Non-targeted analysis of per- and polyfluorinated substances in consumer food packaging.

This study sought to develop a non-targeted workflow using high-resolution mass spectrometry (HRMS) to investigate previously unknown PFAS in consumer food packaging samples. Samples composed of various materials for different food types were subjected to methanolic extraction, controlled migration with food simulants and total oxidizable precursor (TOP) assay. The developed HRMS workflow utilized many signatures unique to PFAS compounds: negative mass defect, diagnostic breakdown structures, as well as retention time prediction. Potential PFAS features were identified in all packaging studied, regardless of food and material types. Five tentatively identified compounds were confirmed with analytical standards: 6:2 fluorotelomer phosphate diester (6:2 diPAP) and one of its intermediate breakdown products 2H-perfluoro-2-octenoic acid (6:2 FTUCA), perfluoropentadecanoic acid (PFPeDA), perfluorohexadecanoic acid (PFHxDA) and perfluorooctadecanoic acid (PFOcDA). Longer perfluorocarboxylic acids including C17 and C19 to C24 were also found present within a foil sample. Concentrations of 6:2 FTUCA ranged from 0.78 to 127 ng g-1 in methanolic extracts and up to 6 ng g-1 in food simulant after 240 h migration test. These results demonstrate the prevalence of both emerging and legacy PFAS in food packaging samples and highlight the usefulness of non-targeted tools to identify PFAS not included in targeted methods.

Evaluating contamination of seafood purchased from U.S. retail stores by persistent environmental pollutants, pesticides and veterinary drugs.

Studies have reported health risks associated with seafood contamination, but few data exist on levels in commercially available seafood in the US. To better understand, the magnitude of foodborne exposure and identify vulnerable populations in the US, we measured concentrations of veterinary drugs, persistent organic pollutants (POPs) (polycyclic aromatic hydrocarbons [PAHs], polybrominated diphenyl ethers [PBDEs] and polychlorinated biphenyls [PCBs]), and legacy and current-use pesticides in 46 seafood samples purchased from retail outlets. Measured levels were used to estimate risk based on available maximum residue limits (MRLs) and toxic equivalence (TEQ) factors for analytes. Only seventeen of the 445 analytes were detected, at low substance frequencies. However, half of the samples tested positive for one or more analyte, with total concentrations ranging from below the limit of detection (LOD) to as high as 156 µg/kg wet weight. Based on the risk assessment for individual pesticides and veterinary drugs, the hazard quotients (HQ) were all <1, indicating no risk. However, for the sum of PCB126 and PCB167, two dioxin-like PCBs detected in our samples, the TEQ was nearly two orders of magnitude higher than the WHO limits in one catfish sample. Moreover, vulnerable groups with higher rates of consumption of specific fish types may face higher risks.

Per- and polyfluoroalkyl substances (PFAS) measured in seafood from a cross-section of retail stores in the United States.

Seafood is a dominant source of human exposure to per- and polyfluoroalkyl substances (PFAS). Existing studies on foodborne PFAS exposure have focused on only a subset of these compounds. Here, we conducted a pilot study to screen 33 PFAS in 46 seafood samples from a cross-section of national and local stores in the US. Low levels of 8 PFAS were measured in 74% of the samples, predominated by PFHxS (59%). Total PFAS ranged between 0.12 and 20 ng/g; highest levels were measured in Estonia-sourced smelt. The highest median levels were of PFOA (0.84 ng/g) with elevated concentrations found in Chinese clams (2.4 ng/g), which exceeds the EU established maximum limits (MLs). Measured levels of PFHxS, PFOA, and PFNA also exceeded MLs in 24%, 7%, and 5% of the samples, respectively. For average consumption levels, exposures were below the EU established tolerable weekly intakes (TWIs). However, for more frequent consumption of flounder, catfish, and cod, exposures exceeded regulations, which warrants identifying vulnerable high seafood consuming populations. Accidental PFBS cross contamination from sample storage bags resulted in 100% detection in samples, highlighting the problem with post-purchase food handling practices such as storage and cooking that could also have a substantial impact on human exposure, potentially in larger amounts than the (sea)food itself.

Assessing per- and polyfluoroalkyl substances in globally sourced food packaging.

