Validation of an UHPLC-MS/MS Method for Screening of Antimicrobial Residues in Eggs and Their Application to Analyses of Eggs from Laying Hens Subjected to Pharmacological Treatment (original) (raw)
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Quantification of antibiotic residues in raw and layers hen eggs by rp-hplc
International journal of health sciences
Antibiotics (Oxytetracycline & Enrofloxacin) residual levels in raw and layered samples are to be quantified using RP-HPLC. The antibiotic (oxytetracycline and enrofloxacin) residual levels in raw and layers hen egg samples were evaluated qualitatively by high performance liquid chromatography in this investigation, which used one hundred randomly selected samples (HPLC). C18 (Hypersil ODS-BPS, 250 4.6mm; 5) was used for the separation at a flow rate of 1 ml/min in a mobile phase of 0.1 percent formic acid: acetonitrile (50:50, v/v). At a detection wavelength of 350 nm, the residues were measured. Found oxytetracycline residues, which indicate widespread usage of antibiotics on farms and a lack of application of required withdrawal durations, were detected in samples. Because of this, the antibiotic residues found in broiler hen eggs have been reduced. Restrictive methods and stricter restrictions should be implemented to prevent the presence of abuse residues prior to marketing, ac...
Journal of Chromatography A, 2011
A liquid chromatography tandem-mass spectrometry (LC-MS/MS) method has been developed for the simultaneous determination of 22 veterinary drug residues, belonging to eight classes (coccidiostats, lincosamides, macrolides, tetracyclines, sulfonamides, benzimidazoles, diterpenes, and diaminopyrimidines), in eggs. Chromatographic separations were achieved on an XBridge BEH C18 column (150 × 2.1 mm, 3.5 μm, Waters, USA) maintained at 35°C. The mobile phase was eluted at 400 μL min −1 in gradient mode between water and methanol/acetonitrile (20:80 v/v), both containing 0.1 % formic acid. The samples were prepared by protein precipitation with acetonitrile without additional cleanup steps. The method was successfully validated according to the Commission Decision 2002/657/EC and was demonstrated to be highly selective and free of matrix and residual effects. The method presented low limits of detection (0.37 to 7.5 μg kg −1) and quantification (1.25 to 20 μg kg −1). For banned substances, the decision limit values (CCα) and detection capability (CCβ) were 0.62-7.5 and 0.65-8.1 μg kg −1 , respectively. For substances with a maximum residue limit, the CCα and CCβ values were 2.15-1061.5 and 2.3-1135.9 μg kg −1 , respectively. All calibration curves showed excellent correlation (r ≥ 0.99). The recovery of the analytes and internal standards (49.0-103.7 %) was reached with high precision (RSD <8 %). At different concentration levels, the variations in precision and accuracy, in terms of repeatability and in-laboratory reproducibility, were <11 %. The new method is applicable to the routine analysis of commercial egg samples.
Journal of Chromatography A, 2009
a b s t r a c t A simple and rapid method able to determine residues of erythromycin A, tylosin and tilmicosin in whole eggs is presented here. The analytical protocol involves a one-step extraction followed by liquid chromatography (LC)-tandem mass spectrometry. Analytes were extracted from 1 g of egg spiked with an internal standard (josamycin) with acetonitrile. In terms of accuracy, matrix effect and ion signal stability, no extract cleanup was found to be necessary. After partial solvent removal, the final extract was injected into the LC column. Extraction was effective, since absolute recovery of the analyte in egg at their maximum residue limit (MRL) level was 85-102%. Estimated limits of quantification (S/N = 10) were 0.2-0.5 ng/g. Based on the EU Commission Decision 2002/657/EC, the method was in-house validated in terms of ruggedness, specificity, linearity, within-laboratory reproducibility, decision limit (CC˛) and detection capability (CCˇ). The within-laboratory reproducibility, expressed as RSD (n = 18 at the MRL levels), was not higher than 13%. After validation, a short study on EA depletion in eggs was conducted after administration of this drug to laying hens.
