ABSTRACT
In this paper, we present analytical methodologies for the determination of the thiazolidine fungicide flutianil (trade name GATTEN) and its primary metabolite OC56635 in hemp cannabis matrices. A total of nine crop matrices were tested: whole seed, fiber, flower buds, hemp hearts, hemp seed oil, hemp meal, hemp flour, ethanol extracted CBD resin (CBD-E), and supercritical CO2 extracted CBD resin (CBD-C). Processing of the CBD-E and CBD-C crop fractions was carried out in-house using methods detailed herein. Field sample analysis utilized sequential extractions, stacked solid phase extraction (SPE) column cleanups, and evaporation to prepare the samples for LC-MS/MS quantitation. Method validations for each fraction were carried out using untreated hemp matrices over a minimum of three levels, with lowest levels of method validation (LLMV) of 0.010 µg/g for all fractions except the CBD resins, for which LLMV was 0.020 µg/g. Flutianil-treated samples from nine field sites were collected from several crop production regions and analyzed to determine the distribution of incurred flutianil and OC56635 residues within the different hemp matrices. This data was generated in support of nationwide registration with the United States Environmental Protection Agency (USEPA).
ABSTRACT
Beer has over 600 flavor compounds and creates a positive tasting experience with acceptable sensory properties, which are essential for the best consumer experience. Spontaneous and mixed-culture fermentation beers, generally classified as sour beers, are gaining popularity compared to typical lager or ale styles, which have dominated in the USA for the last few decades. Unique and acceptable flavor compounds characterize sour beers, but some unfavorable aspects appear in conjunction. One such unfavorable flavor is called "mousy". This description is usually labeled as an unpleasant odor, identifying spoilage of fermented food and beverages. It is related as having the odor of mouse urine, cereal, corn tortilla chips, or freshly baked sour bread. The main compounds responsible for it are N-heterocyclic compounds: 2-acetyltetrahydropyridine, 2-acetyl-1-pyrroline, and 2-ethyltetrahydropyridine. The most common beverages associated with mousy off-flavor are identified in wines, sour beers, other grain-based beverages, and kombucha, which may contain heterofermentative lactic acid bacteria, acetic acid bacteria, and/or yeast/fungus cultures. In particular, the fungal species Brettanomyces bruxellensis are associated with mousy-off flavor occurrence in fermented beverages matrices. However, many factors for N-heterocycle formation are not well-understood. Currently, the research and development of mixed-cultured beer and non/low alcohol beverages (NABLAB) has increased to obtain the highest quality, sensory, functionality, and most notably safety standards, and also to meet consumers' demand for a balanced sourness in these beverages. This paper introduces mousy off-flavor expression in beers and beverages, which occurs in spontaneous or mixed-culture fermentations, with a focus on sour beers due to common inconsistency aspects in fermentation. We discuss and suggest possible pathways of mousy off-flavor development in the beer matrix, which also apply to other fermented beverages, including non/low alcohol drinks, e.g., kombucha and low/nonalcohol beers. Some precautions and modifications may prevent the occurrence of these off-flavor compounds in the beverage matrix: improving raw material quality, adjusting brewing processes, and using specific strains of yeast and bacteria that are less likely to produce the off-flavor. Conceivably, it is clear that spontaneous and mixed culture fermentation is gaining popularity in industrial, craft, and home brewing. The review discusses important elements to identify and understand metabolic pathways, following the prevention of spoilage targeted to off-flavor compounds development in beers and NABLABs.
Subject(s)
Beer , Lactobacillales , Alcoholic Beverages , Bacteria , Beer/analysis , Fermentation , Saccharomyces cerevisiae/metabolism , Wine/analysisABSTRACT
Applications of systemic pesticides can have unexpected direct and indirect effects on nontarget organisms, producing ecosystem-level impacts. We investigated whether a systemic insecticide (imidacloprid) could be absorbed by a plant pathogenic fungus infecting treated plants and whether the absorbed levels were high enough to have detrimental effects on the survival of a mycophagous beetle. Beetle larvae fed on these fungi were used to assess the survival effects of powdery mildew and imidacloprid in a factorial design. Fungal conidia were collected from treated and untreated plants and were tested for the presence and concentration of imidacloprid. The survival of beetles fed powdery mildew from imidacloprid-treated leaves was significantly lower than that of the beetles from all other treatments. Imidacloprid accumulated in fungal conidia and hyphae was detected at levels considered lethal to other insects, including coccinellid beetles. Water-soluble systemic insecticides may disrupt mycophagous insects as well as other nontarget organisms, with significant implications for biodiversity and ecosystem function.
