Development and validation of a solid-phase extraction method coupled to high-performance liquid chromatography with ultraviolet-diode array detection for the determination of sulfonylurea herbicide residues in bovine milk samples

This study proposes a fast, simple and sensitive liquid chromatography diode array detector (LC/UV-DAD)-based method for the simultaneous determination of eight sulfonylurea herbicides (bensulfuron methyl, chlorsulfuron, metsulfuron methyl, primisulfuron methyl, rimsulfuron, thifensulfuron methyl, triasulfuron and tribenuron methyl) in bovine whole milk at concentrations lower than the default limit of 0.01 mg kg(-1) allowed by current legislation (Regulation EC/396/2005 and following Annexes). An effective one-step solid phase extraction (SPE) and clean up procedure was defined with use of Chem Elut cartridges, providing good recoveries for all the analytes tested and with no matrix effects affecting method accuracy. Separation of herbicides was obtained on a C(18) column by acetonitrile- water gradient elution. Method validation has been performed according to European Commission Decision 2002/657/EC criteria, in terms of linearity, recovery, precision, specificity, decision limit (CC(α)) and detection capability (CC(β)). Typical recoveries ranged between 78.4% and 99.7%, at the maximum residue limits (MRLs) levels established by Regulation EC/396/2005, with relative standard deviations (RSD) no larger than 10%.     

Optimization of a pressurized liquid extraction method by experimental design methodologies for the determination of fluoroquinolone residues in infant foods by liquid chromatography

In the present study, we have developed a method based on pressurized liquid extraction (PLE) and liquid chromatography with fluorescence detection (LC-FLD) for the determination of residues of fluoroquinolones (FQs) in infant food products. PLE extraction has been optimized by the application of experimental design methodologies. Initially, a fractional factorial design (FFD) was used to screen the significance of four extraction parameters: solvent composition, temperature, pressure and number of cycles. The most significant factors, identified by ANOVA analysis, were the solvent composition, temperature and pressure, which were further optimized with the aid of a face centred design (FCD) and the desirability function. The optimized operating PLE conditions were as follows: ACN/o-phosphoric acid 50 mM pH 3.0 (80:20, v/v), 80 °C, 2000 psi and three extraction cycles of 5 min. Under these conditions, recoveries of the target FQs varied between 69% and 107% with RSDs below 9%. The whole method was validated according to the Commission Decision 2002/657/EC guidelines. The proposed method has been successfully applied to the analysis of different infant food products bought in local supermarkets and pharmacies. The results showed the presence of residues of enrofloxacin in a non-compliant baby food sample corresponding to a chicken-based formulation, which were also confirmed and quantified by LC–MS/MS analysis.     

Use of solid-phase extraction and high-performance liquid chromatography for the determination of triazine residues in water: validation of the method

A method for determination of some triazine residues in water has been developed. The method involves concentration with C18 solid-phase extraction cartridges followed by high-performance liquid chromatographic analysis using a C18 column with UV detection at 230 nm, a mobile phase of methanol-water (60:40, v/v) at pH 4.6 (phosphoric acid) and a flow-rate of 0.8 ml/min. After optimization of the extraction and separation conditions, the method was validated. The method can be used for determination of atrazine, simazine, cyanazine and ametryn in water, within the international limits of 0.1 microg/l.     

Core-shell magnetic porous organic polymer for magnetic solid-phase extraction of fluoroquinolone antibiotics in honey samples followed by high-performance liquid chromatography with fluorescence detection

