ß-Cyclodextrin Polymerized in Cross-Flowing Channels of Biomass Sawdust for Rapid and Highly Efficient Pharmaceutical Pollutants Removal from Water.

Water pollution arising from pharmaceuticals has raised great concerns about the potential risks for biosphere and human health. However, rapid and efficient removal of pharmaceutical contaminants from water remains challenging. Wood sawdust, a byproduct of the wood-processing industry, is an abundant, cost-effective, and sustainable material with a unique hierarchically porous microstructure. These features make wood sawdust quite interesting as a filtration material. Here, we report a novel cross-flow filtration composite based on ß-cyclodextrin-polymer-functionalized wood sawdust (ß-CD/WS) in which the pharmaceutical contaminant water flows through the sawn-off vessel channels and the micropores on the surface of the cell walls, generating the turbulence. Such water flow characteristics ensure full contact between pharmaceutical pollutants and ß-CD grafted on the cellulose backbone of wood sawdust, thereby enhancing the water treatment efficiency. Consequently, the ß-CD/WS filter device shows a high removal efficiency of over 97.5% within 90 s for various pharmaceutical contaminants including propranolol, amitriptyline, chlortetracycline, diclofenac, and levofloxacin, and a high saturation uptake capacity of 170, 156, 257, 159, and 185 mg g-1, respectively. The high-performance wood-sawdust-based cross-flow filtration opens new avenues for solving the global water pollution issues, especially those caused by pharmaceutical contaminants.

Simultaneous determination of olaquindox, oxytetracycline and chlorotetracycline in feeds by high performance liquid chromatography with ultraviolet and fluorescence detection adopting online synchronous derivation and separation.

Olaquindox, oxytetracycline and chlorotetracycline were widely used in feed as antibiotics and growth promoter to improve feed conversion efficiency and increase the rate of weight gain for animals. However, the use of these antibiotics in feed was gradually prohibited because of concerns about contamination and resistance in animals. A quantitative and confirmatory method for determining the presence of olaquindox, oxytetracycline and chlorotetracycline in feed by high performance liquid chromatography equipped with ultraviolet detector in series with fluorescence detector (HPLC-UVD-FLD) was developed, optimized, and validated in three different matrices (compound, concentrated and premix feed). The analytes extraction was performed with a mixture of acetonitrile and 0.1 mol/L ethylenediamine tetraacetic acid disodium-Mcllvaine buffer (1:4, v/v) by one step sample preparation procedure. The validated method presented a broad linear range and good linearity with weighted least square method. The decision limit of the analytes ranged from 0.61 to 0.77 mg/kg for olaquindox, 0.90 to 1.2 mg/kg for oxytetracycline and 1.3 to 2.0 mg/kg for chlorotetracycline. The average recovery values found in intermediate precision conditions were ranged from 88.0 to 99.7% for olaquindox with RSD lower than 11.1%, from 84.4 to 99.0% for oxytetracycline with RSD lower than 9.6%, from 83.8 to 97.5% for chlorotetracycline with RSD lower than 10.0%. By Youden test and bottom-up method, the method was proved to be sufficiently robust and had a small uncertainty for different concentration levels. The developed method was successfully utilized for commercial feed samples to monitor complex cross contamination and residue conditions. Online synchronous derivation and separation using ultraviolet detector in series with fluorescence detector can effectively prevent false positive of chlorotetracycline in feed caused by vegetable meal. Since olaquindox, oxytetracycline and chlorotetracycline are widely used in feed, the developed method provide an important and analytical tool for the simultaneous identification and quantification of them in feed to monitor its risk of cross contamination and excessive content.

Quantification of Veterinary Antibiotics in Pig and Poultry Feces and Liquid Manure as a Non-Invasive Method to Monitor Antibiotic Usage in Livestock by Liquid Chromatography Mass-Spectrometry.

