Dispersive Solid-phase Extraction Clean-up Research Articles

An evaluation method for determination of non-polar pesticide residues in animal fat samples by using dispersive solid-phase extraction clean-up and GC-MS

A rapid and easy method has been proposed, optimized and evaluated for quantitative determination at trace level of a representative group of non-polar pesticides in fat samples. The method includes n-hexane-saturated acetonitrile extraction, fat precipitation by cooling pre clean-up followed by dispersive solid-phase extraction (d-SPE) based on QuEChERS procedure clean-up. Determination was performed by gas chromatography-mass spectrometry (GC-MS) in selected ion monitoring (SIM) mode. Efficiency of the d-SPE clean-up step was evaluated by comparison with fat oxidation treatment and gel permeation chromatography. Different combinations of d-SPE extraction reagents and sample amounts were tested in order to minimize matrix co-extractives and interferences. Best recoveries were obtained with 1200 mg of MgSO(4), 400 mg of end-capped C(18), 400 mg of PSA and 1 g of sample amount. SIM method, matrix effect, precision, and accuracy were evaluated with spiked pork fat samples for 38 representative pesticides. Results of this study showed that this technique is applicable in routine analysis for its application into monitoring programs. It simplifies time-consuming clean-up steps and allows a satisfactory long-term chromatographic performance.

Large-scale pesticide testing in olives by liquid chromatography–electrospray tandem mass spectrometry using two sample preparation methods based on matrix solid-phase dispersion and QuEChERS

In this work we have evaluated the performance of two sample preparation methodologies for the large-scale multiresidue analysis of pesticides in olives using liquid chromatography–electrospray tandem mass spectrometry (LC–MS/MS). The tested sample treatment methodologies were: (1) liquid–liquid partitioning with acetonitrile followed by dispersive solid-phase extraction clean-up using GCB, PSA and C 18 sorbents (QuEChERS method – modified for fatty vegetables) and (2) matrix solid-phase dispersion (MSPD) using aminopropyl as sorbent material and a final clean-up performed in the elution step using Florisil. An LC–MS/MS method covering 104 multiclass pesticides was developed to examine the performance of these two protocols. The separation of the compounds from the olive extracts was achieved using a short C 18 column (50 mm × 4.6 mm i.d.) with 1.8 μm particle size. The identification and confirmation of the compounds was based on retention time matching along with the presence (and ratio) of two typical MRM transitions. Limits of detection obtained were lower than 10 μg kg −1 for 89% analytes using both sample treatment protocols. Recoveries studies performed on olives samples spiked at two concentration levels (10 and 100 μg kg −1) yielded average recoveries in the range 70–120% for most analytes when QuEChERS procedure is employed. When MSPD was the choice for sample extraction, recoveries obtained were in the range 50–70% for most of target compounds. The proposed methods were successfully applied to the analysis of real olives samples, revealing the presence of some of the target species in the μg kg −1 range. Besides the evaluation of the sample preparation approaches, we also discuss the use of advanced software features associated to MRM method development that overcome several limitations and drawbacks associated to MS/MS methods (time segments boundaries, tedious method development/manual scheduling and acquisition limitations). This software feature recently offered by different vendors is based on an algorithm that associates retention time data for each individual MS/MS transition, so that the number of simultaneously traced transitions throughout the entire chromatographic run (dwell times and sensitivity) is maximized.

Evaluation of two sample treatment methodologies for large-scale pesticide residue analysis in olive oil by fast liquid chromatography–electrospray mass spectrometry

In this study, a comprehensive evaluation of two simple sample treatment methodologies has been carried out for the development of large-scale multi-residue methods for pesticide testing in olive oil. The proposed methodologies are based on (a) liquid–liquid partitioning with acetonitrile followed by dispersive solid-phase extraction clean-up using graphitized carbon black, primary-secondary amine and C 18 sorbents; (b) liquid partitioning with acetonitrile saturated with petroleum ether followed by matrix solid-phase dispersion (MSPD) using aminopropyl as sorbent material and a Florisil cartridge for final clean-up in the elution step. To evaluate the proposed sample treatment methodologies, 105 representative multi-class pesticides were studied using fast liquid chromatography–electrospray time-of-flight mass spectrometry (LC–TOFMS). For validation purposes, recoveries studies were carried out at 10 and 100 μg kg −1 levels, yielding recovery rates in the range 70–130% for 72% of analytes using liquid–liquid procedure and for 57% analytes using MSPD procedure. The LC–MS method provided good linearity, precision and accuracy. The limits of detection obtained were lower than 10 μg kg −1 for more than 85% analytes using both sample treatment methodologies. In addition, minor matrix effects (i.e. signal suppression or enhancement ≤20%) were observed in ca. 70% of the studied compounds. Data obtained shows that both sample treatment methodologies proposed can be successfully applied for large-scale pesticide testing in olive oil samples, showing the ability to quickly detect trace amount of over one hundred target species with different physicochemical properties, without requiring expensive instrumentation for sample treatment step and involving relatively low amounts of solvent consumption and waste generation.