The purpose of this study was to investigate the presence and levels of per- and polyfluoroalkyl substances (PFAS) in food packaging originating from different geographic locations. Food packaging samples were extracted and analyzed by targeted analysis with liquid chromatography-mass spectrometry (LC-MS/MS) before and after a total oxidizable precursor (TOP) assay. Additionally, full-scan high resolution MS (HRMS) was used to screen for PFAS not included in the targeted list. Of the 88 food packaging samples, 84% had detectable levels of at least one PFAS prior to oxidation with a TOP assay, with 6:2 fluorotelomer phosphate diester (6:2 diPAP) found most frequently and at the highest levels (224 ng/g). Other frequently detected substances (in 15-17% of samples) were PFHxS, PFHpA and PFDA. Shorter chain perfluorinated carboxylic acids PFHpA (C7), PFPeA (C5) and PFHxS (C6) were present at levels up to 51.3, 24.1 and 18.2 ng/g, respectively. Average ∑PFAS levels were 28.3 ng/g and 381.9 ng/g before and after oxidation with the TOP assay. The 25 samples with highest frequency of detection and amounts of measured PFAS were selected for migration experiments with food simulants to better understand potential dietary exposure. PFHxS, PFHpA, PFHxA and 6:2 diPAP were measured in the food simulants of five samples at concentrations ranging from 0.04 to 12.2 ng/g and at increasing concentrations over the 10-day migration period. To estimate potential exposure to PFAS that had migrated from food packaging samples, weekly intake was calculated and ranged from 0.0006 ng/kg body weight/week for PFHxA exposure in tomato packaging to 1.1200 ng/kg body weight/week for PFHxS exposure in cake paper. These values were below the established EFSA maximum tolerable weekly intake (TWI) of 4.4 ng/kg body weight/week for the sum of PFOA, PFNA, PFHxS and PFOS.

Investigating migration potential of a new rechargeable antimicrobial coating for food processing equipment.

Antimicrobial coatings are designed to inhibit the growth of pathogens and have been used to reduce foodborne illness bacteria on food processing equipment. Novel N-halamine based antimicrobial coatings are highly advantageous due to their unique properties and low cost, and are being investigated for applications in food safety, health care, water and air disinfection, etc. In this study, we evaluated the chemical safety of a novel N-halamine antimicrobial polymer coating (Halofilm) for use on food processing equipment. Migration tests were performed on stainless steel tiles prepared with four different treatment groups: negative control, positive control, Halofilm coating without chlorination, and Halofilm coating with chlorination. An LC-MS/MS method was developed and validated for four formulation components: polyethylenimine (PEI), Trizma® base, hydantoin acrylamide (HA) and dopamine methacrylamide (DMA), followed by stability and recovery tests. Migration tests were conducted at 40 °C with three food simulants (10, 50 and 95% ethanol/water) to mimic various food properties, and aliquots of migration extracts were analyzed at 2, 8, 72, 240 and 720 h. In general, measured concentration levels were consistent among simulant types for the four tested chemicals. Chlorinated tiles had non-detects for three analytes (PEI, HA and DMA), and less than 0.05 mg/kg of HA migration over 30 days. A chlorination step could possibly change the measured mass (m/z) hence leading to non-detects in targeted LC-MS/MS. In non-chlorinated tiles, all four compounds were detected during the migration test. This suggests that addition of the chlorination step may have a stabilizing effect on the polymer. Additionally, full scan high resolution mass spectrometry (HRMS) analysis was employed to screen for migration of other extractable and leachable (E&L) chemicals, which led to the identification of eight common E&L chemicals. To our knowledge, this is the first report evaluating chemical migration from an N-halamine antimicrobial polymer coating product.

Comparison and validation of the QuEChERSER mega-method for determination of per- and polyfluoroalkyl substances in foods by liquid chromatography with high-resolution and triple quadrupole mass spectrometry.

