José F. Huertas-Pérez

Determination of N-methylcarbamate pesticides in water and vegetable samples by HPLC with post-column chemiluminescence detection using the luminol reaction

In this paper we proposed a reverse high performance liquid chromatography method for the simultaneous determination of three N-methylcarbamates (NMCs) named carbofuran, carbaryl and methiocarb, using the post-column chemiluminescence (CL) detection with the luminol reaction. This method is based on the enhancing effect of these analytes on the CL emission generated by the oxidation of luminol with potassium permanganate in alkaline medium. The separation was reached in less than 14 min using a C18 column and an isocratic binary mobile phase consisting of acetonitrile:water (50:50, v/v) pumped at a flow rate of 1 mL min(-1). CL reagents (luminol and KMnO(4)) were incorporated by means of a peristaltic pump and were firstly mixed using a three-way connector. Then this stream was mixed with the eluate using another three-way connector just before reaching the detection cell. The optimization of variables affecting the CL reaction (reaction medium, concentration, flow rate of reagents and distance between both connectors) were optimized by means of experimental designs. Ethiofencarb, a NMC which has nowadays fallen into disuse was used as internal standard. For the analysis of theses pesticides in real water samples a pre-treatment step consisting of solid phase extraction (SPE) was conducted in order to reach sensitivity levels below the legal maximum concentration permitted. In the case of vegetable sample, SPE was used for matrix cleaning purpose.

A new approach in sample treatment combined with UHPLC-MS/MS for the determination of multiclass mycotoxins in edible nuts and seeds

A sensitive, simple and rapid method for the determination of fourteen mycotoxins in nuts and seeds (including almonds, peanuts, sunflower seeds, pumpkin seeds, walnuts, macadamia nuts, pistachios, hazelnuts and pine nuts) has been developed using ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS). The sample treatment comprises a first step based on QuEChERS procedure for the determination of fumonisin B1, fumonisin B2, deoxynivalenol, fusarenon-X, T-2 and HT-2 toxin, citrinin, sterigmatocystin, zearalenone and ochratoxin A. A subsequent clean-up step based on the dispersive liquid-liquid microextraction (DLLME) was necessary for the determination of aflatoxins (B1, B2, G1 and G2), since their determination was not possible applying only the QuEChERS-based extraction. The method was validated for peanuts as representative matrix and was subsequently evaluated for the other eight matrices. Quantification limits obtained for aflatoxins, the unique mycotoxins legislated on these matrices, were lower than the maximum levels allowed by the current legislation, while quantification limits obtained for the other mycotoxins were lower than the limits usually permitted by the legislation in other food matrices. Precision of the method was always lower than 11%, and recoveries ranged between 60.7% and 104.3%.

Determination of carbamates in edible vegetable oils by ultra-high performance liquid chromatography-tandem mass spectrometry using a new clean-up based on zirconia for QuEChERS methodology.

In this study a fast, selective and sensitive multiresidue method based on QuEChERS methodology has been evaluated and validated for the determination of carbamate pesticides, in edible vegetable oils by UHPLC-MS/MS. A new clean-up sorbent, Supel(TM) QuE Z-Sep(+), has been successfully applied in vegetable oil extracts. Z-Sep(+) was compared with other sorbents (i.e. mixture of C18 and PSA) previously used for dispersive solid phase extraction of these matrices, reducing more effectively matrix effects without a significant decrease of analyte recoveries. Matrix effect was studied in different matrices (extra-virgin olive, sunflower, maize, linseed and sesame oil) being ≤│30│% for most of the studied pesticides. Under optimum conditions, recoveries ranged from 74% to 101%, with relative standard deviations lower than 10%. Limits of quantification ranged from 0.09 to 2.0 µg kg(-1), allowing their determination at the low concentration levels demanding by current legislation.

Chemiluminescence determination of carbofuran at trace levels in lettuce and waters by flow-injection analysis

A simple, fast chemiluminescence (CL) flow-injection (FI) method based on the reaction of luminol with KMnO(4) in alkaline medium has been described for the direct determination of carbofuran. The method is based on the enhancing effect in the emission light from the oxidation of luminol produced in presence of carbofuran. The optimisation of instrumental and chemical variables influencing the CL response of the method has been carried out by applying experimental design, using the proposed flow-injection manifold. Under the optimal conditions, the CL intensity was linear for a carbofuran concentration over the range of 0.06-0.5mugml(-1), with a detection limit of 0.02mugml(-1). The method has been successfully applied to the determination of carbofuran residues in spiked water and lettuce samples.

Method optimization and validation for the determination of eight sulfonamides in chicken muscle and eggs by modified QuEChERS and liquid chromatography with fluorescence detection.

A simple, effective and reliable method for the determination of eight sulfonamide antibiotics (sulfadiazine, sulfapiridine, sulfamerazine, sulfamethazine, sulfachloropiridazine, sulfamethoxazole, sulfadoxine, sulfadimethoxin) in chicken muscle and eggs by liquid chromatography and fluorescence detection has been developed and validated. Sulfonamides do not present native fluorescence, however their direct determination was achieved by on-line post-column photochemical derivatization by UV irradiation. Sample treatment was based on QuEChERS with several modifications depending on the matrix. Egg extracts were cleaned-up using PSA for the dispersive solid phase extraction step. On the other hand, a new clean-up sorbent, Supel™ QuE Z-Sep(+), has been successfully applied in chicken muscle extract and has proved to be effective for interference removal from this matrix. Under optimum conditions, recoveries from 65.9 to 88.1%, relative standard deviations lower than 10% (except for sulfachloropiridazine), and limits of quantification (LOQs) from 14 to 85 μg kg(-1) were achieved. Thus, the method complies with current European requirements.

