Optimization of ultrasound-assisted extraction based on response surface methodology using HPLC-DAD for the analysis of red clover (Trifolium pretense L.) isoflavones and its anti-inflammatory activities on LPS-induced 3D4/2 cell.

Introduction: Trifolium pratense L. has anti-inflammatory, antioxidant, cardiovascular disease prevention, and estrogen-like effects. The existing method for the assay of effective components is commonly based on a spectrophotometer, which could not meet the requirement of quality control. Furthermore, although there have been many studies on the anti-inflammation effect of red clover, a few have been reported on the regulatory effect of red clover isoflavones (RCI) on lipopolysaccharide (LPS)-induced inflammatory response in porcine alveolar macrophages (3D4/2 cells), and its mechanism of action is still unclear. Methods: The main components of RCI including daidzein, genistein, and biochanin A were accurately quantified by high-performance liquid chromatography coupled with diode array detection (HPLC-DAD) after optimizing the extraction process through response surface methodology. The anti-inflammatory potential of RCI was carried out by detecting the level of inflammatory cytokines and mRNA expression of related genes. Furthermore, its anti-inflammatory mechanism was explored by investigating two signaling pathways (NF-κB and MAPK). Results: The optimal extraction conditions of RCI were as follows: the concentration of ethanol is 86% and the solid-liquid ratio is 1:29, with the herb particle size of 40 mesh sieve. Under the optimal conditions, the total extraction of target components of RCI was 2,641.469 µg/g. The RCI could significantly suppress the production and expression of many pro-inflammatory cytokines. The results of the Western blot revealed that RCI dramatically reduced the expression of p65, p-p65, IκB-α, p38, and p-p38. These results are associated with the suppression of the signal pathway of p38 MAPK, and on the contrary, activating the NF-κB pathway. Collectively, our data demonstrated that RCI reversed the transcription of inflammatory factors and inhibited the expression of p65, p-p65, IκB-α, and p38, indicating that RCI had excellent anti-inflammatory properties through disturbing the activation of p38 MAPK and NF-κB pathways. Conclusion: The extraction conditions of RCI were optimized by HPLC-DAD combined with response surface methodology, which will contribute to the quality control of RCI. RCI had anti-inflammatory effects on the LPS-induced 3D4/2 cells. Its mechanism is to control the activation of NF-κB and p38 MAPK pathways, thereby reducing the expression of inflammatory-related genes and suppressing the release of cytokines.

Analysis of authorized coccidiostats in chicken feces and environmental water by ultra-performance liquid chromatography-tandem mass spectrometry based on dispersive solid-phase extraction and lyophilization.

A simple, sensitive, and efficient method based on ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was developed for the determination of 8 coccidiostats in chicken feces and environmental water (including sewage, pond water, and lake water) surrounding the farm. Target analytes in chicken feces were extracted with 2% acetic acid in acetonitrile solution, followed by a dispersive solid-phase extraction (DSPE) cleanup step using the mixture of PSA and C18 adsorbents. Environmental water samples were pretreated using a lyophilization approach. Analysis was carried out on a UPLC-MS/MS with the combination of methanol and 0.1% formic acid aqueous solution as the mobile phase under multiple reaction monitoring in positive and negative ionization modes. Results showed that 8 coccidiostats were linear with correlation coefficients higher than 0.99. Method validation was performed using fortified samples, reaching satisfactory recoveries of 75.9%-97.8% in chicken feces and 71.9%-108.2% in environmental water. Limits of detection for 8 analytes in chicken feces and environmental water were 0.03∼2 µg/kg and 0.005∼1 µg/L, respectively. Matrix effects were calculated and strong signal suppression (>50%) for some coccidiostats was observed. The developed method was successfully applied to analyze coccidiostats in chicken feces and environmental water collected from local chicken farms.

Cu(II) removal from water by trimercapto-s-triazine trisodium salt modified alkaline lignin.

