On-Chip Purification of Tetracyclines Based on Copper Ions Interaction
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Abstract:
Antibiotics are widely used to both prevent and treat bacterial diseases as well as promote animal growth. This massive use leads to the presence of residual antibiotics in food with severe consequences for human health. Limitations and regulations on the tolerated amount of antibiotics in food have been introduced and analytical methods have been developed. The bioanalytical methods usually employed to detect antibiotic residues, however, are time-consuming, expensive and laboratory-based. Novel methods with improved rapidity, portability and cost that are easy-to-use and sustainable are therefore highly desirable. In the attempt to fulfill this need, a microfluidic system was set up herein for the purification and pre-concentration of tetracyclines from raw milk selected as the case-study. The system includes a polymeric microfluidic chip containing magnetic beads loaded with copper to exploit the preferential interaction of tetracycline with divalent ions. The microfluidic system was demonstrated to efficiently pre-concentrate tetracycline, oxytetracycline and chlortetracycline with similar performances and efficiently purify tetracycline from raw milk without any pre-treatment. The simplified method described in this paper could be easily integrated in a compact and portable device for the in-field detection of tetracyclines, with the economic advantage of preventing food wastes and guaranteeing food safety.Keywords:
Chlortetracycline
Microfluidic chip
Tetracycline antibiotics
Lab-on-a-Chip
The aim of this study was to detect the presence of antibiotic residues in foods of animal origin, including 42 pieces of chicken gizzard and 46 pieces of bovine kidney and 102 chicken eggs belonging to various brands. These samples were gathered from December 2020 to April 2021 in the Aegean province of Turkey. A sensitive, simple, rapid, experimentally convenient and cost effective RP-LC method with high recovery output was developed. The method was thoroughly validated for the optimized parameters and produced satisfactory results. The analysis of bovine offal by the developed RP-LC method showed the presence of oxytetracycline, tetracycline, and chlortetracycline residues in 14 (30.43%) kidney samples. Chlortetracycline was detected in 7 (16.67%) chicken gizzard samples. In addition, the analysis of chicken eggs revealed the presence of oxytetracycline and tetracycline residues in nine egg samples (8.82%). Since, the amount of antibiotic residues in these samples was below the detection limit, quantification could not be carried out. Only one (0.98%) of the 102 egg samples exceeded the MRL (267.1 mg/kg) for oxytetracycline concentration. According to the study's overall findings, it is recommended that tetracycline antibiotics should be regularly checked in a variety of foods made from animals because they were found in 32 out of 190 analysed samples. Tetracycline residues may pose dangers to human health, so it's important to conduct further research and more information should be given for both producers and consumers.
Chlortetracycline
Tetracycline antibiotics
Gizzard
Maximum Residue Limit
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A method for simultaneous determination of oxytetracycline,tetracycline and chlortetracycline in seafood was successfully developed using high performance liquid chromatography.The mobile phase was composed of solium dihydrogen phosphate and acetonitrile.The detection wavelength was set at 356 nm and 371 nm.The results showed that three tetracycline antibiotics were well separated,and the limits of detection for three antibiotics were 14 ng for oxytetracycline,11 ng for tetracycline,and 34 ng for chlortetracycline.The recoveries of standard antibiotics were 12.61% for oxytetracycline,11.18% for tetracycline,and 28.10% for chlortetracycline.The relative standard deviation was 0.34%~2.79% for oxytetracycline,0.53%~3.36% for tetracycline,0.46%~5.45% for chlortetracycline.The method is rapid,sensitive,accurate and reliable,and can be used in the analysis of antibiotics residues in seafood.
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Tetracycline antibiotics
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Tissue samples from 623 hogs suspected of having received antibiotic treatment were collected at federally-inspected abattoirs in Manitoba and submitted for analysis from October 1987 to March 1988. Initially, samples were screened by a multi-residue thin layer chromatography/bio-autography method, but, during the final two months of the investigation, analysis was restricted to a quantitative high performance liquid chromatography method for tetracycline antibiotics only. Sixteen animals (3%) were found with detectable tissue levels of chlortetracycline ranging from 0.10 to 1.38 ppm. One hundred thirty-eight animals (22%) were found with detectable tissue levels of oxytetracycline ranging from 0.05 to 2.07 ppm. Comparison of these levels with previously published values suggested that some producers were administering tetracyclines at therapeutic levels without observing the seven-day withdrawal period. The data generated by this study provided information for use by regulatory officials at Health and Welfare Canada in establishing specific tolerance levels for residues of the tetracycline antibiotics in animal tissue; previously, a zero tolerance had been in effect. During the study, only one hog had tetracycline residues in muscle which would have exceeded the current tolerance levels.
