A precise and reliable analytical method of high performance liquid chromatography-tandem mass spectrometry(HPLCMS/MS)was developed to measure trace levels of enrofloxacin(ENR)and its major metabolite ciprofloxacin(CI...A precise and reliable analytical method of high performance liquid chromatography-tandem mass spectrometry(HPLCMS/MS)was developed to measure trace levels of enrofloxacin(ENR)and its major metabolite ciprofloxacin(CIP)in carp tissues.Optimized chromatographic separation was obtained on a Waters Xterra MS C_(18) reversed-phase column using gradient elution with methanol and 0.1%formic acid aqueous solution including 5mmolL^(-1) of ammonium acetate.The established method was applied to study the pharmacokinetics and distribution of ENR and CIP in tissues of carp following a single oral administration in feed at a dosage of 40mgkg^(-1) bw(body weight).Data were analyzed using DAS 2.0 dynamics software,and the experimental results suggest that ENR was rapidly absorbed and extensively distributed in carp tissues through systemic circulation,and the pharmacokinetic characteristics can be described with a two-compartment model.The elimination half-lives(t_(1/2β))from muscle,liver,gill,plasma and skin were 131,160,104,132 and 310 h,respectively.The areas under the drug concentration-time curves(AUC)for these tissues were 491,972,750,249 and 706hmgkg^(-1),respectively.The maximum concentration(C_(max))values were 13,29,37,9 and 5mgkg^(-1) with peak times(t_(max))of 8,4,4,2 and 4 h,respectively.Ciprofloxacin,the active metabolite of ENR,was also detected in carp tissues,indicating that only 1.54%of de-ethylation of ENR occurs in carp.At a water temperature of 18℃,the drug withdrawal time was determined to be no less than 24 d while the carp was fed at a single dosage of 40mgkg^(-1).展开更多
Turbot Scophthalmus maximus, an important aquaculture species in China, currently suffers from epizootic diseases because of high density aquaculture. Enrofloxacin has been used to treat various systemic bacterial fis...Turbot Scophthalmus maximus, an important aquaculture species in China, currently suffers from epizootic diseases because of high density aquaculture. Enrofloxacin has been used to treat various systemic bacterial fish infections. However, studies concerning the pharmacokinetics of enrofloxacin in turbot are limited. In this study, the pharmacokinetics of enrofloxacin and its metabolite ciprofloxacin, were investigated in the turbot following intravenous and oral administration at 10 mg enrofloxacin/kg body weight, at 16℃ and 10℃ water temperatures. The concentrations of enrofloxacin and ciprofloxacin in the main tissues （plasma, muscle, liver and kidney） were detected by HPLC. The results show that the plasma concentration-time data for enrofloxacin were best described as a two-compartment open model after intravenous and oral administration. Three pharmacokinetic equations were established between the concentrations and temperatures. The kinetic profile of enrofloxacin was temperature dependent. The absorption half-life of enrofloxacin was 1.99 h and 2.17 h after oral administration, whereas the elimination half-life of the drug was 98.63 h and 136.59 h at 16℃ and 10℃, respectively. The peak concentration of enrofloxacin in plasma and tissues was higher at 16℃ than that at 10℃, and the peak plasma concentration time in the liver was the shortest at both temperatures among those of other tissues. The plasma ℃/MIC ratio varied between 11.08 and 5 540.00 at 16℃; and between 7.92 and 3 960.00 at 10℃. The AUC/MIC ratio was 467.82-280 690.00 at 16℃, and 359.48-215 690.00 at 10℃. These ratios indicate that it is possible to obtain therapeutic efficacy. Very low levels of ciprofloxacin were detected. The AUC ratios of ciprofloxacin and enrofloxacin in plasma suggest that plasma ciprofloxacin might play a minor role in enrofloxacin treatment for turbot.展开更多
Adenosine triphosphate-binding cassette transporter breast cancer resistance protein(BCRP) exists highly in the apical membranes of epithelia, and is involved in drug availability. Ko143 is a typical inhibitor of BCRP...Adenosine triphosphate-binding cassette transporter breast cancer resistance protein(BCRP) exists highly in the apical membranes of epithelia, and is involved in drug availability. Ko143 is a typical inhibitor of BCRP in rodents. The synthetic antibacterial agent enrofloxacin(ENRO) is a fluoroquinolone employed as veterinary and aquatic medicine, and also a substrate for BCRP. BCRP gene highly expressed in the hepatopancreas and intestine of Exopalaemon carinicauda as was determined with real-time quantitative reverse transcription-polymerase chain reaction(RT-q PCR) method. The effects of Ko143 on the abundance of BCRP m RNA and ENRO pharmacokinetics in