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Incomplete mineralization of antibiotics in biological sludge systems poses a risk to the environment. In this study, the toxicity associated with ciprofloxacin (CIP) biodegradation in activated sludge (AS), anaerobic methanogenic sludge (AnMS), and sulfur-mediated sludge (SmS) systems was examined via long-term bioreactor tests and a series of bioassays. The AS and AnMS systems were susceptible to CIP and its biotransformation products (TPs) and exhibited performance deterioration, while the SmS system exhibited high tolerance against the toxicity of CIP and its TPs along with excellent pollutant removal. Up to 14 TPs were formed via piperazinyl substituent cleavage, defluorination, decarboxylation, acetylation, and hydroxylation reactions in AS, AnMS, and SmS systems. Biodegradation of CIP in the AS, AnMS, and SmS systems, however, could not completely eliminate its toxicity as evident from the inhibition of Vibrio fischeri luminescence along with Escherichia coli K12 and Bacillus subtilis growth . The anaerobic systems (AnMS and SmS) were more effective than the aerobic AS system at CIP biodegradation, significantly reducing the antibacterial activity of CIP and its TPs in the aqueous phase. In addition, the quantitative structure-activity relationship analysis indicated that the TPs produced via decarboxylation and hydroxylation (TP2 and TP4) as well as by cleavage of piperazine (TP12, TP13, and TP14) exhibited higher toxicity than CIP. The findings of this study provide insights into the toxicity and possible risks associated with CIP biodegradation in biological wastewater treatment.

Emerging contaminants, including pharmaceutical compounds, are receiving research attention as a result of their widespread presence in effluents and wastewater treatment plants (WWTPs). The antibiotic ciprofloxacin (CIP) is extensively employed to treat infections in animal and human medicine. Both CIP and its metabolites are common contaminants found in WWTPs. In this study, a label-free ultrasensitive U-bent optical fiber-based immunosensor for the determination of CIP in wastewater samples was developed using the properties of the conducting polymer polyaniline (PANI). The anti-CIP immunoglobulin G (IgG) was deposited on a silica optical fiber surface previously functionalized with PANI. Scanning electron microscopy and micro-Raman spectroscopy were used to investigate the surface of the immunosensor. The analysis of CIP in wastewater was performed without the use of an organic solvent or sample preparation steps, with only the sample dilution in saline buffer (pH = 7.4). The linear range for CIP was from 0.01 to 10,000 ng L -1 . The detection limit was 3.30 × 10 -3 ng L -1 and the quantification limit was 0.01 ng L -1 . The immunosensor provided a high average recovery of 91% after spiking wastewater samples with CIP at a concentration of 9,100 ng L -1 . The method was applied in triplicate to wastewater samples from Quebec (Canada), obtaining concentrations of 549 and 267 ng L -1 . A comparison with a reference method showed no significant difference ( t -test at 95% confidence). The new technique developed is selective, allowing a quantitative analysis of CIP in wastewater.

Herein, a fluorescence turn off-on nanosensor has been successfully developed using functionalized N-doped carbon dots (N-CDs) as the label-free sensing probe for the ultrasensitive detection of Cu 2+ ions first and then ciprofloxacin (CIP), one of the most commonly used antibiotics for disease control, in the presence of bipyridine. The homogeneous and narrowly distributed N-CDs with a mean size of 5.7 nm and a high quantum yield of 84% are fabricated via the hydrothermal method in the presence of citric acid and ethylenediamine as the carbon and nitrogen sources, respectively. The Cu 2+ ions serve as both analyte and fluorescence quenchers in the sensing platform of N-CDs, and a good linear response to Cu 2+ in the range of 0.01-0.35 μM with a limit of detection (LOD) of 0.076 nM is observed. Then, 0.35 μM Cu 2+ is used as the fluorescence quencher of N-CDs to build up the fluorescence turn off-on sensing probe for the detection of CIP using bipyridine (bipy) as the linker for CIP and Cu 2+ ions. The addition of CIP to the bipy-Cu@N-CD composites triggers the formation of CIP-bipy-Cu conjugate as well as the release of N-CDs, resulting in the recovery of fluorescence intensity after 6 min of incubation. The sensing probe exhibits a two-phase linear response to CIP in the concentration range of 0.05-1 and 1-50 μM with a LOD of 0.4 nM. In addition, the bipy-Cu@N-CD probe shows high sensitivity toward CIP over the 19 other interferences. Good recovery of 96-110% is also observed when 0.1-0.9 μM CIP is spiked into the real samples. Results obtained in this study clearly demonstrate a newly developed sensing platform with rapid detection of metal ions and antibiotics, which can open an avenue to develop highly efficient and robust sensing probes for the detection of metal ions, organic metabolites, and biomarkers in biological applications.