To overcome the poor removal ability of alkaline lignin (AL) toward heavy metals, trimercapto-s-triazine trisodium salt (TMT) was selected as the modifying agent to introduce reaction groups. The Fourier transform infrared (FT-IR) spectra and the scanning electron microscopy (SEM) suggested that -SNa, C-N, and C = N groups were successfully introduced. Copper (II) was applied to evaluate the uptake performance of the adsorbent (AL-TMT). Adsorbent dosage and solution pH were taken into account to study their effects in the batch experiments. The pseudo-second-order dynamics and Langmuir models better described the experimental data. Nitrogen (N) and carbon (C) functional groups in thiotriazinone carried by AL-TMT were determined to be the primary uptake sites through X-ray photoelectron spectroscopy (XPS), FT-IR, and electrostatic potential (ESP). The selective experiments of AL-TMT toward Cd(II), Cu(II), Pb(II), Zn(II), Co(II), and Mg(II) were performed. It showed that AL-TMT possessed better adsorption selectivity toward Cu(II) than others. Furthermore, the density functional theory (DFT) calculations of thiotriazinone in AL-TMT also exhibited the lowest binding energy toward Cu than toward others. This work may provide a theoretical basis to facilitate the extraction of specific heavy metals from water or wastewater by using such modified alkaline lignin.

Response Surface Optimization of Dispersive Solid-Phase Extraction Combined with HPLC for the Rapid Analysis of Multiple Coccidiostats in Feed.

Since antimicrobials were banned as feed additives, coccidiostats with favorable anticoccidial action and growth promotion have been widely used in the breeding industry. The monitoring of coccidiostats in feed is necessary, while the current methods based on mass-spectrometer analysis have limited applicability and matrix effects could interfere with the results. Accordingly, in the present paper, a rapid analytical strategy for the simultaneous determination of six synthetic coccidiostats in feed using high-performance liquid chromatography coupled with diode-array detection was developed. Coccidiostats in chicken feeds were extracted with the trichloroacetic acid-acetonitrile solution. The cleanup was performed by dispersive solid-phase extraction after the optimization of the response surface methodology. The method exhibited good linearity for target coccidiostats within the range of 0.05~20 µg/mL. Recoveries for six compounds in fortified feed samples were from 67.2% to 107.2% with relative standard deviations less than 9.6%. The limit of detection was 0.2~0.3 mg/kg. The successful application of the method in commercial feed verified that it is effective and sensitive for the rapid determination of multiple coccidiostats in chicken feeds.

Surface molecularly imprinted solid-phase extraction for the determination of vancomycin in plasma samples using HPLC-MS/MS.

Vancomycin is a glycopeptide antibiotic used to treat infections caused by Gram-positive bacteria. Due to the narrow therapeutic index of vancomycin, it is necessary to develop a sensitive and reliable analytical method to monitor the drug concentration in plasma. A novel method based on surface molecularly imprinted solid-phase extraction combined with liquid chromatography-tandem mass spectrometry for the determination of vancomycin in plasma sample was developed. The plasma sample was cleaned up through the solid-phase extraction process before the analysis. The calibration standard of vancomycin in plasma ranged between 1 and 100 ng/mL, and the correlation coefficient (r) was 0.9993. The average recoveries were from 94.3 to 104.0%, and the precision was less than 10.5%. The limit of detection and limit of quantification were 0.5 ng/mL and 1 ng/mL, respectively. The method validated was successfully used for the detection of vancomycin in mice after oral administration.

Determination of polypeptide antibiotics in animal tissues using liquid chromatography tandem mass spectrometry based on in-line molecularly imprinted solid-phase extraction.

Effective purification and enrichment of polypeptide antibiotics in animal tissues is always a challenge, due to the co-extraction of other endogenous peptides which usually interfere their final determination. In this study, a molecularly imprinted column was prepared by packing polymyxin E-imprinted particles into a 100 mm × 4.6 mm i.d. HPLC column. The as-prepared imprinted columns were able to tolerate 100% aqueous phase and exhibited good stability and high column efficiency. Polypeptides antibiotics with similar molecular size or spatial structure to polymyxin E were well retained by the imprinted column, suggesting class selectivity. After optimization of mobile phase conditions of imprinted column, polypeptide antibiotics in animal tissue extracts were enriched and cleaned up by in-line molecularly imprinted solid-phase extraction, allowing the screening of target analytes in complex samples at low concentration levels by UV detection. Eluate fraction from the imprinted column was collected, and further dried and re-dissolved with methanol-0.5% formic acid aqueous solution (80:20, v/v) for final LC-MS/MS analysis. Analysis was accomplished using multiple reaction monitoring (MRM) in positive electrospray ionization mode and analytes quantified using the matrix-matched external calibration curves. The results showed high correlation coefficients for target analytes in the linear range of 2 ∼ 200 µg kg-1. At four different concentration levels (limit of quantification, 50, 100 and 200 µg kg-1), recoveries of four polypeptide antibiotics in swine, cattle and chicken muscles ranged from 66.7 to 94.5% with relative standard deviations lower than 16.0%. The limits of detection (LOD) were 2.0 ∼ 4.0 µg/kg, depending upon the analyte and sample. Compared with a conventional pretreatment method, the imprinted column was able to remove more impurities and to significantly reduce matrix effects, allowing the accurate analysis of polypeptide antibiotics.