Benzimidazole carbamate residues in milk: Detection by Surface Plasmon Resonance-biosensor, using a modified QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) method for extraction

A surface plasmon resonance (SPR) biosensor screening assay was developed and validated to detect 11 benzimidazole carbamate (BZT) veterinary drug residues in milk. The polyclonal antibody used was raised in sheep against a methyl 5(6)-[(carboxypentyl)-thio]-2-benzimidazole carbamate protein conjugate. A sample preparation procedure was developed using a modified QuEChERS method. BZT residues were extracted from milk using liquid extraction/partition with a dispersive solid phase extraction clean-up step. The assay was validated in accordance with the performance criteria described in 2002/657/EC. The limit of detection of the assay was calculated from the analysis of 20 known negative milk samples to be 2.7 μg kg −1. The detection capability (CCβ) of the assay was determined to be 5 μg kg −1 for 11 benzimidazole residues and the mean recovery of analytes was in the range 81–116%. A comparison was made between the SPR-biosensor and UPLC–MS/MS analyses of milk samples ( n = 26) taken from cows treated different benzimidazole products, demonstrating the SPR-biosensor assay to be fit for purpose.

Development and validation of a multi-residue method for the determination of pesticides in honeybees using acetonitrile-based extraction and gas chromatography–tandem quadrupole mass spectrometry

An optimized analytical method employing gas chromatography–tandem quadrupole mass spectrometry (GC–MS/MS) has been developed for the simultaneous screening of roughly 150 pesticides in honeybees suspected of poisoning by pesticides during field spraying. In this work, a sample preparation approach based on acetonitrile extraction followed by dispersive solid-phase extraction (d-SPE) cleanup was implemented and validated for pesticides in honeybees for the first time. The procedure involved homogenization of a 2 g sample (23 insects on average) with acetonitrile–water mixture followed by salting out with citrate buffer, magnesium sulphate and sodium chloride. An amount of matrix constituents with limited solubility in acetonitrile was reduced in the extract by precipitation at low-temperature (freezing-out cleanup). Hereafter, d-SPE cleanup was carried out using primary secondary amine (PSA), octadecyl (C18) and graphitized carbon black (GCB). This combination of cleanup steps ensured efficient extract purification. Linearity of the calibration curves was studied using matrix-matched standards in the concentration range between 4 and 500 ng mL −1 (equivalent to 10 and 1250 ng g −1), and coefficients of determination ( R 2) were ≥0.99 for approximately 90% of the targeted compounds. The recovery data were obtained by spiking honeybees samples free of pesticides at three concentration levels of 10, 50, and 500 ng g −1 (approximately 0.9, 4.3, 43.5 ng per bee). At these spiking levels 47, 77 and 92% of the targeted compounds were recovered, respectively. Generally the recoveries were in the range between 70 and 120% with precision values, expressed as relative standard deviation (RSD) ≤ 20%. The expanded uncertainty was estimated following a “top down” empirical model as being 28% on average (coverage factor k = 2, confidence level 95%). Preliminary results from practical application to analysis of real samples are presented. A total of 25 samples of honeybees from suspected pesticides poisoning incidents were analyzed, in which 10 different pesticides were determined.

Rapid analysis of organic farming insecticides in soil and produce using ultra-performance liquid chromatography/tandem mass spectrometry

A new method for the analysis of three ecological insecticides, namely azadyrachtin, spinosad (sum of spinosyn A and spinosyn D) and rotenone, in produce and soil samples is presented. Investigated compounds are one of the most significant insecticides authorized for organic farming crop protection in many countries. Extraction of the pesticides from plant and soil matrices was performed by using a modified quick, easy, cheap, effective, rugged, and safe (QuEChERS) method. The method entailed a single extraction of the investigated compounds with acidified acetonitrile followed by a dispersive solid-phase extraction cleanup step prior to the final determination by reverse-phase ultra-performance liquid chromatography/tandem quadrupole mass spectrometry (UPLC-MS/MS). Validation studies were carried out on cabbage, tomato and soil samples. Recoveries of the spiked samples were in the range between 67% and 108%, depending on the matrix and the spiking level. Relative standard deviations for all matrix-compound combinations did not exceed 12%. The limits of quantification were < or = 0.01 mg kg(-1) in all cases, except for azadirachtin. The developed method was applied to the analysis of real samples originating from organic farming production.

