Study of pesticide transformation processes in different wheat varieties and their effects on plant metabolism.

BACKGROUND: This study aims to obtain systematic understanding of the way by which pesticides are metabolized in plants and the influence of this process on plants' metabolism as this process has a key impact on plant-based food safety and quality. The research was conducted under field conditions, which enabled to capture metabolic processes taking place in plants grown under multihectare cultivation conditions. RESULTS: Research was conducted on three wheat varieties cultivated under field conditions and treated by commercially available preparations (fungicides, herbicides, insecticides, and growth regulator). Plant tissues with distinctions in roots, green parts, and ears were collected periodically during spring-summer vegetation period, harvested grains were also investigated. Sample extracts were examined by chromatographic techniques coupled with tandem mass spectrometry for: dissipation kinetics study, identification of pesticide metabolites, and fingerprint-based assessment of metabolic changes. CONCLUSION: Tissue type and wheat varieties influenced pesticide dissipation kinetics and resulting metabolites. Metabolic changes of plants were influenced by type of applied pesticide and its concentration in plants tissues. Despite differences in plant metabolic response to pesticide stress during cultivation, grain metabolomes of all investigated wheat varieties were statistically similar. 4-[cyclopropyl(hydroxy)methylidene]-3,5-dioxocyclo-hexanecarboxylic acid and trans-chrysantemic acid - metabolites of crop-applied trinexapac-ethyl and lambda-cyhalothrin, respectively, were identified in cereal grains. These compounds were not considered to be present in cereal grains up to now. The research was conducted under field conditions, enabling the measurement of metabolic processes taking place in plants grown under large-scale management conditions. © 2024 Society of Chemical Industry.

Stepping-up accurate quantification of chlorinated paraffins: Successful certification of the first matrix reference material.

BACKGROUND: Chlorinated paraffins (CPs) are industrial chemicals categorised as persistent organic pollutants because of their toxicity, persistency and tendency to long-range transport, bioaccumulation and biomagnification. Despite having been the subject of environmental attention for decades, analytical methods for CPs still struggle reaching a sufficient degree of accuracy. Among the issues negatively impacting the quantification of CPs, the unavailability of well-characterised standards, both as pure substances and as matrix (certified) reference materials (CRMs), has played a major role. The focus of this study was to provide a matrix CRM as quality control tool to improve the comparability of CPs measurement results. RESULTS: We present the process of certification of ERM®-CE100, the first fish reference material assigned with certified values for the mass fraction of short-chain and medium-chain chlorinated paraffins (SCCPs and MCCPs, respectively). The certification was performed in accordance with ISO 17034:2016 and ISO Guide 35:2017, with the value assignment step carried out via an intercomparison of laboratories of demonstrated competence in CPs analysis and applying procedures based on different analytical principles. After confirmation of the homogeneity and stability of the CRM, two certified values were assigned for SCCPs, depending on the calibrants used: 31 ± 9 µg kg-1 and 23 ± 7 µg kg-1. The MCCPs certified value was established as 44 ± 17 µg kg-1. All assigned values are relative to wet weight in the CRM that was produced as a fish paste to enhance similarity to routine biota samples. SIGNIFICANCE AND NOVELTY: The fish tissue ERM-CE100 is the first matrix CRM commercially available for the analysis of CPs, enabling analytical laboratories to improve the accuracy and the metrological traceability of their measurements. The certified CPs values are based on results obtained by both gas and liquid chromatography coupled with various mass spectrometric techniques, offering thus a broad validity to laboratories employing different analytical methods and equipment.

Unveiling the potential of proteomics in addressing food and feed safety challenges.