Two series of amphiphilic antimicrobial copolymers containing ciprofloxacin (CPF) and amine functional groups have been synthesized via free-radical copolymerization. The chemical structures of the different amine groups and the copolymer compositions have been systematically varied to study how the structure of the copolymer exerts an influence on the antibacterial activity. The viability of Escherichia coli in the presence of antimicrobial copolymers was observed by confocal laser-scanning microscopy (CLSM). CLSM as well as atomic-force microscopy (AFM) were applied to visualize changes in morphology of bacteria treated with antimicrobial copolymers and elucidate the antimicrobial mechanism of the antimicrobial copolymers. Morphological changes of bacteria observed via AFM and CLSM demonstrated that the antibacterial mechanism was due to the disruption of the bacterial membrane. The destruction of the cell membrane was also confirmed by the leakage of intracellular components, which had a strong absorbance at 260 nm. The inhibitory process was monitored by UV absorption dynamically.

Abuse of antibiotics results in a large number of antibiotics residues in the environment and even causes the problem of "super bacteria". Therefore, it is crucial to develop a powerful analytic method to monitor antibiotics quickly and simply. Photonic crystal (PC), as a sensing material, has promising application prospects. Herein, we try to use PC to realize pretreatment-free and label-free detection of Ciprofloxacin (CF) through Michael addition reaction. The recognition process is carried out by the Michael addition reaction between the piperazine group of CF and the o-benzoquinone group on the PC. The monodisperse microspheres with o-benzoquinone groups are prepared by polymerization and oxidation and then stacked to form PC. During the detection, the peak intensity of the PC decreases with the increasing CF concentration, and the linear range is from 2 to 512 μg/L. The limit of detection (LOD) is 0.76 μg/L. Furthermore, the PC retains 97% of the initial response after storage in a Petri dish at room temperature for 1 month, which shows that it has good stability. Moreover, CF in fish farming water can be detected directly without any pretreatment and label, and the results are in good accordance with the LC-MS-MS results. This Michael-addition-mediated PC is accurate, easily prepared, cost-efficient, and long-term stable. In addition, it is environmentally friendly, because little organic solvent is needed during both the preparation and the detection.

This study focuses on the development of electrochemical sensors for the detection of Ciprofloxacin (CFX) in natural waters and wastewater effluents. The sensors are prepared by depositing a layer of multiwalled carbon nanotubes (MWCNTs) dispersed in a porous Nafion film on to a boron-doped diamond (BDD) electrode substrate. The porous-Nafion-MWCNT/BDD electrode enhanced detection of CFX due to selective adsorption, which was accomplished by a combination of electrostatic attraction at -SO3(-) sites in the porous Nafion film and the formation of charge assisted hydrogen bonding between CFX and -COOH MWCNT surface functional groups. By contrast, the bare BDD electrode did not show any activity for CFX oxidation. The sensors were selective for CFX detection in the presence of other antibiotics (i.e., amoxicillin) and other nontarget water constituents (i.e., Cl(-), Ca(2+), humic acid, sodium dodecylbenzenesulfonate, salicylic acid, 4-aminobenzoic acid, and 4-hydroxybenzoic acid). A limit of detection of 5 nM (S/N = 5.04 ± 0.26) in a 0.1 M KH2PO4 supporting electrolyte (pH = 4.5) was obtained using differential pulse voltammetry. The linear dynamic ranges with respect to CFX concentration were 0.005-0.05 μM and 0.05-10 μM, and the sensitivities were 41 ± 5.2 μA μM(-1) and 2.1 ± 0.22 μA μM(-1), respectively. Sensor fouling was observed at high concentrations of some organic compounds such as 1 mM 4-aminobenzoic acid and 4-hydroxybenzoic acid. However, a short cathodic treatment fully restores sensor response. The results indicate that these sensors have application in detecting CFX in natural waters and wastewater effluents.

