Dispersive SPE Cleanup Research Articles

Effects of carotenoid pigmentation in salmon on antibiotic extraction recovery, matrix effects and accuracy of quantification by ultrahigh performance liquid chromatography coupled to tandem mass spectrometry

Carotenoid pigmentation in salmon may interfere with the accuracy of antibiotic analysis with ultra-high pressure liquid chromatography coupled to tandem spectrometry (UPLC-MS/MS) by causing matrix effects or affecting the recovery of compounds during extraction. In the present study, we used both pigmented and non-pigmented salmon to understand the role pigments play on antibiotic analysis, and tested whether clean-up of the extract with dispersive solid phase extraction (dSPE) or hydrophilic-lipophilic balance (HLB) SPE clean-up reduces matrix effects. Thirty antibiotics and their respective class-specific surrogate standards were measured in Sockeye (pigmented), King (pigmented) and Ivory King (non-pigmented) salmon extracted using the QUEChERS method, or a modified QUEChERS method involving dSPE or HLB SPE clean-up (for Sockeye salmon only). Significant matrix effects and lower percent recoveries of spiked antibiotics were observed in pigmented versus non-pigmented salmon extracted with the QUEChERS method. Dispersive SPE clean-up did not improve extraction recoveries or matrix effects. However, SPE clean-up with HLB columns improved matrix effects for several antibiotics but reduced the percent recovery to < 30%. Across all types of salmon analyzed, the accuracy of quantitation was minimally impacted, likely due to similar behavior of the surrogate standards tagged to each antibiotic class during extraction. Our results demonstrate that carotenoids in salmon are associated with significant matrix effects and low extraction recoveries, but do not impact accuracy.

Determination of multi-pesticide residues in honey with a modified QuEChERS procedure followed by LC-MS/MS and GC-MS/MS

This paper presents a modified QuEChERS method to determine 346 pesticide residues in honey by LC/MS-MS and GC/MS-MS. Acetate buffered acetonitrile extraction followed by dispersive SPE clean-up with C18-E, PSA and MgSO4 sorbents were used for sample preparation. This method was validated according to the European Union SANTE/2019/12682 guidelines. Acceptable values were obtained for the linearity, the limit of detection (0.001–0.003 mg/kg), limit of quantification (0.002–0.008 mg/kg), trueness (for the four tested levels, the recovery assays values were between 76 and 112%), precision (RSD ≤20.0%) and measurement uncertainty tests (<50%). The method was used for the analysis of 100 honey samples from Northern Cyprus and residues of 4 pesticides (coumaphos, thiamethoxam, N-(2,4-dimethylphenyl)formamide, piperonyl butoxide) were quantified in 42 honey samples. Coumaphos (29%) and N-(2,4-dimethylphenyl)formamide (11%) were the residues frequently detected in the samples. N-(2,4-dimethylphenyl)formamide concentrations (0.56 and 0.84 mg/kg) were determined above the MRL limits (0.2 mg/kg) in two samples. This study shows that LC/MS-MS and GC/MS-MS in combination with modified QuEChERS can be used to routinely scan pesticide residues in honey samples and direct contamination with acaricides/veterinary drugs applied in the treatment of plagues in the hive during honey harvest is the main source of honey pollution in Northern Cyprus.

Development of a method for assessing the accumulation and metabolization of antidepressant drugs in zebrafish (Danio rerio) eleutheroembryos

Antidepressant drugs are widely used for the treatment of common mental or other psychiatric disorders such as depression, which affect about 121 million people worldwide. This widespread use has contributed to the input of these pharmaceuticals and their metabolites into the environment. The aim of this work was to develop an analytical method to quantify the most widely used antidepressant drugs, selective serotonin reuptake inhibitors (SSRI), and their main metabolites in the environment. For this, a new and reliable miniaturized extraction method based on dispersive SPE cleanup procedure for extraction of SSRI followed by derivatization with n-heptafluorobutyrylimidazole, and detection by GC-MS was developed. The methodology, including a first-order one-compartment model, was then applied to a bioconcentration study in zebrafish (Danio rerio) eleutheroembryos. The results showed low bioaccumulation of these compounds; however, a biotransformation evidence of the parent compounds into their metabolites was observed after 6 h of exposure. These results indicate the need to integrate metabolic transformation rates to fully model and understand the bioaccumulation patterns of SSRI and their metabolites.Graphical abstract