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Tetracycline antibiotics
Residue (chemistry)
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Tetracycline antibiotics, including tetracycline (TC), chlortetracycline (CTC), oxytetracycline (OTC) and doxycycline (DOX), are commonly used in veterinary medicine because of their broad-spectrum activity, cost effectiveness and certain favourable pharmacokinetic properties. The use of antibiotics in feed for non-medicinal purposes has been prohibited in the European Union. In Poland and other countries of the European Union, there are official national programmes for controlling the use of TCs and all other antibacterials in feed. This paper presents the pharmacological properties of TCs as well as the main reasons for and consequences of their use in feed. It also presents a chromatographic method for detection of TCs in animal feeds. The method is used in routine testing of food for the presence of tetracycline antibiotics. In the years 2015-2017, tetracycline antibiotics were detected in 31 out of 126 feed samples analysed by this method.
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Tetracycline antibiotics
Animal Feed
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Toxicity of tetracyclines was studied experimentally on different species of laboratory animals. It was shown that tetracycline, oxytetracycline and chlortetracycline were close by their chemical structure and physico-chemical properties, as well as by the main toxicity parameters, i.e. acute toxicity, cumulative activity, skin-irritating and sensitizing effect. Under the conditions of subacute experiments the above 3 antibiotics induced evenly pronounced one direction changes in animals. The data obtained during the experiments provided recommendation of the level of 0.1 mg/m3 as the maximum allowable concentration (MAC) of oxytetracycline and chlortetracycline, i.e. the same level as the previously recommended for tetracycline.
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Tetracycline antibiotics
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A method for simultaneous determination of oxytetracycline,tetracycline,chlortetracycline in animal tissue was successfully established by HPLC-ESI MS/MS.The tetracycline antibiotics in animal tissue were extracted by liquid-solid extraction and cleaned up by SPE.The mass spectrometer was operated in the positive-ion mode using selected reaction monitoring(SRM).The results showed that three tetracycline antibiotics were well separated under the optimal conditions.The detection limits of oxytetracycline,tetracycline and chlortetracycline were 0.16 ng/g,0.1 ng/g and 0.1 ng/g,respectively.The spiked recoveries were range of 84%-94% and the relative standard deviations were 2.4%-2.9%.
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Solid phase extraction
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The concentration and distribution of four tetracycline antibiotics(TCs) including tetracycline(TC),oxytetracycline(OTC),chlortetracycline(CTC) and deoxytetracycline(DC) in soils from 24 typical vegetable fields in 18 towns of Dongguan city using high-performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS) was discussed.The total concentrations of four tetracycline antibiotics(∑TCs) ranged from 0(below LOQ) to 97.04μg/kg,which predominated with under 10μg/kg and averaged at 13.42μg/kg.The concentrations of four tetracycline compounds detected in 59~89% of the samples ranged from 0(below LOQ) to 75.29μg/kg which falls mostly bellow 10μg/kg.Oxytetracycline which averaged at 6.51μg/kg was dominant compound,followed by chlortetracycline and deoxytetracycline which averaged at 2.95μg/kg and 2.92μg/kg respectively.The concentration and constitute of tetracycline compounds in soils from different vegetable fields varied greatly,consisting mainly of four models,i.e.the first one dominated with oxytetracycline,the second one dominated with chlortetracycline,the third one dominated with deoxytetracycline,and the last one dominated with both oxytetracycline and deoxytetracycline.However,the concentrations and constitute of tetracycline compounds in soils grown different cultivars within the same vegetable fields varied weakly.The concentrations of tetracycline antibiotics in soils from vegetable fields of Dongguan city were generally lower and less ecotoxic risk for soil ecosystem.But it should not be ignored as to combined toxic effect and resistance of various compounds.
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Tetracycline antibiotics
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Chlortetracycline
Tetracycline antibiotics
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A method was validated for analysis of oxytetracycline (OTC), tetracycline (TC), and chlortetracycline (CTC) in fortified salmon muscle tissue. Recoveries of OTC were 100 +/- 6, 86 +/- 6, and 82 +/- 5% (n = 6) at fortification levels of 1.0, 0.5, and 0.2 ppm, respectively. Recoveries of TC were 68 +/- 4, 65 +/- 6, and 66 +/- 7%; recoveries of CTC were 45 +/- 9, 48 +/- 8, and 0%, respectively. Detection limits for OTC and TC were 0.08 and 0.09 ppm, respectively.
Chlortetracycline
Tetracycline antibiotics
Muscle tissue
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Chlortetracycline
Tetracycline antibiotics
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