Comparison of Sample Preparation and Determination of 60 Veterinary Drug Residues in Flatfish Using Liquid Chromatography-Tandem Mass Spectrometry.

This study was performed to optimize the analytical method for multi-residues of 60 compounds in flatfish samples. Three sample preparation methods were tested to identify the optimal recovery conditions for target analytes. As a result, 10 mL of water/acetonitrile (1:4, v/v) was used to extract analytes from fish samples. For purification, C18 and 10 mL of acetonitrile saturated hexane were used to treat the samples. After evaporation and reconstitution, the fish samples were analyzed by ultra-performance liquid chromatography-tandem mass spectrometry. The proposed method was validated according to the CODEX guidelines (CAC/GL-71). Our results showed the recoveries of 73.2%-115% and coefficients of variation of 1.6%-22.1%. The limit of quantification was 0.0005-0.005 mg/kg in the fishery products. In analysis of real samples, no samples exceeded the limit of quantification. This analytical method can be used for multi-residue screening and confirmation of the residues of veterinary drugs in fishery products.

Multiresidue Screening of Veterinary Drugs in Meat, Milk, Egg, and Fish Using Liquid Chromatography Coupled with Ion Trap Time-of-Flight Mass Spectrometry.

New approaches to veterinary drug screening based on liquid chromatography-mass spectrometry (LC-MS/MS) and time-of-flight mass spectrometry (ToF/MS) are rapid and have high selectivity and sensitivity. In this study, we developed a multiresidue method for screening over 100 veterinary drug residues using ion trap (IT)-ToF/MS. The screened compounds comprised major drug classes used in veterinary practice, representing the following: amphenicols, anthelmintics, benzimidazoles, ß-lactams, coccidiostats, ionophores, macrolides, non-steroidal anti-inflammatory drugs, quinolones, sulfonamides, tetracyclines, and tranquilizers. The method was developed based on chromatographic retention time, specific accurate mass, isotope distribution, and fragment data. Each compound was validated at three levels, and the mass accuracy, accuracy, and repeatability were calculated. All parameters showed acceptable values and conformed to the Commission Decision 2002/657/EC criteria. This screening method can simultaneously analyze over 100 veterinary drugs in meat, milk, eggs, and fish in a single analytical run.

Veterinary drug residues in domestic and imported foods of animal origin in the Republic of Korea.

The Korean National Residue Programme comprises three different approaches for evaluating domestic and imported foods of animal origin: monitoring, surveillance/enforcement and an exploratory test programme. Monitoring and surveillance/enforcement testing programmes are routinely implemented by 17 Provincial Veterinary Services for domestic products and regional offices of the Animal and Plant Quarantine Agency (QIA) for imported products. The exploratory project conducted at QIA headquarters is designed to test substances that are not included in monitoring and enforcement testing programmes. Here, we carried out exploratory testing for determining the presence of 42 veterinary drugs that have no established Korean maximum residue limits and analysed their levels simultaneously, in a total of 3108 samples of domestic and imported animal-origin foods. Of the tested drugs, acetylsalicylic, paracetamol, clopidol, diclazuril, amprolium, toltrazuril and its metabolites (toltrazuril sulphone and toltrazuril sulphoxide) and phenylbutazone and its metabolites (oxyphenylbutazone) were detected.

Chemical residues and contaminants in foods of animal origin in Korea during the past decade.

Residues of veterinary drugs, pesticides, and environmental contaminants in domestic and imported foods of animal origin were monitored by the National Residue Program and inspection service in Korea in the past decade. In all, 134 substances were analyzed in the monitoring plan; 35 substances were examined in the surveillance and enforcement testing program, and 27 substances were investigated in exploratory projects. The overall trend of violation rates gradually decreased over the past decade. Pesticides were not found in any domestic samples of animal origin. The violation rates of chlortetracycline and oxytetracycline decreased, but quinolone and penicillin detections increased in Korea. Several kinds of residue violations of veterinary drugs, endosulfan, or dioxins were found in the imported products each year. In an example event in 2008, the Korea monitoring plan contributed globally to investigate the dioxin contamination from Chilean pork. Continuous monitoring based on internationally harmonized standards and methods provides the essential scientific basis to manage and ensure food safety.