Analysis of pharmaceuticals in fish using ultrasound extraction and dispersive spe clean-up on que Z-Sep/C18 followed by LC-QToF-MS detection

The presence of pharmaceutically active compounds (PhACs) in aquatic biota has been received much less attention than their presence in surface or waste water, and it was not until the mid-2000s, this gap started to be addressed. Here, we present SQUEEZe (Solid-liQuid Ultrasound Extraction with QuE Z-Sep/C18 as dispersive clean-up): a fast method for analysis of the trace 47 PhACs in fish muscle. Compared to our previously reported method [1], it offers alternatives with improvements in recoveries, number of analytes, sample volume and solvent used.The key aspects of this method are:•The ultrasound extraction was performed with acetonitrile/isopropanol 0.1% V/V formic acid. A clean-up step using QuE Z-Sep/C18 sorbents was employed to reduce lipid content of the extracts and further matrix effects in the detection of the analytes.•A HPLC separation with a Kinetex EVO C18 packed column in 11 min was optimized. MS and MS/MS data were collected using SWATH acquisition on the SCIEX X500R QTOF in (+)-ESI mode.•The method validated at 3 different concentrations levels: 5, 25 and 50 ng/g fish. It presented good intraday/interday reproducibility and absolute recoveries ≥ 60% for majority of analytes in composite homogenate muscle matrix of Squalius cephalus.•10 out 47 compounds were detected in fish samples.

Multi-residue methodology for the determination of 16 coccidiostats in animal tissues and eggs by hydrophilic interaction liquid chromatography – Tandem mass spectrometry

A simple, sensitive and efficient confirmatory method was developed and validated for the determination of 16 coccidiostats in animal tissues and eggs using hydrophilic interaction liquid chromatography–tandem mass spectrometry (HILIC-MS/MS). The sample preparation consisted of a solid-liquid extraction with ACN and dispersive SPE cleanup with MgSO4 and C18. Analysis was realized in an Acquity BEH HILIC silica column, in SRM mode. Both positive and negative ionization was performed, using polarity switching. Isocratic elution was used with a mobile phase of ACN: aqueous ammonium formate 1 mM with 0.1% formic acid (80:20, v/v). Method validation was performed in eggs, poultry, bovine, ovine, porcine and rabbit tissue and exceptionally low LODs were achieved, varying from 0.004 μg kg−1 (decoquinate in porcine tissue) to 0.560 μg kg−1 (halofuginone in eggs). The developed methodology was applied in 82 muscle and egg samples through the Greek National Residue Control Plan for coccidiostats.

Development and validation of a multiclass method for the determination of antibiotic residues in honey using liquid chromatography-tandem mass spectrometry

ABSTRACTA new, simple and fast method was developed for the determination of multi-class antibiotic residues in honey (sulfonamides, tetracyclines, macrolides, lincosamides and aminoglycosides). Separation and determination were carried out by LC-MS/MS. During sample preparation, various parameters affecting extraction efficiency were examined, including the type of solvent, pH, efficiencies of cleavage of N-glycosidic linkages by hydrochloric acid, ultrasonic extraction and its duration compared with shaking, along with dispersive SPE clean-up. Experiments with fortified samples demonstrated that 10 min of ultrasonic treatment with acidified methanol (HCl 2 M) followed by dispersive SPE clean-up with 50 mg PSA gave an effective sample preparation method for several classes of antibiotics in honey. Anhydroerythromycin A, erythromycin A enol ether and desmycosin were used as markers for the presence of erythromycin A and tylosin A in honey samples. The method was validated according to European Commission Decision (EC) No. 2002/657. The recoveries of analytes ranged from 85% to 111%. Repeatability and intra-laboratory reproducibility were < 20.6% and 26.8%, respectively. Decision limit (CCα) and detection capability (CCβ) ranged from 6 to 9 µg kg–1 and from 7 to 13 µg kg–1, respectively, except for streptomycin and neomycin, which showed slightly higher CCα at 25 µg kg–1 and CCβ at 34 µg kg–1. Finally, the method was applied to the honey test material 02270 through a FAPAS proficiency test (PT) for the determination of tetracyclines. PT results were obtained within a z-score range of ±2, proving that the validated method is suitable for routine analysis to ensure the quality of honey.

