Preparation of pH-responsive block copolymers for separation of cephalosporin antibiotics by open-tubular capillary electrochromatography.

Stimuli-responsive block copolymers have exhibited their feasibility for drug delivery and analysis of biomolecules. However, study of the electrophoretic behavior of antibiotics by open tubular capillary electrochromatography (OT-CEC) based on smart block copolymers coatings is still a substantial challenge. Herein, we reported an OT-CEC protocol for analysis of cephalosporin antibiotics with pH-responsive block copolymers as coatings. By using the reversible addition-fragmentation chain-transfers radical polymerisation technique, the smart poly(styrene-maleic anhydride-acrylic acid) (P(St-MAn-AA)) was synthesized and subsequently chemical bonded onto the inner walls of amino-grafted capillaries. The pH induced changes in the stretch/curl states of P(St-MAn-AA) chains were used to generate an adjustable hydrophobic/hydrophilic interaction and hydrogen bonds between the polymer coatings and the analytes. The OT-CEC performance was evaluated by varying the monomer ratios, polymer coating amounts and layers, buffer concentrations and pH values. Baseline separation of the three-test antibiotics was achieved at pH 8.0. The proposed OT-CEC technique was further applied to the determination of rat serum antibiotics in the metabolic processes. The present work demonstrates an enhancement in antibiotics separation efficiency, and shows a great potential for the preparation of stimuli-responsive block copolymers coatings and in OT-CEC analysis of real samples in living bio-systems.

Development and Validation of Liquid Chromatography-Tandem Mass Spectrometry Methods for the Quantification of Cefquinome, Ceftiofur, and Desfuroylceftiofuracetamide in Porcine Feces with Emphasis on Analyte Stability.

Cefquinome and ceftiofur are ß-lactam antibiotics used for the treatment of bacterial infections in swine. Although these antimicrobials are administered intramuscularly, the exposure of the gut microbiota to these cephalosporins is not well described. This exposure can contribute to the emergence and spread of antimicrobials in the environment and to the possible spread of antimicrobial resistance genes. To assess the impact of drug administration on the intestinal excretion of these antimicrobials it is essential to measure the amounts of native compound and metabolites in feces. Two (ultra)-high-performance liquid chromatography-tandem mass spectrometry ((U)HPLC-MS/MS) methods were developed and validated, one for the determination of cefquinome and ceftiofur and the other for the determination of ceftiofur residues, measured as desfuroylceftiofuracetamide, in porcine feces. The matrix-based calibration curve was linear from 5 ng g-1 to 1000 ng g-1 for cefquinome (correlation coefficient (r) = 0.9990 ± 0.0007; goodness of fit (gof) = 3.70 ± 1.43) and ceftiofur (r = 0.9979 ± 0.0009; gof = 5.51 ± 1.14) and quadratic from 30 ng g-1 to 2000 ng g-1 for desfuroylceftiofuracetamide (r = 0.9960 ± 0.0020; gof = 7.31 ± 1.76). The within-day and between-day precision and accuracy fell within the specified ranges. Since ß-lactam antibiotics are known to be unstable in feces, additional experiments were conducted to adjust the sampling protocol in order to minimize the impact of the matrix constituents on the stability of the analytes. Immediately after sampling, 500 µL of an 8 µg mL-1 tazobactam solution in water was added to 0.5 g feces, to reduce the degradation in matrix.

Ceftobiprole medocaril for the treatment of community-acquired pneumonia.

INTRODUCTION: Ceftobiprole is a novel broad-spectrum cephalosporin with excellent activity against a broad range of pathogens that are important in community-acquired pneumonia (CAP), including drug-resistant pneumococci, methicillin-resistant Staphylococcus aureus, and Pseudomonas aeruginosa. Areas covered: This article reviews the spectrum of activity, the main pharmacological and pharmacodynamic characteristics of ceftobiprole as well its clinical efficacy and safety in the treatment of CAP in adult patients. Expert opinion: Taking into account that the current treatment guidelines for CAP recommend the use of an adequate empirical therapy to improve its prognosis, ceftobiprole shows a profile of antimicrobial activity that would cover most etiological agents in patients with risk factors for infection caused by multidrug resistant organisms. The results of the pivotal clinical trial of patients hospitalized with CAP treated with ceftobiprole showed a high rate of clinical cure. The clinical tolerance of ceftobiprole in clinical trials was generally very good. These findings make ceftobiprole a good parenteral therapeutic alternative for the empirical treatment of CAP that requires hospitalization, especially in patients with risk factors for CAP caused by resistant microorganisms.