The metal-organic coordination polymers with tunable structures and properties have been rapidly emerging as very important functional materials. In this work, we prepared terbium (Tb(3+))-based coordination polymer nanoparticles (CPNPs) by employing adenine (Ad) as bridging ligands. The CPNPs was further used as a receptor reagent for ciprofloxacin (CF) detection in aqueous solution. Addition of CF induces a typical emission of Tb(3+) due to the formation of Ad/Tb-CF complex and the sensitization of CF. The fluorescent intensity of Tb(3+) was enhanced linearly with increasing the CF concentration from 60 nM to 14 μM. The detection limit for CF in aqueous solution is 60 nM. The Ad/Tb CPNPs was successfully applied to detect CF in tablet and urine samples and showed a satisfactory result. Compared with other methods, the proposed method is advantageous because that it provides a very simple strategy for CF detection, which does not require complicated sample pretreatment processes or special reaction media. The proposed strategy could be contributed to expand the potential applications of lanthanide coordination polymers in biological and environmental fields.

A luminescence method was developed to screen residues of enrofloxacin (ENRO) and its metabolite, ciprofloxacin (CIPRO), in swine liver. Target analytes were extracted in acetonitrile/1.5% trifluoroacetic acid/NaCl, cleaned up by dispersive liquid-liquid microextraction (DLLME), and finally detected by terbium-sensitized luminescence (TSL) using a time-resolved luminescence photometer. CIPRO yielded slightly lower TSL response than ENRO, so a common threshold was derived from CIPRO and applied to both fluoroquinolones. Among 37 samples randomly spiked with CIPRO or ENRO up to 1 μg/g, all 19 samples spiked above the 500 ng/g tolerance were correctly screened as positive with no false negatives, but 3 of 18 samples spiked below 500 ng/g were classified as positive. This method minimized the use of chlorinated solvents and significantly improved sample throughput.

Land application of solid wastes leads to the accumulation of both metals and antimicrobials in soils. To understand the effects of metal and antibiotic interaction on their accumulation by the earthworm Eisenia fetida, uptake and elimination kinetics and subcellular distribution of cadmium (Cd) and ciprofloxacin (CIP) were determined. The kinetics was accurately described by a one-compartment first-order kinetic model. Bioaccumulation kinetics and subcellular distribution of CIP were not affected by Cd addition. However, Cd exhibited different metabolic and subcellular distribution patterns. With CIP, Cd uptake flux and elimination rate constants were about 2.2 and 9.8 times, respectively, those without CIP. In the presence of CIP, Cd redistributed from fractions D (associated with granules) and E (associated with tissue fragments and cell membranes) to fraction C (associated with cytosol). Without CIP, Cd in fraction C could not be excreted, whereas with CIP, Cd in fraction C was significantly excreted, and the excretion rate constant was consistent with that of CIP. A good relationship was found between CIP and Cd in earthworms during uptake and elimination periods (p < 0.01). Our results indicated that the Cd-CIP complex may be taken up, stored, and eliminated by earthworms.

Recent work demonstrates that three widely administered antibiotics (ciprofloxacin, lincomycin, and trimethoprim) are oxidized by potassium permanganate [KMnO(4), Mn(VII)] under conditions relevant to water treatment operations. However, tests show that little to no mineralization occurs during reactions with Mn(VII), so studies were undertaken to characterize the reaction products and pathways and to assess the effects of Mn(VII)-mediated transformations on the antibacterial activity of solutions. Several oxidation products were identified for each antibiotic by liquid chromatography-tandem mass spectrometry (LC-MS/MS). For ciprofloxacin, 12 products were identified, consistent with oxidation of the tertiary aromatic and secondary aliphatic amine groups on the piperazine ring and the cyclopropyl group. For lincomycin, seven products were identified that indicate structural changes to the pyrrolidine ring and thioether group. For trimethoprim, seven products were identified, consistent with Mn(VII) reaction at C═C double bonds on the pyrimidine ring and the bridging methylene group. Oxidation pathways are proposed based on the identified products. Bacterial growth inhibition bioassays (E. coli DH5α) show that the mixture of products resulting from Mn(VII) reactions with the antibiotics collectively retain negligible antibacterial potency in comparison to the parent antibiotics. These results suggest that permanganate can be an effective reagent for eliminating the pharmaceutical activity of selected micropollutants during drinking water treatment.