Development and Application of Dispersion Solid-Phase Extraction for Estrogens in Aquatic Animal Samples

A simple, fast, robust, sensitive, and specific method for the confirmation and quantification of four kinds of estrogens, including 17β-estradiol (E2), estriol (E3), 4-hydroxyestradiol (4-OHE2), and 2-methoxyestradiol (2-MeOE2), in fortified and incurred fish and shrimp flesh samples was developed. The method used simple extraction of a sample with acetonitrile, dispersive solid-phase extraction (dispersive SPE) cleanup, and high-performance liquid chromatography (HPLC) coupled with a fluorescence detector for confirmation and quantitation. Compared with conventional SPE cleanup, dispersive SPE greatly simplified and accelerated sample cleanup and was less time-consuming. Average recoveries of fortified samples were more than 80% for all estrogens except 4-OHE2, which had an average recovery of 60%. The results showed good repeatability that the variation coefficients of the instrument precision and the method were less than 8% and 13%, respectively. The relative standard deviation (RSD) of intraday repeatability tests was generally less than 12%.This method was able to demonstrate quantitative recoveries and provide confirmatory data for unambiguous analyte identification.

Multiresidue Determination of 156 Pesticides in Watermelon by Dispersive Solid Phase Extraction and Gas Chromatography/Mass Spectrometry

A rapid multiresidue method was developed for the simultaneous determination of 156 pesticides in commercial watermelon. The method involves a liquid-liquid extraction using acetonitrile coupled with dispersive solid phase extraction cleanup. The extracted elution of pesticides was determined by gas chromatography with electron impact mass spectrometric detection in the selected ion monitoring mode (GC-MS-SIM). Standards were prepared spiking blank watermelon samples to counteract the observed matrix effect. The method was validated by fortified at the level 0.020-0.120 mg/kg in watermelon. The average recoveries of all analytes were between 70% and 121%, and standard deviations were below 16%. The limit of quantitation (LOQ) for most compounds was below 0.005 mg/kg, which were lower than the maximum residue levels established by Korean legislations. The proposed method has been applied to the analysis of the 156 pesticide residues in commercial watermelon samples.

Determination of the Food Colorant, Chrysoidine, in Fish by GC–MS

A method has been developed for confirmatory determination of chrysoidine in fish by GC–MS. Samples were extracted with methanol, and an aliquot was subjected to dispersive solid-phase extraction clean-up with octadecyl sorbent. After centrifuging and filtering, extracts were evaporated to dryness and derivatized with acetic anhydride. The detection and quantification limits were 2.3 and 7.7 μg kg−1 respectively, demonstrating the potential of the method for quality control of food.

Streamlining methodology for the multiresidue analysis of β-lactam antibiotics in bovine kidney using liquid chromatography–tandem mass spectrometry

A previously reported multiresidue method for the analysis of 11 important β-lactams (amoxicillin, ampicillin, cefazolin, cephalexin, cloxacillin, desfuroylceftiofur cysteine disulfide (DCCD), deacetylcephapirin, dicloxacillin, nafcillin, oxacillin, and penicillin G) in bovine kidney has been further streamlined. The method is based on a simple extraction using acetonitrile–water (4:1, v/v), followed by dispersive solid-phase extraction clean-up with C 18 sorbent, concentration of an extract aliquot, and filtration of the final extracts using syringeless filter vials, which are used for the sample introduction in the liquid chromatographic–tandem mass spectrometric (LC–MS/MS) analysis. The recoveries have been improved by adding the internal standard [ 13C 6]sulfamethazine to the homogenized sample before the extraction step, which enabled a proper control of the volume changes during the sample preparation. Average recoveries of fortified samples were 87–103% for all β-lactams, except for DCCD, which had an average recovery of 60%. Based on the results of the stability study and LC mobile phase tests, methanol has been eliminated from the entire method, including the LC–MS/MS analysis. The best overall LC–MS/MS (electrospray positive ionization) performance was achieved by using 0.1% formic acid as an additive in both parts of the mobile phase, in water and in acetonitrile. To prevent carry-over in the LC–MS/MS analysis, the LC method was divided into two parts: one serving as an analytical method for injection of the sample and elution of the analytes and the other one, starting at a highly organic mobile phase composition, being dedicated for injection of a solvent, washing of the system, and equilibration of the column to the initial conditions of the analytical method. In this way, a blank solvent is injected after each sample, but these in-between injections contribute minimally to the overall sample throughput.

