Pesticide residues in fresh and processed edible mushrooms from Czech markets.

The occurrence of pesticide residues, their metabolites and degradation products in 49 edible mushrooms (fresh, dried, canned and frozen) was investigated. Using QuEChERS extraction, followed by liquid and gas chromatography coupled with tandem mass spectrometry, the samples collected from Czech markets were tested for 427 analytes. A total of 21 pesticide residues, their metabolites and pesticide synergists were found in quantifiable concentrations. The most frequently detected pesticide residues included prochloraz and its metabolites, metrafenone and carbendazim. Two mushroom samples were found to be in violation of the maximum residue level. In addition, the fate of 13 pesticides during the processing of fresh mushrooms was investigated. The processing factors calculated from the data ranged from 6.0 to 12.3 for drying and from 0.3 to 1.3 for deep-freezing. The results showed that most of the pesticide residues tested were stable under the conditions of mushroom processing.

POPs and PAHs in fish oil-based food supplements at the Czech market.

The objectives of this study were to assess concentrations of three groups of persistent organic pollutants (POPs) and polycyclic aromatic hydrocarbons (PAHs) in 44 fish oil-based food supplements, to estimate their daily intake by consumers and, to evaluate the compliance of the oil samples with the oil origin declarations (cod liver oil or fish oil). The concentrations of ∑PCBs (7 congeners), OCPs (19 compounds, represented mainly by ∑DDTs), ∑PBDEs (10 congeners), and ∑PAHs (16 compounds) found in samples ranged between 0.15-55.7 µg kg-1, 0.93-72.8 µg kg-1, 0.28-27.5 µg kg-1, and 0.32-51.9 µg kg-1, respectively. Besides, the authenticity of the oils was assessed based on the fingerprints obtained by DART-HRMS, an ambient mass spectrometry technique. Four samples declared as fish oil were probably prepared from cod liver oil, which is much cheaper. Furthermore, these samples contained elevated concentrations of halogenated POPs when compared to supplements produced from fish oil.

Detection of microscopic filamentous fungal biofilms - Choosing the suitable methodology.

Microscopic filamentous fungi are ubiquitous microorganisms that adapt very easily to a variety of environmental conditions. Due to this adaptability, they can colonize a number of various surfaces where they are able to start forming biofilms. Life in the form of biofilms provides them with many benefits (increased resistance to desiccation, UV radiation, antimicrobial compounds, and host immune response). The aim of this study is to find a reliable and reproducible methodology to determine biofilm growth of selected microscopic filamentous fungi strains. Several methods (crystal violet staining, MTT assay, XTT assay, resazurin assay) for the determination of total biofilm biomass and its metabolic activity were tested on four fungi - Alternaria alternata, Aspergillus niger, Fusarium culmorum and Fusarium graminearum, and their biofilm was also imaged by spinning disc confocal microscopy using fluorescent dyes. A reproducible biofilm quantification method is essential for the subsequent testing of the biofilm growth suppression using antifungal agents or physical methods. Crystal violet staining was found to be a suitable method for the determination of total biofilm biomass of selected strains, and the MTT assay for the determination of metabolic activity of the biofilms. Calcofluor white and Nile red fluorescent stains successfully dyed the hyphae of microscopic fungi.

Critical Assessment of Clean-Up Techniques Employed in Simultaneous Analysis of Persistent Organic Pollutants and Polycyclic Aromatic Hydrocarbons in Fatty Samples.

Interference of residual lipids is a very common problem in ultratrace analysis of contaminants in fatty matrices. Therefore, quick and effective clean-up techniques applicable to multiple groups of analytes are much needed. Cartridge and dispersive solid-phase extraction (SPE and dSPE) are often used for this purpose. In this context, we evaluated the lipid clean-up efficiency and performance of four commonly used sorbents-silica, C18, Z-Sep, and EMR-lipid-for the determination of organic pollutants in fatty fish samples (10%) extracted using ethyl acetate or the QuEChERS method. Namely, 17 polychlorinated biphenyls (PCBs), 22 organochlorine pesticides (OCPs), 13 brominated flame retardants (BFRs), 19 per- and polyfluoroalkyl substances (PFAS), and 16 polycyclic aromatic hydrocarbons (PAHs) were determined in this study. The clean-up efficiency was evaluated by direct analysis in real time coupled with time-of-flight mass spectrometry (DART-HRMS). The triacylglycerols (TAGs) content in the purified extracts were significantly reduced. The EMR-lipid sorbent was the most efficient of the dSPE sorbents used for the determination of POPs and PAHs in this study. The recoveries of the POPs and PAHs obtained by the validated QuEChERS method followed by the dSPE EMR-lipid sorbent ranged between 59 and 120%, with repeatabilities ranging between 2 and 23% and LOQs ranging between 0.02 and 1.50 µg·kg-1.

