Laser induced fluorescence coupled to capillary electrophoresis for the determination of fluoroquinolones in foods of animal origin using molecularly imprinted polymers

A method for the simultaneous determination of four fluoroquinolones of veterinary use (ciprofloxacin, danofloxacin, enrofloxacin and sarafloxacin) in two complex matrixes, such as bovine raw milk and pig kidney, has been established and validated. The method is based on the use of capillary electrophoresis (CE) coupled with a very sensitive detection mode, such as laser induced fluorescence (LIF) detection, due to the fact the all the compounds selected show native fluorescence. In order to achieve high selectivity in the sample treatment procedure, a commercially available molecularly imprinted polymer has been used for the solid phase extraction of the analytes. Once the retention and elution processes were optimized, the final extract was analyzed by CE-LIF using a 325 nm He–Cd laser. Optimum separation was obtained in a 70 cm × 75 μm capillary using a 125 mM phosphoric acid solution at pH 2.8 with 36% methanol as background electrolyte. The method provided very low detection limits, ranging from 0.17 to 0.98 μg/kg for milk and 1.10 to 10.5 μg/kg for kidney, with acceptable precision and satisfactory recoveries.     

Determination of the Alternaria mycotoxin tenuazonic acid in cereals by high-performance liquid chromatography–electrospray ionization ion-trap multistage mass spectrometry after derivatization with 2,4-dinitrophenylhydrazine

Tenuazonic acid (TA) is a major Alternaria mycotoxin. In the present work a novel approach for the detection of TA in cereals by liquid chromatography–ion-trap multistage mass spectrometry after derivatization with 2,4-dinitrophenylhydrazine is described. The product of the derivatization reaction and its major MS² fragments were characterised by Fourier transform-ion cyclotron resonance tandem mass spectrometry. Without preconcentration, the established method features a limit of detection of 10 μg/kg using 2 g of sample in a rapid workup procedure. Accuracy, precision and linearity were evaluated in the working range of 50–5000 μg/kg. TA was detected in 13 and quantified in 3 out of 27 cereal samples obtained from a local supermarket, the average content being 49 μg/kg (highest incidence: 851 ± 41 μg/kg).     

Development and validation of two multiresidue liquid chromatography tandem mass spectrometry methods based on a versatile extraction procedure for isolating non-steroidal anti-inflammatory drugs from bovine milk and muscle tissue

The main difficulties in analysing non-steroidal anti-inflammatory drugs (NSAIDs) in food and biological samples are due to the tight non-covalent interactions established with matrix proteins and the amount of occurring fatty material. The present paper describes an effective extraction procedure able to isolate fifteen NSAIDs (acetaminophen, salicylic acid, ibuprofen, diclofenac, flunixin and its metabolite 5-hydroxy-flunixin, nimesulide, phenylbutazone, meclofenamic acid, tolfenamic acid, meloxicam, carprofen, ketoprofen, naproxen and etodolac) from bovine milk and muscle tissue through two succeeding steps: (a) deproteinisation/extraction with organic solvent, essential to lower the medium dielectric constant and, therefore, to release the analytes from matrix; (b) SPE clean-up on OASIS cartridges. Lipids were easily removed during low-temperature centrifugations. The advantages of the developed procedure pertain to the efficient removal of the fat substances (very low matrix effect and high recovery yields) and its versatility, since it can be applied both to milk and muscle with few adjustments due to the diversity of the two matrices. Ion-pairing reversed-phase chromatography combined with the negative electrospray detection was able to achieve low detection capabilities (CCβs) for all analytes and, in particular, for diclofenac whose Maximum Residue Limit (MRL) in milk is 0.1 μg kg(-1). The methods were validated according to the guidelines of the Commission Decision 2002/657/EC and then applied for a small monitoring study. A number of samples showed traces of salicylic acid (SA), but its occurrence was not ascribed to a misuse of drugs (aspirin, salicylic acid) since SA, accumulating in plants in response to a pathogen attack, may be introduced into the food chain.     