The food and feed sector in Europe is rapidly evolving to address contemporary challenges, striving for fairer, safer, greener and more sustainable food systems. This includes the exploration of new protein sources for human consumption and animal feed such as protein derived from insects, algae or novel plant-derived proteins, and the re-evaluation of existing sources like processed animal protein (PAP). To generate reliable data on the diverse array of emerging protein sources for future food and feed safety assessments, a growing demand for the development and implementation of advanced analytical techniques exists. New approach methodologies (NAMs) including, mass spectrometry (MS)-based proteomics methods have been emerging as valuable techniques which potentially can be implemented in regulatory laboratory settings to complement conventional approaches in this realm. These MS-driven strategies have already proven their utility in diverse applications, including the detection of prohibited substances in feed, identification of allergens, differentiation of fish species in complex mixtures for fraud detection and the verification of novel foods and alternative protein sources. This EU-FORA programme was focused on three core objectives namely: (i) the training of the fellow in utilising MS-based proteomics for food and feed safety analyses, (ii) the involvement of the fellow in the development of standardised operating procedures (SOP) for targeted and non-targeted proteomic MS-based workflows for species and tissues specific PAP identification in a national reference laboratory (NRL) and (iii) the transfer and implementation of MS-based approaches and standardised protocols for PAP analysis at the fellow's home institution. Altogether, this programme facilitates the broadening and diversification of use of MS-based proteomic methodologies for reinforcing their significance within the domains of food and feed safety research and regulatory science applications.

Yeast-based system for in vivo evaluation of alleles of the anthocyanin production pathway.

Plants produce anthocyanins to incite the pollination and seed dispersion performed by pigment-attracted animals. These natural blue-to-red-coloured pigments can be used as food colourants and antioxidants. For this purpose, microbial bioproduction of anthocyanins has become of industrial interest in recent years. 20 new alleles of anthocyanin production pathway genes were extracted and characterised for protein expression level and stability using a developed single-PCR product gene-entry system for tagged protein synthesis in yeast S. cerevisiae. Enzymatic activities of these proteins in the episomally complemented in vivo systems were compared by HPLC-MS analysis. Results show that the codon optimisation of the anthocyanin pathway genes is not essential for the effective heterologous expression in yeast. Elevating the cellular abundance of CHS and F3H enzymes can increase anthocyanidin production from supplemented precursors. New alleles VmF3Hv1 and VuCHS were shown to have the best performance in the analysed system. System complementation with flavonoid 3',5'-hydroxylase substantially increases total anthocyanidin production. The described single-entry yeast episomal complementation system is a convenient and rapid tool for the complex evaluation of new alleles in vivo.

The metabolic processes of selected pesticides and their influence on plant metabolism. A case study of two field-cultivated wheat varieties.

Pesticides that are absorbed by plants undergo biotransformation and might affect plant metabolic processes. The metabolisms of two cultivated wheat varieties, Fidelius and Tobak, treated with commercially available fungicides (fluodioxonil, fluxapyroxad, and triticonazole) and herbicides (diflufenican, florasulam, and penoxsulam) were studied under field conditions. The results provide novel insights regarding the effects of these pesticides on plant metabolic processes. Plants (roots and shoots) were sampled six times during the six-week experiment. Pesticides and pesticide metabolites were identified using GC-MS/MS, LC-MS/MS, and LC-HRMS, while root and shoot metabolic fingerprints were determined using non-targeted analysis. Fungicide dissipation kinetics were analyzed according to the quadratic mechanism (R2: 0.8522-0.9164) for Fidelius roots, and zero-order for Tobak roots (R2: 0.8455-0.9194); shoot dissipation kinetics were analyzed according to first-order (R2: 0.9593-0.9807) and quadratic (R2: 0.8415-0.9487) mechanisms for Fidelius and Tobak, respectively. The fungicide degradation kinetics were different compared to reported literature values, most likely due to differences in pesticide application methods. The following metabolites were respectively identified in shoot extracts of both wheat varieties for fluxapyroxad, triticonazole, and penoxsulam: 3-(difluoromethyl)-N-(3',4',5'-trifluorobiphenyl-2-yl)-1H pyrazole-4-carboxamide, 2-chloro-5-{(E)-[2-hydroxy-3,3-dimethyl-2-(1H-1,2,4-triazol-1-ylmethyl)-cyclopentylidene]-methyl}phenol, and N-(5,8-dimethoxy[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)-2,4-dihydroxy-6 (trifluoromethyl)benzene sulfonamide. Metabolite dissipation kinetics varied depending on the wheat variety. These compounds were more persistent than parent compounds. Despite having the same cultivation conditions, the two wheat varieties varied in their metabolic fingerprints. The study revealed that pesticide metabolism has a greater dependence on plant variety and method of administration compared to the physicochemical properties of the active substance. This highlights the necessity of conducting research on pesticide metabolism under field conditions.