Instances of food contamination with per- and polyfluoroalkyl substances (PFAS) continue to occur globally, but sample preparation and analytical methods are quite limited and often monitor for a small percentage of known PFAS. This study aimed to evaluate, validate, and compare performance of two instruments with the recently developed "quick, easy, cheap, effective, rugged, safe, efficient, and robust" (QuEChERSER) sample preparation mega-method - a method developed to monitor chemicals over a broad range of physicochemical properties. Initial evaluation of the QuEChERSER mega-method for determination of PFAS in food demonstrated recoveries, matrix interferences, and co-extractive removal comparable to (or better than) US Food and Drug Administration (FDA) and USDA Food Safety and Inspection Service (FSIS) methods. Subsequent validation of QuEChERSER in beef, catfish, chicken, pork, liquid eggs, and powdered eggs on a high-resolution mass spectrometer achieved acceptable recoveries (70-120%) and precision (RSDs ≤20%) for all 33 target analytes at the 1 and 5 ng g-1 levels and 67-88% of analytes at the 0.1 ng g-1 level, depending on the matrix. Additional validation was performed by tandem mass spectrometry on a triple quadrupole instrument. This approach provided no non-detects and better recoveries at the 0.1 ng g-1 level than the HRMS method but exhibited more variability at 1 and 5 ng g-1 spiking levels. Analysis of NIST SRMs 1946 and 1947 gave accuracies of 70-117%. These results demonstrate the capability of combining PFAS analysis with a mega-method previously validated for 350 analytes, while collecting non-target data for future retrospective analysis of emerging alternatives with a high-resolution mass spectrometry method.

Analysis of pesticides, veterinary drugs, and environmental contaminants in goat and lamb by the QuEChERSER mega-method.

Analysis of chemical residues in foods is a big challenge for developing countries due to lack of financial and professional resources needed to meet international quality standards for trade. However, the implementation of simple multiclass, multi-residue methods in monitoring programs can provide significant benefits to save cost, time, and labor. The aim of this project was to investigate the "quick, easy, cheap, effective, rugged, safe, efficient, and robust" (QuEChERSER) mega-method for the fatty muscle matrices of goat and lamb. To achieve wide analytical scope covering pesticides, environmental contaminants, and veterinary drugs, extracts were analyzed by both ultrahigh-performance liquid and low-pressure gas chromatography (UHPLC and LPGC) coupled with tandem mass spectrometry (MS/MS). The QuEChERSER mega-method was validated in ovine (goat) and caprine (lamb) muscles at four different spiking levels with 10 replicates per level for a total of 330 analytes and metabolites, consisting of 225 pesticides, 89 veterinary drugs, and 16 polychlorinated biphenyls (PCBs). In the case of LPGC-MS/MS (preceded by automated "instrument-top sample preparation"), 92% and 82% of the analytes met the data quality objectives of 70-120% recovery and <20% RSD for goat and lamb, respectively. For UHPLC-MS/MS, 95% and 92% of the analytes met the acceptable validation criteria in goat and lamb, respectively. Thus, the QuEChERSER mega-method has been demonstrated to be a useful streamlined approach to more efficiently replace multiple methods currently used to cover the same analytical scope.

Non-targeted analysis with liquid chromatography - high resolution mass spectrometry for the identification of food packaging migrants.

Potential contamination of food with chemicals migrating from food packaging is an important, yet under-investigated area of food safety. In this study, we examined chemicals migrating from common paper-based food packaging: pizza boxes and pizza box liners, butcher paper and liquid egg containers. Migration tests were conducted with a food simulant for 10 days, and migrated chemicals were identified with liquid chromatography (LC) - high resolution mass spectrometry (HRMS) with mass error < 3 ppm. HRMS identification was based on spectra and/or structure matching against commercial databases (MzCloud, ChemSpider, and Extractable and Leachable high resolution accurate mass (HRAM) database). Following HRMS identification, orthogonal LC retention information was utilized to further refine the data and reduce false positive findings. A model for calculating retention times (tR) based on octanol-water partition coefficient (log Kow) values was evaluated and applied for HRMS data refining. Using this approach, 153 migrated chemicals were identified, of which five were further confirmed with reference analytical standards. Additionally, amounts of bisphenol A and bisphenol S, the chemicals of toxicological concerns, were measured at the levels below the established regulatory limits for migration, indicating no/low risk to consumer's health. This study demonstrated the utility of LC-HRMS for confident identification of food packaging migrants.

Assessing Chemical Migration from Plastic Food Packaging into Food Simulant by Gas and Liquid Chromatography with High-Resolution Mass Spectrometry.

