). The decision limits (CC␣) range from 0.5 to 1.6 ng mL −1 and the detection capabilities (CC), range from 0.8 to 2.6 ng mL −1. The results of the interassay study, which was performed by fortifying bovine plasma samples (n = 18) on three separate days, show the accuracy calculated for the various analytes range between 101% and 108%. The precision of the method, expressed as CV% values for the inter-assay variation of each analyte at the three levels of fortification (3, 4.5 and 6.0 ng mL −1), ranged between 4.9% and 15.2%. A day 4 analysis was carried out to examine species variances in animals such as avian, ovine, porcine and equine.
Analytica Chimica Acta, 2007
Nitroimidazoles (Ronidazole, Dimetridazole, Metronidazole, Ipronidazole) and their hydroxy metabolites are banned substances with antibiotic and anticoccidial activity. They are suspected to be carcinogenic and mutagenic. Since nitroimidazoles showed an inhomogeneous distribution and a rapid degradation in incurred muscle samples, plasma is the preferred target matrix for residue analysis. The analytical method of Polzer et al. [J. Polzer, C. Stachel, P. Gowik, Anal. Chim. Acta 521 (2004) 189] was adapted for liquid chromatography-tandem mass spectrometry detection and was validated in house according to the Commission Decision 2002/657/EC. The method is specific for all nitroimidazole except for Ipronidazole and its metabolite, due to interferences at their retention times in chromatograms of blank plasma and reagents samples. The absence of a matrix effect enables the use of a (linear) calibration curve in solution for quantitation. The apparent recovery (obtained after correction with a deuterated internal standard) is between 93% and 123%, except for the metabolite of Metronidazole (58-63%). The repeatability (CVr = 2.49-13.39%) and intralaboratory reproducibility (CVRW = 2.49-16.38%) satisfy the Horwitz equation. The obtained values for the detection capacity (CC) range from 0.25 to 1 g L−1, while values obtained for the decision limit (CC␣) are below CC. Summary of the procedure followed to determine all validation parameters. Each sample was performed in triplicate (replicate # 1, 2 and 3) at each concentration level. All samples analyzed on the same day (day # 1, 2 or 3) were injected with a solution standard calibration curve and with a week in between Day #1 Day #2 Day #3 1/4 "MRPL" Replicate #1 Replicate #4 Replicate #7 Replicate #2 Replicate #5 Replicate #8 Replicate #3 Replicate #6 Replicate #9 ½ "MRPL" Replicate #1 Replicate #4 Replicate #7 Replicate #2 Replicate #5 Replicate #8 Replicate #3 Replicate #6 Replicate #9 1× "MRPL" Replicate #1 Replicate #4 Replicate #7 Replicate #2 Replicate #5 Replicate #8 Replicate #3 Replicate #6 Replicate #9 1.5× "MRPL" Replicate #1 Replicate #4 Replicate #7 Replicate #2 Replicate #5 Replicate #8 Replicate #3 Replicate #6 Replicate #9 2× "MRPL" Replicate #1 Replicate #4 Replicate #7 Replicate #2 Replicate #5 Replicate #8 Replicate #3 Replicate #6 Replicate #9
Food Control, 2013
A simple and inexpensive sample preparation method based on solvent extraction, followed by low temperature cleanup, was demonstrated to be applicable for the determination of avermectin and milbemycin residues in bovine muscle by liquid chromatography-tandem mass spectrometry (LC-MS/ MS) and liquid chromatography with fluorescence (LC-FL) detection. The analytical methodology was validated according to the Commission Decision 2002/657/EC, using LC-MS/MS for confirmatory and LC-FL for quantitative purposes. Mean recovery was between 88.9 and 100.7% in three distinct concentrations. The coefficient of variation for repeatability and within-laboratory reproducibility ranged from 0.78 to 5.1% and from 0.28 to 9.0%, respectively. Method precision led to satisfactory values of decision limits (CCa) and detection capabilities (CCb). The proposed method has been applied in the Brazilian National Residue Control Plan since 2010 for the determination of avermectins and milbemycin residues in bovine muscle samples. A total of 760 samples were analyzed and none of them presented residues at concentrations above the permitted levels established by the more recently applied directives.
Journal of pharmaceutical and biomedical analysis, 2017
Veterinary drugs containing synthetic anabolic steroid and nitroimidazole active agents are not allowed for their applications in livestock of the European Union (EU). This paper presents analyses of twelve selected steroids and six nitroimidazole antibiotics at low levels (1.56μg/L-4.95μg/L and 0.17μg/kg-2.14μg/kg, respectively) in body fluids and egg incurred samples. Analyses involved clean-up procedures, high performance liquid chromatography (HPLC) separation, and tandem mass spectrometric screening and confirmatory methods. Target steroids and nitroimidazoles in samples were cleaned by two independent supported liquid extraction and solid phase extraction procedures. Separation of the selected compounds was conducted on Kinetex XB C-18 HPLC column using gradient elution. The screening methods utilised supported liquid extraction that enabled fast and cost effective clean-up. The confirmatory methods were improved by extending the number of matrices and compounds, and by introd...
Meat science, 2014
In contrast with the information of the inspection body concerning the use of ronidazole, several non compliant muscle samples were detected using a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method in accordance with confirmation criteria of Decision 2002/657/EC. This led to the suspicion that non compliance could be due to false positive results. In this context, a liquid chromatography-high resolution mass spectrometry (LC-HRMS) method was developed and sample extracts were re-analyzed, resolving the co eluting isobaric interfering peak, which also has an interfering product ion with the transition product (m/z 201 N 140).