Antibiotics are active substances frequently used to treat and prevent diseases in animal husbandry, especially in swine and poultry farms. The use of manure as a fertilizer may lead to the dispersion of antibiotic residue into the environment and consequently the development of antibiotic-resistant bacteria. Most pharmaceutical active ingredients are excreted after administration, in some cases up to 90% of the consumed dose can be found in the feces and/or urine as parent compound. Therefore, due to antibiotic metabolism their residues can be easily detected in manure. This article describes a method for simultaneous analysis of ciprofloxacin, chlortetracycline, doxycycline, enrofloxacin, lincomycin, oxytetracycline, tetracycline, tiamulin, trimethoprim and tylosin in feces, liquid manure and digestate. Antibiotics were extracted from the different matrices with McIlvaine-Na2EDTA buffer solution and the extract was purified by the use two techniques: d-SPE and SPE (Strata-X-CW cartridges) and final eluent was analyzed by LC-MS and LC-MS/MS. The European Commission Decision 2002/657/EC was followed to conduct the validation of the method. Recoveries obtained from spiked pig and poultry feces and liquid manures samples ranged from 63% to 93% depending on analytes. The analysis of 70 samples (feces, liquid manure and digestate) revealed that 18 samples were positive for the presence of doxycycline, oxytetracycline, tetracycline, chlortetracycline, enrofloxacin, tiamulin and lincomycin. The results obtained in the presented study demonstrated that animal feces can be used as a non-invasive method detection antibiotic usage in animal production.

Removal of chlortetracycline by nano- micro-electrolysis materials: Application and mechanism.

Nano micro-electrolysis materials (nMETs) have been used to degrade refractory pollutants in batch experiments. The reasonable formation mechanism of nMETs was given through DMXY digital biomicroscopy. Based on the kinetic data of Chlortetracycline (CTC) removal by nMETs in batch experiments, combined with the binomial distribution equation of CTC reduction by nano materials an experimental-scale fluidized bed (ESFB) was designed. The effects of CTC removal performance, pH and iron ion concentration were investigated. Under pure CTC solution environment, the experimental data showed that the average removal rates of CTC by nMET and nano micro-electrolysis material with loading copper (Cu-nMET) are 90.0% and 95.7% in ESFB, respectively. In the presence of nitrate, although the consumption of two kinds of nano-materials increased, their removal efficiencies of pollutants have 2.2%, 0.2% increase compared with the nitrate-free ESFB. At the same time, the CTC degradation pathway and the enhanced removal mechanism by Cu-nMET was proposed. Through microelectrolysis reaction, complexation reaction and the active substances produced, the intermediate products can be degraded completely to NH4+, CO2, H2O and so on. This study aims to provide a theoretical basis for the environmental application of nMETs.

Evaluating the efficiency of different natural clay sediments for the removal of chlortetracycline from aqueous solutions.

Natural clay sediments were collected from ten different localities in Saudi Arabia (S-1 from eastern, S-2 to S-4 from middle and S-5 to S-10 from western regions), characterized and evaluated for their efficiency towards chlortetracycline (CTC) removal from aqueous solutions. Sediment S-4 exhibited highest surface area (288.5 m2 g-1), followed by S-5, S-9, and S-1 (252.1, 249.6, and 110.4 m2 g-1, respectively). Sediments S-5, S-9, S-2, and S-4 showed the highest cation exchange capacities (CEC) (62.33, 56.54, 52.72, and 46.85 cmol kg-1, respectively). The pH range of 3.5-5.5 was optimum for the highest CTC removal. Freundlich model was best fitted to CTC sorption data (R2 = 0.96-0.99), followed by Dubinin-Radushkevich model (R2 = 0.89-0.97). The sediments S-4, S-5, and S-9 exhibited the highest CTC removal efficiency (98.80-99.05%), which could be due to higher smectite and kaolinite contents, CEC, surface area and layered structure. Post-sorption XRD patterns shown new peaks and peak shifts confirming the sorption of CTC. Electrostatic interactions, interlayer sorption and H-π bonding were the potential CTC sorption mechanisms. Therefore, natural clay sediments with high sorption capacities could efficiently remove CTC from contaminated aqueous media.

Adsorption properties, kinetics & thermodynamics of tetracycline on carboxymethyl-chitosan reformed montmorillonite.