Some components of plastic food packaging can migrate into food, and whereas migration studies of known components are required and relatively straightforward, identification of nonintentionally added substances (NIAS; unknowns) is challenging yet imperative to better characterizing food safety. To this aim, migration was investigated across 24 unique plastic food packaging products including plastic wrap, storage bags, vacuum bags, and meat trays. Gas and liquid chromatography separation systems coupled with Orbitrap mass analyzers were used for comprehensive nontargeted screening of migrants. Tentative identifications of features were assigned by searching commercial databases (e.g., NIST, MZCloud, ChemSpider, Extractables and Leachables) and filtering results based on mass accuracy, retention time indices, and mass spectral patterns. Several migrants showed elevated levels in specific food packaging types, particularly meat trays and plastic wrap, and varying degrees of migration over the 10 days. Eleven putative migrants are listed as substances of potential concern or priority hazardous substances. Additionally, migration amounts of an Irgafos 168 degradation product determined by semiquantitation exceeded proposed theoretical maximum migration values.

Validation of the QuEChERSER mega-method for the analysis of pesticides, veterinary drugs, and environmental contaminants in tilapia (Oreochromis Niloticus).

Diverse food safety programmes around the world are designed to help ensure production of safe food. To meet this need, the development and implementation of more efficient and effective analytical methods to monitor residues (pesticides and veterinary drugs) and contaminants in food is important. In this study, we report the validation results for a simple high-throughput mega-method for residual analysis of 213 pesticides and veterinary drugs, including 15 metabolites, plus 12 environmental contaminants (polychlorinated biphenyls) in tilapia muscle for implementation in routine laboratory analyses. The generic sample preparation method and analytical approach are known as QuEChERSER (more than QuEChERS). A small portion of the initial extract (204 µL) is taken for analysis by ultrahigh-performance liquid chromatography (UHPLC) tandem mass spectrometry (MS/MS) covering 145 analytes, and the remaining extract undergoes a salting out step followed by an automated robotic instrument top sample preparation (ITSP) cleanup, also known as micro-solid-phase extraction (µSPE), plus fast low-pressure gas chromatography LPGC-MS/MS for 134 analytes (66 pesticides are targeted in both UHPLC-MS/MS and LPGC-MS/MS). The mega-method was validated in spiked tilapia samples at 5, 10, 15, and 20 ng/g with 10 replicates per level over two days (n = 80 overall), and 70-140% recoveries with RSDs ≤20% were achieved for 92% of the analytes in LC and 82% in GC. No significant matrix effects were observed for the analytes in LPGC-MS/MS, and only 5% of the analytes exceeded ±20% matrix effect in UHPLC-MS/MS. Analysis of standard reference materials (NIST SRMs 1946 and 1947) for contaminants in freeze-dried fish showed acceptable results, further demonstrating that the QuEChERSER mega-method can be implemented to expand analytical scope and increase laboratory efficiency compared to the QuEChERS method.

Screening of chemicals migrating from plastic food contact materials for oven and microwave applications by liquid and gas chromatography - Orbitrap mass spectrometry.

Contamination of food with chemicals migrating from food contact materials (FCMs) is an important area of food safety. This study was aimed to investigate migration of chemicals from plastic FCMs used for microwave and conventional oven heating. Migration tests were conducted for samples of microwave trays, microwave oven bags, and oven bags. GC- and LC-Orbitrap mass spectrometry (MS) was used for non-targeted screening and identification of chemicals with mass error <5 ppm. A non-targeted identification approach was validated with isotopically labeled chemicals to establish acceptable criteria for identification of migrated compounds. A total of 74 migrated compounds were tentatively identified: 24 chemicals by GC-Orbitrap MS with electron ionization (EI), plus 35 and 19 by LC-Orbitrap MS electrospray ionization (ESI) with positive and negative polarities, respectively. Four migrated chemicals were identified by more than one instrumental analysis. Both intentionally added substances (IAS), i.e. additives used in the production of polymeric materials and plastics, and non-intentionally added substances (NIAS), i.e. derivatives and degradation/oxidation products of IAS, were identified among the migrated chemicals. The levels of 25 migrated chemicals were significantly different (p < 0.05) between microwave treatments and conventional oven treatments, where 20 migrants had higher levels for microwave compared with 5 for conventional oven treatments. For several identified chemicals, no previous reports on their migration from FCMs were found.