Antimicrobial activity of crude extracts from actinomycetes against mastitis pathogens.

The emergence of antimicrobial resistance to commonly used antibiotics has necessitated the development of new antimicrobial products. Crude extracts produced by actinomycetes contain antimicrobial metabolites that can inhibit bacterial growth. The objective of our study was to evaluate the antimicrobial activity of crude extracts (Caat1-54 and CaatP5-8) produced by actinomycetes against isolates of Staphylococcus aureus, Staphylococcus chromogenes, Streptococcus dysgalactiae, and Streptococcus uberis, which were obtained from the milk of cows affected by mastitis in 23 dairy herds. Twenty isolates of each bacterial species were used to define minimum inhibitory concentrations (MIC) of both crude extracts and ceftiofur (positive control). The MIC50 and MIC90 were defined at the concentration required to inhibit the growth of 50 and 90% of bacterial isolates tested, respectively. The MIC results were evaluated by survival analysis. Staphylococcus aureus isolates presented MIC90 of Caat 1-54 ≥6.25 µg/mL, ceftiofur ≥12.5 µg/mL, and Caat P5-8 ≥100 µg/mL. Streptococcus uberis presented MIC90 of ceftiofur ≥0.39 µg/mL, Caat 1-54 ≥50 µg/mL, and Caat P5-8 ≥100 µg/mL. Staphylococcus chromogenes isolated from subclinical mastitis presented MIC90 of Caat 1-54 ≥0.78 µg/mL and ceftiofur and Caat P5-8 of ≥6.25 and ≥100 µg/mL, respectively. Streptococcus dysgalactiae isolated from clinical mastitis presented similar MIC90 values between antimicrobials tested (ceftiofur, Caat 1-54, and Caat P-58), but these values (≥100 µg/mL) were higher than the values obtained from other pathogens evaluated in the present study. Our results indicate that Caat 1-54 and Caat P5-8 crude extracts present in vitro antimicrobial activity against isolates of Staph. aureus, Staph. chromogenes, Strep. dysgalactiae, and Strep. uberis isolated from clinical and subclinical mastitis.

Isolation, identification and in silico toxicity predictions of two isomers from cefotiam hydrochloride.

Two structural isomers of cefotiam in cefotiam hydrochloride for injection were observed, and the structures of the isomers were determined by mass spectrometry and various 1D and 2D NMR techniques. The thermo-isomerization mechanism of cefotiam was also discussed. Thermo-isomerization occurred not only in cefotiam but also in cephalosporins containing a 1-alkyl-1H-tetrazole-5-thiol side chain at C-3. Furthermore, the toxic effects of the two impurities of cefotiam hydrochloride were predicted and it is thought that they could be more toxic than cefotiam. The results reported in this article may be important for quality control and stability studies of this class of drugs.

Online fully automated SPE-HPLC-MS/MS determination of ceftiofur in bovine milk samples employing a silica-anchored ionic liquid as sorbent.

Solid-phase extraction coupled online with high performance liquid chromatography and tandem mass spectrometry was successfully applied to determine low concentrations of ceftiofur antibiotic in bovine milk samples. A silica-anchored ionic liquid was applied as sorbent material to be used as extraction phase in the proposed online system. The material was characterized by Fourier transform infrared spectroscopy and scanning electron microscopy. In order to improve the system reproducibility, the following experimental parameters were optimized: organic solvent percentage, time and sample loading flow rate. Subsequently, the method was validated presenting satisfactory results as adequate selectivity, good linearity and correlation coefficient higher than 0.98. The limit of detection and quantification were 0.1 and 0.7 µg/L, respectively. The precision of the methodology was evaluated as repeatability and intermediate precision, with relative standard deviation values lower than 15%. The accuracy of the method ranged from 72.8 to 137% and the minimum and maximum recovery values were 73.4 and 111.3%, respectively. After the validation, seven milk samples were analyzed and although ceftiofur was not detected in any of them the method was demonstrated to be efficient when applied to the analysis of milk samples fortified with the pollutant of interest.

Development of pH-responsive polymer and citrate aqueous two-phase system for extractive bioconversion of cefprozil.