Analyte quantitation can be achieved from second-order data in the presence of uncalibrated components using multivariate calibration methods such as partial least-squares with residual bilinearization. However, the latter fails under conditions of identical profiles for interfering agents and calibrated components in one of the data dimensions. To overcome this problem, a new residual bilinearization procedure for linear dependency is here introduced. Simulated data show that the new model can conveniently handle the studied analytical problem, with a success comparable to multivariate curve resolution-alternating least-squares and also comparable to a version of parallel factor analysis adapted to cope with linear dependencies. The new approach has also been applied to two experimental examples involving the determination of the antibiotic ciprofloxacin in (1) urine samples from lanthanide-sensitized excitation-time decay matrixes and (2) serum samples from a novel second-order signal based on the time evolution of chemiluminescence emission. The results indicate good analytical performance of the new procedure toward the analyte in comparison with the classical approaches.

This paper describes the construction of an impedimetric immunosensor for the label-free detection of ciprofloxacin, an antibiotic belonging to synthetic fluoroquinolones. A poly(pyrrole-N-hydroxysuccinimide) film was electrogenerated onto electrodes and then used for the reagentless covalent binding of a fluoroquinolone model bearing an amino group. The resulting electrodes were utilized to immobilize a layer of anticiprofloxacin antibody onto the polymer surface by immunoreaction. In presence of ciprofloxacin, the antibody was displaced in solution inducing marked changes in the impedance of the sensor electrodes. These phenomena were detected and characterized by electrochemical impedance spectroscopy allowing the selective detection of extremely low ciprofloxacin concentration, namely, 1 x 10(-12) g mL(-1) or 3 pmol L(-1). Sensors exposed to ciprofloxacin showed a decrease in the sum of the interfacial resistances with the increase in ciprofloxacin concentration from 1 x 10(-12) to 1 x 10(-6) g mL(-1).

A new spectrofluorimetric method has been developed for the quantification of danofloxacin (DANO) and difloxacin (DIFLO), in the presence of the primary metabolite of difloxacin, with sarafloxacin (SARA) as interference, in chicken tissue samples. The method is based on second-order multivariate calibration, applying parallel factor analysis (PARAFAC), to the excitation-emission matrices (EEMs) of these compounds. High overlapping of the signals and influence of matrix effects were observed. To solve the problem, the standard addition method was used. Chemical variables were optimized. The measured EEMs of the analytes, as analytical signals, allowed their quantification in chicken tissue samples. Solid phase extraction was used for the extraction of the analytes in real samples. The range of concentration examined varied from 30 to 100 ng g(-1) for danofloxacin, and from 100 to 200 ng g(-1) for difloxacin. Both analytes can be analyzed individually, and the binary mixture can be resolved, with recoveries comprising between 88.7 and 106.6%.

This paper describes the construction of a labeless immunosensor for the antibiotic ciprofloxacin in milk and its interrogation using an ac impedance protocol. Commercial screen-printed carbon electrodes were used as the basis for the sensor. Polyaniline was electrodeposited onto the sensors and then utilized to immobilize a biotinylated antibody for ciprofloxacin using classical avidin-biotin interactions. Antibody loaded electrodes were exposed to solutions of antigen in milk and interrogated using an ac impedance protocol. The faradaic component of the impedance of the electrodes was found to increase with increasing concentration of antigen. Control samples containing a nonspecific IgG antibody were also studied but were found to display large nonspecific responses, probably due to the antibody binding some of the large number of components found in milk. Control sensors could, however, be fabricated using antibodies specific for species not found in milk. Calibration curves could be obtained by subtraction of the responses for specific and control antibody-based sensors, thereby eliminating the effects of nonspecific adsorption of antigen. Sensors exposed to ciprofloxacin in milk gave increases in impedance whereas ciprofloxacin in phosphate buffer led to decreases, indicating the possibility of developing sensors which can both detect and differentiate between free and chelated antigen.