By monomer-mediated in-situ growth synthesis strategy, with hydroquinone and 1,3,5-tris(4-aminophenyl)benzene as monomers, a core-shell magnetic porous organic polymer was synthesized through a simple azo reaction. Based on this, a magnetic solid-phase extraction–high-performance liquid chromatography–fluorescence detection method was proposed for the analysis of fluoroquinolones in a honey sample. With ofloxacin, ciprofloxacin, enrofloxacin, lomefloxacin, and difloxacin as target analytes, factors affecting the extraction efficiency had been optimized. The LODs were 1.5–5.4 ng/L (corresponding to 0.23–0.81 μg/kg in honey). The linear range was 0.005–20 μg/L for difloxacin, 0.01–20 μg/L for ofloxacin, ciprofloxacin and lomefloxacin, and 0.02–20 μg/L for enrofloxacin. The enrichment factor was 84.4–91.7-fold with a high extraction efficiency of 84.4–91.7%. The method was assessed by the analysis of target fluoroquinolones in honey samples, and the recoveries for the spiked samples were 79.3–95.8%. The results indicated that the established magnetic solid-phase extraction–high-performance liquid chromatography–fluorescence detection method is efficient for the analysis of trace fluoroquinolones in honey.     

Determination of amoxicillin residues in animal tissues by solid-phase extraction and liquid chromatography with fluorescence detection

Trace levels of amoxicillin residues were determined in animal tissues by liquid chromatography (LC) with fluorescence detection. An improved solid-phase extraction (SPE) procedure requiring less flammable solvent (diethyl ether) was developed for sample preparation. Muscle samples of beef, pork, chicken, and tilapia were extracted with a phosphate buffer followed by the modified SPE procedure for cleanup and concentration prior to the LC-fluorescence analysis. Average recoveries of fortified amoxicillin at 5, 10, and 20 micrograms/kg ranged from 83.9 to 85.8% in beef, 86.1 to 88.1% in pork, 81.7 to 82.9% in chicken, and 92.5 to 95.4% in tilapia. Relative standard deviations were < 4%.     

Development of an analytical method for eight fluoroquinolones using solid-phase extraction and liquid chromatography with fluorescence detection

In this study, a practicable and effective analytical method based on solid-phase-extraction and reversed-phase liquid chromatography with fluorescence detection (SPE-LC-FLD) was developed and partially validated for routine analysis of eight FQs in wastewater at the trace level. Different SPE materials, pH conditions and eluents were modified to find an economic and effective SPE conditions. In our work, it is the first time that well-known commercially available SPE sorbent are compared to ‘generic’ cheap SPE sorbent. Aqueous samples (pH 2–3) were extracted using Anpel? MEP cartridges where they were subsequently eluted by 6?mL of 2% formic acid in MeOH. The aqueous extracts were analysed by gradient elution LC-FLD, whose initial mobile phase was composed of ACN and 10?mmol?L^?1 tetrabutyl ammonium bromide (4/96, v/v, pH 3). The LODs and LOQs of the wastewater were as low as 0.32–2.12?ng?L^?1 and 1.07–7.07?ng?L^?1, respectively. The precisions of the overall method (RSD, n?=?3) using wastewater were below 10%. The method was used to quantify FQs in influents and effluents of several typical sewage treatment plants (STPs) in Shanghai. The extraction recoveries of 100?mL influent, 500?mL effluent and 500?mL of river water samples were between 88.6 and 102.6%, 79.2 and 109.2%, 80.0 and 105.5% and 87.4 and 99.4%, respectively. FQs of interest except sarafloxacin were identified in the influents, effluents and river waters with concentrations varying from 0.012–1.163?µg?L^?1, 0.003–0.291?µg?L^?1, and 0.002–0.040?µg?L^?1, respectively. The method can serve as a tool to obtain detailed information on occurrence, behaviour and fate of FQs in the aquatic environment. Occurrence of FQs detected in summer is higher than in spring at STPs, and those detected in the suburban area are less than those in the urban area. Complete removal of FQs is not achieved from the STPs, indicating domestic wastewater and STP discharge is the source of FQs in the surface water.     

Optimization of high-performance liquid chromatography and solid-phase extraction for determination of organophosphorus pesticide residues in environmental samples

High-performance liquid chromatography with solid-phase extraction (HPLC-SPE) was optimized for the analysis of three organophosphorus pesticide residues in water, apples and vegetable samples. Octadecylsilica disks (47-mm diameter) were used for solid-phase extraction. The parameters that affect both separation and extraction of methyl parathion, parathion and phoxim, such as mobile-phase composition, ionic strength, temperature, pH, and breakthrough volume, were investigated. The application of optimized HPLC-SPE to environmental samples gave reproducible results with low detection limits of 5 µg L^?1 for methyl parathion and parathion and 2.5 µgL^?1. Precisions of less than 8, 9 and 12% were obtained for water, spinach and apple samples, respectively.     