QuEChERS 및 LC-MS/MS를 이용한 토양 중 124종 잔류농약다성분 분석법

토양환경 중 잔류농약 동시분석을 위해 124종의 농약을 대상으로 QuEChERS 전처리와 LC-MS-MS를 이용한 분석법을 개발하고자 하였다. QuEChERS 전처리 과정은 토양시료에 0.2N <TEX>$NH_4Cl$</TEX> 용액 침지하고 1% acetic acid 포함 acetonitrile 용매로 추출 후 <TEX>$MgSO_4$</TEX> 및 NaOAc로 salting out하고 PSA 50 mg, <TEX>$C_{18}$</TEX> 50 mg, <TEX>$MgSO_4$</TEX> 150 mg를 이용해 정제하였다. 분석법의 검증을 위해 양토 및 사양토 두 토양에서 0.005, 0.01, 0.02, 0.1 mg/kg 4수준으로 처리 후 회수율 실험을 실시하였다. 회수율 실험결과 Benfuracarb, Ethiofencarb, Pymetrozine, Pyrethrin 성분을 제외하고 나머지 성분들은 사양토와 양토 모두 70~120%의 적정 회수율을 나타내었으며, 상대표준편차도 20% 이내를 양호한 결과를 나타내었다. 따라서 QuEChERS 전처리 및 LC-MS를 이용한 분석법은 토양 중 잔류농약 다성분동시분석에 적용이 가능할 것으로 판단되었다. A QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) multiresidue method was developed for the simultaneous analysis of 124 pesticides in soil by LC-MS. The procedure involved liquid extraction of soil immersed with 0.2N <TEX>$NH_4Cl$</TEX> by acetonitrile with 1% acetic acid, followed by anhydrous <TEX>$MgSO_4$</TEX> and sodium acetate, and dispersive SPE cleanup with <TEX>$MgSO_4$</TEX>, primary secondary amine (PSA) and <TEX>$C_{18}$</TEX>. The extracts were analyzed with LC-MS/MS in ESI positive mode. Standard calibration curves were made by matrix matched standards and their correlation coefficients were higher than 0.99. Recovery studies for the validation were carried out using two type soils, loam and sandy loam, at four concentration levels (0.005, 0.01, 0.02, and 0.1 mg/kg). The recoveries of pesticides were in the range of 70-120% with < 20% RSD except 4 pesticides, Benfuracarb, Ethiofencarb, Pymetrozine, and Pyrethrin. This result indicated that the method using QuEChERS and LC-MS/MS could be applied for the simultaneous determination of pesticide residues in soils.

Development, optimization and validation of a multimethod for the determination of 36 mycotoxins in wines by liquid chromatography–tandem mass spectrometry