Assessment of pesticide residues in citrus fruit on the Czech market.

When assessing citrus fruit quality, besides natural health-promoting compounds, attention also has to be paid to residues of chemicals used to protect fruit against various pests. A set of 49 samples of different types of citrus fruits collected at the Czech market were analysed for 460 pesticide residues using LC-MS/MS and GC-MS/MS methods. While no residues were detected in citruses from organic farming, altogether 38 various pesticide residues were detected in conventional production samples. Buprofezin in two grapefruit samples and fenbutatin oxide in one tangerine sample exceeded maximum residue limits (MRLs). Depending on the pesticide group, 10-70% of residues were found in pulp, this means that their processing factors calculated for peeling are in the range of 0.02-0.76. In the case of a beverage prepared from unpeeled lemon slices, the transfer of residues from contaminated fruit into infusion was, depending on the beverage type and processing conditions, in the range of 8-61%.

Food fraud in oregano: Pesticide residues as adulteration markers.

Oregano, a widely used and popular herb, is particularly vulnerable to fraud. Less valued plants, adulterants that are often used for dilution, may introduce into this commodity additional contaminants such as pesticide residues. In this study, more than 400 pesticides were screened in a representative set of 42 genuine and 34 adulterated dried oregano samples collected from various locations across Europe. The results obtained by advanced mass spectrometry-based methods, showed, that some pesticide residues could be detected in virtually all tested samples, nevertheless, on average, higher contamination was found in the adulterated oregano samples. Increased incidence of insecticides such as cyfluthrin, permethrin and cyhalothrin was typical for these samples, moreover, pyriproxyfen was detected exclusively in adulterated samples. Thus, based on a critical assessment of pesticide profiles, suspected adulterated oregano samples can be selected for follow up authenticity testing.

Assessment of pesticide residues in strawberries grown under various treatment regimes.

The dynamics of pesticide residues in strawberries that involved quantification of pesticide residues in ripe fruits after model treatment was evaluated in repeated field trials conducted over 3 years. Sixteen commercial pesticide formulations in various combinations were employed in applications from 7 to 44 days before harvest. Altogether 21 active ingredients and some of their metabolites were determined in treated strawberries using LC-MS and GC-MS methods. Except for propargite, the concentrations of all active ingredients declined below the respective MRLs (Regulation (EC) No. 396/2005); nevertheless, most of the tested fungicides often persisted above the 0.01 mg kg⁻¹ limit required by baby food producers to avoid the risk of exceeding the 'baby food limit' established in Commission Directive 2006/141/EC. On the other hand, residues of the majority of tested insecticides, namely spinosad, pymetrozine, deltamethrin, lambda-cyhalothrin and azadirachtin, declined below this limit.

Gas chromatography-triple quadrupole tandem mass spectrometry: a powerful tool for the (ultra)trace analysis of multiclass environmental contaminants in fish and fish feed.

A new method for rapid determination of 73 target organic environmental contaminants including 18 polychlorinated biphenyls, 16 organochlorinated pesticides, 14 brominated flame retardants and 25 polycyclic aromatic hydrocarbons in fish and fish feed using gas chromatography coupled with triple quadrupole tandem mass spectrometry (GC-MS/MS) was developed and validated. GC-MS/MS in electron ionization mode was shown to be a powerful tool for the (ultra)trace analysis of multiclass environmental contaminants in complex matrices, providing measurements with high selectivity and sensitivity. Another positive aspect characterizing the newly developed method is a substantial simplification of the sample preparation, which was achieved by an ethyl acetate QuEChERS (quick, easy, cheap, effective, rugged and safe) based extraction followed by silica minicolumn clean-up. With use of this sample preparation approach the sample laboratory throughput was increased not only because six samples may be prepared in approximately 1 h, but also because all the above-mentioned groups of contaminants can be determined in a single GC-MS/MS run. Under the optimized conditions, the recoveries of all target analytes in both matrices were within the range from 70 to 120% and the repeatabilities were 20% or less. The method quantification limits were in the range from 0.005 to 1 µg kg(-1) and from 0.05 to 10 µg kg(-1) for fish muscle tissue and fish feed, respectively. The developed method was successfully applied to the determination of halogenated persistent organic pollutants and polycyclic aromatic hydrocarbons in fish and fish feed samples.