The pharmacokinetic and residue depletion study of eugenol in carp (Cyprinus carpio).

Introduction: The pharmacokinetic profile and residue depletion of eugenol in carp (Cyprinus carpio) tissues and plasma were performed by a convenient and reliable high-performance liquid chromatography (HPLC) method. Methods: The eugenol in carp tissues and plasma was extracted with a mixed solution of acetonitrile and methanol. N-hexane was used to remove lipid impurities. The method was successfully applied to the pharmacokinetic and residue elimination of eugenol in carp after the carp was administered a medicated bath. Results: The average recoveries of eugenol in tissues and plasma fortified with four concentration levels were 69.0-106.6% and 80.0-86.7%, respectively. The relative standard deviations were < 8.9%. The limit of detection (LOD) was 0.01 µg/g in tissue and 0.008 µg/ml in plasma, respectively. The pharmacokinetic parameter of Cmax for eugenol in plasma at the concentrations of 20, 35, and 75 mg/L were 10.86, 17.21, and 37.32 mg/L, respectively. The t1/2 values were 3.68, 4.22, and 9.31 h. After the investigation of the anesthetic effect, 35 mg/L of eugenol was the optimal concentration for anesthesia. The highest accumulation concentration of eugenol in carp is in the liver and the lowest is in the muscle. In addition, the eugenol in tissue was eliminated rapidly and at a lower level than the LOD at 48 h. According to the residue elimination, the withdrawal time of eugenol was suggested at 5.2 days. Discussion: These results indicate that the developed method had good linearity and accuracy, and is sensitive enough for the monitoring of eugenol residue in carp. The half-life of eugenol decreased with the increase in drug concentration and the eugenol was eliminated rapidly in carp tissues. 35 mg/L eugenol was recommended as an anesthetic in carp due to its favorable anesthetic effect and no mortality. This study will contribute to the establishment of MRL regulation and setting a withdrawal period.

Simultaneous Determination of Multiple Polypeptide Antibiotics Residues in Lake Water by Lyophilization Combined with Liquid Chromatography-Tandem Mass Spectrometry.

It is significant to develop a method for the simultaneous determination of multiple polypeptide antibiotics residues in lake water because of the emergence of multidrug-resistant microorganisms in water. A sensitive, eco-friendly and simple method was developed for the determination of multiple polypeptide antibiotics, including vancomycin, teicoplanin, polymyxin B, colistin and bacitracin A in lake water using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Water samples were lyophilized to enrich them after adjusting the pH to 3. Then, 80% methanol in water containing 0.1% formic acid was used to reconstitute the residues for LC-MS/MS analysis. The results showed that target compounds were well separated and detected under the optimum instrumental conditions. The limits of detection and the limits of quantification of polypeptide antibiotics were in the range of 0.01 - 0.1 and 0.02 - 0.2 ng mL-1, respectively. The matrix-matched calibration curves of all compounds were linear in the calibration range of 1 - 200 ng mL-1. At three spiked levels of 0.2 (0.04), 0.4 (0.1) and 1.0 (0.2) ng mL-1 in lake water, the average recoveries of analytes were higher than 70%, except for teicoplanin, with relative standard deviations of less than 20%. Compared with other common sample pretreatment methods, the lyophilization process is simpler and more eco-friendly, achieving the simultaneous detection of multiple polypeptide antibiotics in lake water. The developed method is successfully applied to the routine monitoring of polypeptide antibiotics residues in lake water.

Preparation of surface molecularly imprinted polymer and its application for the selective extraction of teicoplanin from water.