A fast and efficient multimethod for the determination of 36 mycotoxins in wine, using ultra-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS), was developed, optimized, validated and implemented in routine analysis. A simplified, quick extraction was performed with acetonitrile, derived from the QuEChERS (quick, easy, cheap, effective, rugged and safe) approach, which was traditionally developed for pesticides analysis. This study aimed at a single extraction and chromatographic separation for 36 mycotoxins.Optimization tests were performed to find the proper ratio of wine: water and extraction solvent and the need for an additional buffering step with ammonium formate/formic acid and a dispersive SPE cleanup with various sorbents. The dSPE steps did not show significant improvement in analysis results, therefore, it was not applied in the final method to be validated. The mycotoxins were separated and detected on a UPLC–MS/MS system, used in the ESI positive ionization mode. The various mycotoxins were divided in three different concentration level groups, according to their sensitivity in UPLC–MS/MS. The validation was performed by analyzing recovery samples at three different spike levels with six replicates (n=6) at each level. Linearity (r2) of calibration curves, accuracy (recovery %), instrument limits of detection and method limits of quantification (LOD and LOQ), precision (RSD%) and matrix effects (%) were determined for each individual mycotoxin. From the 36 mycotoxins analyzed by UPLC–MS/MS (ESI+), 35 showed average recoveries in the range 70–120%, and 86% of these with a RSD≤20% at the lowest spike level (for Group I, II and III, respectively, 1, 50 and 10µgkg−1). The higher spike levels showed even better results. Only nivalenol could not be quantified at any concentration level. The method LOQ for 86% of the mycotoxins studied was the lowest spike level tested. The matrix effect observed was low for most mycotoxins analyzed and had no significant influence on the analytical results obtained. The developed procedure was applied successfully in routine analysis in a survey of wine samples originating from different countries.

Optimisation of a sample preparation method for the determination of fumonisin B1 in rice

A simple, rapid and cost-effective sample preparation method for the determination of fumonisin B1 in rice was optimised using a strategy of sequential experimental designs. Initially, a Plackett–Burman design was applied to select the statistically significant variables for the determination of fumonisin B1, and then, a central composite rotatable design was used to define the optimal conditions of these variables. The method involves extraction with a 50% acetonitrile aqueous solution and glacial acetic acid, liquid–liquid partitioning with addition of anhydrous sodium sulphate and sodium chloride, followed by dispersive SPE clean-up with diatomaceous earth. The final extract was analysed by HPLC-FLD after precolumn derivatisation with ortho-phthaldialdehyde. The optimised method was validated for selectivity, linearity, matrix effect, limits of detection and quantification, trueness, and precision, and then applied to commercial samples of polished rice. This is the first report of the occurrence of fumonisin B1 in commercial samples of polished rice from the Southeast region of Brazil.

Determination of Pesticide Residues in Honeybees using Modified QUEChERS Sample Work-Up and Liquid Chromatography-Tandem Mass Spectrometry

Increasing emissions of chemical compounds to the environment, especially of pesticides, is one of factors that may explain present honeybee colony losses. In this work, an analytical method employing liquid chromatography-tandem mass spectrometry (LC-MS/MS) was optimized for the simultaneous screening of 19 pesticides which have not been yet determined in honeybee samples from northern Poland (Pomerania). The sample preparation, based on the QuEChERS method combining salting-out liquid-liquid extraction to acetonitrile and a dispersive-SPE clean-up, was adjusted to honeybee samples by adding a small amount of hexane to eliminate beeswax. The recovery of analytes ranged from 70% to 120% with relative standard deviation ≤20%. The limits of detection were in the range of 0.91–25 ng/g. A total of 19 samples of honeybees from suspected pesticide poisoning incidents were analyzed, in which 19 different pesticides were determined.

Analytical performance of two miniaturised extraction methods for triclosan and methyltriclosan, in fish roe and surimi samples

A new and reliable miniaturised QuEChERS-based extraction method combined with a dispersive SPE cleanup procedure for extracting triclosan and methyltriclosan from fish roe and surimi samples was proposed. The effectiveness of different extraction/partition conditions for QuEChERS method was systematically investigated, and the use of acetonitrile extraction solvent and MgSO4, PSA, C18 and Florisil as cleanup reagents was recommended in the final method. Other method based on ultrasonic extraction with ethylacetate and clean-up with SPE was also evaluated for these samples. Different polymeric and silica sorbents for clean up were tested and the combination of Florisil and PSA was finally selected. The performance of these miniaturised sample preparation methods combined with GC–MS with quadrupole detection were compared. Extraction efficiency as well as cleaning effectiveness, laboriousness and speed were taken as criteria for method evaluation. Satisfactory validation parameters, such as linearity, recovery, precision and LODs and LOQs for both developed analytical methods were obtained from fish roe and surimi samples. Finally, both methods were applied to real samples. The sensitivity of the proposed methods was good enough to ensure reliable determination of target analytes at concentration levels commonly found in this kind of samples.