This paper describes a modification method of Na-montmorillonite (Na-Mt) with carboxymethyl-chitosan (CMC). The as-prepared samples were analyzed by Fourier transform infrared (FT-IR), X-ray diffraction (XRD), Brunauer-Emmett-Teller (BET) surface analyzer and thermogravimetric analysis (TGA). Two common tetracycline antibiotics, tetracycline (TET) and chlortetracycline (CTC), were selected as the represented pollutants and adsorbed by CMC-Mt under different experimental conditions. The intercalation of CMC obviously amplified the basal spacing of the interlayers confirmed by XRD measurements and improved the adsorption capacities of montmorillonite to some degree. The results showed that the tetracycline antibiotic sorption onto CMC-Mt was mainly dependent on pH and was not affected by temperature. Besides, the removal of TET and CTC rapidly attained an equilibrium within 2 h of contact time. The kinetic data of adsorption was determined by first-order, second-order kinetics and intraparticle diffusion models. The kinetic study indicates that the TET and CTC adsorption processes obeyed the second-order kinetics. The Freundlich isotherm study was in agreement with the practical data, suggesting a heterogeneous sorption process. Furthermore, the thermodynamic studies revealed that the removal process was more spontaneous at a lower temperature, implying it an exothermic reaction. The synthesized adsorbent CMC-Mt can be widely used in the treatment of wastewater.

Contrastive removal of oxytetracycline and chlortetracycline from aqueous solution on Al-MOF/GO granules.

The presence of tetracycline antibiotics (TCS) in the water and wastewater has raised growing concern due to its potential environmental impacts; thus, their removal is of high importance. In this study, a novel aluminum-based MOF/graphite oxide (Al-MOF/GO) granule was prepared as an adsorbent for the removal of TCS including oxytetracycline (OTC) and chlortetracycline (CTC). The adsorbent was characterized via XRD, FTIR, BET, SEM, and XPS methods. The granules exhibited similar crystal structure and some new mesopores appearing compared to the parent Al-MOF/GO powder. In addition, the adsorption behavior of OTC and CTC on samples was explored as a function of initial concentration, contact time, pH, and ionic strength by means of batch experiments. The adsorption capacity reached to 224.60 and 240.13 mg·L-1 for OTC and CTC, at C0 = 60 mg·L-1 as well as ambient temperature respectively. Moreover, the adsorption process of OTC and CTC on Al-MOF/GO samples can be better delineated by pseudo-second-order kinetics and Freundlich isotherm models. Besides, the adsorption mechanism over Al-MOF/GO granules was proposed, which could be ascribed to π-π interaction, cation-π bonding, and hydrogen bond. Finally, the great water stability, separation performance, and regeneration efficiency of these novel granules indicated their potential application in the OTC and CTC removals from aqueous solution.

[Removal of Typical Antibiotics During Aerobic Composting of Human Feces].

Aerobic composting experiments were conducted under three different temperatures (55℃, 35℃, and non-temperature-controlled) with human feces and sawdust as the compost material and bulky matrix respectively. Attention was paid to the effects of temperature on the removal of four typical antibiotics (tetracycline, chlortetracycline, sulfadiazine, and ciprofloxacin) during human feces aerobic composting. Furthermore, three specific experiments were conducted to identify the possible antibiotic degradation mechanisms in aqueous solution and during composting with moist sterile sawdust but without feces and composting with feces and moist sterile sawdust. The most effective removal of the four antibiotics was achieved through thermophilic composting (55℃). At 55℃, more than 90% of all the antibiotics were removed. The degradation of tetracycline and chlortetracycline was more temperature-dependent than that of sulfadiazine and ciprofloxacin. Moreover, tetracycline and chlortetracycline removal was primarily through hydrolysis in moist environments. However, sulfadiazine and ciprofloxacin removal was mainly attributed to adsorption by sawdust particles. The contribution of microbial action to antibiotic degradation was insignificant for all the antibiotics except ciprofloxacin. Up to 20% of this antibiotic was degraded by microbial action.

Activation of persulfate by homogeneous and heterogeneous iron catalyst to degrade chlortetracycline in aqueous solution.

This study investigates the removal of chlortetracycline (CTC) antibiotic using sulfate radical-based oxidation process. Sodium persulfate (PS) was used as a source to generate sulfate radicals by homogeneous (Fe2+) and heterogeneous (zero valent iron, ZVI) iron as a catalyst. Increased EDTA concentration was used to break the CTC-Fe metal complexes during CTC estimation. The influence of various parameters, such as PS concentration, iron (Fe2+ and ZVI) concentration, PS/iron molar ratio, and pH were studied and optimum conditions were reported. CTC removal was increased with increasing concentration of PS and iron at an equal molar ratio of PS/Fe2+ and PS/ZVI processes. PS/Fe2+ and PS/ZVI oxidation processes at 1:2 (500 µM PS and 1000 µM) molar ratio showed 76% and 94% of 1 µM CTC removal in 2 h. Further increased molar ratio 1:2 onwards, PS/Fe2+ process showed a slight increase in CTC degradation whereas in PS/ZVI process showed similar degradation to 1:2 (PS/Fe) ratio at constant PS 500 µM concentration. Slower activation of persulfate which indirectly indicates the slower generation of sulfate radicals in PS/ZVI process showed higher degradation efficiency of CTC. The detected transformation products and their estrogenicity results stated that sulfate radicals seem to be efficient in forming stable and non-toxic end products.