Fluoroquinolone antibiotic determination in bovine,ovine and caprine milk using solid-phase extraction and high-performance liquid chromatography-fluorescence detection with ionic liquids as mobile phase additives

This paper describes the use of 1-ethyl-3-methylimidazolium tetrafluoroborate (EMIm-BF₄) as mobile phase additive for the analysis by high-performance liquid chromatography with fluorescence detection of a group of seven basic fluoroquinolone antibiotics (i.e. fleroxacin, ciprofloxacin, lomefloxacin, danofloxacin, enrofloxacin, sarafloxacin and difloxacin) in different milk samples. EMIm-BF₄ was found superior to 1-butyl-3-methylimidazolium tetrafluoroborate for the separation of the analytes from chromatographic interferences of the sample matrix. The optimized method was applied to the analysis of ovine, caprine and bovine milk, in the last case in either skimmed, semi-skimmed and full-cream milk after suitable acidic deproteination followed by a solid-phase extraction procedure. Recovery values between 73% and 113% were obtained for the three types of bovine milk samples, as well as for ovine and caprine milk (RSDs below 16% in all cases), which clearly demonstrates the applicability of the method to the three types of milk irrespective of the fat content of the samples. Limits of detection were in the range of 0.5–8.1 μg/L (approximately 0.5–25.9 μg/kg), well below the maximum residue limits established for these compounds by the current European legislation. A screening study of 24 different milk samples was also developed. In none of the samples, residues of the selected antibiotics were found.     

Ultra-high-pressure liquid chromatography–tandem mass spectrometry for the analysis of six resorcylic acid lactones in bovine milk

This work reports a rapid, reliable and sensitive multi-residue method for the simultaneous determination of six resorcylic acid lactones in bovine milk by ultra-high-pressure liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS). The resorcylic acid lactones were extracted, purified, and concentrated from milk samples in one step using a solid-phase extraction (SPE) cartridge that contained a polymeric mixed-mode anion-exchange sorbent. The analysis was performed on a Waters Acquity BEH C₁₈ column utilizing a gradient elution profile. Each LC run was completed in 3.5 min. The analytes were detected by multiple reaction monitoring (MRM) using electrospray ionization (ESI) negative mode. Mean recoveries from fortified samples ranged from 92.6% to 112.5%, with relative standard deviations lower than 11.4%. Using 5 mL bovine milk, the limits of detection and quantification for resorcylic acid lactones were in the ranges of 0.01–0.05 and 0.05–0.2 μg/L, respectively. The application of this newly developed method was demonstrated by analyzing bovine milk samples from markets.     

Development and validation of a confirmatory method for the determination of 12 non steroidal anti-inflammatory drugs in milk using liquid chromatography-tandem mass spectrometry

A rapid and reliable LC-MS/MS method for the simultaneous confirmation of twelve non steroidal anti-inflammatory drugs (NSAIDs) in bovine milk was developed and fully validated in accordance with the European Commission Decision 2002/657/EC. The validation scheme was built in accordance with the MRLs or target analytical levels (EU-CRL recommended concentrations and detection capabilities) of the analytes, except for diclofenac for which the lower level of validation achieved was 0.5 μg kg(-1) whereas its MRL is 0.1 μg kg(-1). The NSAIDs investigated were as follows: phenylbutazone (PBZ), oxyphenylbutazone (OPB), naproxen (NP), mefenamic acid (MF), vedaprofen (VDP), flunixin (FLU), 5-hydroxyflunixin (FLU-OH), tolfenamic acid (TLF), meloxicam (MLX), diclofenac (DC), carprofen (CPF) and ketoprofen (KTP). Several extraction procedures had been investigated during the development phase. Finally, the best results were obtained with a procedure using only methanol as the extraction solvent, with an evaporation step included and no further purification. Chromatographic separation was achieved on a C18 analytical column and the run was split in 2 segments. Matrix effects were also investigated. Data acquisition implemented for the confirmatory purpose was performed by monitoring 2 MRM transitions per analyte under the negative electrospray mode. Mean relative recoveries ranged from 94.7% to 110.0%, with their coefficients of variation lying between 2.9% and 14.7%. Analytical limits expressed in terms of decision limits (CCα) were evaluated between 0.69 μg kg(-1) (FLU) and 27.54 μg kg(-1) (VDP) for non-MRL compounds, and at 0.10 (DC), 15.37 (MLX), 45.08 (FLU-OH), and 62.96 μg kg(-1) (TLF) for MRL compounds. The validation results proved that the method is suitable for the screening and confirmatory steps as implemented for the French monitoring plan for NSAID residue control in bovine milk.     