Multi-residue analysis of pesticide residues in mangoes using solid-phase microextraction coupled to liquid chromatography and UV-Vis detection

A sensitive and efficient solid‐phase microextraction method, based on liquid chromatography and UV–Vis detection, was developed and validated as an alternative method for sample screening prior to LC‐MS analysis. It enables the simultaneous determination of ten pesticides in mango fruits. The fiber used was polydimethylsiloxane while optimum SPME conditions employed have been developed and optimized in a previous work. The desorption process was performed in static mode, using acetonitrile as a solvent. The results indicate that the DI‐SPME/HPLC/UV–Vis procedure resulted in good linear range, accuracy, precision and sensibility and is adequate for analyzing pesticide residues in mango fruits. The limits of detection (0.6–3.3 μg/kg) and quantification (2.0–10.0 μg/kg) were achieved with values lower than the maximum residue levels (MRLs) established by Brazilian legislation for all pesticides in this study. The average recovery rates obtained for each pesticide ranged from 71.6 to 104.3% at three fortification levels, with the relative standard deviation ranging from 4.3 to 18.6%. The proposed method was applied for the determination of the aforementioned compounds in commercial mango samples and residues of azoxystrobin, fenthion, permethrin, abamectin and bifenthrin were detected in the mango samples, although below the MRLs established by Brazilian legislation.     

Simultaneous determination of phenolic xenoestrogens by solid-phase extraction and high-performance liquid chromatography with fluorescence detection.

A highly sensitive and selective method for simultaneous determination of some hydroxyl group-containing endocrine disruptors, including bisphenol A (BPA), bisphenol B (BPB), bisphenol E (BPE), bisphenol F (BPF) and 4-nonylphenol (4-NP), was developed. The method consists of precolumn derivatization of the analytes, solid-phase extraction (SPE) and subsequent chromatographic analysis by high-performance liquid chromatography (HPLC) with fluorescence detection. 4,4'-Cyclohexylidenebisphenol (BPZ) was used as an internal standard. Derivatization was carried out using 4-(4,5-diphenyl-1H-imidazol-2-yl)benzoyl chloride (DIB-Cl) as a label. Parameters of the derivatization reaction (temperature, time, concentration of reagent, stability, etc.) and of the solid-phase extraction (recovery, solvent, etc.) were studied in detail. Detection limits of compounds studied in standard solutions ranged from 0.08-1.3 ppb (ng/ml). The proposed method was successfully applied to plastic samples; BPA was found in both polycarbonate and polyvinyl chloride plastics, while 4-NP was found in plastics made of polyvinyl chloride and another polymer.     

On-fibre solid-phase microextraction coupled to conventional liquid chromatography versus in-tube solid-phase microextraction coupled to capillary liquid chromatography for the screening analysis of triazines in water samples

This paper compares the advantages and disadvantages of two different configurations for the extraction of triazines from water samples: (1) on-fibre solid-phase microextraction (SPME) coupled to conventional liquid chromatography (LC); and (2) in-tube SPME coupled to capillary LC. In-tube SPME has been effected either with a packed column or with an open capillary column. A critical evaluation of the main parameters affecting the performance of each method has been carried out in order to select the most suitable approach according to the requirements of the analysis. In the on-fibre SPME configuration the fibre coating was polydimethylsiloxane (PDMS)-divinylbenzene (DVB). The limits of detection (LODs) obtained with this approach under the optimized extraction and desorption conditions were between 25 and 125 microg/L. The in-tube SPME approach with a C18 packed column (35 mm x 0.5 mm I.D., 5 microm particle size) connected to a switching micro-valve provided the best sensitivity; under such configuration the LODs were between 0.025 and 0.5 microg/L. The in-tube SPME approach with an open capillary column coated with PDMS (30 cm x 0.25 mm I.D., 0.25 microm of thickness coating) connected to the injection valve provided LODs between 0.1 and 0.5 microg/L. In all configurations UV detection at 230 nm was used. Atrazine, simazine, propazine, ametryn, prometryn and terbutryn were selected as model compounds.     