Determination of Chlortetracycline Residues, Antimicrobial Activity and Presence of Resistance Genes in Droppings of Experimentally Treated Broiler Chickens.

Tetracyclines are important antimicrobial drugs for poultry farming that are actively excreted via feces and urine. Droppings are one of the main components in broiler bedding, which is commonly used as an organic fertilizer. Therefore, bedding becomes an unintended carrier of antimicrobial residues into the environment and may pose a highly significant threat to public health. For this depletion study, 60 broiler chickens were treated with 20% chlortetracycline (CTC) under therapeutic conditions. Concentrations of CTC and 4-epi-CTC were then determined in their droppings. Additionally, this work also aimed to detect the antimicrobial activity of these droppings and the phenotypic susceptibility to tetracycline in E. coli isolates, as well as the presence of tet(A), tet(B), and tet(G) resistance genes. CTC and 4-epi-CTC concentrations that were found ranged from 179.5 to 665.8 µg/kg. Based on these data, the depletion time for chicken droppings was calculated and set at 69 days. All samples presented antimicrobial activity, and a resistance to tetracyclines was found in bacterial strains that were isolated from these samples. Resistance genes tet(A) and tet(B) were also found in these samples.

Magnetic chitosan-based adsorbent prepared via Pickering high internal phase emulsion for high-efficient removal of antibiotics.

A novel magnetic chitosan-g-poly(2-acrylamide-2-methylpropanesulfonic acid) (CTS-g-AMPS) porous adsorbent was prepared by grafting the AMPS onto the CTS in the Fe3O4 stabilized Pickering high internal phase emulsions (Pickering-HIPEs) and used for the adsorptive removal of the antibiotics tetracycline (TC) and chlorotetracycline (CTC). The results of the structure characterization showed that porous structure of the adsorbent can be tuned easily by altering amount of Fe3O4-MNPs-M and the electrostatic attraction of between SO3- and CTC, TC was the main adsorption driving force. The adsorption capacities of the adsorbent for TC and CTC can be reached to 806.60 and 876.60mg/g in a wide pH ranged from 3.0 to 11.0, respectively. And the adsorption equilibrium can be reached within 90min for TC and 50min for CTC. The magnetic porous adsorbent had good reusability, which can still attain a high adsorption capacity of 759.82 and 842.99mg/g for TC and CTC after five consecutive adsorption cycles, respectively. Therefore, the as-prepared CTS-g-AMPS magnetic adsorbent is potential to be used for adsorption removal of antibiotics from water.

An Unprecedented Blue Chromophore Found in Nature using a "Chemistry First" and Molecular Networking Approach: Discovery of Dactylocyanines A-H.

Guided by a "chemistry first" approach using molecular networking, eight new bright-blue colored natural compounds, namely dactylocyanines A-H (3-10), were isolated from the Polynesian marine sponge Dactylospongia metachromia. Starting from ilimaquinone (1), an hemisynthetic phishing probe (2) was prepared for annotating and matching structurally related natural substances in D. metachromia crude extract network. This strategy allowed characterizing for the first time in Nature the blue zwitterionic quinonoid chromophore. The solvatochromic properties of the latter are reported.

[Removal of Chlortetracycline and Morphological Changes in Heavy Metals in Swine Manure Using the Composting Process].

The widespread use of antibiotics and heavy metals in livestock farms results in large residues of antibiotics and heavy metals in the livestock manure. Composting technology can biodegrade residual antibiotics and solidify heavy metals. A pilot composting reactor was used to analyze the characteristics of chlortetracycline (CTC) removal at different antibiotic concentrations[0 mg·kg-1 (CK), 10 mg·kg-1 (T1), and 50 mg·kg-1 (T2)]. Moreover, the morphological changes in heavy metals during the composting process were analyzed. After composting, no chlortetracycline was detected in the CK group and the antibiotics degradation rates of T1 and T2 groups reached 96.31% and 97.32%, respectively. The chlortetracycline degradation fits the pseudo-first-order kinetics model. Heavy metals can be solidified during the composting; thus, the bioavailable state of Cu and Zn (exchangeable state, reducible state) changed into the oxidation state and residues with apparent passivation formed. The correlation analysis showed that the removal of CTC showed strong positive correlations with the biological available Cu and Zn.