Qualitative screening and quantitative determination of pesticides and contaminants in animal feed using comprehensive two-dimensional gas chromatography with time-of-flight mass spectrometry

A method has been developed for the target analysis of over 100 pesticides and contaminants in a complex feed matrix. The method is based on extraction with ethyl acetate, cleanup by gel permeation chromatography (GPC) and dispersive solid-phase extraction (SPE) with primary secondary amine phase (PSA), and analysis by comprehensive two-dimensional gas chromatography with full scan time-of-flight mass spectrometric detection (GC × GC–TOF-MS). Parameters studied during method development included a dispersive SPE cleanup step after GPC, large volume injection into the GC system and the GC × GC separation. Qualitative and quantitative performance of the GC × GC system was evaluated by analyzing spiked extracts in the range equivalent to 1–100 μg/kg in feed. At levels of 50 μg/kg and higher, all compounds targeted for could be identified fully automatically by the software based on their mass spectra. At lower levels the hit rate decreased with the concentration. System linearity was excellent in solvent and only slightly affected by matrix (correlation coefficients r ≥ 0.995 for 90% of the compounds). Limits of quantification were in the 1–20 μg/kg range for most compounds. The overall method was validated for 106 compounds at the 10 and 100 μg/kg level. Recoveries between 70% and 110% and RSDs below 20% were obtained for the majority of the compounds.

Multicommuted fluorescence based optosensor for the screening of bitertanol residues in banana samples

A multicommuted flow through optosensor is developed in order to determine bitertanol in banana samples, by measuring the pesticide native fluorescence at 261/326 nm. The solid support used in the flow-through cell, C 18, allows both high selectivity and sensitivity necessary for the screening of the pesticide at residue levels. These characteristics of optosensing are implemented with low sample consumption and automation intrinsic properties of multicommutation, providing a useful tool for pesticide residue routine determination in foods. An acetonitrile extraction/partitioning and dispersive solid-phase extraction clean-up procedure proved to be useful; an additional cleaning step with C 18 solid phase extraction (SPE) cartridges was carried out, eliminating all matrix limitations. Recovery experiments performed on banana samples at different concentrations provided recoveries between 81% and 115%. A detection limit of 0.014 mg kg −1 was obtained. This makes the method suitable for screening bitertanol in banana samples, fulfilling the maximum residue levels of 3 mg kg −1 established by the European Union.

Confirmatory and Quantitative Analysis of β-Lactam Antibiotics in Bovine Kidney Tissue by Dispersive Solid-Phase Extraction and Liquid Chromatography−Tandem Mass Spectrometry

A simple, rapid, rugged, sensitive, and specific method for the confirmation and quantitation of 10 beta-lactam antibiotics in fortified and incurred bovine kidney tissue has been developed. The method uses a simple solvent extraction, dispersive solid-phase extraction (dispersive-SPE) cleanup, and liquid chromatography-tandem mass spectrometry (LC/MS/MS) for confirmation and quantitation. Dispersive-SPE greatly simplifies and accelerates sample cleanup and improves overall recoveries compared with conventional SPE cleanup. The beta-lactam antibiotics tested were as follows: deacetylcephapirin (an antimicrobial metabolite of cephapirin), amoxicillin, desfuroylceftiofur cysteine disulfide (DCCD, an antimicrobial metabolite of ceftiofur), ampicillin, cefazolin, penicillin G, oxacillin, cloxacillin, naficillin, and dicloxacillin. Average recoveries of fortified samples were 70% or better for all beta-lactams except DCCD, which had an average recovery of 58%. The LC/MS/MS method was able to demonstrate quantitative recoveries at established tolerance levels and provide confirmatory data for unambiguous analyte identification. The method was also tested on 30 incurred bovine kidney samples obtained from the USDA Food Safety and Inspection Service, which had previously tested the samples using the approved semiquantitative microbial assay. The results from the quantitative LC/MS/MS analysis were in general agreement with the microbial assay for 23 samples although the LC/MS/MS method was superior in that it could specifically identify which beta-lactam was present and quantitate its concentration, whereas the microbial assay could only identify the type of beta-lactam present and report a concentration with respect to the microbial inhibition of a penicillin G standard. In addition, for 6 of the 23 samples, LC/MS/MS analysis detected a penicillin and a cephalosporin beta-lactam, whereas the microbial assay detected only a penicillin beta-lactam. For samples that do not fall into the "general agreement" category, the most serious discrepancy involves two samples where the LC/MS/MS method detected a violative level of a cephalosporin beta-lactam (deacetylcephapirin) in the first sample and a possibly violative level of desfuroylceftiofur in the second, whereas the microbial assay identified the two samples as having only violative levels of a penicillin beta-lactam.