A new method for the determination of ciprofloxacin, the major metabolite of enrofloxacin, for concentrations between 20 and 200 ng/mL by means of matrix isopotential synchronous fluorescence spectrometry and derivative techniques is proposed. This new method is useful for the determination of compounds in samples with unknown background fluorescence, such as ciprofloxacin in whey, without the need of tedious preseparation. The determination was performed in an ethanol/water medium (20% v/v) at pH 4.8, provided by adding a sodium acetate/acetic acid buffer solution. Since enrofloxacin is widely used as an antibacterial agent in veterinary medicine, the method was successfully applied to the determination of its main metabolite in milk. An exhaustive statistical analysis has been developed to all calibration graphs. This treatment includes robust regression such as least median of squares, which also detects outliers and leverage points. The overall least-squares regression has been applied to find the more exact straight line that fits the experimental data. The error propagation has been considered to calculate the detection limit and the repeatability of the method.

The on-line nanoscale coupling of a surface plasmon resonance (SPR)-based inhibition biosensor immunoassay (iBIA) for the screening of low molecular weight molecules with nano-liquid-chromatography electrospray ionization time-of-flight mass spectrometry (nano-LC ESI TOF MS) for identification is described. The interface is based on a reusable recovery chip (RC) that contains a nanoscale biosorbent composed of a hydrogel layer modified with antibodies raised against the analyte featuring the unique possibility of performance characterization using the SPR biosensor. Various hydrogel chemistries were evaluated, and the standard Biacore CM5 chip showed the highest capture capacity in combination with affinity-purified polyclonal antibodies. The procedure has four stages: the samples are prepared (1) and screened using a screening chip (SC) in the iBIA (2). Suspected noncompliant samples as being noncompliant are reinjected over the RC, and the analyte is captured at subnanogram level (3). The captured analyte is released, and the eluate is analyzed with nano-LC ESI TOF MS via a loop-type interface (4). The coupling of the technologies proved effective for screening enrofloxacin, a model compound, in incurred chicken muscle samples followed by identity confirmation in suspected noncompliant samples. Ciprofloxacin, a known metabolite of enrofloxacin, was identified as well in incurred chicken samples. This demonstrates the potential of the technologies coupled by means of a RC for the rapid screening and identification of known as well as unknown compounds. Finally, we demonstrate the feasibility of combining the two biosensor chips (SC and RC) with a robust chip-based nano-LC chip TOF MS system, thus providing a robust alternative triple-chip system.

A competitive indirect enzyme-linked immunosorbent assay (ciELISA) using monoclonal antibodies (Mabs) having broad specificity for fluoroquinolone (FQ) antibiotics is described. Four FQs, ciprofloxacin (CIP), enrofloxacin (ENR), norfloxacin (NOR), and ofloxacin (OFL), were conjugated to bovine serum albumin for immunogens and to ovalbumin for coating antigens. A Mab C4A9H1 raised against the CIP hapten exhibited high cross-reactivity (35-100%) with 12 of 14 FQs and detected these FQs in a ciELISA below their maximum residue levels (MRLs) with good sensitivity at 50% binding inhibition (IC50). The quantitative structure-activity relationship (QSAR) between Mab C4A9H1 and various FQs by comparative molecular field analysis (CoMFA) showed a high predictive ability with a cross-validation q2 value of 0.866. Using a simple purification process and the broad-specificity ciELISA adapted for analysis of FQs in chicken muscle, chicken liver, honey, shrimp, and whole egg samples demonstrated recoveries of 60-93% for CIP, ENR, NOR, OFL, flumequine, and danofloxacin.