Degradation of chlortetracycline in wastewater sludge by ultrasonication, Fenton oxidation, and ferro-sonication.

Residual emerging contaminants in wastewater sludge remain an obstacle for its wide and safe applications such as landfilling and bio-fertilizer. In this study, the feasibility of individual ultrasonication (UlS) and Fenton oxidation (FO) and combined, Ferro-sonication processes (FO) on the degradation of chlortetracycline (CTC) in wastewater sludge was investigated. UlS parameters such as amplitude and sonication time were optimized by response surface methodology (RSM) for further optimization of FS process. Generation of highly reactive hydroxyl radicals in FO and FS processes were compared to evaluate the degradation efficiency of CTC. Increasing in the ratio of hydrogen peroxide and iron concentration showed increased CTC degradation in FO process; whereas in FS, an increase in iron concentration did not show any significant effect (p>0.05) on CTC degradation in sludge. The estimated iron concentration in sludge (115mg/kg) was enough to degrade CTC without the addition of external iron. The only adjustment of sludge pH to 3 was enough to generate in-situ hydroxyl radicals by utilizing iron which is already present in the sludge. This observation was further supported by hydroxyl radical estimation with adjustment of water pH to 3 and with and without the addition of iron. The optimum operating UlS conditions were found to be 60% amplitude for 106min by using RSM. Compared to standalone UlS and FO at 1:1 ratio, FS showed 15% and 8% increased CTC degradation respectively. In addition, UlS of sludge increased estrogenic activity 1.5 times higher compared to FO. FS treated samples did not show any estrogenic activity.

Chlortetracycline removal by using hydrogen based membrane biofilm reactor.

In the last years, increasing attention has been paid on the presence of antibiotics in aqueous environments due to their ecological damage and potential adverse effects on organisms. Membrane biofilm reactors (MBfR) have been gained a significant popularity as an advanced wastewater treatment technology in removing of organic micro-pollutants. In this study, the performance of H2-MBfR for simultaneous removal of nitrate and chlortetracycline, formation of transformation products and community analysis of the biofilm grown on the gas permeable hollow fiber membranes was evaluated by considering effect of the hydraulic retention time, surface loadings of target pollutants and H2 pressure. The results showed that the simultaneous chlortetracycline (96%) and nitrate removal (99%) took placed successfully under the conditions of 5h HRT and 2psi H2 pressure. It has been determined that the main elimination process was biodegradation and Betaproteobacteria species was responsible for chlortetracycline degradation.

Quantitative analysis of antibiotics in aquifer sediments by liquid chromatography coupled to high resolution mass spectrometry.

A highly effective analytical method for multi-residue determination of antibiotics in aquifer sediments was first established in this study. Microwave-assisted solvent extraction (MASE) and solid-phase extraction were used for sample pre-concentration and purification, ultra-high performance liquid chromatography coupled to hybrid quadrupole-high resolution Orbitrap mass spectrometry (UHPLC-Q-Orbitrap) was applied for detection. For high resolution mass spectrometry (HRMS), the target compounds were tentatively identified by retention time and accurate mass which was measured with precursor ions in Target-SIM scan, and then confirmed by the monitoring of daughter ion fragments which were generated in dd-MS(2) scan. The results provided good mass accuracy with mass deviations below 2ppm (except norfloxacin with -2.3ppm) for quantitative analysis of the compounds by HRMS. Reasonable recoveries of all analytes were obtained more than 60% (except doxytetracycline) in fortification samples at concentrations higher than 10µgkg(-1). Relative standard deviations of repeatability and inter-day precision were below 21% and 11%. Limits of detection (LOD) ranged from 0.1 to 3.8µgkg(-1), whereas limits of quantification (LOQ) were established between 0.3-9.0µgkg(-1). The method was applied to analyze real aquifer sediment samples in different aquifer depth of 4.0, 7.5, 13.0 and 18.0m. Chlorotetracycline and ofloxacin were observed at relative high concentrations of 53 and 19µgkg(-1) respectively in 18.0m deepness. The exposure to low doses of these compounds in subsurface environment increases concerns on long-term ecological security of underground system.