E. carinicauda were studied. The m RNA abundance of BCRP decreased significantly in hepatopancreas and intestine(P < 0.05) after Ko143 treatment. Co-administration of Ko143 significantly changed the pharmacokinetics of orally administered enrofloxacin, which was supported by higher distribution half-life(t_(1/2α)), elimination half-life(t_(1/2β)), area under the curve up to the last measurable concentration(AUC_(0-t)), peak concentration(C_(max)) and lower clearance(CL/F). These findings revealed that Ko143 downregulates BCRP expression in hepatopancreas and intestine, thus affects the pharmacokinetics of orally administered enrofloxacin in E. carinicauda. The drug-drug interaction can be caused by the change in BCRP activity if ENRO is used in combination with other drugs in shrimp.展开更多
A two-step zero-length cross-linking procedure using active esters was successfully adopted for conjugating en-rofloxacin (EF) to human serum albumin (HSA). The derived conjugate was characterized by UV spectrum and t...A two-step zero-length cross-linking procedure using active esters was successfully adopted for conjugating en-rofloxacin (EF) to human serum albumin (HSA). The derived conjugate was characterized by UV spectrum and then usedfor immunization of BALB/C mice. In enzyme-linked immunosorbent assay (ELISA) and competitive inhibition ELISA ex-periments, the derived antiserum exhibited high antibody titer (greater than 1: 250 000) as well as varied cross-reactivity(from 97.8% to 161.7%) to three analogs of EF belonging to fluoroquinolones family. But over the concentration rangestudied, no significant cross-reactivity was observed to other group of antibiotics (chloramphenicol, oxytetracycline, sul-phamethoxazole and nysfungin). It was confirmed that the synthesized immunogen was highly antigenic and elicited specificantibody responses in BALB/C mice against EF.展开更多
This review outlines the current knowledge on the use of enrofloxacin in veterinary medicine from biochemical mechanisms to the use in the field conditions and even resistance and ecotoxicity. The basics of biochemist...This review outlines the current knowledge on the use of enrofloxacin in veterinary medicine from biochemical mechanisms to the use in the field conditions and even resistance and ecotoxicity. The basics of biochemistry, the mechanisms of action and resistance and pharmacokinetics are presented. Then an overview of available veterinary products, their efficacy and their toxicity against target species, human and environment is provided.展开更多
An ester activation method was employed to couple enrofloxacin(ENFX) to the carrier proteins BSA and OVA. The conjugates ENFX-BSA and ENFX-OVA were identified with an UV spectrophotometer and amino acid automation a...An ester activation method was employed to couple enrofloxacin(ENFX) to the carrier proteins BSA and OVA. The conjugates ENFX-BSA and ENFX-OVA were identified with an UV spectrophotometer and amino acid automation analysis instrument, and resulted in conjugates with 48 ENFX molecules per carrier molecule(BSA). Splenocytes from mice immunized with ENFX-BSA were fused with SP2/0 myeloma cells and hybridomas secreting antibodies against enrofloxacin were selected and cloned. Two stable monoclonal antibodies, 2C5, 5D5 of the subclass IgG2a, were isolated. Using antibody 5D5, an indirect competitive inhibition enzyme-linked immunosorbent assay (Ci-ELISA) was developed for the quantitative detection of enrofloxacin and its metabolites. The IC50 of the standard curve was 21.67 ng mL-1 and the limit of detection for enrofloxacin was 0.13 ng mL-1. This method was sensitive and had a linear range from 0.13 to 10 000 ng mL-1 (r= -0.9782). Monoclonal antibody 5D5 exhibited high relative affinity to enrofloxacin, and the cross-reactivities with ciprofloxacin, marbofloxacin, sarafloxacin and danorfloxacin were 110.8, 27.40, 71.05 and 37.41%, respectively. Three non-fluoroquinolones of cefadroxil, chloramphenicol, sulfadimethoxine were tested and there was no cross-reaction between them.展开更多
Aim of this research was to find out the concentration of enrofloxacin residue in tilapia meat for several weeks after antibiotic treatment. Twenty seven tilapia fishes were divided into three groups. The first group ...Aim of this research was to find out the concentration of enrofloxacin residue in tilapia meat for several weeks after antibiotic treatment. Twenty seven tilapia fishes were divided into three groups. The first group was not infected and treated, the second group was infected with A. salmonicida subsp, smithia and the third group was infected with A. salmonicida subsp. achromogenes intramuscularly. Six days after infection, treatment was carried out using Baytril administered orally for the second group and