Targeted and non-targeted analysis for the investigation of pesticides influence on wheat cultivated under field conditions.

A comprehensive approach was applied to evaluate the effects of pesticides on the metabolism of wheat (Triticum aestivum L). The application of commercially available pesticide formulations under field cultivation conditions provided a source of metabolic data unlimited by model conditions, representing a novel approach to study the effects of pesticides on edible plants. Gas and liquid chromatography coupled to tandem mass spectrometry were employed for targeted and non-targeted analysis of wheat roots and shoots sampled six times during the six-week experiment. The applied pesticides: prothioconazole, tebuconazole, fluoxastrobin, diflufenican, florasulam, and penoxulam were found at concentrations ranging 0.0070-25.20 mg/kg and 0.0020-2.2 mg/kg in the wheat roots and shoots, respectively. The following pesticide metabolites were identified in shoots: prothioconazole-desthio (prothioconazole metabolite), 5-(4-chlorophenyl)-2,2-dimethyl-3-(1,2,4-triazol-1-ylmethyl)pentane-1,3-diol (tebuconazole metabolite), and N-(5,8-dimethoxy[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)-2,4-dihydroxy-6-(trifluoromethyl)benzene sulphonamide (penoxulam metabolite). The metabolic fingerprints and profiles changed during the experiment, reflecting the cumulative response of wheat to both its growth environment and pesticides, as well as their metabolites. Approximately 15 days after the herbicide treatment no further changes in the plant metabolic profiles were observed, despite the presence of pesticide and their metabolites in both roots and shoots. This is the first study to combine the determination of pesticides and their metabolites plant tissues with the evaluation of plant metabolic responses under field conditions. This exhaustive approach contributes to broadening the knowledge of pesticide effects on edible plants, relevant to food safety.

Procyanidins from Cranberry Press Residues-Extraction Optimization, Purification and Characterization.

Procyanidins are a polyphenolic group that can be found in a variety of foods such as chocolate, tea, cranberries and others. Type A procyanidins can be found in a handful of sources and one of the richest sources are American cranberries. These compounds possess antioxidative, anticancer and anti-inflammatory activities and are most widely used as prevention for urinary tract infections. Cranberries are utilized for jam and juice production, and the latter produces industrial food waste press residues. Press residues contain free and bound procyanidins which can be extracted for use as nutraceuticals. In this study, the extraction of cranberry press residues has been optimized using RSM and the resulting extracts have been purified and fractionated. The obtained procyanidin fractions have been investigated for their antioxidative potential and analyzed using LC-ESI-FTICR-HRMS to determine individual procyanidins. The optimization showed that the optimal extraction can be conducted using acetone in a concentration of 53% without the addition of an acidifying agent. Strong correlation was observed for procyanidin contents and their antioxidative activity using DPPH, ABTS and FRAP methods. The purified fractions contained 78 individual (65 Type A) procyanidins with the degree of polymerization of up to 9.

Qualitative fingerprinting of psychoactive pharmaceuticals, illicit drugs, and related human metabolites in wastewater: A year-long study from Riga, Latvia.