Elimination patterns of worldwide used sulfonamides and tetracyclines during anaerobic fermentation.

Antibiotics such as sulfonamides and tetracyclines are frequently used in veterinary medicine. Due to incomplete absorption in the animal gut and/or unmetabolized excretion, the substances can enter the environment by using manure as soil fertilizer. The anaerobic fermentation process of biogas plants is discussed as potential sink for antibiotic compounds. However, negative impacts of antibiotics on the fermentation process are suspected. The elimination of sulfadiazine, sulfamethazine, tetracycline and chlortetracycline in semi-continuous lab-scale fermenters was investigated. Both biogas production and methane yield were not negatively affected by concentrations up to 38 mg per kg for sulfonamides and 7 mg per kg for tetracyclines. All substances were partly eliminated with elimination rates between 14% and 89%. Both matrix and structure of the target molecule influenced the elimination rate. Chlortetracycline was mainly transformed into iso-chlortetracycline. In all other cases, the elimination pathways remained undiscovered; however, sorption processes seem to have a negligible impact.

Hybrid intelligent control of substrate feeding for industrial fed-batch chlortetracycline fermentation process.

The lack of accurate process models and reliable online sensors for substrate measurements poses significant challenges for controlling substrate feeding accurately, automatically and optimally in fed-batch fermentation industries. It is still a common practice to regulate the feeding rate based upon manual operations. To address this issue, a hybrid intelligent control method is proposed to enable automatic substrate feeding. The resulting control system consists of three modules: a presetting module for providing initial set-points; a predictive module for estimating substrate concentration online based on a new time interval-varying soft sensing algorithm; and a feedback compensator using expert rules. The effectiveness of the proposed approach is demonstrated through its successful applications to the industrial fed-batch chlortetracycline fermentation process.

Biodegradation of three tetracyclines in swine wastewater.

Tetracyclines (TCs), including tetracycline (TC), oxytetracycline (OTC), and chlortetracycline (CTC), are amongst the most common antibiotics used in animal husbandry. Residual amounts of these antibiotics in the environment are a concern because they contribute to selection of resistant bacteria. In this study, we investigated the biodegradation of three TCs in swine wastewater. In batch experiments, OTC and CTC were completely degraded at d 18 and 20, respectively, but TC was remained at 7.1% after 20 d incubation. The degradation rates of TCs in the wastewater were in the order of OTC > CTC > TC. Degradation of the TCs was enhanced by the addition of enzyme extract from spent mushroom compost (SMC) of Pleurotus eryngii. The degradation rates were higher with the addition of extract-containing microcapsules than suspended enzyme extract in swine wastewater. In the bioreactor experiment, the addition of extract-containing microcapsules enhanced the removal rates of the three TCs, and adding TCs twice maintained enzyme activity in the swine wastewater. Of the microorganism strains isolated from the wastewater samples, strain HL2 (identified as Xanthobacter flavus) showed the best degrading ability.

Zeolite imidazolate framework-8 as sorbent for on-line solid-phase extraction coupled with high-performance liquid chromatography for the determination of tetracyclines in water and milk samples.

Zeolite imidazolate framework-8 (ZIF-8) was used as the novel sorbent for on-line solid-phase extraction coupled with high-performance liquid chromatography (HPLC) for the determination of oxytetracycline (OTC), tetracycline (TC) and chlorotetracycline (CTC) in water and milk samples. 390mg of ZIF-8 was packed into a stainless steel column (3cm long×4.6mm i.d.) which was mounted on the HPLC injector valve to replace the sample loop. On-line solid-phase extraction of OTC, TC and CTC was achieved by loading sample solution at a flow rate of 3.0mLmin(-1) for 10min with the aid of a flow-injection system. The extracted analytes were subsequently eluted into a C18 analytical column (25cm long×4.6mm i.d.) for HPLC separation under isocratic condition with a mobile phase (10% MeOH-20% ACN-70% 0.02molL(-1) oxalic acid solution) at a flow rate of 1.0mLmin(-1). Under optimized conditions, the developed method gave the enhancement factors of 35-61, the linearity range of 5-1000µgL(-1), the detection limits of 1.5-8.0µgL(-1), quantification limits of 5.0-26.7µgL(-1), uncertainties of 0.9-1.1µgL(-1), and the sample throughput of 4 samples h(-1). The recoveries of OTC, TC and CTC at 50µgL(-1) in water and milk samples ranged from 70.3% to 107.4%.