Multiresidue Pesticide Analysis of Dried Botanical Dietary Supplements Using an Automated Dispersive SPE Cleanup for QuEChERS and High-Performance Liquid Chromatography–Tandem Mass Spectrometry

An automated dispersive solid phase extraction (dSPE) cleanup procedure as part of the Quick, Easy, Cheap, Effective, Rugged, and Safe (QuEChERS) method, coupled with liquid chromatography-tandem mass spectrometry using electrospray ionization in positive mode, was used for the simultaneous analysis of 236 pesticides in three dried powdered botanical dietary supplements (ginseng, saw palmetto, and gingko biloba). The procedure involved extraction of the dried powdered botanical samples with salt-out acetonitrile/water extraction using anhydrous magnesium sulfate and sodium chloride, followed by an automated dSPE cleanup using a mixture of octadodecyl- (C18) and primary-secondary amine (PSA)-linked silica sorbents and anhydrous MgSO4 and online LC-MS/MS analysis. Dynamic multiple-reaction monitoring (DMRM) based on the collection of two precursor-to-product ion transitions with their retention time windows was used for all of the targeted pesticides and the internal standard. Matrix-matched calibration standards were used for quantitation, and standard calibration curves showed linearity (r(2) > 0.99) across a concentration range of 0.2-400 ng/mL for the majority of the 236 pesticides evaluated in the three botanical matrices. Mean recoveries (average %RSD, n = 4) were 91 (6), 93 (4), 96 (3), and 99 (3)% for ginseng, 101 (9), 98 (6), 99 (4), and 102 (3)% for gingko biloba, and 100 (9), 98 (6), 96 (4), and 96 (3)% for saw palmetto at fortification concentrations of 25, 100, 250, and 500 μg/kg, respectively. The geometric mean matrix-dependent instrument detection limits were 0.17, 0.09, and 0.14 μg/kg on the basis of the studies of 236 pesticides tested in ginseng roots, gingko biloba leaves, and saw palmetto berries, respectively. The method was used to analyze incurred ginseng samples that contained thermally labile pesticides with a concentration range of 2-200 μg/kg, indicating different classes of pesticides are being applied to these botanicals other than the traditional pesticides that are commonly used and analyzed by gas chromatography techniques. The method demonstrates the use of an automated cleanup procedure and the LC-MS/MS detection of multiple pesticide residues in dried, powdered botanical dietary supplements.

Carryover of Perfluorooctanoic Acid (PFOA) and Perfluorooctane Sulfonate (PFOS) from Soil to Plant and Distribution to the Different Plant Compartments Studied in Cultures of Carrots (Daucus carota ssp. Sativus), Potatoes (Solanum tuberosum), and Cucumbers (Cucumis Sativus)

A vegetation study was carried out to investigate the carryover of Perfluorooctanoic Acid (PFOA) and Perfluorooctane Sulfonate (PFOS) from soil mixed with contaminated sewage sludge to potato, carrot, and cucumber plants. Analysis was done by liquid-extraction using acetonitrile with dispersive SPE cleanup and subsequent HPLC-MS/MS. In order to assess the transfer potential from soil, transfer factors (TF) were calculated for the different plant compartments: TF = [PFC](plant (wet substance))/[PFC](soil (dry weight)). The highest TF were found for the vegetative plant compartments with average values for PFOS below those for PFOA: cucumber, 0.17 (PFOS), 0.88 (PFOA); potato, 0.36 (PFOS), 0.40 (PFOA); carrot, 0.38 (PFOS), 0.53 (PFOA). Transfer of PFOA and PFOS into potato peelings (average values of TF: PFOA 0.03, PFOS 0.04) exceeded the carryover to the peeled tubers (PFOA 0.01, PFOS < 0.01). In carrots, this difference did not occur (average values of TF: PFOA 0.04, PFOS 0.04). Transfer of PFOS into the unpeeled cucumbers was low and comparable to that of peeled potatoes (TF < 0.01). For PFOA, it was higher (TF: 0.03).