A pH-responsive aqueous two-phase system (pH-ATPS) has been developed by sodium citrate and a recyclable pH-responsive polymer PADB6.8 that can response to the change of pH values. Phase separation mechanism is studied through Low field-NMR. All variables affecting the phase separation are evaluated. Phase characteristics (viscosity, density, interfacial tension) and phase separation kinetic are studied for understanding of separation process and operational parameters in applications. This pH-ATPS has the characters of low interfacial tension, high recovery leading efficient mass transfer and low cost. The proposed system can be used as a mild medium for extractive bioconversion with low cost. We applied this pH-ATPS in extractive bioconversion of cefprozil. Cefprozil is partitioned towards the polymer-rich phase while the substrates tended to be partitioned in the salt-rich phase. Extractive bioconversion of cefprozil in this pH-ATPS can improve yield of the enzymatic process and reduce the product hydrolysis in optimal conditions. The maximal conversion yield of cefprozil in the studied system is 91.0%.

Removal of cephalosporin antibiotics 7-ACA from wastewater during the cultivation of lipid-accumulating microalgae.

The aim of this study is to evaluate the feasibility of using lipid-accumulating microalgae to remove cephalosporin antibiotics 7-amino cephalosporanic acid (7-ACA) from wastewater with the additional benefit of biofuels production. Three isolated microalgal strains (namely, Chlorella sp. Cha-01, Chlamydomonas sp. Tai-03 and Mychonastes sp. YL-02) were cultivated under 7-ACA stress and their biomass productivity, lipid production and N-NO3- consumption were monitored. It was found that 7-ACA had slight inhibition effects on the microalgal growth at the ratio of 12.0% (Cha-01), 9.6% (YL-02), 11.7% (Tai-03). However, lipid accumulation in the three microalgae was not influenced by the presence of 7-ACA. The investigation on the 7-ACA removal mechanisms during microalgal growth shows that 7-ACA was mainly removed by microalgae adsorption as well as hydrolysis and photolysis reactions. This study demonstrates that using microalgae to treat antibiotic-containing wastewater is promising due to the potential of simultaneous antibiotic removal and biofuel production.

Mixed hemimicelles solid-phase extraction of cephalosporins in biological samples with ionic liquid-coated magnetic graphene oxide nanoparticles coupled with high-performance liquid chromatographic analysis.

A novel mixed hemimicelles solid phase extraction based on magnetic graphene oxide (Fe3O4/GO) and ionic liquid (IL) was developed for the simultaneous extraction and determination of trace cephalosporins in spiked human urine. The high surface area and excellent adsorption capacity of the graphene oxide after modification with1-hexadecyl-3-methylmidazoliumbromide(C16mimBr) were utilized adequately in the solid phase extraction(SPE) process. A comprehensive study of the parameters affecting the extraction recovery, such as the zeta-potential of magnetic graphene oxide, amounts of magnetic graphene oxide and surfactant, pH of solution, ionic strength, extraction time, and desorption condition were optimized. A comparative study on the use of different surfacant-coated Fe3O4/GO NPs as sorbents was presented. Good linearity (R(2)>0.9987) for all calibration curves was obtained. The LODs were ranged between 0.6 and 1.9ng mL(-1) for the cephalosporins and the LOQs were 1.5 to 5.5, respectively. Satisfactory recoveries(84.3% to 101.7%)and low relative standard deviations from 1.7% to 6.3% in biological matrices were achieved. The mixed hemimicelles magnetic SPE (MSPE) method based on ILs and Fe3O4/GO NPs magnetic separation has ever been successfully used for pretreatment of complex biological samples.

Feasibility study of recycling cephalosporin C fermentation dregs using co-composting process with activated sludge as co-substrate.

Composting is a potential alternative for cephalosporin C fermentation dregs (CCFDs) compared with incineration process or landfill because of its advantage of recovering nutrients. In this research, CCFDs and activated sludge (AS) were co-composted to analyze the feasibility of recycling the nutrients in CCFDs. A pilot-scale aerobic composting system with an auto-control system was used in this research, and the maturity and security of the compost product were evaluated. The temperature of the composting mixtures was maintained above 55°C for more than 3 days during the composting, indicating that co-composting of CCFDs and AS could reach the compost maturity standard, and the seeds germination index (GI) increased from 17.61% to 68.93% by the end of the composting process (28 days). However, the degradation rate of cephalosporin C (CPC) was only 6.58% during the composting process. Monitoring the quality of antibiotic resistance genes (ARGs) in the composts showed that the log copy of blaTEM in the composts increased from 2.15 in the initial phase to 6.37 after 28 days. Long-term investigation of CPC degradation and ARGs variation was conducted for the composts; CPC could still be detected after the maturity phases. A removal efficiency of 49.10% could be achieved in 110 days, while the log copy of ARGs increased to 7.93. Although a higher GI value (>80.00%) was observed, the risk of recycling the CCFDs compost product into land is still high.