Electromembrane extraction (EME) and HPLC determination of non-steroidal anti-inflammatory drugs (NSAIDs) in wastewater samples

In this paper, an electromembrane extraction (EME) combined with a HPLC procedure using diode array (DAD) and fluorescence detection (FLD) has been developed for the determination of six widely used non-steroidal anti-inflammatory drugs (NSAIDs): salicylic acid (SAC), ketorolac (KTR), ketoprofen (KTP), naproxen (NAX), diclofenac (DIC) and ibuprofen (IBU). The drugs were extracted from basic aqueous sample solutions, through a supported liquid membrane (SLM) consisting of 1-octanol impregnated in the walls of a S6/2 Accurel^® polypropylene hollow fiber, and into a basic aqueous acceptor solution resent inside the lumen of the hollow fiber with a potential difference of 10 V applied over the SLM. Extractions that were carried out in 10 min using a potential of 10 V from pH 12 NaOH aqueous solutions shown concentration enrichments factors of 28-49 in a pH 12 NaOH aqueous acceptor solution. The proposed method was successfully applied to urban wastewaters. Excellent selectivity was demonstrated as no interfering peaks were detected. The procedure allows very low detection and quantitation limits of 0.0009-9.0 and 0.003-11.1 μg L⁻¹, respectively.     

Trace determination of nine haloacetic acids in drinking water by liquid chromatography–electrospray tandem mass spectrometry

A simple, fast and sensitive liquid chromatography–electrospray tandem mass spectrometry method was established for trace levels of nine haloacetic acids (HAAs) in drinking water. Water samples were removed of residual chlorine by adding l-ascorbic acid, and directly injected after filtered by 0.22 μm membrane. Nine HAAs were separated by liquid chromatography in 7.5 min, and the limits of detection were generally between 0.16 and 0.99 μg/L except for chlorodibromoacetic acid (1.44 μg/L) and tribromoacetic acid (8.87 μg/L). The mean recoveries of nine target compounds in spiked drinking water samples were 80.1–108%, and no apparent signal suppression was observed. Finally, this method was applied to determine HAAs in the tap water samples collected from five waterworks in Shandong, China. Nine HAAs except for monochloroacetic acid, monobromoacetic acid, dibromochloroacetic acid and tribromoacetic acid were detected, and the total concentrations were 7.79–36.5 μg/L. The determination results well met the first stage of the Disinfectants/Disinfection By-Products (D/DBP) Rules established by U.S.EPA and Guidelines for Drinking-water Quality of WHO.     

The Use of Bovine Serum Albumin as a Ligand in Affinity Chromatographic Clean-up of Non-Steroidal Anti-Inflammatory Drugs from Bovine Plasma

The European Union regulates the use of non-steroidal anti-inflammatory drugs (NSAID) in animal production and official national analytical monitoring programmes have been set up in each member state to detect residues of the drugs in bovine plasma. In this work, we describe the development and application of affinity chromatography for molecular recognition of NSAID in bovine plasma. Bovine serum albumin (BSA), the plasma protein which binds such drugs, was covalently bound to a polymeric support via its glycosyl moieties. Loading capacities and specificity were tested both on standard solutions and on spiked bovine plasma for nine drugs—ketoprofen, naproxen, phenylbutazone, oxyphenylbutazone, acetylsalicylic acid, salicylic acid, tolfenamic acid, diclofenac, and nimesulide—chosen as being the most representative of the different chemical sub-classes of NSAID. The BSA columns could bind up to 150 × 10^–6 g total of a mixture of these nine NSAID, with mean recoveries ranging from 74.0% (tolfenamic acid) to 96.0% (nimesulide). HPLC separation on an RP C₁₈ column with a linear gradient enabled resolution of the drugs; identification was achieved by coupling with photodiode array detection (DAD) operated at 240 and 280 nm. Results showed that all the compounds except acetylsalicylic acid were bound effectively by the BSA affinity columns. When plasma samples were spiked at 2.5 g mL^–1 with a mixture of the NSAID the chromatographic profile and UV spectra recorded (in the range 220–350 nm) indicated the procedure was sufficiently selective for identification of the drugs at the concentrations expected according to their pharmacokinetics. No memory effects and matrix interferences were observed.Revised: 18 December 2003 and 16 April 2004     

Trace determination of 10 β-lactam antibiotics in environmental and food samples by capillary liquid chromatography