intramuscularly for the third group during five days. At the 1 st, 4th and 8th week after the treatment, Three fish were taken from each group to be analyzed for its concentration of enrofloxacin residue by diffusion on Mueller Hinton Agar （MHA） method and quantitatively using high performance liquid chromatography （HPLC） method. The MHA test showed the formation of inhibition zone, at the 1 st week and 4th week after the treatment, while at 8th week after treatment did not show inhibition zone. The HPLC test on enrofloxacin residual concentration in tilapia infected with A. salmonicida subsp, smithia （second group） at the 1st, 4th and 8th week after treatment showed the average of 33.0, 6.10 and 0.0021 μg/g of enrofloxacin residue level. While in tilapia infected with A. salmonicida subsp, achromogenes and treated with enrofloxacin intramuscularly （third group） showed the average of residue level 35.79, 2.18 and 0.00065 μg/g. In conclusion, the residue of enrofloxacin was still high concentration until the fourth week after treatment in the second and third groups. Based on Indonesian National Standards and Rules, the maximum limit of enrofloxacin residue is 0.01 μg/g. The concentration of enrofloxacine residue was very low and the concentration of enrofloxacin residue collected from tilapia using orally and intramuscularly method of treatment was not different.展开更多
MXenes,a new family of two-dimensional transition metal carbides or nitrides,have attracted tremendous attention for various applications due to their unique properties such as good electrical conductivity,hydrophilic...MXenes,a new family of two-dimensional transition metal carbides or nitrides,have attracted tremendous attention for various applications due to their unique properties such as good electrical conductivity,hydrophilicity,and ion intercalability.In this work,Ti_(3)C_(2) MXene,or MX,is converted to MX-TiO_(2) composites using a simple and rapid microwave hydrothermal treatment in HCl/NaCl mixture solution that induces formation of fine TiO_(2) particles on the MX parent structure and imparts photocatalytic activity to the resulting MX-TiO_(2) composites.The composites were used for enrofloxacin(ENR),a frequently found contaminating antibiotic,removal from water.The relative amount of the MX and TiO_(2) can be controlled by controlling the hydrothermal temperature resulting in composites with tunable adsorption/photocatalytic properties.NaCl addition was found to play important role as composites synthesized without NaCl could not adsorb enrofloxacin well.Adding NaCl into the hydrothermal treatment causes sodium ions to be simultaneously intercalated into the composite structure,improving ENR adsorption greatly from 1 to 6 mg ENR/g composite.It also slows down the MX to TiO2 conversion leading to a smaller and more uniform distribution of TiO_(2) particles on the structure.MX-TiO_(2)/NaCl composites,which have sodium intercalated in their structures,showed both higher ENR adsorption and photocatalytic activity than composites without NaCl despite the latter having higher TiO2 content.Adsorbed ENR on the composites can be efficiently degraded by free radicals generated from the photoexcited TiO2 particles,leading to high photocatalytic degradation efficiency.This demonstrates the synergetic effect between adsorption and photocatalytic degradation of the synthesized compounds.展开更多
Modified 1-ethyl-3-(3-dimethylaminopropy) carbodiimide(EDC) method was employed to synthesize the artificial antigen of enrofloxacin(ENR),and New Zealand rabbits were used to produce anti-ENR polyclonal antibody(pAb)....Modified 1-ethyl-3-(3-dimethylaminopropy) carbodiimide(EDC) method was employed to synthesize the artificial antigen of enrofloxacin(ENR),and New Zealand rabbits were used to produce anti-ENR polyclonal antibody(pAb).Based on the checkerboard titration,an indirect competitive enzyme-linked immunosorbent assay(ELISA) standard curve was established.This assay was sensitive and had a linear range from 0.6 to 148.0 μg/kg(R2=0.9567),with the half maximal inhibitory concentration(IC50) and limit of detection(LOD) values of 9.4 μg/kg and 0.2 μg/kg,respectively.Of all the competitive analogues,the produced pAb exhibited a high cross-reactivity to ciprofloxacin(CIP)(87%),the main metabolite of ENR in tissues.After optimization,the matrix effects can be ignored using a 10-fold dilution in beef and 20-fold dilution in pork.The overall recoveries and coefficients of variation(CVs) were in the ranges of 86%-109% and 6.8%-13.1%,respectively.It can be concluded that the established ELISA method is suitable for simultaneous detection of ENR and CIP in animal tissues.展开更多
基金supported by the Central Public-Interest Scientific Institution Basal Research Fund,CAFS(No.2020TD71).