Determination of niacin in infant formula by solid-phase extraction/liquid chromatography: peer-verified method performance-interlaboratory validation

This paper reports the results of the interlaboratory peer validation study of AOAC Peer-Verified Method (PVM) 1:2,000 for the determination of niacin in infant formula by solid-phase extraction/liquid chromatography. We have used a Data Quality Objectives (DQO) approach to address not only method variability and robustness but also accuracy of data through the use of an appropriate reference material in conjunction with the interlaboratory validation study. Our DQO included the following: (1) statistical agreement of analytical results and quantitative recovery between 2 collaborating laboratories; (2) the repeatability relative standard deviation (RSDr) values and the HORRAT (Horwitz ratio) obtained (1.07), which satisfied the criteria of the Horwitz "limits of acceptability" at the analyte level present; (3) validation of lack of interference; and (4) accuracy agreement within assigned values for a certified reference material. National Institute of Standards and Technology Standard Reference Material (NIST SRM) 1846 Infant Formula, with a certified value of 63.3 +/- 7.6 microg/g for niacin content, was used as a test material for collaborative study and accuracy assessment. Niacin values obtained by the originating laboratory were 59.7 +/- 4.0 microg/g (95% confidence interval [CI] = 1.4 microg/g with a relative standard deviation [RSD] of 6.7%) and by the peer laboratory were 56.6 +/- 6.6 microg/g (95% CI = 4.1 microg/g, with an RSD of 11.7%). Statistical evaluation using the means equivalence test showed that nicotinic acid values obtained by the peer laboratory were equivalent to those values obtained by the originating laboratory. Linear calibration curves and quantitative recovery were obtained. Integration of the PVM process with a readily available certified reference material gives the user confidence in the accuracy of the data generated by the method through traceability to the reference material used.     

Development and validation of a microsomal online cytochrome P450 bioreactor coupled to solid-phase extraction and reversed-phase liquid chromatography

The development and validation of an online cytochrome P450 (CYP)-based bioreactor coupled to automated solid-phase extraction (SPE) and gradient HPLC separation is described. The analytical method was checked on intra- and inter-day repeatability of the ethoxyresorufin-O-demethylation (EROD) reaction with CYP 1Al/1A2 containing beta-NF induced rat liver microsomes as an enzyme source. These experiments showed that CYP activity was linearly decreased with 16% over an 11 h period. Inter-day measurements had a CV of 9.1%. Furthermore, Km and Vmax values of the EROD reaction, measured with the bioreactor, were 2.72 +/- 0.46 microM and 7.9 +/- 0.5 nmol/min/mg protein, respectively. These were in good correspondence with Km and Vmax values, measured with standard batch assay, which amounted 0.66 +/- 0.08 microM and 6.4 +/- 0.2 nmol/min/mg protein respectively. In conclusion the newly developed analytical method can be used effectively and at a microliter scale for online generation, extraction and separation of metabolites.     

Matrix solid-phase dispersion extraction and determination by high-performance liquid chromatography with fluorescence detection of residues of glyphosate and aminomethylphosphonic acid in tomato fruit

A method based on matrix solid-phase dispersion (MSPD) is described for the quantitative extraction of glyphosate and its major metabolite aminomethylphosphonic acid (AMPA) from tomato fruit. After application of 120 microL of HNO3 1M to the sample, the dispersion column was packed with 0.5 g of sample blended into 1 g of NH2-silica. Two aqueous fractions were obtained. First, AMPA was eluted from the column using deionized water (F1), and then a NaH2PO4 0.005 M solution was used for the elution of glyphosate (F2). Cleanup of F1 and F2 was made by ion exchange chromatography on a SAX anion exchange silica. Determination was done by HPLC with fluorescence detection after precolumn derivatization with 9-fluorenylmethylchloroformate (FMOC-Cl). Mean recoveries calculated at fortification levels of 0.5 microg/g for glyphosate and 0.4 microg/g for AMPA were 87% and 78%, respectively. The relative standard deviations (n=7) for the total procedure were 10% and 16%. Detection limits were 0.05 microg/g for glyphosate and 0.03 microg/g for AMPA.     