We investigated ciprofloxacin (a fluoroquinolone antibiotic) speciation as a function of pH in aqueous solution and in the presence of dissolved ferric ions and goethite using ATR-FTIR and UV-vis spectroscopy. The presence of dissolved and surface bound ferric species induced the deprotonation of the ciprofloxacin carboxylic acid group at pH < pKa1. The resultant ciprofloxacin zwitterions appeared to interact via both carboxylate oxygens to form bidentate chelate and bridging bidentate complexes within colloidal iron oxide-ciprofloxacin precipitates and bidentate chelates on the goethite surface. However, the structure of the aqueous ferric-ciprofloxacin complexes remains unclear. Our evidence for bidentate chelates (involving only the carboxylate oxygens) on the goethite surface was distinct from previous IR studies of fluoroquinolone sorption to metal oxides that have proposed surface complexes involving both the keto and the carboxylate groups. We find that the distinct ciprofloxacin surface complex proposed at the goethite-water interface may be a result of differences in metal oxide mineralogy or assignment of the carboxylate antisymmetric stretch in the metal oxide-fluoroquinolone spectra.

A study of the depletion of enrofloxacin residues in eggs was carried out using a diphasic dialysis procedure for the extraction of fluoroquinolone residues from the matrix. Enrofloxacin was administered to laying hens through the intramuscular route (15 mg/day) and orally (12 mg/day). After daily collection, the egg albumen and the egg yolk were separated, and the residue levels were determined using an HPLC-MS (API-ESI) method. The enrofloxacin and ciprofloxacin peaks gradually increased until the fifth day, because the drug was employed for 5 days. However, differences were observed in the depletion curves of enrofloxacin and its metabolite when both parts of the egg and the mode of administration were considered.

A method for the quantitative determination of humanuse fluoroquinolone antibacterial agents (FQs) ciprofloxacin and norfloxacin in sewage sludge and sludge-treated soil samples was developed. The accelerated solvent extraction was optimized with regard to solvents and operational parameters, such as temperature, pressure, and extraction time. A 50 mM aqueous phosphoric acid/ acetonitrile mixture (1:1) was found to be optimum in combination with an extraction temperature of 100 degrees C at 100 bar, during 60 and 90 min for sewage sludge and sludge-treated soil samples, respectively. A cleanup step using solid-phase extraction substantially improved the selectivity of the method. Overall recovery rates for FQs ranged from 82 to 94% for sewage sludge and from 75 to 92% for sludge-treated soil, with relative standard deviations between 8 and 11%. Limits of quantification were 0.45 and 0.18 mg/kg of dry matter for sewage sludge and sludge-treated soils, respectively. The presented method was successfully applied to untreated and anaerobically digested sewage sludges and sludge-treated soils. Ciprofloxacin and norfloxacin were determined in sewage sludges from several wastewater treatment plants with concentrations ranging from 1.40 to 2.42 mg/kg of dry matter. Therefore, contrary to what may be expected for human-use pharmaceuticals, FQs may reach the terrestrial environment as indicated by the occurrence of FQs in topsoil samples from experimental fields, to which sewage sludge had been applied.

The mass flows of fluoroquinolone antibacterial agents (FQs) were investigated in the aqueous compartments of the Glatt Valley Watershed, a densely populated region in Switzerland. The major human-use FQs consumed in Switzerland, ciprofloxacin (CIP) and norfloxacin (NOR), were determined in municipal wastewater effluents and in the receiving surface water, the Glatt River. Individual concentrations in raw sewage and in final wastewater effluents ranged from 255 to 568 ng/L and from 36 to 106 ng/L, respectively. In the Glatt River, the FQs were present at concentrations below 19 ng/L. The removal of FQs from the water stream during wastewater treatment was between 79 and 87%. During the studied summer period, FQs in the dissolved fraction were significantly reduced downstream in the Glatt River (15-20 h residence time) (66% for CIP and 48% for NOR). Thus, after wastewater treatment, transport in rivers causes an additional decrease of residual levels of FQs in the aquatic environment. Refined predicted environmental concentrations for the study area compare favorably with the measured environmental concentrations (MEC) obtained in the monitoring study. Total measured FQ concentrations occurring in the examined aquatic compartments of the Glatt Valley Watershed were related to acute ecotoxicity data from the literature. The risk quotients obtained (MEC/PNEC < 1) following the recommendations of the European guidelines or draft documents suggest a low probability for adverse effects of the occurring FQs, either on microbial activity in WWTPs or on algae, daphnia, and fish in surface waters.