In this study, a new surface molecularly imprinted polymer (SMIP) of teicoplanin (TEC) was prepared in an aqueous solution using amino-modified silica gel as a carrier. The molar ratio of the template molecule, functional monomer and cross-linker in the optimized synthesis system was 1 : 15 : 40. The structure and morphology of SMIP were characterized by Fourier-transform infrared spectra and scanning electron microscopy, respectively. It was shown that the silica gel modified with different active groups; the type and structure of functional monomers have a great influence on the specificity of SMIP. The SMIPs synthesized from a series of methacrylic acid and its hydroxylalkyl esters as functional monomers have good specificity for TEC. The results of static adsorption experiments showed that the adsorption capacity of SMIP was 6.5 times that of non-molecularly imprinted polymer, which were 152.6 mg g-1 and 23.6 mg g-1, respectively, indicating that SMIP had a larger affinity for TEC. Finally, the SMIP was successfully used as a dispersive solid-phase extraction adsorption material to selectively extract and enrich TEC from the water sample. The limit of detection of the proposed liquid chromatographic method for TEC was 5 µg L-1.

Synthesis of Molecularly Imprinted Polymers for the Selective Extraction of Polymyxins from Environmental Water Samples.

The authors wish to make a change to the published paper [...].

Synthesis of Molecularly Imprinted Polymers for the Selective Extraction of Polymyxins from Environmental Water Samples.

The emergence of colistin resistance gen has aroused public concern. It is significant to assess the concentrations of polymyxins residues in aquatic environment since resistant bacteria carrying colistin resistance gen are frequently isolated from wastewater; surface water and ground water. However; no literature on the determination of polymyxins in water is available; probably due to the absence of an efficient extraction method. Accordingly; molecularly imprinted polymers were synthesized by precipitation polymerization with colistin as the template. The polymers were successfully used as sorbents for the determination of polymyxins from water based on molecularly imprinted solid-phase extraction and high-performance liquid chromatography-ultraviolet detection. The molecularly imprinted cartridge showed excellent affinity and cross-reactivity to analytes in aqueous media. Recoveries obtained from water samples were between 65.9% and 90.1%, with relative standard deviations lower than 10.2%. Limits of detection were between 1.0 and 2.0 µg L-1 concentration levels. Compared with C18 cartridge; the molecularly imprinted cartridge could remove more interference from co-extracted matrices. This method is practical for the routine monitoring of polymyxin residues in environmental water; which will benefit studies on drug-resistance and occurrence of polymyxins in the environment.

Analysis of Nosiheptide in Food Animal Tissues via Its Unique Degradation Product by Liquid Chromatography-Tandem Mass Spectrometry after Alkaline Hydrolysis.

Very weak signals of fragment ions of nosiheptide could be observed using liquid chromatography-tandem mass spectrometry. The preparation of 4-hydroxymethyl-3-methyl-1H-indole-2-carboxylic acid (HMIA), a specific fragment of nosiheptide, by alkaline hydrolysis is described. HMIA showed a good mass spectrometric signal in negative electrospray ionization mode. In the new method, the nosiheptide residue in muscle tissue was hydrolyzed with sodium hydroxide aqueous solution; this was followed by cleanup using mixed mode cartridges. Identification and quantification of nosiheptide were carried out by analyzing HMIA in hydrolysate of muscles. Nosiheptide showed a good linear relationship (r > 0.996) in the calibration range of 2-500 µg/kg, and a low limit of quantification of 2 µg/kg was obtained in swine, chicken, and fish muscles. Recoveries of nosiheptide from spiked muscle samples were 85-108% with relative standard deviations less than 10%. The proposed method was successfully applied for the detection of the nosiheptide residue in medicated animal tissues samples.

Preparation and Application of Molecularly Imprinted Monolithic Extraction Column for the Selective Microextraction of Multiple Macrolide Antibiotics from Animal Muscles.

This study aimed to prepare a molecularly imprinted monolithic extraction column (MIMC) inside a micropipette tip by situ polymerization with roxithromycin as the dummy template. The polymers possessed excellent adsorption capacity and class-specificity to multiple macrolide drugs. MIMC was directly connected to a syringe for template removal and for the optimization of extraction conditions without any other post-treatment of polymers. A liquid chromatography-tandem mass spectrometric method was developed for the selective microextraction and determination of macrolide antibiotics in animal muscles based on MIMC. High recoveries of 76.1-92.8% for six macrolides were obtained with relative standard deviations less than 10.4%. MIMC exhibited better retention ability and durability when compared with the traditional C18 and HLB cartridges. The proposed method shows a great potential for the analysis of macrolide drugs at the trace level in animal foodstuffs.