On-line solid-phase extraction coupled to ultra-performance liquid chromatography with tandem mass spectrometry detection for the determination of benzotriazole UV stabilizers in coastal marine and wastewater samples

Benzotriazoles are a group of UV absorbing compounds considered emerging contaminants that are used in different personal care products, and therefore, it is of high interest to develop sensitive and fast methods for investigating their presence in the environment. In this work, we present the development and application of a novel method based on on-line solid-phase extraction coupled to ultra-performance liquid chromatography with tandem mass spectrometry detection (SPE-UPLC-MS/MS) for the determination of seven benzotriazole UV stabilizers (BUVSs) in coastal marine and wastewater samples. This process is compared with a conventional off-line SPE procedure followed by UPLC-MS/MS. The parameters affecting the performance of the sample preparation and determination processes were evaluated. The results indicate that the on-line procedure provides for better sensitivity and reproducibility and is faster and easier than the off-line procedure. The detection limits and quantification limits achieved were in the range of 0.6–4.1 ng∙L⁻¹ and 2.1–14 ng∙L⁻¹ and relative standard deviation between 6.2 and 10 %. The developed method was applied to coastal marine and wastewater samples from Gran Canaria Island (Spain). All of the BUVSs studied were detected in the samples from wastewater treatment plants and two were found in the seawater samples (UV P in the range of 2.8–4.4 ng∙L⁻¹ and UV 360 between 3.6 and 5.2 ng∙L⁻¹).     

Study of semi-automated solid-phase extraction for the determination of acaricide residues in honey by liquid chromatography

A solid-phase extraction (SPE) method followed by a reversed-phase high-performance liquid chromatography (HPLC) procedure is reported for the assay of a wide polarity range acaricide residues in honey. After selection of suitable chromatographic and detection conditions, most steps of the SPE procedure that may affect to the recovery were investigated. Honey sample was buffered at pH 6 and then applied to the preconditioned C₁₈ sorbent. A washing step was performed with 1 ml of a mixture of tetrahydrofuran (THF)–phosphate buffer (10:90, v/v) and finally, the analytes were eluted with 1 ml of THF. The extract was evaporated to dryness, reconstituted in mobile phase and chromatographed on a reversed-phase C₁₈ column with diode array detection. The recoveries of the more polar acaricides were higher than 80% and 60–70% for the more apolar ones. Limits of detection obtained ranged from 1 to 200 ng/g.     

Rapid determination of fluoroquinolone residues in honey by a microbiological screening method and liquid chromatography

A rapid and efficient method was developed for the simultaneous determination of seven fluoroquinolone (FQ) residues: norfloxacin, ciprofloxacin, danofloxacin, enrofloxacin, orbifloxacin, sarafloxacin, and difloxacin in honey. The samples were first screened with a microbiological method by using test plates made from metal-free purified agar seeded with Bacillus subtilis BGA. When a sample was found to contain FQ residues by using the microbiological method, it was analyzed by LC with fluorescence detection (LC/FL). FQs were extracted with Na2EDTA-McIlvaine buffer and purified by a dual SPE method in which a cation-exchange cartridge was connected to an anion-exchange cartridge. The overall recoveries of the seven FQs ranged from 70.0 to 92.1%. The intra-assay and interassay CVs were < or = 7.8 and < or = 5.1%, respectively. For the microbiological method, the LOD values ranged from 2 to 9 microg/kg. For LC/FL, the LOQ values ranged from 2 to 7 microg/kg. The developed method was used to analyze 70 honey samples. In 14 samples in which the microbiological method detected the presence of FQ residues, norfloxacin, ciprofloxacin, and enrofloxacin were identified by LC/FL.     