Application and optimization of organic-inorganic hybrid monolithic capillary electrochromatography for in vivo cefdinir determination with microdialysis.

In this study, an organic-inorganic hybrid monolithic capillary column was applied and optimized for the determination of cefdinir in plasma, and the electro-osmotic flow that usually hinders migration in reverse polarity became a driving force. The Sample used for pharmacokinetic research was collected by microdialysis using phosphate buffer (pH 7.4) as perfusate, and a volume of 60 µL fluid was mixed with 140 µL of acetonitrile. By using a silica-allyldimethyldodecylammonium monolithic column (100 µm inner diameter, 21 cm effective length and 31.2 cm total length), and a mobile phase consisting of phosphate and acetonitrile (pH 4.5, 50:50, v/v), at a voltage of 20 kV, the analytes were successfully separated with the background within 2.5 min. The detection wavelength was 214 nm. The calibration curve showed a good linearity (r(2) = 0.9994) over the concentration range of 0.2-50 µg/mL. The proposed method showed good specificity, linearity, sensitivity, precision and recovery, and the introduction of field amplified sample stacking helped to improve the low recovery caused by microdialysis. This method was successfully applied to quantify cefdinir in rat plasma to support a pre-clinical pharmacokinetic trial.

High-throughput method for the determination of residues of ß-lactam antibiotics in bovine milk by LC-MS/MS.

This study describes the development and validation procedures for scope extension of a method for the determination of ß-lactam antibiotic residues (ampicillin, amoxicillin, penicillin G, penicillin V, oxacillin, cloxacillin, dicloxacillin, nafcillin, ceftiofur, cefquinome, cefoperazone, cephapirine, cefalexin and cephalonium) in bovine milk. Sample preparation was performed by liquid-liquid extraction (LLE) followed by two clean-up steps, including low temperature purification (LTP) and a solid phase dispersion clean-up. Extracts were analysed using a liquid chromatography-electrospray-tandem mass spectrometry system (LC-ESI-MS/MS). Chromatographic separation was performed in a C18 column, using methanol and water (both with 0.1% of formic acid) as mobile phase. Method validation was performed according to the criteria of Commission Decision 2002/657/EC. Main validation parameters such as linearity, limit of detection, decision limit (CCα), detection capability (CCß), accuracy, and repeatability were determined and were shown to be adequate. The method was applied to real samples (more than 250) and two milk samples had levels above maximum residues limits (MRLs) for cloxacillin - CLX and cefapirin - CFAP.

Silica-based 2-(N,N-dimethylamino)-1,3-propanediol hydrophilic interaction liquid chromatography stationary phase for separating cephalosporins and carbapenems.

A silica-based stationary phase bearing both hydrophilic hydroxyl and amino groups was developed by covalently bonding a small molecular N,N-dimethylamino 1,3-propanediol moiety onto silica beads via copper(I)-catalyzed Huisgen azide-alkyne 1,3-dipolar cycloaddition (CuAAC). This new stationary phase showed good HILIC characteristics and high column efficiency (the theoretical plate number is up to 37000 plates m(-1) in the case of inosine) in the separation of polar compounds, such as nucleosides and bases, organic acids, cephalosporins, and carbapenems.

Quantification of piperacillin, tazobactam, cefepime, meropenem, ciprofloxacin and linezolid in serum using an isotope dilution UHPLC-MS/MS method with semi-automated sample preparation.