Polycyclic aromatic hydrocarbons and halogenated persistent organic pollutants in canned fish and seafood products: smoked versus non-smoked products.

In this study, levels of several groups of environmental contaminants represented by PAHs, PCBs, organochlorine pesticides and polybrominated diphenyl ethers were determined in various types of canned smoked and non-smoked fish and seafood products (54 samples) obtained from the Czech market. PAHs were detected in all of the studied samples, and at least one of the target halogenated persistent organic pollutants was present above the LOQ in 85% of the samples. The levels of PAHs, PCBs, organochlorine pesticides (mainly DDTs) and polybrominated diphenyl ethers found in the canned products varied in the range of 1.4-116 µg kg(-1), 0.6-59.6 µg kg(-1), 0.6-82.7 µg kg(-1) and 0.1-2.1 µg kg(-1) can content, respectively. Smoked sprats were the most contaminated fish product (n = 12) in which the highest levels of both PAHs and persistent organic pollutants were found. In 67% of the samples of smoked sprats in oil, the level of benzo[a]pyrene exceeded the maximum level of 5 µg kg(-1) established for smoked fish by European Union legislation. The distribution of target analytes between oil and fish fractions was also assessed. Significantly higher levels of PAHs were measured in the oil fraction.

Rapid determination of polycyclic aromatic hydrocarbons (PAHs) in tea using two-dimensional gas chromatography coupled with time of flight mass spectrometry.

A simple, fast, and cost effective sample preparation procedure has been developed and validated for the determination of 15+1 European Union Polycyclic Aromatic Hydrocarbons (15+1 EU PAHs) in dried tea leave samples. Based on a critical assessment of several sample extraction/clean-up approaches, the method based on the ethyl acetate extraction followed by the use of PAHs dedicated cartridges with molecularly imprinted polymers (MIPs) has been found as an optimal alternative in terms of time demands and obtained good extract purity. For the final identification/quantification of target PAHs, two dimensional gas chromatography coupled to a time-of-flight mass spectrometry (GC×GC-TOFMS) was used. The performance characteristics of the overall analytical method for individual PAHs determined at three spiking levels (0.5, 2.5 and 5 µg kg(-1)) were in following ranges: limits of quantitation (LOQs) 0.05-0.2 µg kg(-1), repeatabilities 2-9%, and recoveries 73-103%. The recoveries achieved by the newly developed sample preparation procedure when employed for naturally contaminated sample ("incurred" PAHs) were comparable to those obtained by other routinely used approaches employing sonication and/or pressurised liquid extraction for sample analytes isolation. The validated method was subsequently used for the determination of selected genotoxic PAHs in 36 samples of black and green tea obtained from the Czech retail market. The levels of ΣPAH4 (sum of benzo[a]anthracene (BaA), chrysene (CHR), benzo[b]fluoranthene (BbFA) and benzo[a]pyrene (BaP)) in black and green tea leaves ranged from 7.4 to 700 µg kg(-1) and from 4.5 to 102 µg kg(-1), respectively. Contamination of tested tea samples by BaP was in the range of 0.2-152 µg kg(-1).

Streamlining sample preparation and gas chromatography-tandem mass spectrometry analysis of multiple pesticide residues in tea.