ABSTRACT
Harmful cyanobacterial blooms compromise human and environmental health, mainly due to the cyanotoxins they often produce. Microcystins (MCs) are the most commonly measured group of cyanotoxins and are hepatotoxic, neurotoxic, and cytotoxic. Due to MCs ability to covalently bind to proteins, quantification in complex matrices is difficult. To analyze bound and unbound MCs, analytical methods were optimized for analysis in sediment and clam tissues. A clean up step was incorporated to remove lipids, improving percent yield. This method was then applied to sediment and clam samples collected from the Sacramento-San Joaquin River Delta (Delta) in the spring and fall of 2017. Water samples were also tested for intracellular and extracellular MCs. These analyses were used to quantify the partitioning of MCs among sediment, clams, and water, and to examine whether MCs persist during non-summer months. Toxin analysis revealed that multiple sediment samples collected in the Delta were positive for MCs, with a majority of the positive samples from sites in the San Joaquin River, even while water samples from the same location were below detection limit. These data highlight the importance of analyzing MCs in complex matrices to accurately evaluate environmental risk.
Subject(s)
Bivalvia/chemistry , Environmental Monitoring/methods , Geologic Sediments/chemistry , Microcystins/analysis , Rivers/chemistry , Water Pollutants, Chemical/analysis , Animals , Gas Chromatography-Mass Spectrometry , Geologic Sediments/microbiology , Harmful Algal Bloom , Limit of Detection , Oxidation-Reduction , Reproducibility of Results , Rivers/microbiology , Solid Phase ExtractionABSTRACT
An analytical method was developed for the determination of morpholine on apples and citrus. The method utilized acidified methanol extraction, centrifugation, and determination by hydrophilic interaction liquid chromatography with electrospray ionization and tandem mass spectrometry (HILIC-ESI-MS/MS). Validation of the method occurred at the Pacific Agricultural Laboratory (PAL, Portland, OR, USA) and the Trace Analytical Laboratory (TAL, UC Davis, CA, USA). Method validation recoveries from control apple, orange, lemon, and grapefruit samples ranged from 84 to 120% over three levels of fortification (0.01, 0.04, and 0.2 µg/g). The limit of quantitation (LOQ) for all commodities was 0.01 µg/g, and the calculated method detection limit (MDL) ranged from 0.0010 to 0.0040 µg/g.
Subject(s)
Chromatography, Liquid/methods , Food Contamination/analysis , Fruit/chemistry , Morpholines/analysis , Pesticide Residues/analysis , Tandem Mass Spectrometry/methods , Citrus/chemistry , Malus/chemistryABSTRACT
This study was conducted to evaluate the effect of molinate on retinoids homeostasis in rat testis. Molinate was administrated to male Sprague-Dawley rats (200 mg kg(-1) in corn oil, ip). Retinoid measurements were made at 6, 12, 48 and 168 h time points after administration. Testis levels of retinoic acid decreased (32 %) in a statistically significant manner at the 12 and 48 h time points. However, retinol and retinaldehyde were not significantly affected by molinate. These results suggest that molinate affects retinoic acid synthesis in testis and could contribute to understanding the molecular mechanism of molinate involved testicular toxicity.