BACKGROUND: Recent studies have demonstrated highly variable blood concentrations of piperacillin, tazobactam, cefepime, meropenem, ciprofloxacin and linezolid in critically ill patients with a high incidence of sub-therapeutic levels. Consequently, therapeutic drug monitoring (TDM) of these antibiotics has to be considered, requiring robust and reliable routine analytical methods. The aim of the present work was to develop and validate a multi-analyte ultra high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method for the simultaneous quantification of the above mentioned antibiotics. METHODS: Sample preparation included a manual protein precipitation step followed by two-dimensional ultra high performance liquid chromatography (2D-UHPLC). Corresponding stable isotope-labeled substances were used as internal standards for all of the analytes, with the exception of tazobactam. The injected sample volume was 7 µL. The run time was 5.0 min. RESULTS: Inaccuracy was ≤8% and imprecision coefficient of variation (CV) was <9% for all analytes. Only minor matrix effects and negligible carry-over was observed. The method was found to be robust during the validation period. CONCLUSIONS: We were able to develop a reliable 2D-UHPLC-MS/MS method addressing analytes with highly heterogeneous physico-chemical properties. The novel assay may be an efficient tool for an optimized process workflow in clinical laboratories for important antibiotics in regards to TDM.

Click N-benzyl iminodiacetic acid: novel silica-based tridentate zwitterionic stationary phase for hydrophilic interaction liquid chromatography.

Iminodiacetic acid (IDA) is dicarboxylic acid amine, which may produce stronger interaction with polar or charged compounds than bidentate α,ß-amino acid. In this article, a novel type of tridentate zwitterionic HILIC stationary phase was prepared by covalently bonding N-benzyl IDA on silica gel via copper(I) catalyzed Huisgen azide-alkyne 1,3-dipolar cycloaddition (CuAAC). The structure of this stationary phase and all related intermediates was confirmed by NMR, FT-IR, MS spectrum and elemental analysis. The new stationary phase showed good HILIC characteristics and high column efficiency (the theoretical plate number is up to 44000 plates m(-1) in the case of guanosine) in the application of separation of polar compounds, including organic acids, organic bases, as well as highly polar and hydrophilic compounds, such as cephalosporins and carbapenems. Most of them displayed good peak shape and selectivity.

Performance and model of a novel multi-sparger multi-stage airlift loop membrane bioreactor to treat high-strength 7-ACA pharmaceutical wastewater: effect of hydraulic retention time, temperature and pH.

In this study, three novel multi-sparger multi-stage airlift loop membrane bioreactors (Ms(2)ALMBRs) were set up in parallel for treating synthetic high-strength 7-ACA pharmaceutical wastewater under different HRTs, temperatures and pHs, respectively. During the 200-day operating time, average COD removal efficiencies were 94.96%, 96.05% and 93.9%. While average 7-ACA removal efficiencies were 66.44%, 59.04% and 59.60%, respectively. The optimal conditions were 10h, 15-35°C and 7-9 for HRT, temperature and pH, respectively. Moreover, the sludge characteristics and microorganism drug-resistances were explored. Results showed that different temperatures and pHs influenced contaminant removals by affecting MLSS concentration and ß-lactamase activity significantly. In addition, mathematical statistical models, built on the polynomial and linear regression techniques, were developed for exploring the inner relationships between HRT, temperature and pH changes and MLSS concentrations, ß-lactamase activities and contaminant removals of the Ms(2)ALMBR system.

Multiclass method for the determination of quinolones and ß-lactams, in raw cow milk using dispersive liquid-liquid microextraction and ultra high performance liquid chromatography-tandem mass spectrometry.

An analytical method based on a sample treatment by dispersive liquid-liquid microextraction (DLLME) followed by ultra high performance liquid chromatography-tandem mass spectrometry analysis (UHPLC-MS/MS) for the determination of 17 quinolones and 14 ß-lactams (penicillins and cephalosporins) in raw cow milk, was validated according to the European Commission guidelines as cited in the Decision 2002/657/EC. The extraction efficiency of the DLLME depends on several parameters such as the nature and volumes of extractant and dispersive solvents, pH, concentration of salt, shaking time and time of centrifugation. These variables were accurately optimized using multivariate optimization strategies. A Plackett-Burman design to select the most influential parameters and a Doehlert design to obtain the optimum conditions have been applied. Two different pH values were used for the extraction of compounds (pH 3 for acidic quinolones and ß-lactams and pH 8 for amphoteric quinolones). The method was validated using matrix-matched standard calibration followed by a recovery assay with spiked samples. The limits of quantification found ranged from 0.3 ng g(-1) for amoxicillin to 6.6 ng g(-1) for ciprofloxacin, and the precision was lower than 15% in all cases as is required by the European Regulation. The decision limits (CCα) ranged between 4.1 and 104.8 ng g(-1), while detection capabilities (CCß) from 4.2 to 109.7 ng g(-1). These values were very close to the corresponding maximum residue limits (MLRs) for the studied antibiotics. Recoveries between 72 and 110% were also obtained. Finally, in order to evaluate the applicability of the method, 28 raw cow milk samples were analysed and it was observed that 28% of the samples were positive. However, only 11% were considered non-compliant with the current EU legislation (Commission Regulation 37/2010), due to some milk samples corresponded to treated cows with these antibiotics.