A liquid chromatographic method was developed for the determination of ciprofloxacin, enrofloxacin, and sarafloxacin at 10-200 ppb in both egg yolk and egg albumen of laying hens. Egg yolk or albumen was acidified with 1 M phosphoric acid followed by deproteination with acetonitrile and centrifugation. The supernate was pipetted out, and the remaining protein pellet was extracted three times with acetonitrile. The supernates were combined and concentrated at 50 degrees C to <0.7 mL. The final volume was adjusted to 2 mL with 0.02 M potassium phosphate buffer, pH 2.5. Separation of the analytes was achieved using reversed-phase HPLC with fluorometric detection. The recoveries were >80% and coefficients of variation <20%. After validation, the method was applied for use in a national survey for fluoroquinolones in table eggs. Of the 276 eggs assayed, none was found positive for fluoroquinolones. The findings suggest that illegal use of fluoroquinolones in laying hens is not widespread.

Fluoroquinolones (FQs) are among the most important antibacterial agents (synthetic antibiotics) used in human and veterinary medicine. An analytical method based on reversed-phase liquid chromatography with fluorescence detection was developed and validated for the simultaneous determination of nine FQs and the quinolone pipemidic acid in urban wastewater. Aqueous samples were extracted using mixed-phase cation-exchange disk cartridges that were subsequently eluted by ammonia solution in methanol. Recoveries were above 80% at an overall precision of better than 10%. Instrumental quantification limits varied between 150 and 450 pg injected. The presented method was successfully applied to quantify FQs in effluents of urban wastewater treatment plants. The two most abundant human-use FQs, ciprofloxacin and norfloxacin, occurred in primary and tertiary waste-water effluents at concentrations between 249 and 405 ng/L and from 45 to 120 ng/L, respectively. The identity of FQs in urban wastewater was confirmed by recording full fluorescence spectra and liquid chromatography directly coupled to tandem mass spectrometry. These results indicate that conventional environmental risk assessment overestimates FQ concentrations in surface waters by 1 to 2 orders of magnitude.

An indirect competitive enzyme-linked immunosorbent assay (ELISA) was developed to detect ciprofloxacin (CPFX) in food animal edible tissues. CPFX was converted by an active ester method into conjugates CPFX-bovine serum albumin (CPFX-BSA) and CPFX-human serum albumin (CPFX-HSA), which both allowed production of CPFX-specific rabbit antisera. In the ELISA, CPFX-HSA was coated onto the microtiter plate, followed by incubation with standard CPFX and anti-CPFX antibody. The indirect competitive ELISA revealed that the antisera have no cross-reactivity with penicillin, gentamicin, neomycin, sulfadiazine, and chlortetracycline. The antisera cross-reacted with enrofloxacin and norfloxacin about 69.8 and 44.6% as much as they did with CPFX. This ELISA was highly sensitive (0.32 ng/mL) to CPFX determination. Recovery of CPFX at 40 microg/kg was 75.58% in pork, 81.29% in chicken, and 84.30% in milk. The coefficients of variation varied from 3.7 to 9.2% over the range of CPFX concentrations studied. The linear detection range was between 1.6 and 1000 ng/mL. The results suggest that this ELISA is a specific, accurate, and convenient method for the detection of CPFX residues in food animal edible tissues.

Isolation of sarafloxacin (SAR) from fortified and incurred chicken eggs was done by a combination of liquid-liquid extraction and aqueous on-line microdialysis performed on an automated trace enrichment of dialysates (ASTED) system. The ASTED system coupled a sample cleanup procedure with HPLC and programmable fluorescence detection. Overall recoveries of 87-102% for SAR were obtained from samples fortified over a range of 1-100 ng/g. The relative standard deviation values ranged from 22 to 26% for samples fortified between 1 and 5 ng/g and from 2 to 12% for samples fortified between 10 and 100 ng/g. The limits of detection and quantitation were 0.2 and 1 ng/g, respectively. Eggs containing incurred SAR, which were collected over a 3-day dosing period and for 5 consecutive days thereafter, also were analyzed by using this technique. Because the method is automated, 35 samples can be processed within a 24-h period, which enables large data sets to be acquired over a short time period.