Determination of phthalate esters in water samples using Nylon6 nanofibers mat-based solid-phase extraction coupled to liquid chromatography

A new method was developed for simultaneous determination of five phthalate esters by a combination of mat-based solid-phase extraction (SPE) with high-performance liquid chromatography. The mat is composed of Nylon6 nanofibers. Dimethyl phthalate (DMP), diethl phthalate (DEP), di-n-butyl phthalate (DBP), di-(2-ethylhexyl) phthalate (DEHP) and dioctyl phthalate (DOP) were successfully separated on a RP-C18 column. Under optimized conditions, the detection limits found for DMP, DEP, DBP, DEHP and DOP were 3, 2, 6, 10 and 33 pg mL^?1, respectively. The method was applied to the analysis of various water samples. Spiked samples gave recoveries in the range from 86.9 to 101.9%, with relative standard deviations below 7.0%. A comparison of Nylon6 nanofibers mat as sorbents, and C18 cartridges and other kinds of SPE sorbents was carried out with respect to recovery, sensitivity, and reproducibility. The results indicated that the Nylon mat is a viable material for the enrichment and determination of phthalate esters in environmental water samples.     

Magnetic mixed hemimicelles solid-phase extraction of xanthohumol in beer coupled with high-performance liquid chromatography determination

In this study, silica-coated magnetic nanoparticles (Fe(3)O(4)/SiO(2) NPs) modified by cetyltrimethylammonium bromide (CTAB) were synthesized. They were successfully applied for extraction of xanthohumol in beer based on magnetic mixed hemimicelles solid-phase extraction (MMHSPE) coupled with high-performance liquid chromatography-ultraviolet determination. The main factors influencing the extraction efficiency including the surfactant amount, the beer pH, the extraction time, the desorption condition and the maximum extraction beer volume were optimized. Under the optimized conditions, a concentration factor of 60 was achieved by extracting 120 mL beer sample using MMHSPE and the detection limit of xanthohumol is 0.0006 mg/L. The proposed method was successfully applied for determination of xanthohumol in various beer samples with the xanthohumol contents in the range of 0.031-0.567 mg/L. The satisfactory recoveries (90-103%) were obtained in analyzing spiked beer samples.     

Determination of quinolones in water samples by solid-phase extraction and liquid chromatography with fluorimetric detection

A method is reported for the determination, in water samples, of 10 quinolones which are used as veterinary drugs. Analytes are isolated from samples by solid-phase extraction (SPE) and analysed by reversed-phase high-performance liquid chromatography using fluorimetric detection. A solid-phase extraction procedure based on retention on HBL OASIS cartridges and elution with a mixture of acetonitrile-water in basic medium is suitable for pre-concentration of the analytes. Pre-concentration factors up to 250 can be obtained. The quinolones are separated with an octyl silica-based column and mobile phases consisting of aqueous oxalic acid solutions and acetonitrile mixtures. The attained detection limits of the whole process are in the ng l(-1) level when 250 ml of water sample is processed. Recovery rates, from natural water samples spiked at 2060 ng l(-1) level, range from 70 to 100% and common standard deviation are about 6-12%.     

Determination of quinolone residues in infant and young children powdered milk combining solid-phase extraction and ultra-performance liquid chromatography-tandem mass spectrometry

The present work describes a method based on solid-phase extraction (SPE) and ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) for the simultaneous determination of three quinolones (pipemidic acid, oxolinic acid and flumequine) and twelve fluoroquinolones (marbofloxacin, fleroxacin, pefloxacin, levofloxacin, norfloxacin, ciprofloxacin, enrofloxacin, danofloxacin, lomefloxacin, difloxacin, sarafloxacin, and moxifloxacin) in different infant and young children powdered milks. After suitable deproteination of the reconstituted powdered samples, a SPE procedure was developed providing recovery values higher than 84% (RSDs lower than 13%) for all the analytes, with limits of detection between 0.04 and 0.52 μg/kg. UPLC-MS/MS analyses were carried out in less than 10 min. Sixteen infant and young children powdered milk samples of different origin, type and composition bought at Spanish markets were analyzed. Residues of the selected antibiotics were not detected in any of the analyzed samples.