Non-targeted screening of chemicals migrating from paper-based food packaging by GC-Orbitrap mass spectrometry.

The aim of this study was to identify chemicals migrating from paper-based food packaging materials by GC-Orbitrap mass spectrometry (MS). Migration tests were conducted with samples of butcher paper, liquid egg containers, pizza boxes and pizza box liners. A new approach for an automated data processing of GC-Orbitrap EI data was developed and evaluated with Compound Discoverer™ software. A total of 35 migrated chemicals were putatively identified. They represented natural compounds, fatty acid ethyl esters, FDA substances added to food and Indirect Additives in Food Contact Substances, common phthalates, alkanes, siloxanes, ketones, and chemicals used as additives in packaging. Additionally, some unexpected compounds suspected to be degradation products of additives were identified. Levels of 23 identified chemicals significantly (p < 0.05) changed over the 10-day migration test. For two identified migrants, dibutyl phthalate and benzophenone, calculated specific migration limits (SMLs) were below the established regulatory levels. The information on the identity of chemicals migrating from food packaging materials is important to inform regulatory agencies and to develop risk management strategies.

High-Throughput Mega-Method for the Analysis of Pesticides, Veterinary Drugs, and Environmental Contaminants by Ultra-High-Performance Liquid Chromatography-Tandem Mass Spectrometry and Robotic Mini-Solid-Phase Extraction Cleanup + Low-Pressure Gas Chromatography-Tandem Mass Spectrometry, Part 1: Beef.

In this work, a new mega-method of sample preparation called "QuEChERSER" (more than QuEChERS) is being presented for the first time. Fast, efficient, and cost-effective analysis of chemical contaminants in meat is useful for international trade, domestic monitoring, risk assessment, and other purposes. The goal of this study was to develop and validate a simple high-throughput mega-method for residual analysis of 161 pesticides, 63 veterinary drugs, 24 metabolites, and 14 legacy environmental contaminants (polychlorinated biphenyls) in bovine muscle for implementation in routine laboratory analyses. Sample preparation of 2 g test portions entailed QuEChERS-based extraction with 10 mL of 4:1 (v/v) acetonitrile/water, and then 204 µL was taken, diluted, and ultracentrifuged prior to analysis of veterinary drugs and pesticides by ultra-high-performance liquid chromatography-tandem mass spectrometry. The remaining extract was salted out with 4:1 (w/w) anhydrous MgSO4/NaCl, and 1 mL was transferred to an autosampler vial for automated mini-cartridge solid-phase extraction (Instrument Top Sample Preparation) cleanup with immediate injection using fast low-pressure gas chromatography-tandem mass spectrometry analysis. The automated cleanup and both instruments were all operated in parallel in 13-15 min cycle times per sample. Method validation according to United States Department of Agriculture requirements demonstrated that 221 (85%) of the 259 analytes gave average recovery between 70 and 120% and interday relative standard deviation of ≤25%. Analysis of a certified reference material for veterinary drugs in freeze-dried bovine muscle was also very accurate, further demonstrating that the QuEChERSER mega-method can be implemented to save time, labor, and resources compared to current practices to use multiple methods to cover the same analytical scope.

High-Throughput Mega-Method for the Analysis of Pesticides, Veterinary Drugs, and Environmental Contaminants by Ultra-High-Performance Liquid Chromatography-Tandem Mass Spectrometry and Robotic Mini-Solid-Phase Extraction Cleanup + Low-Pressure Gas Chromatography-Tandem Mass Spectrometry, Part 2: Catfish.