A sensitive and reliable method using capillary HPLC with UV-diode array detection (DAD) has been developed and validated for the trace determination of residues of 10 β-lactam antibiotics of human and veterinary use, in milk, chicken meat and environmental water samples. The analytes included ampicillin, amoxicillin, penicillin V, penicillin G, cloxacillin, oxacillin, dicloxacillin, nafcillin, piperacillin and clavulanic acid. Legal levels are regulated by the EU Council regulation 2377/90 in animal edible tissues for these compounds. For food analysis, a solid-phase extraction (SPE) procedure consisting in a tandem of Oasis HLB and Alumina N cartridges was applied for off-line preconcentration and cleanup. For water analysis, the first step was only necessary. The limits of detection for the studied compounds were between 0.04–0.06 μg l⁻¹ for water samples and 0.80–1.40 μg l⁻¹ (or μg kg⁻¹) in the case of foods derived from animals. Average recoveries for fortified samples at different concentration levels ranged between 82.9% and 98.2%, with relative standard deviations (RSDs) lower than 9%. The method showed the advantages of capillary HPLC for the detection of these widely applied antibiotics in different samples at very low concentration levels.     

Novel high-sensitive fluorescent detection of deoxyribonuclease I based on DNA-templated gold/silver nanoclusters

Herein, fluorescent DNA-templated gold/silver nanoclusters (DNA-Au/Ag NCs) are presented as a novel probe for sensitive detection of deoxyribonuclease I (DNase I). The procedure is based on quenching fluorescence of DNA-Au/Ag NCs by DNase I digestion of the DNA (5′-CCCTTAATCCCC-3′) template. This decrease in fluorescence intensity permitted sensitive detection of DNase I in a linear range of 0.013–60 μg mL⁻¹, with a detection limit of 3 ng mL⁻¹ at a signal-to-noise ratio of 3. Furthermore, the practicality of this probe for detection of DNase I in human serum and saliva samples was validated, demonstrating its advantages of simplicity, selectivity, sensitivity and low cost. Importantly, satisfactory agreement between results obtained by the fluorescent method described here and high performance liquid chromatography (HPLC) further confirmed the reliability and accuracy of this approach.     

Optimization of the derivatization reaction and the solid-phase microextraction conditions using a D-optimal design and three-way calibration in the determination of non-steroidal anti-inflammatory drugs in bovine milk by gas chromatography-mass spectrometry

An experimental design optimization is reported of an analytical procedure used in the simultaneous determination of seven non-steroidal anti-inflammatory drugs (NSAIDs) in bovine milk by gas chromatography with mass spectrometry detection (GC-MS). This analytical procedure involves a solid-phase microextraction (SPME) step and an aqueous derivatization procedure of the NSAIDs to ethyl esters in bovine milk. The following NSAIDs are studied: ibuprofen (IBP), naproxen (NPX), ketoprofen (KPF), diclofenac (DCF), flufenamic acid (FLF), tolfenamic acid (TLF) and meclofenamic acid (MCL). Three kinds of SPME fibers - polyacrylate (PA), polydimethylsiloxane/divinylbenzene (PDMS/DVB) and polydimethylsiloxane (PDMS) - are compared to identify the most suitable one for the extraction process, on the basis of two steps: to determine the equilibrium time of each fiber and to select the fiber that provides the best figures-of-merit values calculated with three-way PARAFAC-based calibration models at the equilibrium time. The best results were obtained with the PDMS fiber. Subsequently, 8 experimental factors (related to the derivatization reaction and the SPME) were optimized by means of a D-optimal design that involves only 14 rather than 512 experiments in the complete factorial design. The responses used in the design are the sample mode loadings of the PARAFAC decomposition which are related to the quantity of each NSAID that is extracted in the experiment. Owing to the fact that each analyte is unequivocally identified in the PARAFAC decomposition, a calibration model is not needed for each experimental condition. The procedure fulfils the performance requirements for a confirmatory method established in European Commission Decision 2002/657/EC.     

The derivatisation of avermectins and milbemycins in milk: new insights and improvement of the procedure

Derivatisation of the avermectines ivermectin (IVM), doramectin (DOR), abamectin (ABA) and eprinomectin (EPR), and the milbemycin moxidectin (MOX) to fluorescent derivatives is commonly used for quantitative analysis at relevant levels using high performance liquid chromatography (HPLC) with fluorescence detection. Problems associated with the differences in reactivity towards derivatisation (EPM) and limited stability of the derived products (IVM, DOR, ABA) may seriously hamper the applicability of the method and the reliability of the obtained results. A study was performed to obtain more insight in this derivatisation process from an organic chemistry point of view. This study demonstrated the occurrence of two main fluorescent derivatives: the trifluoroacetyl esters (flu-TFA) and the derivatives with a free hydroxy group at the glycosidic ring (flu-OH). Optimisation of the derivatisation conditions resulted in a fast and reproducible formation of the fluorescent derivatives for all analytes including EPM. The improved procedure involves the addition of 1-methylimidazole (MI), trifluoroacetic anhydride (TFAA), triethylamine (TEA) and trifluoroacetic acid (TFA) with a subsequent incubation for 30 min at 70 °C. With this procedure for IVM, DOR and ABA flu-TFA derivatives are obtained instead of flu-OH derivatives as generally described in literature. The derivatisation is reproducible in different milk samples and the derivatives proved to be stable for at least 80 h at room temperature. Using the optimised procedure a limit of detection (LoD) of 0.1 μg kg⁻¹ in milk was readily obtained.     