Determination of Antibacterial Drug Residues in Commercial Eggs Distributed in Urmia, Iran
Journal of food quality and hazards control, 2015
Background: The presence of antibacterial drug residues in food caused many concerns for consumers. This study was designed to determine of antibacterial drug residues in commercial eggs distributed in Urmia, Iran. Methods: Disc diffusion microbial inhibition test was performed using three media seeded with Bacillus subtilis at different pH values (6, 7.2 or 8) and a fourth medium seeded with Micrococcus luteus. Two hundred commercial eggs were collected randomly and examined for antibacterial drug residues. Standard solutions of antibiotics were used as control. Results: The results of this study revealed that twenty five samples (12.5%) of the prepared eggs were positive for antibacterial substances and all of them were related to macrolides group. Conclusion: The presence of these residues in eggs could cause health hazards like hypersensitivity reaction, development of resistant organisms to these antibacterial agents, destruction of gastrointestinal natural flora, etc. In conclusion, the antibiotic residues in commercial eggs have to be monitored as routine test due to their side effects on human health.
Novel Rapid Screening Technique for Antibiotic Residues in Desi-Chicken Eggs
Journal of emerging technologies and innovative research, 2018
Extensive use of antibiotics in chickens causes a serious public health issue because of reasonable alarm regarding harmful concentrations of residues to be present in the edible animal products (meat, milk and eggs). Currently, the trend is increasing towards consumption of desi chicken eggs because of awareness about the presence of antibiotic residues in commercial eggs. However, there is a possibility that some small-scale poultry farmers who are rearing desi chicken for the commercial purpose may also use antibiotics. Therefore, this study was carried out to determine the antimicrobial drug residues in desi chicken eggs obtained from Tamil Nadu. Microbial tube test method was performed using the bacterial strain Bacillus subtilis and Sixty desi eggs were collected randomly and examined for antimicrobial drug residues. The results of this study revealed that no egg samples were positive for antimicrobial residues related to any class of antibiotics.
A survey of antibiotic residues in commercial eggs in Kermanshah, Iran
The Iranian Journal of Veterinary Science and Technology, 2015
Poultry nutrition or inappropriate treatments by antibiotics can lead to production of eggs with antibiotic residues. The aim of the present study is to assess the prevalence of drug residues in eggs presented in Kermanshah, Iran. For this purpose, 120 eggs with the average weight of 60 grams were collected randomly from the supermarkets in Kermanshah and later were tested for antibiotic residues using the Four-Plate Test (FPT) method. The results showed that 3.3% of the tested eggs had antibiotic residues. The main residues in egg yolk was due to tetracycline. Aminoglycosides were also detected as minor contaminants. Accordingly, antibiotic residues in eggs presented in Kermanshah are a threat for human consumption and therefore strong supervision of the authorities is essential.
Animals
Eggs are the main product generated from backyard poultry production systems (BPS) because they can quickly be consumed and sold to meet essential family needs. Nevertheless, antimicrobial residues can accumulate in this product. The objective of this study was to evaluate the presence of antimicrobial residues in eggs produced by poultry kept in BPS in central Chile. To assess this, eggs were obtained from 83 BPS and analysed to evaluate the presence of antibiotic residues (families: tetracyclines, beta-lactams, aminoglycosides and macrolides), using a Four-Plate Test screening method for the detection, based on a bacterial growth inhibition method. Results show a lack of biosecurity procedures at BPS level, making these systems susceptible to the dissemination of antimicrobial residues. These include intensive animal production units in the proximity, and the presence of shared watercourses with other farms. Furthermore, 66% of the surveyed owners are indicated as giving pharmacol...