The goal of this study was to develop and validate a new method for simultaneous determination of 106 veterinary drugs and 227 pesticides and their metabolites plus 16 polychlorinated biphenyls (PCBs) at and below their regulatory levels established for catfish muscle in the European Union and U.S.A. To do this, two different QuEChERS-based methods for veterinary drugs and pesticides and PCBs were modified and merged into a single mega-method dubbed "QuEChERSER" (more than QuEChERS), which is presented here for the first time. The mega-method was validated in catfish at four different spiking levels with 10 replicates per level. Sample extraction of 2 g test portions was made with 10 mL of 4:1 (v/v) acetonitrile/water, and then an aliquot was taken for ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) analysis of 106 veterinary drugs and 125 pesticides, including metabolites. The remaining extract after salting out was subjected to automated mini-solid-phase extraction cleanup (Instrument Top Sample Preparation) for immediate injection in low-pressure gas chromatography-tandem mass spectrometry (LPGC-MS/MS). The cleanup was conducted in parallel with the 10 min LPGC-MS/MS analysis for 167 PCBs, pesticides, and metabolites, which was conducted in parallel with the 10 min UHPLC-MS/MS analysis for 231 analytes to increase sample throughput (49 analytes were included in both techniques). In MS/MS, three ion transitions were monitored for nearly all targeted analytes to provide unambiguous identification as well as quantification. Satisfactory recoveries (70-120%) and relative standard deviations of ≤20% were achieved for 98 (92%) of the veterinary drugs and their metabolites and for 222 (91%) of pesticides, metabolites, and PCBs, demonstrating that the developed method is applicable for the analysis of these contaminants in fish as part of regulatory monitoring programs and other purposes.

Spatial and seasonal occurrence of semi-volatile organic compounds (SVOCs) in fish influenced by snowmelt and municipal effluent discharge.

In the present study we examined spatial and seasonal trends in the levels of a wide suite of semi-volatile organic compounds (SVOCs) in brown trout (Salmo trutta) and mottled sculpin (Cottus bairdii) in East Canyon Creek, Utah, USA, an effluent-dominated stream during summer months. Fish samples were collected from four sampling sites, including one reference site upstream, and three sites at incremental distances downstream of the effluent discharge over multiple seasons. The samples were analyzed for 218 lipophilic contaminants, including pesticides and their metabolites, polychlorinated biphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs), and polybrominated diphenyl ethers (PBDEs) and other flame retardants. Some PAHs, pesticides and their metabolites, PCBs, PBDEs and other flame retardants were measured in mottled sculpin (11 analytes) and brown trout (17 analytes). Hexachlorobenzene (HCB), p,p'-DDE, BDE-47 and triphenyl phosphate (TPHP) were the most frequently detected contaminants in mottled sculpin and brown trout, while BDE-47 and p,p'-DDE were measured at the highest concentrations, reaching up to 73 and 19 ng/g wet weight, respectively. Our results indicated that snowmelt did not alter accumulation of the examined lipophilic contaminants, and no consistent seasonal differences were observed in their accumulation. A spatial pattern was observed for PBDE congeners, where lowest levels were measured in fish tissues from a reference site, and highest concentrations were measured in fish collected downstream of the effluent discharge, indicating that municipal effluent discharge contributes to the elevated PBDE levels in fish residing in this effluent-dominated stream. We further calculated screening level consumption risks following United States Environmental Protection Agency (EPA) methods, and identified the importance of considering discharge gradients in effluent-dominated systems during bioaccumulation assessments.

Semi-automated high-throughput method for residual analysis of 302 pesticides and environmental contaminants in catfish by fast low-pressure GC-MS/MS and UHPLC-MS/MS.

A fast, fit-for-purpose, high-throughput method was developed and validated for the analysis of 302 targeted contaminants in catfish muscle. Targeted contaminants included pesticides and metabolites with US regulatory levels, plus other lipophilic pesticides and environmental contaminants: PAHs, PCBs, PBDEs and other flame retardants. The sample preparation was based on QuEChERS extraction. The extract was split and analyzed by UHPLC-MS/MS for 128 analytes after filtering and by low pressure (LP) GC-MS/MS for 219 analytes after an automated robotic micro-SPE cleanup. The method was validated at four spiking levels (5, 10, 20 and 40 ng/g) with 10 replicates per level, and satisfactory recoveries (70-120%) and RSDs (≤20%) were achieved for 80% of the analytes. The method was further validated with NIST SRMs 1946 and 1947 and applied for the analysis of 22 market fish samples. Six pesticides and one PAH were found in the market fish samples, all below US regulatory levels.

Analysis and Occurrence of Organophosphate Esters in Meats and Fish Consumed in the United States.