Simple and efficient methodology to determine mycotoxins in cereal syrups.

Consumption of cereal syrups is increasing nowadays. Mycotoxins may be found in syrups resulting from the use of contaminated raw material or invading microorganisms in the final manufactured product. However, these matrices have been scarcely explored regarding their mycotoxin content. A sensitive, simple and rapid method for the determination of ten mycotoxins (ochratoxin A, fumonisin B1, fumonisin B2, deoxynivalenol, fusarenon-X, T-2 and HT-2 toxin, citrinin, sterigmatocystin and zearalenone) in cereal syrups (rice, wheat and barley) has been developed and characterised using ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) and a sample treatment based on QuEChERS procedure. Matrix-matched calibration curves were established and limits of quantification were below the limits usually established by current legislation in different foodstuff. The relative standard deviation of the whole analytical method was lower than 12% in all cases, while recoveries ranged from 70.2% to 100.6%, therefore fulfilling the current requirements for mycotoxins analysis.

Ultrasound-assisted surfactant-enhanced emulsification microextraction for the determination of carbamates in wines by ultra-high performance liquid chromatography–tandem mass spectrometry

A new sensitive multiresidue method based on ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) has been developed for the detection, confirmation and quantification of twenty five carbamates in wine samples. The separation was achieved in 5.5 min, using a Zorbax Eclipse plus RRHD C18 column (50 mm×2.1 mm, 1.8 μm), with a mobile phase of water and methanol, both of them with 0.01% formic acid. The analytes were detected in positive mode with multiple reaction monitoring mode. Ultrasound-assisted surfactant-enhanced emulsification microextraction (UASEME), using a low-density extraction solvent has been optimized for the satisfactory extraction of carbamates and clean-up of extracts. The matrix effect was studied, showing that the proposed procedure provides very clean extracts. Under optimum conditions, recoveries for fortified wine samples ranged from 74 to 102%, with relative standard deviations lower than 6%. Limits of quantification ranged from 0.15 to 0.92 μgl(-1), showing the high sensitivity of this fast and simple method and its compliance with current requirements.

Vortex-assisted surfactant-enhanced emulsification liquid-liquid microextraction for the determination of carbamates in juices by micellar electrokinetic chromatography tandem mass spectrometry.

A new method based on vortex-assisted surfactant-enhanced-emulsification liquid-liquid microextraction has been developed for the extraction of carbamate pesticides in juice samples prior to their determination by micellar electrokinetic chromatography coupled to tandem mass spectrometry. This sample treatment allowed the satisfactory extraction and the extract clean-up of 25 carbamates from different fruit and vegetal juices (banana, tomato, and peach). In this study, the addition of ammonium perfluorooctanoate in the aqueous sample in combination with vortex agitation, provided very clean extracts with short extraction times. Under optimized conditions, recoveries of the proposed method for these pesticides from fortified juice samples ranged from 81% to 104%, with relative standard deviations lower than 15%. Limits of quantification were between 2.3µgkg(-)(1) and 4.7µgkg(-)(1), showing the high sensitivity of this fast and simple method.

High-throughput determination of citrinin in rice by ultra-high-performance liquid chromatography and fluorescence detection (UHPLC-FL)

A rapid, simple and effective method for the determination of citrinin in rice by UHPLC coupled to fluorescence detection has been developed and validated. Extraction of citrinin from rice samples was achieved by applying a QuEChERS-based extraction/partitioning process with water and acetonitrile, without the need of further extract clean-up. The method was fully validated for white rice, and its applicability in other rice matrices, such as brown and red rice, was confirmed by recovery experiments. Under optimum conditions, recoveries ranged from 72.5% to 92.8%, with relative standard deviations (RSDs) lower than 7.1%. Detection and quantification limits were estimated to be 1.5 and 5.0 µg kg−1, respectively. Finally, 21 organic rice samples were analysed, but none was contaminated with citrinin above the detection limit of the method. The method proved to be fast and non-laborious.

A high-throughput method for the determination of quinolones in different matrices by ultra-high performance liquid chromatography with fluorescence detection

Ultra-high performance liquid chromatography coupled to fluorescence detection has been proposed for the determination of thirteen quinolones of human and veterinary use (danofloxacin, sarafloxacin, difloxacin, flumequine, norfloxacin, pipemidic acid, enoxacin, lomefloxacin, marbofloxacin, ciprofloxacin, enrofloxacin, moxifloxacin and oxolinic acid). Sample treatment consisted of a modified method based on salting-out assisted liquid–liquid extraction, which involves the use of magnesium sulphate and sodium chloride as salting-out agents. To demonstrate the applicability of the method, it was characterized for three different matrices of interest (milk, urine and environmental water), obtaining very low limits of quantification (0.2–192 μg L−1). The precision of the method was evaluated in terms of repeatability and intermediate precision, and the results were acceptable in all cases (relative standard deviations lower than 11%). Recovery studies were performed on the three matrices, obtaining values between 71% and 104%.