Simultaneous determination of thirty non-steroidal anti-inflammatory drug residues in swine muscle by ultra-high-performance liquid chromatography with tandem mass spectrometry

An ultra-high-performance liquid chromatography with tandem mass spectrometric detection (UHPLC–MS/MS) method was established for the simultaneous determination of residues of thirty non-steroidal anti-inflammatory drugs (NSAIDs) in swine muscle. The samples were extracted with acetonitrile and phosphoric acid. The extracts were defatted with n-hexane, and then purified by HLB solid-phase extraction cartridge. Analysis was carried out on UHPLC–ESI-MS/MS working with multiple reaction monitoring mode with polarity switching. Limits of detection were between 0.4 μg/kg and 2.0 μg/kg, and limits of quantification were between 1.0 μg/kg and 5.0 μg/kg. The recoveries of NSAIDs were between 61.7% and 125.7% at spiked levels of 1.0–500 μg/kg. The repeatability was less than 8% and the within-laboratory reproducibility was not more than 12.3%. The method was reliable, convenient and sensitive.     

High through-put aflatoxin determination in plant material by automated solid-phase extraction on-line coupled to laser-induced fluorescence screening and determination by liquid chromatography–triple quadrupole mass spectrometry

A screening test based on laser-induced fluorescence (LIF) and a method for individual identification – quantitation of aflatoxins (AFs) in olive leaves and drupes, based on chromatographic separation and triple-quad mass-spectrometry detection with electrospray ionization in positive mode, is here reported. The sensitivity and selectivity of both methods are enhanced by a preconcentration–cleanup step developed by a Prospekt station. The analysis frequency is at least 3.5 samples/h. The screening test makes able to detect the target analytes at concentrations of 0.7 μg/kg without “false negatives”. The LC–MS/MS method provides limits of detection (LOD) and quantification (LOQ) ranging between 0.01–0.03 and 0.03–0.11 μg/kg, respectively. The linear dynamic range is between LOQ–50 μg/kg. The between-day precision, expressed as relative standard deviation, ranges between 0.97–2.86% and the within laboratory reproducibility, also expressed as RSD, between 1.63% and 4.84%.     

Dynamic supported liquid membrane tip extraction of glyphosate and aminomethylphosphonic acid followed by capillary electrophoresis with contactless conductivity detection

A dynamic supported liquid membrane tip extraction (SLMTE) procedure for the effective extraction and preconcentration of glyphosate (GLYP) and its metabolite aminomethylphosphonic acid (AMPA) in water has been investigated. The SLMTE procedure was performed in a semi-automated dynamic mode and demonstrated a greater performance against a static extraction. Several important extraction parameters such as donor phase pH, cationic carrier concentration, type of membrane solvent, type of acceptor stripping phase, agitation and extraction time were comprehensively optimized. A solution of Aliquat-336, a cationic carrier, in dihexyl ether was selected as the supported liquid incorporated into the membrane phase. Quantification of GLYP and AMPA was carried out using capillary electrophoresis with contactless conductivity detection. An electrolyte solution consisting of 12 mM histidine (His), 8 mM 2-(N-morpholino)ethanesulfonic acid (MES), 75 μM cetyltrimethylammonium bromide (CTAB), 3% methanol, pH 6.3, was used as running buffer. Under the optimum extraction conditions, the method showed good linearity in the range of 0.01–200 μg/L (GLYP) and 0.1–400 μg/L (AMPA), acceptable reproducibility (RSD 5–7%, n = 5), low limits of detection of 0.005 μg/L for GLYP and 0.06 μg/L for AMPA, and satisfactory relative recoveries (90–94%). Due to the low cost, the SLMTE device was disposed after each run which additionally eliminated the possibility of carry-over between runs. The validated method was tested for the analysis of both analytes in spiked tap water and river water with good success.     