Rapid multiresidue analysis of authorized/banned cyclopolypeptide antibiotics in feed by liquid chromatography-tandem mass spectrometry based on dispersive solid-phase extraction.

An effective analytical strategy was developed for simultaneous determination of seven cyclopolypeptide antibiotics (vancomycin, polymyxin B, polymyxin E, teicoplanin A2, bacitracin A, daptomycin and virginiamycin M1) in feed using liquid chromatography-tandem mass spectrometry. The extraction of sample was based on acidified methanol-aqueous solution, followed by a simply dispersive solid-phase extraction for further purification with primary secondary amine as an adsorbent. The method showed good linearity for target analytes in the experimental concentration ranges. At three spiked concentration levels of 50, 100 and 200 µg/kg, the average recoveries for target compounds were in the range of 63.1%-107.5% with the relative standard deviations less than 14.7%. Matrix effects in different feeds were evaluated and the strong signal suppression (>50%) was observed. Compared with the corresponding methods in literature, the developed method is more economical, more sensitive and involves the detection of authorized/banned cyclopolypeptides in animal feed. The proposed method was successfully applied to the routine supervision and rapid screening of cyclopolypeptide antibiotics in complete and premix feed.

Rapid determination of nosiheptide in feed based on dispersive SPE coupled with HPLC.

A novel, simple, and reliable method based on high-performance liquid chromatography coupled with fluorescence detection has been developed for the determination of nosiheptide in feed. The feed samples were extracted with acetonitrile 0.1% formic acid aqueous solution and then purified via a dispersive solid-phase extraction procedure using silica gel powder as the sorbent. Using a mixture of acetonitrile and 5 mM ammonium acetate solution (containing 0.1% formic acid) as the mobile phase, good separation and peak shape were obtained for nosiheptide on a Poroshell C8 column (250 × 4.6 mm id, 4 µm) via the isocratic elution program. The resulting calibration curve shows high levels of linearity (r2  > 0.999) for nosiheptide concentrations of 50-1000 µg/L. At three spiked levels, i.e., 0.500, 2.50 and 5.00 mg/kg, the intra- and interday recoveries of nosiheptide in five types of feed ranged from 78.5-96.8 and 84.9-94.2%, respectively. The intra- and interday relative standard deviations were less than 10.8%. The limits of quantification for nosiheptide in complete feed and premixes were measured as 50 and 100 µg/kg, respectively. Compared with other common adsorbents, silica gel presents stronger recovery and purification results for feed samples during the dispersive solid-phase extraction process.

Determination of Ten Macrolide Drugs in Environmental Water Using Molecularly Imprinted Solid-Phase Extraction Coupled with Liquid Chromatography-Tandem Mass Spectrometry.

With the extensive application of antibiotics in livestock, their contamination of the aquatic environment has received more attention. Molecularly imprinted polymer (MIP), as an eco-friendly and durable solid-phase extraction material, has shown great potential for the separation and enrichment of antibiotics in water. This study aims at developing a practical and economical method based on molecularly imprinted solid phase extraction (MISPE) combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS) for simultaneously detecting ten macrolide drugs in different sources of water samples. The MIP was synthesized by bulk polymerization using tylosin as the template and methacrylic acid as the functional monomer. The MIP exhibited a favorable load-bearing capacity for water (>90 mL), which is more than triple that of non-molecularly imprinted polymers (NIP). The mean recoveries of macrolides at four spiked concentration levels (limit of quantification, 40, 100, and 400 ng/L) were 62.6⁻100.9%, with intra-day and inter-day relative standard deviations below 12.6%. The limit of detection and limit of quantification were 1.0⁻15.0 ng/L and 3.0⁻40.0 ng/L, respectively. Finally, the proposed method was successfully applied to the analysis of real water samples.

Freeze-thaw approach: A practical sample preparation strategy for residue analysis of multi-class veterinary drugs in chicken muscle.