Procedures for Antibiotic Residues in Bovine Muscle Tissues

Acetonitrile extraction followed by primary-secondary amine dispersive SPE cleanup QuEChERS (quick, easy, cheap, effective, rugged, and safe), was compared to pressurized liquid extraction (PLE) using water at 70 degrees C for 10 min at 1500 psi for the determination of 16 veterinary drugs in bovine muscle tissues by LC/MS/MS. PLE was significantly more effective for the extraction of veterinary drugs (ranging from 69 to 103% with RSD < or = 18%) than QuEChERS (ranging from 19 to 89% with RSD < or = 19%). Linearity of the calibration curves was obtained over the range considered from 10 microg/kg or LOQ to 1000, microg/kg) with r2 > or = 0.99 for all the analytes by both methods. Although an internal standard was used, matrix effects were corrected using matrix- matched standards. LODs were from 5 to 30 microg/kg for PLE and from 10 to 100 microg/kg for QuEChERS. To establish and assess the most efficient conditions for each extraction method, statistical parametric and nonparametric tests were used. PLE with water almost eliminates the use or generation of hazardous wastes. The two methods were applied successfully in a routine analysis during surveys in 2008.

Optimization of Detection Conditions and Single-Laboratory Validation of a Multiresidue Method for the Determination of 135 Pesticides and 25 Organic Pollutants in Grapes and Wine by Gas Chromatography Time-of-Flight Mass Spectrometry

This paper describes single-laboratory validation of a multiresidue method for the determination of 135 pesticides, 12 dioxin-like polychlorinated biphenyls, 12 polyaromatic hydrocarbons, and bisphenol A in grapes and wine by GC/time-of-flight MS in a total run time of 48 min. The method is based on extraction with ethyl acetate in a sample-to-solvent ratio of 1:1, followed by selective dispersive SPE cleanup for grapes and wine. The GC/MS conditions were optimized for the chromatographic separation and to achieve highest S/N for all 160 target analytes, including the temperature-sensitive compounds, like captan and captafol, that are prone to degradation during analysis. An average recovery of 80-120% with RSD < 10% could be attained for all analytes except 17, for which the average recoveries were 70-80%. LOQ ranged within 10-50 ng/g, with < 25% expanded uncertainties, for 155 compounds in grapes and 151 in wine. In the incurred grape and wine samples, the residues of buprofezin, chlorpyriphos, metalaxyl, and myclobutanil were detected, with an RSD of < 5% (n = 6); the results were statistically similar to previously reported validated methods.

Analysis of pesticide residues in sugarcane juice using QuEChERS sample preparation and gas chromatography with electron capture detection

QuEChERS sample preparation was used for the determination of 7 pesticides residues in 80 samples of sugarcane juice collected from two Brazilian cities, in two different periods. The method involved extraction with acetonitrile, liquid–liquid partition with addition of MgSO4 and NaCl followed by dispersive SPE cleanup with PSA sorbent and the analyses were carried out with a GC–ECD equipment. The method was validated using sugarcane juice spiked at 0.025, 0.10 and 0.20mg/L and the average recovery by the method varied from 62.9% to 107.5% with RSDs<18%. The method showed good linearity and the LODs for the pesticides studied ranged from 0.003 to 0.04mg/L. No pesticide residue was detected (>LOD) amongst the 80 samples analysed.