Subject(s)
Azepines/toxicity , Herbicides/toxicity , Testis/drug effects , Thiocarbamates/toxicity , Vitamin A/metabolism , Animals , Male , Rats , Rats, Sprague-Dawley , Retinaldehyde/metabolism , Testis/metabolismABSTRACT
One of the most serious diseases in potato cultivars is caused by the pathogen Phytophthora infestans, which affects leaves, stems and tubers. Metalaxyl is a fungicide that protects potato plants from Phytophthora infestans. In Mexico, farmers apply metalaxyl 35 times during the cycle of potato production and the last application is typically 15 days before harvest. There are no records related to the presence of metalaxyl in potato tubers in Mexico. In the present study, we evaluated the effect of Acinetobacter sp on metalaxyl degradation in potato seedlings. The effect of bacteria and metalaxyl on the growth of potato seedlings was also evaluated. A metabolite profile analysis was conducted to determine potential molecular biomarkers produced by potato seedlings in the presence of Acinetobacter sp and metalaxyl. Metalaxyl did not affect the growth of potato seedlings. However, Acinetobacter sp strongly affected the growth of inoculated seedlings, as confirmed by plant length and plant fresh weights which were lower in inoculated potato seedlings (40% and 27%, respectively) compared to the controls. Acinetobacter sp also affected root formation. Inoculated potato seedlings showed a decrease in root formation compared to the controls. LC-MS/MS analysis of metalaxyl residues in potato seedlings suggests that Acinetobacter sp did not degrade metalaxyl. GC-TOF-MS platform was used in metabolic profiling studies. Statistical data analysis and metabolic pathway analysis allowed suggesting the alteration of metabolic pathways by both Acinetobacter sp infection and metalaxyl treatment. Several hundred metabolites were detected, 137 metabolites were identified and 15 metabolic markers were suggested based on statistical change significance found with PLS-DA analysis. These results are important for better understanding the interactions of putative endophytic bacteria and pesticides on plants and their possible effects on plant metabolism.
Subject(s)
Acinetobacter/physiology , Alanine/analogs & derivatives , Metabolome , Seedlings/metabolism , Seedlings/microbiology , Solanum tuberosum/metabolism , Solanum tuberosum/microbiology , Acinetobacter/genetics , Acinetobacter/growth & development , Acinetobacter/isolation & purification , Alanine/metabolism , Alanine/pharmacology , Biodegradation, Environmental/drug effects , Chromatography, Liquid , DNA, Ribosomal/genetics , Discriminant Analysis , Galactose/metabolism , Internet , Least-Squares Analysis , Mass Spectrometry , Metabolic Networks and Pathways/drug effects , Metabolome/drug effects , Phenylalanine/metabolism , Phylogeny , Plant Roots/cytology , Plant Roots/drug effects , Plant Roots/microbiology , Rhizosphere , Seedlings/drug effects , Solanum tuberosum/drug effects , Statistics as TopicABSTRACT
An analytical method was developed for the determination of eleven agrochemicals [abamectin (as B1a), bifenazate, bifenthrin, carfentrazone-ethyl, cymoxanil, hexythiazox, imidacloprid, mefenoxam, pymetrozine, quinoxyfen, and trifloxystrobin] in dried hops. The method utilized polymeric and NH2 solid phase extraction (SPE) column cleanups and liquid chromatography with mass spectrometry (LC-MS/MS). Method validation and concurrent recoveries from untreated dried hops ranged from 71 to 126% for all compounds over three levels of fortification (0.10, 1.0, and 10.0 ppm). Commercially grown hop samples collected from several field sites had detectable residues of bifenazate, bifenthrin, hexythiazox, and quinoxyfen. The control sample used was free of contamination below the 0.050 ppm level for all agrochemicals of interest. The limit of quantitation and limit of detection for all compounds were 0.10 and 0.050 ppm, respectively.
Subject(s)
Chromatography, Liquid , Humulus/chemistry , Pesticides/analysis , Tandem Mass Spectrometry , Food Contamination/analysis , Reproducibility of ResultsABSTRACT
An analytical method was developed for the determination of the neo-nicotinoid insecticide flonicamid ( N-cyanomethyl-4-trifluoromethylnicotinamide) and its metabolites N-(4-trifluoronicotinoyl) glycine (TFNG), 4-trifluoronicotinic acid (TFNA), and 4-trifluoromethylnicotinamide (TFNA-AM) in dried hops. The method utilized C18 and polymeric solid phase extraction (SPE) column cleanups, liquid-liquid partitioning, and liquid chromatography (LC) with mass spectrometry (MS/MS). Method validation and concurrent recoveries from untreated dried hops ranged from 66 to 119% for all compounds over five levels of fortification (0.005, 0.02, 0.2, 2.0, and 4.0 ppm). Flonicamid-treated hop samples collected from three field sites had the following residues: flonicamid levels of 0.561-2.85 ppm, TFNA levels of 0.302-0.470 ppm, TFNA-AM levels of 0.038-0.177 ppm, and TFNG levels of 0.098-0.204 ppm. Untreated hop samples from all fields had residues <0.005 ppm for flonicamid, TFNA, TFNA-AM, and TFNG. The limit of quantitation and limit of detection for all compounds were 0.005 and 0.0025 ppm, respectively.