The COVID-19 pandemic has become an unprecedented public health emergency causing immense societal and socio-economic consequences. Multiple studies have outlined that interventions to curb the spread of the virus are likely to have an effect on substance use patterns. In this study, we explored the presence of psychoactive pharmaceuticals, illicit drugs and related human metabolites in 24-h composite wastewater samples that were collected weekly in 2021 from the central WWTP of Riga, Latvia. The analysis was performed via suspect screening approach using three separate high-resolution mass spectrometry (HRMS) workflows, which relied on reversed-phase liquid chromatography (RPLC), hydrophilic interaction liquid chromatography (HILIC) and direct infusion HRMS. In total, 39 out of 149 substances were detected throughout the sampling period. These include pharmaceuticals (mainly antiepileptics, antidepressants and antipsychotics), illicit drugs (e.g., MDMA, MDEA, cocaine, etc.) and new psychoactive substances (alpha-PVP). The results were evaluated in relation to COVID-19 incidence rate and the severity of containment and closure policies. For some compounds we observed temporal changes that may be potentially linked to the state of the pandemic. For instance, higher detection rates were observed for several illicit drugs during periods, when restrictions on public events were relaxed. Meanwhile, some psychoactive pharmaceuticals and drugs used to treat upper respiratory tract infections displayed increased prevalence in weeks when the national COVID-19 incidence rates were higher. However, without baseline reference data from previous years, it is difficult to discern how much of the relationships seen are linked to pandemic progression and seasonal variability.

Consumption trends of pharmaceuticals and psychoactive drugs in Latvia determined by the analysis of wastewater.

Wastewater-based epidemiology (WBE) was applied to evaluate seasonal variations of the consumption of pharmaceuticals (i.e. antibiotics, NSAIDs, antiepileptics, antihypertensives and others), caffeine, alcohol and nicotine in Latvia throughout 2021. In addition, weekly variation of caffeine, nicotine and alcohol consumption was investigated. Pronounced seasonality was observed in the consumption of antibiotics and decongestants, as well as caffeine, nicotine and alcohol. Correlation with COVID-19 statistics was observed in the case of macrolide antibiotics and antiasthmatic salbutamol. Comparison of the estimated consumption values obtained using the WBE approach and the statistics revealed that the majority of compounds data are in good agreement except angiotensin II receptor blocker group antihypertensives where the most overestimated consumption values were observed.

Amino Acid Metabolism is Significantly Altered at the Time of Admission in Hospital for Severe COVID-19 Patients: Findings from Longitudinal Targeted Metabolomics Analysis.

The heterogeneity in severity and outcome of COVID-19 cases points out the urgent need for early molecular characterization of patients followed by risk-stratified care. The main objective of this study was to evaluate the fluctuations of serum metabolomic profiles of COVID-19 patients with severe illness during the different disease stages in a longitudinal manner. We demonstrate a distinct metabolomic signature in serum samples of 32 hospitalized patients at the acute phase compared to the recovery period, suggesting the tryptophan (tryptophan, kynurenine, and 3-hydroxy-DL-kynurenine) and arginine (citrulline and ornithine) metabolism as contributing pathways in the immune response to SARS-CoV-2 with a potential link to the clinical severity of the disease. In addition, we suggest that glutamine deprivation may further result in inhibited M2 macrophage polarization as a complementary process, and highlight the contribution of phenylalanine and tyrosine in the molecular mechanisms underlying the severe course of the infection. In conclusion, our results provide several functional metabolic markers for disease progression and severe outcome with potential clinical application. IMPORTANCE Although the host defense mechanisms against SARS-CoV-2 infection are still poorly described, they are of central importance in shaping the course of the disease and the possible outcome. Metabolomic profiling may complement the lacking knowledge of the molecular mechanisms underlying clinical manifestations and pathogenesis of COVID-19. Moreover, early identification of metabolomics-based biomarker signatures is proved to serve as an effective approach for the prediction of disease outcome. Here we provide the list of metabolites describing the severe, acute phase of the infection and bring the evidence of crucial metabolic pathways linked to aggressive immune responses. Finally, we suggest metabolomic phenotyping as a promising method for developing personalized care strategies in COVID-19 patients.

Two-dimensional liquid chromatography - mass spectrometry as an effective tool for assessing a wide range of pharmaceuticals and biomarkers in wastewater-based epidemiology studies.