文摘A precise and reliable analytical method of high performance liquid chromatography-tandem mass spectrometry(HPLCMS/MS)was developed to measure trace levels of enrofloxacin(ENR)and its major metabolite ciprofloxacin(CIP)in carp tissues.Optimized chromatographic separation was obtained on a Waters Xterra MS C_(18) reversed-phase column using gradient elution with methanol and 0.1%formic acid aqueous solution including 5mmolL^(-1) of ammonium acetate.The established method was applied to study the pharmacokinetics and distribution of ENR and CIP in tissues of carp following a single oral administration in feed at a dosage of 40mgkg^(-1) bw(body weight).Data were analyzed using DAS 2.0 dynamics software,and the experimental results suggest that ENR was rapidly absorbed and extensively distributed in carp tissues through systemic circulation,and the pharmacokinetic characteristics can be described with a two-compartment model.The elimination half-lives(t_(1/2β))from muscle,liver,gill,plasma and skin were 131,160,104,132 and 310 h,respectively.The areas under the drug concentration-time curves(AUC)for these tissues were 491,972,750,249 and 706hmgkg^(-1),respectively.The maximum concentration(C_(max))values were 13,29,37,9 and 5mgkg^(-1) with peak times(t_(max))of 8,4,4,2 and 4 h,respectively.Ciprofloxacin,the active metabolite of ENR,was also detected in carp tissues,indicating that only 1.54%of de-ethylation of ENR occurs in carp.At a water temperature of 18℃,the drug withdrawal time was determined to be no less than 24 d while the carp was fed at a single dosage of 40mgkg^(-1).
基金Supported by the National Natural Science Foundation of China(Nos.30700617,31101298)the National Special Research Fund for NonProfit Sector(No.nyhyzx07-046)
文摘Turbot Scophthalmus maximus, an important aquaculture species in China, currently suffers from epizootic diseases because of high density aquaculture. Enrofloxacin has been used to treat various systemic bacterial fish infections. However, studies concerning the pharmacokinetics of enrofloxacin in turbot are limited. In this study, the pharmacokinetics of enrofloxacin and its metabolite ciprofloxacin, were investigated in the turbot following intravenous and oral administration at 10 mg enrofloxacin/kg body weight, at 16℃ and 10℃ water temperatures. The concentrations of enrofloxacin and ciprofloxacin in the main tissues （plasma, muscle, liver and kidney） were detected by HPLC. The results show that the plasma concentration-time data for enrofloxacin were best described as a two-compartment open model after intravenous and oral administration. Three pharmacokinetic equations were established between the concentrations and temperatures. The kinetic profile of enrofloxacin was temperature dependent. The absorption half-life of enrofloxacin was 1.99 h and 2.17 h after oral administration, whereas the elimination half-life of the drug was 98.63 h and 136.59 h at 16℃ and 10℃, respectively. The peak concentration of enrofloxacin in plasma and tissues was higher at 16℃ than that at 10℃, and the peak plasma concentration time in the liver was the shortest at both temperatures among those of other tissues. The plasma ℃/MIC ratio varied between 11.08 and 5 540.00 at 16℃; and between 7.92 and 3 960.00 at 10℃. The AUC/MIC ratio was 467.82-280 690.00 at 16℃, and 359.48-215 690.00 at 10℃. These ratios indicate that it is possible to obtain therapeutic efficacy. Very low levels of ciprofloxacin were detected. The AUC ratios of ciprofloxacin and enrofloxacin in plasma suggest that plasma ciprofloxacin might play a minor role in enrofloxacin treatment for turbot.