Organophosphate esters (OPEs) are chemicals extensively used as plasticizers and flame retardants in commercial and consumer products. In this study, we developed and validated a method for the analysis of 13 common OPEs in meat (chicken, pork, and beef) and fish (catfish and salmon) to study their occurrence in those foods in the United States. The method was based on QuEChERS extraction with acetonitrile and automated robotic cleanup of the extracts, followed by low pressure gas chromatography-tandem mass spectrometry (GC-MS/MS) analysis for 8 of the OPEs and ultrahigh-pressure liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) analysis for 13 OPEs, including 8 overlapping OPEs. The developed method was validated in the muscle tissues at four spiking levels (5, 10, 20, and 40 ng/g). The background levels in the laboratory environment and materials presented a challenge for accurate quantification at low ng/g levels. UHPLC-Q-Orbitrap MS analysis was utilized to pinpoint the source of their contamination. OPEs were found in the water used in the liquid chromatography (LC) mobile phase, and flow injection analysis with organic mobile phase was suggested as an alternative to avoid OPEs contamination in LC-MS/MS analysis. The validated method was applied to the analysis of 68 real-world meat and fish samples from the U.S. markets by three instrumental methods. Tris(2-chloro-isopropyl) phosphate (TCPP), tri-n-butyl phosphate (TnBP), and triphenyl phosphate (TPP) were found in meat, and TCPP and TPP were measured in fish samples. The sum of median OPE concentrations (averaged for the three instrumental methods) measured in the meat and fish samples were 6.2 and 8.7 ng/g wet weight, respectively. No regulations on the maximum residue levels of OPEs permitted in food were found for the U.S. or other countries.

Characterization of MS/MS Product Ions for the Differentiation of Structurally Isomeric Pesticides by High-Resolution Mass Spectrometry.

Structural isomeric pesticides are used in agriculture and may be challenging to differentiate for accurate identification in pesticide monitoring programs. Due to structural similarity, isomeric pesticides are difficult to separate chromatographically, and thus, their accurate identification may rely solely on mass spectrometric analysis (MS). In this study, we challenged the ability of high-resolution quadrupole-orbitrap (Q-Orbitrap) mass spectrometry to produce and evaluate the tandem mass spectrometry (MS/MS) product ions for the selected five pairs of isomeric pesticides from different classes: Pebulate and vernolate, methiocarb and ethiofencarb, uniconazole and cyproconazole, sebuthylazine and terbuthylazine, and orbencarb and thiobencarb. The use of Q-Orbitrap instrument with a mass error <3 ppm allowed proposed elucidation of the product ion structures with consideration of the ion formulae, data interpretation, and literature searches. Product ions unique to pebulate, vernolate, methiocarb, ethiofencarb, and uniconazole were observed. Elucidation of the observed MS/MS product ion structures was conducted, and the fragmentation pathways were proposed. This information is valuable to increase selectivity in MS/MS analysis and differentiate isomeric pesticides, and thereby reduce the rates of false positives in pesticide monitoring programs.

High-throughput analytical method for 265 pesticides and environmental contaminants in meats and poultry by fast low pressure gas chromatography and ultrahigh-performance liquid chromatography tandem mass spectrometry.

A high throughput method for simultaneous determination of 200 pesticides and 65 environmental contaminants: polychlorinated biphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs), polybrominated diphenyl ethers (PBDEs) and other flame retardants, was developed and validated in cattle, swine and poultry muscle tissues. The method was based on sample preparation with QuEChERS extraction followed by an automated robotic sample cleanup with solid phase extraction (SPE) mini-columns using Instrument Top Sample Preparation (ITSP). The automated robotic ITSP was integrated with instrumental analysis (low pressure (LP)GC-MS/MS) to conduct sample cleanup and analysis in parallel. Another aliquot of the initial QuEChERS extract was subjected to UHPLC-MS/MS analysis, and 53 overlapping pesticides were analyzed by both LPGC- and UHPLC-MS/MS (10 min each). Fit-for-purpose LOQs<5 ng/g were achieved for ≥90% of the analytes. The method was validated in cattle, swine and chicken muscles at three spiking levels at and below the US regulatory levels of concern, and satisfactory recoveries (70-120%) and RSDs ≤ 20% were demonstrated for 219 analytes. Matrix effects were within ±20% for 78-97% of LC- and GC-amenable analytes. The validated method was successfully applied for the analysis of real-world incurred meat and poultry samples, further demonstrating its applicability to real-world samples, and potential for implementation in routine monitoring of contaminants.