The biomarkers used in wastewater-based epidemiology studies have a wide range of physicochemical properties, making simultaneous analysis extremely challenging or even impossible. So far, the majority of analytical procedures employed in this field have focused on specific classes or a limited number of analytes. The heart-cutting two-dimensional liquid chromatography tandem mass spectrometry technique (2D-LC-MS/MS) has enabled the incorporation of compounds with significantly different polarities in a single analytical method. In this study, a 2D-LC-MS/MS method has been developed and optimized for the simultaneous analysis of various biomarkers in wastewater, providing relevant information about lifestyle habits (the usage of alcohol, tobacco, and caffeine), health and well-being (pharmaceuticals), and a population size biomarker (5-HIAA) that is useful for correct calculation of human population during wastewater-based epidemiology studies. A single-laboratory validation procedure showed satisfactory analytical performance, with the mean trueness for most compounds falling in the range of 80÷110 % and the mean repeatability being less than 20 %. The applicability of the method was evaluated by testing 28 wastewater samples collected at different wastewater treatment plants in Latvia. The obtained results revealed the occurrence of 44 out of 62 selected biomarkers, with the lowest quantified concentration at 1.1 ng L-1 in the case of xylometazoline and up to 148 µg L-1 in the case of metformin and 156 µg L-1 for caffeine.

Legacy and alternative brominated, chlorinated, and organophosphorus flame retardants in indoor dust-levels, composition profiles, and human exposure in Latvia.

Flame retardants (FRs) are additives used in consumer products to reduce flammability, even though they can easily contaminate the indoor environment. Since it is common for people in modern cities to spend up to 85% of time indoors, the quality of the indoor environment is critical for human health. In this study, polybrominated diphenyl ethers (PBDEs), organophosphorus flame retardants (OPFRs), emerging brominated flame retardants (EBFRs), and dechlorane-related compounds (DRCs) were measured in household dust samples (n = 34) from Latvia, followed by human exposure assessment. Among all studied compounds, OPFRs showed the highest concentrations (1380-133,000 ng g-1). Despite the phase-out of PBDEs, they were the second most significant flame retardants in the studied dust samples (468-25,500 ng g-1) and the predominant compound was BDE-209. The concentrations of EBFRs were in the range of 120-7295 ng g-1, with the most abundant contaminant being DBDPE, which is widely used as a substitute for the deca-BDE formulation. DRCs were the least common flame retardants in the Latvian indoor environments, with concentrations ranging 22.4-192 ng g-1. Although the concentrations of specific FRs are known to vary between different countries, the levels and patterns observed in dust samples from Latvia were similar to those reported from Central Europe. Human exposure was evaluated as the estimated daily intake (EDI). The calculated exposure to most of the FRs was several orders of magnitude lower than the available reference dose (RfD) values.

Rapid determination of pharmaceuticals in wastewater by direct infusion HRMS using target and suspect screening analysis.

A wide-scope screening of active pharmaceutical ingredients (APIs) and their transformation products (TPs) in wastewater can yield valuable insights and pinpoint emerging contaminants that have not been previously reported. Such information is relevant to investigate their occurrence and fate in various environmental compartments. In this study, we explored the applicability of direct infusion high resolution mass spectrometry (DI-HRMS) for comprehensive and rapid detection of APIs and their TPs in wastewater samples. The method was developed using a Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS) system and incorporated both wide-scope suspect screening and semi-quantitative determination of selected analytes. The identification strategy was based on the following criteria: narrow accurate mass window (±1.25 ppm) for two most abundant full-MS signals, isotopic pattern fit and additional confirmation on the basis of MS2 spectra at three fragmentation levels. The tentative identification of suspects and target compounds relied on an in-house database containing more than 500 different APIs and TPs. The measured fragment spectra were matched against experimental MS2 patterns obtained from a publicly available spectral library (MassBank of North America) and in-silico generated fragmentation features (from the CFM-ID algorithm). In total, 79 suspects were identified and 24 target compounds were semi-quantified in 72 wastewater samples. The highest detection frequencies in treated wastewater effluents were observed for diclofenac, metoprolol and telmisartan, while hydroxydiclofenac, dextrorphan, and carbamazepine metabolites were the most frequently detected TPs. The obtained API profiles were in accordance with the national consumption statistics and the origin of wastewater samples. The developed method is suitable for rapid screening of APIs in wastewater and can be used as a complementary tool to characterize API emissions from wastewater treatment facilities and to identify problematic compounds that require more rigorous monitoring.