Subject(s)
Chromatography, Liquid/methods , Humulus/chemistry , Insecticides/analysis , Mass Spectrometry , Niacinamide/analogs & derivatives , Niacinamide/analysis , Niacinamide/metabolism , Pesticide Residues/analysisABSTRACT
Recent studies indicate that trichloroethylene (TCE) may be a male reproductive toxicant. It is metabolized by conjugation with glutathione and cytochrome p450-dependent oxidation. Reactive metabolites produced along both pathways are capable of forming protein adducts and are thought to be involved in TCE-induced liver and kidney damage. Similarly, in situ bioactivation of TCE and subsequent binding of metabolites may be one mechanism by which TCE acts as a reproductive toxicant. Cysteine-conjugate beta-lyase (beta-lyase) bioactivates the TCE metabolite dichlorovinyl cysteine (DCVC) to a reactive intermediate that is capable of binding cellular macromolecules. In the present study, Western blot analysis indicated that the soluble form of beta-lyase, but not the mitochondrial form, was present in the epididymis and efferent ducts. Both forms of beta-lyase were detected in the kidney. When rats were dosed with DCVC, no protein adducts were detected in the epididymis or efferent ducts, although adducts were present in the proximal tubule of the kidney. Trichloroethylene can also be metabolized and form protein adducts through a cytochrome p450-mediated pathway. Western blot analysis detected the presence of cytochrome p450 2E1 (CYP2E1) in the efferent ducts. Immunoreactive proteins were localized to efferent duct and corpus epididymis epithelia. Metabolism of TCE was demonstrated in vitro using microsomes prepared from untreated rats. Metabolism was inhibited 77% when efferent duct microsomes were preincubated with an antibody to CYP2E1. Dichloroacetyl adducts were detected in epididymal and efferent duct microsomes exposed in vitro to TCE. Results from the present study indicate that the cytochrome p450-dependent formation of reactive intermediates and the subsequent covalent binding of cellular proteins may be involved in the male reproductive toxicity of TCE.
Subject(s)
Cysteine/analogs & derivatives , Cysteine/metabolism , DNA Adducts/biosynthesis , Epididymis/metabolism , Lyases/metabolism , Solvents/metabolism , Transaminases/metabolism , Trichloroethylene/metabolism , Vas Deferens/enzymology , Animals , Biotransformation , Cysteine/toxicity , Cytochrome P-450 CYP2E1/drug effects , Cytochrome P-450 CYP2E1/metabolism , DNA Adducts/drug effects , Dose-Response Relationship, Drug , Environmental Pollutants/metabolism , Environmental Pollutants/toxicity , Epididymis/drug effects , Epididymis/enzymology , Immunohistochemistry , Lyases/drug effects , Male , Protein Binding/drug effects , Rats , Rats, Sprague-Dawley , Solvents/toxicity , Transaminases/drug effects , Trichloroethylene/toxicity , Vas Deferens/cytology , Vas Deferens/drug effectsABSTRACT
The fate of residues of seven agrochemicals (chlorfenapyr, quinoxyfen, tebuconazole, fenarimol, pyridaben, and E- and Z-dimethomorph) from the treatment on hops to the brewing of beer was studied. First, a multi-residue analytical method was developed for the determination of pesticide residues in spent hops, trub, wort, and beer. Each matrix was validated over at least two levels of fortification, for all seven compounds, in the ranges 0.05-5.0, 0.001-1.0, 0.001-0.05, and 0.0005-1.0 ppm for spent hops, trub, wort, and beer, respectively. Recoveries ranged from 73 to 136%. Second, the matrixes prepared from hops, which were treated under commercial practices with each compound, were analyzed using the method developed. The use of treated hops resulted in the carryover of 0.001 ppm of tebuconazole, 0.008 Z-dimethomorph, and 0.005 ppm of E-dimethomorph into the wort. The bulk of the remaining residues of all seven compounds was found on the spent hops. Following fermentation, all compounds were found in levels less than 0.0005 ppm in beer, except Z- (0.006 ppm) and E-dimethomorph (0.004 ppm). Third, when all seven pesticides were spiked prior to the pitching of yeast into clean wort, most of the nonpolar compounds (chlorfenapyr, quinoxyfen, and pyridaben) partitioned into the organic material (trub) which settled to the bottom, while the more polar compounds (fenarimol, tebuconazole, and E- and Z-dimethomorph) were generally distributed evenly between the beer and the trub.