Application of a capillary-assembled microfluidic system for separation of cephalosporins.

This paper demonstrates a simple and easy setting up of a fused-silica capillary-assembled microfluidic system (µCE). This system incorporates a split-flow pressure injection of the sample into a microfluidic system made from PDMS and a short (∼20 cm) length of fused-silica capillary as a separation unit. The on-capillary detection was carried out by fiber optic spectrometry. A mixture of six cephalosporin antibiotics was separated in the µCE system and the obtained results were compared to those achievable by conventional CE. The six components could be separated within 8.5 min with the number of theoretical plates around 10 000.

Non-cephalosporin-susceptible, glucose non-fermentative Gram-negative bacilli meningitis in post-neurosurgical adults: clinical characteristics and therapeutic outcome.

OBJECTIVE: The clinical and laboratory characteristics of non-cephalosporin-susceptible (non-CS) glucose non-fermentative Gram-negative (G(-)) infections in adults with postneurosurgical meningitis are rarely examined solely in the literature. METHODS: The data of 28 post-neurosurgical adults meningitis with glucose non-fermentative G(-) infections, collected during a study period of 5 years (2006-2010), were reviewed. The clinical and laboratory data between the non-cephalosporin-susceptible groups and the cephalosporin-susceptible groups were compared. RESULTS: A total of 30 G(-) strains were collected from the 28 enrolled cases. Among the implicated glucose non-fermentative G(-) strains, 18 strains, belonging to 17 cases, were non-CS. Among the 18 non-cephalosporin-susceptible strains, Acinetobacter spp. (39%, 7/18) was the most common, followed by Pseudomonas spp. (22%, 4/18), Stenotrophomonas maltophilia (22%, 4/18) and Elizabethkingia meningoseptica (11%, 2/18). With a comparative analysis, there were no significant difference between the non-cephalosporin-susceptible and cephalosporin-susceptible glucose non-fermentative G(-) groups. The clinical and laboratory data were also of no statistical significance between the fatal (n=4) and non-fatal (n=13) non-cephalosporin-susceptible groups. CONCLUSION: Sixty percent (18/30) of implicated glucose non-fermentative G(-) strains of post-NS meningitis in adults are non-cephalosporin-susceptible. Among the non-cephalosporin-susceptible glucose non-fermentative G(-) strains, Acinetobacter spp., Pseudomonas spp., S. maltophilia and E. meningoseptica are the commonly implicated pathogens, and their emergence in this specific group of meningitis has caused a therapeutic dilemma. The clinical manifestations of non-cephalosporin-susceptible glucose non-fermentative G(-) meningitis were not unique; therefore, only bacterial culture and antimicrobial susceptibility test are the methods for identification confirmation.

Solid phase extraction using magnetic core mesoporous shell microspheres with C18-modified interior pore-walls for residue analysis of cephalosporins in milk by LC-MS/MS.

A fast and effective extraction method has been developed for measuring the residue of cephalosporins (cefalexin, cefazolin, cefoperazone) in milk by using magnetic core-mesoporous shell microspheres with C18-functionalized interior pore-walls (C18-Fe3O4@mSiO2) as adsorbent. With no need for any protein precipitation procedure, the cephalosporins were directly adsorbed onto the C18-Fe3O4@mSiO2 microspheres through hydrophobic interaction with C18-groups (Octadecyl functional groups) functionalized in the interior walls of mesopore channels while the abundant proteins in milk sample were excluded out of the channel due to the size exclusion effect. Thereafter, the cephalosporins-absorbed C18-Fe3O4@mSiO2 microspheres were rapidly isolated by placing a magnet, and followed by liquid chromatography-tandem mass spectrometry analysis after eluted by methanol. Various parameters which could affect the extraction performance were optimised. The newly developed extraction method was successfully applied in determination of cephalosporin residues in milk samples, offering a valuable alternative to simplify and speed up the sample preparation step.