Non-target and suspected-target screening for potentially hazardous chemicals in food contact materials: investigation of paper straws.

A non-target screening strategy was developed for the safety evaluation of potentially hazardous chemicals in paper food contact materials (FCMs). A tentative list of suspect analytes was generated using publicly available FCM substance inventories, the presence of contaminants in paper straws was confirmed by high-resolution Orbitrap mass spectrometry. Data-independent and data-dependent MS and MS/MS results for candidate compounds were processed using a workflow including peak detection by deconvolution, blank subtraction, retention time alignment, formula assignment and fragmentation spectra search against spectral libraries followed by in silico generated spectra annotation. This workflow allowed for the identification of 74 suspect compounds, of which 40 were assigned a high confidence level of detection. A tentative in silico toxicity evaluation for mutagenic and carcinogenic activities was carried out. Using quantitative structure-activity relationship (QSAR) models it was found that two of the detected compounds tested positive for mutagenicity and three for carcinogenicity.

The impact of baking on chlorinated paraffins: Characterization of C10-C17 chlorinated paraffins in oven-baked pastry products and unprocessed pastry dough by HPLC-ESI-Q-TOF-MS.

This study presents an HPLC-ESI-Q-TOF method for simultaneous quantification of short-chain chlorinated paraffins (SCCPs, C10-13) and an additional characterization of medium-chain chlorinated paraffins (MCCPs, C14-17) in oven-baked pastry products (n = 38) and unprocessed pastry dough material (n = 15). Almost 2 times higher SCCP levels were found in the dough material. ΣSCCP concentrations in products ranged from 0.3 ng g-1 to 23.0 ng g-1 (mean: 6.3 ng g-1), while the results for dough ranged from 5.8 ng g-1 to 22.8 ng g-1 (mean: 12.9 ng g-1). Regardless of the sample matrix, the most abundant CP homologue groups were hepta- and octa-chlorinated undecanes and dodecanes. The average chlorination degree found in dough samples fell within a range of 55-60% (w/w). Meanwhile, a slight decrease of CP chlorination degree was observed for oven-baked products, in particular for C10-C12 SCCPs, thus indicating that thermal decomposition of CPs occurs even under relatively mild conditions.

Development of a Rapid Method for the Determination of Phenolic Antioxidants in Dark Chocolate Using Ultra Performance Liquid Chromatography Coupled to Orbitrap Mass Spectrometry.

Development of a new comprehensive method based on one-step liquid-liquid extraction (LLE) combined with ultra-high-performance liquid chromatography-Orbitrap high resolution mass spectrometry (UHPLC-Orbitrap-HRMS) for the determination of eight phenolic compounds in dark chocolate is demonstrated. The study focuses on the optimization of sample preparation, chromatographic separation and Orbitrap-HRMS detection parameters to adjust selectivity and sensitivity of the method. The LLE mixture of formic acid-water-acetonitrile (2:58:40, v/v) and hexane at the component ratio of 1:1 (v/v) was applied for sample preparation. The inter-day and intra-day precision expressed from the relative standard deviations (RSDs) ranged between 5.50-14.2% and 5.60-16.2%. The quantification limits of the target analytes were between 0.1 and 1.0 mg/kg. The method was applied for the analysis of twenty-six samples of dark chocolate with various cocoa solid contents (30-77%) collected from the markets in Latvia. The total levels of the phenolic contents ranged between 330 and 2631 mg/kg. The study demonstrates advantages of the Orbitrap-HRMS method for the study of phenolic compounds in dark chocolate.

Simultaneous screening and quantification of aminoglycoside antibiotics in honey using mixed-mode liquid chromatography with quadrupole time-of-flight mass spectroscopy with heated electrospray ionization.