In this work, a new rapid method for the determination of 135 pesticide residues in green and black dry tea leaves and stalks employing gas chromatography coupled to tandem mass spectrometry (GC-MS/MS) with a triple quadrupole was developed and validated. A substantial simplification of sample processing prior to the quantification step was achieved: after addition of water to a homogenised sample, transfer of analytes into an acetonitrile layer was aided by the addition of inorganic salts. Bulk co-extracts, contained in the crude organic extract obtained by partition, were subsequently removed by liquid-liquid extraction using hexane with the assistance of added 20% (w/w) aqueous NaCl solution. The importance of matrix hydration prior to the extraction for achieving good recoveries was demonstrated on tea samples with incurred pesticide residues. For most of the analytes, recoveries in the acceptable range of 70-120% and repeatabilities (relative standard deviations, RSDs) ≤20% were achieved for both matrices at spiking levels of 0.01, 0.1 and 1 mg kg(-1). Under optimised GC-MS/MS conditions, most of the analytes gave lowest calibration level ≤0.01 mg kg(-1), permitting the control at the maximum residue levels (MRLs) laid down in Regulation (EC) No 396/2005. The developed method was successfully applied to the determination of pesticide residues in real tea samples.

Implementation of comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry for the simultaneous determination of halogenated contaminants and polycyclic aromatic hydrocarbons in fish.

In the presented study, comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry (GC × GC-TOFMS) was shown to be a powerful tool for the simultaneous determination of various groups of contaminants including 18 polychlorinated biphenyls (PCBs), seven polybrominated diphenyl ethers (PBDEs), and 16 polycyclic aromatic hydrocarbons (PAHs). Since different groups of analytes (traditionally analyzed separately) were included into one instrumental method, significant time savings were achieved. Following the development of an integrated sample preparation procedure for an effective and rapid isolation of several groups of contaminants from fish tissue, the GC × GC-TOFMS instrumental method was optimized to obtain the best chromatographic resolution and low quantification limits (LOQs) of all target analytes in a complex mixture. Using large-volume programmable temperature vaporization, the following LOQs were achieved-PCBs, 0.01-0.25 µg/kg; PBDEs, 0.025-5 µg/kg; PAHs 0.025-0.5 µg/kg. Furthermore, several capillary column combinations (BPX5, BPX50, and Rxi-17Sil-ms in the first dimension and BPX5, BPX50, Rt-LC35, and HT8 in the second dimension) were tested during the experiments, and the optimal separation of all target analytes even of critical groups of PAHs (group (a): benz[a]anthracene, cyclopenta[cd]pyrene and chrysene; group (b): benzo[b]fluoranthene, benzo[j]fluoranthene and benzo[k]fluoranthene; group (c): dibenz[ah]anthracene, indeno[1,2,3-cd]pyrene and benzo[ghi]perylene) was observed on BPX5 × BPX50 column setup. Moreover, since the determination of target analytes was performed using TOFMS detector, further identification of other non-target compounds in real life samples was also feasible.

Simplified and rapid determination of polychlorinated biphenyls, polybrominated diphenyl ethers, and polycyclic aromatic hydrocarbons in fish and shrimps integrated into a single method.

In this study, a new rapid and flexible method for the simultaneous determination of 18 key representatives of polychlorinated biphenyls (PCBs), 7 polybrominated diphenyl ethers (PBDEs), and 32 polycyclic aromatic hydrocarbons (PAHs) in fish and shrimps by gas chromatography coupled to mass spectrometry (GC-MS) was developed and validated. A substantial simplification of sample processing prior to quantification step was achieved: after addition of water to homogenized sample, transfer of hydrophobic analytes into ethyl acetate was supported by added inorganic salts. Bulk fat, contained in crude organic extract obtained by partition, was subsequently removed on a silica minicolumn. This approach enabled to process six samples in less than 1h; moreover, the volume of an extraction solvent and consumption of other chemicals can be significantly reduced compared to, e.g., traditional Soxhlet extraction followed by gel permeation chromatography. The recoveries of target analytes were in the range of 73-120% even at the lowest spiking level (1 µg kg(-1)), repeatabilities (relative standard deviations, RSDs) ranged from 1 to 20%. Under optimized GC-MS conditions (time-of-flight mass analyzer, TOF), the limits of quantification (LOQs) were as follows: PCBs 0.1-0.5 µg kg(-1), PBDEs 0.5 µg kg(-1), and PAHs 0.05-0.25 µg kg(-1). Ambient mass spectrometry employing a direct analysis in real time (DART) ion source was shown as an effective tool for fat control in extract, which is needed during the method development and examination of unknown samples prior to the analysis. Further extension of a method scope by other similar analytes is easily possible.