Evaluation of linear regression statistical approaches for withdrawal time estimation of veterinary drugs.

The safety of foods of animal origin requires the determination of the time at which veterinary drug residues in edible tissues are below a given maximum residue limit (MRL). For this reason, a certain withdrawal time estimate is determined for drugs based on statistical evaluation of concentrations determined by analytical analysis of residues in target organs of healthy animals. The purposes of this paper is to evaluate the linear regression statistical approach for the estimation of withdrawal time of veterinary drugs as recommended by Food and Drug Administration (FDA) and European Union/Committee for Medicinal Products for Veterinary use (EU/CVMP) and compare the application with a real model example. The withdrawal time estimate of the model has shown 2-5 days difference for the increase in tolerance limit from 95% (EU) to 99% (FDA) when calculated including censored data. But when it was excluded the range increased to 2-8 days for the same increase in tolerance. Furthermore, wider range of difference (3-21 days) and variation in significance was observed with inclusion/exclusion of censored data at the same level of tolerance. In conclusion, this study suggests inclusion/exclusion of censored data should be dependent on satisfying the statistical assumptions required rather than always including/excluding.

Translucency and masking ability of various opaque-shade composite resins.

OBJECTIVES: The aims of this study were to evaluate the adequate thickness of opaque resins for situations such as an oral black cavity and discolored tooth structure, as well as the translucency of each opaque material at various thicknesses. METHODS: Six opaque-shade composite resins (Z-350 OA3, Amelogen Universal A2O, Esthet-X A2O, Esthet-X A4O, Charmfil UO, and Aelite Universal OA3) were prepared in metal molds with a hole of 8mm in diameter and various thicknesses (0.5mm, 1.0mm, 1.5mm, 2.0mm, 2.5mm, 3.0mm and 4.0mm). Four backgrounds (white tile, black tile, C4 shade porcelain and opaque resin itself) were used to determine the translucency parameter (TP) (between black and white backgrounds), and to mimic a black oral cavity (between black and opaque resin backgrounds) and a discolored tooth structure (between C4 and opaque resin backgrounds). Color measurements were made by a colorimeter to determine the CIELAB values of each specimen with each background and to calculate the TP and DeltaE* value difference among the specimens on the backgrounds. RESULTS: TP and DeltaE* obtained between black and opaque resin backgrounds decreased in similar pattern as thickness increased. A C4 background was masked by resin thicknesses of 0.5-1.0mm, while a black background required thicknesses of 1.0-2.0mm. SIGNIFICANCE: This study can provide helpful guidelines for the esthetic restoration of discolored tooth structures or 'through and through' class III and IV cavities with composite resin materials.

Multiplex polymerase chain reaction detection of black-pigmented bacteria in infections of endodontic origin.

The purpose of this study was to detect the presence of Porphyromonas endodontalis, P. gingivalis, Prevotella intermedia, P. nigrescens, and P. tannerae from clinical samples using multiplex polymerase chain reactions (PCR). Two different multiplex PCR protocols were used (one for the two Porphyromonas species and the other for the three Prevotella species), each one using a primer pair specific for each target species. The results were compared to those of the conventional culture procedures. Microbial samples were taken aseptically from 40 infected root canals and abscesses from patients. Samples were cultured in an anaerobic condition for conventional identification using a Rapid ID 32 A kit. Multiplex PCR was processed using the DNA extracted from each sample. At least one of the five species of black-pigmented bacteria (BPB) were detected in 65% (26 of 40) of the samples using multiplex PCR, and in 15% (6 of 40) using the conventional culture procedures. Multiplex PCR was more rapid, sensitive, specific, and effective in detecting BPB than the conventional culture procedures.