Seven drugs from different classes, namely, fluoroquinolones (enrofloxacin, ciprofloxacin, sarafloxacin), sulfonamides (sulfadimidine, sulfamonomethoxine), and macrolides (tilmicosin, tylosin), were used as test compounds in chickens by oral administration, a simple extraction step after cryogenic freezing might allow the effective extraction of multi-class veterinary drug residues from minced chicken muscles by mix vortexing. On basis of the optimized freeze-thaw approach, a convenient, selective, and reproducible liquid chromatography with tandem mass spectrometry method was developed. At three spiking levels in blank chicken and medicated chicken muscles, average recoveries of the analytes were in the range of 71-106 and 63-119%, respectively. All the relative standard deviations were <20%. The limits of quantification of analytes were 0.2-5.0 ng/g. Regardless of the chicken levels, there were no significant differences (P > 0.05) in the average contents of almost any of the analytes in medicated chickens between this method and specific methods in the literature for the determination of specific analytes. Finally, the developed method was successfully extended to the monitoring of residues of 55 common veterinary drugs in food animal muscles.

Simultaneous determination of eight cyclopolypeptide antibiotics in feed by high performance liquid chromatography coupled with evaporation light scattering detection.

A high throughput, reliable and reproducible analysis strategy based on high performance liquid chromatography combined to evaporative light scattering detector (HPLC-ELSD) was developed for simultaneous determination of eight cyclopolypeptide antibiotics including vancomycin, polymyxin B (polymyxin B1 and polymyxin B2), polymyxin E (colistin A and colistin B), teicoplanin, bacitracin A, daptomycin and virginiamycin M1 in animal Feed. Feed samples were extracted with methanol-2% formic acid aqueous solution, followed by a solid-phase extraction step using a HLB cartridge. Under the optimum chromatographic conditions and ELSD parameters, target compounds were separated well on a short column filled with biphenyl stationary phase. The method was developed in accordance with pig complete feed and then extended to detect polypeptide antibiotics in piglet premix, pig feed additive, poultry complete feed and fattening pig premix. The results showed that logarithmic calibration curves of eight analytes were linear (r2 > 0.99) within the concentration range of 5-200 mg mL-1. The developed method provided good accuracy and precision for quantification of eight polypeptides in five kinds of feeds with recoveries ranging from 72.0% to 105.4% with relative standard deviations <9.5%. The limits of detection ranged from 2 to 5 mg kg-1. Finally, the method was successfully applied to analyze polypeptide antibiotics in commercial feed.

Determination of macrolide antibiotics residues in pork using molecularly imprinted dispersive solid-phase extraction coupled with LC-MS/MS.

A class-specific macrolide molecularly imprinted polymer was synthesized by precipitation polymerization using tulathromycin as the template and methacrylic acid as the functional monomer. The polymers revealed different specific adsorption and imprinting factor for macrolides with different spatial arrangement of side chains as well as lactonic ring size. And the molecularly imprinted polymer possessed maximum adsorption capacity (54.1 mg/g) and highest imprinting factor (2.4) toward 15-membered ring azithromycin. On the basis of molecularly imprinted polymer dispersive solid-phase extraction, a rapid, selective, and reproducible method for simultaneous determination of seven macrolide antibiotics residues in pork was established by using liquid chromatography with tandem mass spectrometry. At spiking levels of 5, 10, 25, and 100 µg/kg, average recoveries of seven macrolides ranged from 68.6 to 95.5% with intraday and interday relative standard deviations below 8%. The limits of detection and limits of quantification were 0.2-0.5 and 0.5-2.0 µg/kg, respectively.

Molecularly imprinted solid-phase extraction for the determination of ten macrolide drugs residues in animal muscles by liquid chromatography-tandem mass spectrometry.

A simple and sensitive method based on molecularly imprinted solid-phase extraction coupled with liquid chromatography-tandem mass spectrometry was developed for the determination of the residues of ten macrolide drugs in swine, cattle and chicken muscles samples. The molecularly imprinted polymers (MIPs) were synthesized using tylosin as a template and methacrylic acid as a functional monomer. Samples were extracted with sodium borate buffer solution and ethyl acetate, and purified by the MIP cartridge. The results showed that the cartridge exhibited good recognition performance for macrolides, and better purification effect than the traditional solid-phase extraction cartridges. Recoveries of analytes at three spiking levels 1, 5 and 20µgkg(-1) ranged from 60.7% to 100.3% with the relative standard deviations less than 14%. The limits of detection of the method were between 0.1 and 0.4µgkg(-1). The method is useful for the routine monitoring of the residues of macrolide drugs in animal muscles.