Journal of Chromatography A, 2009
a b s t r a c t A simple and rapid method able to determine residues of seven quinolone antibacterials in whole eggs is presented here. This method is based on the matrix solid-phase dispersion technique with hot water as extractant followed by liquid chromatography-tandem mass spectrometry. After depositing 1.5 g of an egg sample containing the analytes and the analyte surrogate (norfloxacin) on sand (crystobalite), this material was packed into an extraction cell. Quinolones were extracted by flowing 6 mL of water acidified with 50 mmol/L formic acid through the cell heated at 100 • C. After pH adjustment and filtration of the extract, 100 L of it was injected into the LC column. MS data acquisition was performed in the multiple reaction monitoring mode, selecting two precursor ion to product ion transitions for each target compound. Hot water appeared an efficient extracting medium, since absolute recoveries of the analyte in egg at the level of 20 ng/g were 89-103%. Estimated limits of quantification (S/N = 10) were 0.2-0.6 ng/g. Based on the EU Commission Decision 2002/657/EC, the method was validated in terms of ruggedness, specificity, linearity, within-laboratory reproducibility, decision limit (CC˛and detection capability (CCˇ). Depending on the particular analyte, CC˛s ranged between 0.41 and 2.6 ng/g, while CCˇs were 0.64-3.7 ng/g. The method was linear in the 3-30 ng/g range, with typical R 2 values higher than 0.97. The within-laboratory reproducibility (n = 21) at 6 ng/g level was in the 9.0-12% range. After validation, a depletion study of enrofloxacin and one of its metabolites, i.e. ciprofloxacin, in eggs was conducted.
Antimicrobial residues in tissues and eggs of laying hens at Chittagong, Bangladesh
Aim: Antimicrobial residue in animal food products is an important index of food safety. Antimicrobial residues could result from chemotherapeutic or chemoprophylactic use of drugs in food animals. This occurrence of residue in animal food products has received enormous worldwide attention from some local, international, and public health agencies. A cross-sectional study was conducted from July to December 2009 to detect the antibiotic residues in tissues and eggs of laying hens at Chittagong of Bangladesh. Materials and Methods: Microbial inhibition test (MIT) and thin layer chromatography (TLC) methods were used to detect antibacterial residues in poultry tissues (liver, kidney, breast, and thigh muscles) and eggs. The bacteria and pH of the MIT method were as follows: Bacillus subtilis on test agar medium with a pH of 7.2, Bacillus cereus with a pH of 6.0, and Escherichia coli at pH with an 8.0. Results: The overall prevalence of antibiotic residues detected by MIT was 64% in liver, 63% in kidney, 56% in breast muscle, 50% in thigh muscle, and 60% in eggs. There was significant variation in results between MIT and TLC (p<0.05). Tetracycline residues were found in 48% in liver, 24% in kidneys, 20% in thigh muscles, 26% in breast muscles, and 36% in eggs. Ciprofloxacin residues were found 46% in liver, 42% in kidneys, 34% in thigh muscles, 30% in breast muscles, and 30% in eggs. Enrofloxacin residues were found 40% in livers, 36% in kidneys, 24% in thigh muscles, 20% in breast muscles, and 26% in eggs. Amoxicillin residues were found 48% in livers, 30% in kidneys, 26% in thigh muscles, 22% in breast muscles, and 24% in eggs. The most frequently detected antibiotic residues by both MIT and TLC were found in liver tissue, tetracycline (48%), ciprofloxacin (46%), enrofloxacin (40%), and amoxicillin (42%) were found in liver. Breast muscle tissue was least likely to contain antibiotic residues (24%). Tetracycline (p=0.01) and amoxicillin (p=0.03) residues had significant variation among the various poultry tissues and eggs. Conclusions: A high percentage of tissues and eggs that could be available for human consumption had antibiotic residues. This study suggests that poultry meat and eggs should not be circulated to markets until the end of the drug's withdrawal period. It is also recommended to observe the withdrawal period of drugs before poultry slaughter or table egg distribution to avoid antimicrobial resistance and to inform both owners and consumers about the risks of antibiotic residues.
Journal of Chromatography A, 2009
A liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for the simultaneous detection and confirmation of halofuginone, robenidine, diclazuril, nicarbazin, monensin, narasin, lasalocid, salinomycin, maduramicin and semduramicin in whole egg has been developed and validated. The anticoccidial residues were extracted by acetonitrile, evaporated and dissolved in a sodium acetate/acetonitrile mixture. Then, the samples were injected on a C8 column in a gradient mode. Diclazuril-bis, DNC-d8 and nigericin were used as internal standards. The results of the full validation in accordance with the guidelines of the Commission Decision no 2002/657/EC are presented. This rapid and sensitive method was found suitable to confirm the anticoccidials at 1 and at 75 g kg −1 for the MRL compound lasalocid.