基金supported by the Natural Science Foundation of Shandong Province,P. R. China (No. ZR2019QC015)the National Key R&D Program of China (No. 2019YFD0900403)+3 种基金the Central Public-Interest Scientific Institution Basal Research Fund,CAFS (Nos. 2019ZD09 03 and 2020TD46)the Marine S&T Fund of Shandong Province for Pilot National Laboratory for Marine Science and Technology (Qingdao)(No. 2018SDKJ0502-2)the Earmarked Fund for Modern Agro-industry Technology Research System (No. CARS-48)the National Natural Science Foundation of China (No. 31873039)。
文摘Adenosine triphosphate-binding cassette transporter breast cancer resistance protein(BCRP) exists highly in the apical membranes of epithelia, and is involved in drug availability. Ko143 is a typical inhibitor of BCRP in rodents. The synthetic antibacterial agent enrofloxacin(ENRO) is a fluoroquinolone employed as veterinary and aquatic medicine, and also a substrate for BCRP. BCRP gene highly expressed in the hepatopancreas and intestine of Exopalaemon carinicauda as was determined with real-time quantitative reverse transcription-polymerase chain reaction(RT-q PCR) method. The effects of Ko143 on the abundance of BCRP m RNA and ENRO pharmacokinetics in E. carinicauda were studied. The m RNA abundance of BCRP decreased significantly in hepatopancreas and intestine(P < 0.05) after Ko143 treatment. Co-administration of Ko143 significantly changed the pharmacokinetics of orally administered enrofloxacin, which was supported by higher distribution half-life(t_(1/2α)), elimination half-life(t_(1/2β)), area under the curve up to the last measurable concentration(AUC_(0-t)), peak concentration(C_(max)) and lower clearance(CL/F). These findings revealed that Ko143 downregulates BCRP expression in hepatopancreas and intestine, thus affects the pharmacokinetics of orally administered enrofloxacin in E. carinicauda. The drug-drug interaction can be caused by the change in BCRP activity if ENRO is used in combination with other drugs in shrimp.
基金supported by the National Natural Science Fundation of China(No.30400336)Science and Technology Project of Qingdao(03-1-NSH-2-2).
文摘A two-step zero-length cross-linking procedure using active esters was successfully adopted for conjugating en-rofloxacin (EF) to human serum albumin (HSA). The derived conjugate was characterized by UV spectrum and then usedfor immunization of BALB/C mice. In enzyme-linked immunosorbent assay (ELISA) and competitive inhibition ELISA ex-periments, the derived antiserum exhibited high antibody titer (greater than 1: 250 000) as well as varied cross-reactivity(from 97.8% to 161.7%) to three analogs of EF belonging to fluoroquinolones family. But over the concentration rangestudied, no significant cross-reactivity was observed to other group of antibiotics (chloramphenicol, oxytetracycline, sul-phamethoxazole and nysfungin). It was confirmed that the synthesized immunogen was highly antigenic and elicited specificantibody responses in BALB/C mice against EF.
文摘This review outlines the current knowledge on the use of enrofloxacin in veterinary medicine from biochemical mechanisms to the use in the field conditions and even resistance and ecotoxicity. The basics of biochemistry, the mechanisms of action and resistance and pharmacokinetics are presented. Then an overview of available veterinary products, their efficacy and their toxicity against target species, human and environment is provided.
基金supported by the Natural Science Foundation of Guangdong Province,China(994162).
文摘An ester activation method was employed to couple enrofloxacin(ENFX) to the carrier proteins BSA and OVA. The conjugates ENFX-BSA and ENFX-OVA were identified with an UV spectrophotometer and amino acid automation analysis instrument, and resulted in conjugates with 48 ENFX molecules per carrier molecule(BSA). Splenocytes from mice immunized with ENFX-BSA were fused with SP2/0 myeloma cells and hybridomas secreting antibodies against enrofloxacin were selected and cloned. Two stable monoclonal antibodies, 2C5, 5D5 of the subclass IgG2a, were isolated. Using antibody 5D5, an indirect competitive inhibition enzyme-linked immunosorbent assay (Ci-ELISA) was developed for the quantitative detection of enrofloxacin and its metabolites. The IC50 of the standard curve was 21.67 ng mL-1 and the limit of detection for enrofloxacin was 0.13 ng mL-1. This method was sensitive and had a linear range from 0.13 to 10 000 ng mL-1 (r= -0.9782). Monoclonal antibody 5D5 exhibited high relative affinity to enrofloxacin, and the cross-reactivities with ciprofloxacin, marbofloxacin, sarafloxacin and danorfloxacin were 110.8, 27.40, 71.05 and 37.41%, respectively. Three non-fluoroquinolones of cefadroxil, chloramphenicol, sulfadimethoxine were tested and there was no cross-reaction between them.