Simultaneous determination of polysorbate 20 and unbound polyethylene-glycol in protein solutions using new core–shell reversed phase column and condensation nucleation light scattering detection

A novel fast and sensitive method has been developed for the specific simultaneous determination of polysorbate 20 (Tween 20) and unbound polyethylene-glycol (PEG) from liquid formulations in the presence of proteins and excipients. The quantitative determination is based on a fast liquid chromatographic (HPLC) separation and condensation nucleation light scattering detection (CNLSD or NQAD™). The method uses a Kinetex core–shell column (100 mm × 3 mm, 2.6 μm) and methanol–water–trifluoroacetic acid mobile phase. The rapid HPLC-CNLSD method presented here is suitable for quantifying polysorbate 20 in the range of 10–60 μg/ml and unbound PEG in the range of 2–40 μg/ml in protein solutions within good manufacturing practices (GMP) of the pharmaceutical industry.     

Development of a rapid,multi-class method for the confirmatory analysis of anti-inflammatory drugs in bovine milk using liquid chromatography tandem mass spectrometry

A rapid liquid chromatography tandem mass spectrometry (LC–MS/MS) method has been developed and validated for the simultaneous identification, confirmation and quantitation of seven licensed anti-inflammatory drugs (AIDs) in bovine milk. The method was validated in accordance with the criteria defined in Commission Decision 2002/657/EC. Two classes of AIDs were investigated, corticosteroids and non-steroidal anti-inflammatory drugs (NSAIDs). The developed method is capable of detecting and confirming dexamethasone (DXM), betamethasone (BTM), prednisolone (PRED), tolfenamic acid (TLF), 5-hydroxy flunixin (5-OH-FLU), meloxicam (MLX) and 4-methyl amino antipyrine (4-MAA) at their associated maximum residue limits (MRLs). These compounds represent all the corticosteroids and NSAIDs licensed for use in bovine animals producing milk for human consumption. These compounds have never been analysed before in the same method and also 4-methyl amino antipyrine has never been analysed with the other licensed NSAIDs. The method can be considered rapid as permits the analysis of up to 30 samples in one day. Milk samples are extracted with acetonitrile; sodium chloride is added to aid partition of the milk and acetonitrile mixture. The acetonitrile extract is then subjected to liquid–liquid purification by the addition of hexane. The purified extract is finally evaporated to dryness and reconstituted in a water/acetonitrile mixture and determination is carried out by LC–MS/MS. Decision limit (CCα) values and detection capability (CCβ) values have been established for each compound.     

Preparation of stir cake sorptive extraction based on polymeric ionic liquid for the enrichment of benzimidazole anthelmintics in water,honey and milk samples

In this work, a new stir cake sorptive extraction (SCSE) using polymeric ionic liquid monolith as sorbent was prepared. The sorbent was obtained by in situ copolymerization of an ionic liquid, 1-allyl-3-methylimidazolium bis[(trifluoro methyl)sulfonyl]imide (AMII) and divinylbenzene (DB) in the presence of N,N-dimethylformamide. The influence of the content of ionic liquid and the porogen in the polymerization mixture on extraction performance was studied thoroughly. The physicochemical properties of the polymeric ionic liquid were characterized by infrared spectroscopy, elemental analysis, scanning electron microscopy and mercury intrusion porosimetry. The usefulness of SCSE–AMIIDB was demonstrated by the enrichment of trace benzimidazole anthelmintics. Several parameters affecting the extraction efficiency were investigated, and under the optimized conditions, a simple and effective method for the determination of trace benzimidazoles residues in water, milk and honey samples was established by coupling SCSE–AMIIDB with high performance liquid chromatography/diode array detection (SCSE–AMIIDB–HPLC/DAD). Results indicated that the limits of detection (S/N = 3) for target compounds were 0.020–0.072 μg L⁻¹, 0.035–0.10 μg L⁻¹ and 0.026–0.076 μg L⁻¹ in water, milk and honey samples, respectively. In addition, an acceptable reproducibility was achieved by evaluating the repeatability and intermediate precision with relative standard deviations (RSD) of less than 9% and 11%, respectively. Finally, the established AMII–SCSE–HPLC/DAD method was successfully applied for the determination of benzimidazoles residues in milk, honey and environmental water samples. Recoveries obtained for the determination of benzimidazole anthelmintics in spiking samples ranged from 70.2% to 117.6%, with RSD below 12% in all cases.