An analytical method based on liquid chromatography with quadrupole time-of-flight mass spectrometry has been developed for the simultaneous determination of six aminoglycoside antibiotics in honey. The sample pretreatment included extraction with aqueous trichloroacetic acid followed by solid-phase extraction on Strata-X polymeric reversed phase cartridges. Liquid chromatography separation was performed on an Obelisc R zwitterionic type mixed-mode column. An ionBooster™ heated electrospray source was used and showed enhanced ionization efficiency in comparison to a conventional electrospray source. The observed signal enhancement ranged from 3- (neomycin) to 16-fold (gentamicin C1). A data-dependent mass spectrometry acquisition approach was employed, in which the full mass spectrometry dataset provided quantification and a scheduled precursor list was used to trigger an alternating data-dependent acquisition of MS2 spectra for confirmation purposes. The described method was validated in accordance to CD 2002/657/EC. Decision limit values were in the range 11.2-33.6 ng/g, and satisfactory performance characteristics were obtained for recovery (65-76%), repeatability (3.8-7.3%), and linearity (≥0.995). The method was applied to the analysis of 49 real honey samples from the country of Georgia. Streptomycin was detected in two samples at 117 and 35 ng/g, and gentamicin C1 was detected in one sample at 32 ng/g.

A comparison of gas chromatography coupled to tandem quadrupole mass spectrometry and high-resolution sector mass spectrometry for sensitive determination of polycyclic aromatic hydrocarbons (PAHs) in cereal products.

Due to the high consumption level, cereals represent one of the major sources of human exposure to PAHs. Taking into account the low maximum permitted level (ML) established in the European Union (EU) for the sum of four priority PAHs (PAH4) in processed cereal-based foods, sensitive and selective analysis for very low concentrations of these carcinogenic compounds was developed and compared by applying two different analytical techniques. Elaborated methods demonstrated appropriate performance in PAHs analysis even at low contamination levels. Limits of detection (LOD) for the PAH4 varied between 0.002µgkg-1 and 0.006µgkg-1, recoveries were in the range of 92-103% for GC-MS/MS and 108-117% for GC-HRMS. Elaborated methods were applied for the analysis of 35 Latvian cereals and bread samples. The concentrations of PAH4 were in a range of 0.22-1.62µgkg-1 with 14% of samples exceeding the current EU maximum permitted levels.

Determination of pharmaceutical residues in wastewater using high performance liquid chromatography coupled to quadrupole-Orbitrap mass spectrometry.

A multi-class method for the determination of 24 emerging pharmaceutical residues has been developed and validated. The method is based on solid-phase extraction of wastewater samples using Strata-X cartridges followed by high performance liquid chromatography coupled to hybrid quadrupole - Orbitrap high resolution mass spectrometry (HPLC-Q-Orbitrap-HRMS). A single-laboratory validation procedure showed satisfactory analytical performance. The analysis of 21 samples collected at the wastewater treatment plant in Riga revealed the occurrence of 20 compounds of different therapeutic classes. The highest concentration was found for the central nervous system stimulator caffeine - up to 12µgL-1, the analgesic acetaminophen up to 4.2µgL-1, the antibiotic ciprofloxacin in the concentration range of 250-400ngL-1, and the non-steroidal anti-inflammatory drug (NSAID) ibuprofen at 100-325ngL-1.

The application of phospholipid removal columns and ultra-high performance liquid chromatography-tandem quadrupole mass spectrometry for quantification of multi-class antibiotics in aquaculture samples.

In this study a robust and sensitive method based on a proposed sample purification procedure, using zirconia-coated Phree™ columns and analysis by ultra-high performance liquid chromatography with triple quadrupole tandem mass spectrometry are presented for the assessment of multi-class antibiotics in farmed fish species. The sample preparation procedure benefited from combined precipitation of proteins and selective removal of phospholipids by Phree™ columns, resulting in a high sensitivity of the method (LOQ 0.3-9mgkg(-1)). The in-house validation results (precision, repeatability, decision limit CCα, detection capability CCß, etc.) indicate that the elaborated method is fully suitable for the analysis of the main classes of antibiotics in accordance with the European Union (EU) Commission Decision 2002/657/EC. The method was applied to the analysis of antibiotics in trout and sturgeon samples obtained from the local inland aquacultures in Latvia. The results revealed the presence of two antibiotics (enrofloxacin and trimethoprim) in 12 out of the 20 analysed fish samples at concentrations (0.33-12.2µgkg(-1)) below the MRLs, thus causing no acute risks to consumers.