文摘Aim of this research was to find out the concentration of enrofloxacin residue in tilapia meat for several weeks after antibiotic treatment. Twenty seven tilapia fishes were divided into three groups. The first group was not infected and treated, the second group was infected with A. salmonicida subsp, smithia and the third group was infected with A. salmonicida subsp. achromogenes intramuscularly. Six days after infection, treatment was carried out using Baytril administered orally for the second group and intramuscularly for the third group during five days. At the 1 st, 4th and 8th week after the treatment, Three fish were taken from each group to be analyzed for its concentration of enrofloxacin residue by diffusion on Mueller Hinton Agar （MHA） method and quantitatively using high performance liquid chromatography （HPLC） method. The MHA test showed the formation of inhibition zone, at the 1 st week and 4th week after the treatment, while at 8th week after treatment did not show inhibition zone. The HPLC test on enrofloxacin residual concentration in tilapia infected with A. salmonicida subsp, smithia （second group） at the 1st, 4th and 8th week after treatment showed the average of 33.0, 6.10 and 0.0021 μg/g of enrofloxacin residue level. While in tilapia infected with A. salmonicida subsp, achromogenes and treated with enrofloxacin intramuscularly （third group） showed the average of residue level 35.79, 2.18 and 0.00065 μg/g. In conclusion, the residue of enrofloxacin was still high concentration until the fourth week after treatment in the second and third groups. Based on Indonesian National Standards and Rules, the maximum limit of enrofloxacin residue is 0.01 μg/g. The concentration of enrofloxacine residue was very low and the concentration of enrofloxacin residue collected from tilapia using orally and intramuscularly method of treatment was not different.
基金National Nanotechnology Center (NANOTEC) (No.P1751698) for financial supportThailand Advanced Institute of Science and Technology and Tokyo Institute of Technology (TAIST-Tokyo Tech) for collaborative program and scholarship supports+1 种基金supported by Thammasat University Research Fund Contract No.TUFT 055/2563Support from Center of Excellence in Functional Advanced Materials Engineering (CoE FAME),SIIT
文摘MXenes,a new family of two-dimensional transition metal carbides or nitrides,have attracted tremendous attention for various applications due to their unique properties such as good electrical conductivity,hydrophilicity,and ion intercalability.In this work,Ti_(3)C_(2) MXene,or MX,is converted to MX-TiO_(2) composites using a simple and rapid microwave hydrothermal treatment in HCl/NaCl mixture solution that induces formation of fine TiO_(2) particles on the MX parent structure and imparts photocatalytic activity to the resulting MX-TiO_(2) composites.The composites were used for enrofloxacin(ENR),a frequently found contaminating antibiotic,removal from water.The relative amount of the MX and TiO_(2) can be controlled by controlling the hydrothermal temperature resulting in composites with tunable adsorption/photocatalytic properties.NaCl addition was found to play important role as composites synthesized without NaCl could not adsorb enrofloxacin well.Adding NaCl into the hydrothermal treatment causes sodium ions to be simultaneously intercalated into the composite structure,improving ENR adsorption greatly from 1 to 6 mg ENR/g composite.It also slows down the MX to TiO2 conversion leading to a smaller and more uniform distribution of TiO_(2) particles on the structure.MX-TiO_(2)/NaCl composites,which have sodium intercalated in their structures,showed both higher ENR adsorption and photocatalytic activity than composites without NaCl despite the latter having higher TiO2 content.Adsorbed ENR on the composites can be efficiently degraded by free radicals generated from the photoexcited TiO2 particles,leading to high photocatalytic degradation efficiency.This demonstrates the synergetic effect between adsorption and photocatalytic degradation of the synthesized compounds.
基金supported by the Henan Innovation Project for University Prominent Research Talents (No. 2010HASTIT026)the Key Scientific & Technological Project of Education Department in Henan Province of China (No. 2011A230003)
文摘Modified 1-ethyl-3-(3-dimethylaminopropy) carbodiimide(EDC) method was employed to synthesize the artificial antigen of enrofloxacin(ENR),and New Zealand rabbits were used to produce anti-ENR polyclonal antibody(pAb).Based on the checkerboard titration,an indirect competitive enzyme-linked immunosorbent assay(ELISA) standard curve was established.This assay was sensitive and had a linear range from 0.6 to 148.0 μg/kg(R2=0.9567),with the half maximal inhibitory concentration(IC50) and limit of detection(LOD) values of 9.4 μg/kg and 0.2 μg/kg,respectively.Of all the competitive analogues,the produced pAb exhibited a high cross-reactivity to ciprofloxacin(CIP)(87%),the main metabolite of ENR in tissues.After optimization,the matrix effects can be ignored using a 10-fold dilution in beef and 20-fold dilution in pork.The overall recoveries and coefficients of variation(CVs) were in the ranges of 86%-109% and 6.8%-13.1%,respectively.It can be concluded that the established ELISA method is suitable for simultaneous detection of ENR and CIP in animal tissues.
