Comparison of polyfunctional thiol, element, and total essential oil contents in 32 hop varieties from different countries.

Volatile thiol 3-mercaptohexan-1-ol (3MH) and particularly 4-mercapto-4-methylpentan-2-one (4MMP) are highly potent flavour compounds in hops. For the determination, a simple and robust stable isotope dilution LC-MS/MS method was developed and applied to 32 hop varieties worldwide from harvest years 2019 and 2020. Limit of detection, precision, and recovery were 0.15 µg/kg, 10%, and 97-108%, respectively. Levels of 3MH and 4MMP ranged from 1.9 to 79.2 µg/kg and from undetectable to 37.1 µg/kg, respectively. Citra, Mosaic, and Strata were rich in both thiols. ICP analyses revealed, that variation of potassium content between the two harvest years was inversely correlated with that of manganese and rubidium (|r| ≥ 0.89) among 12 US varieties excluding Citra and Mosaic. Total essential oil content (0.34-2.7 mL/100 g) was inversely correlated with calcium content (|r| ≥ 0.65). Greatly varying thiol levels depending on variety, region and harvest year might lead to differing flavour results in beer.

Determination of pharmaceuticals and antiseptics in water by solid-phase extraction and gas chromatography/mass spectrometry: analysis via pentafluorobenzylation and stable isotope dilution.

A sensitive yet robust analytical method is presented for the simultaneous determination of 12 human pharmaceuticals (valproic acid, phenytoin, ibuprofen, gabapentin, acetaminophen, gemfibrozil, naproxen, ketoprofen, secobarbital, phenobarbital, 5-fluorouracil, and diclofenac) and 6 antiseptics (biosol, biphenylol, p-chloro-m-cresol, p-chloro-m-xylenol, chlorophene, and triclosan). The method employs solid-phase extraction (SPE) followed by a novel pentafluorobenzylation using a mixture of acetontrile/water (1/1, v/v). The method is simple to perform (derivatization can be completed in a single test tube) and eliminates the need for any solvent/SPE cartridge drying or blow-down. It affords excellent resolution, high sensitivity and reproducibility, and freedom from interference even for matrices as complex as untreated sewage. The method was applied to the analysis of sewage samples using 15 isotopically labeled surrogates, which resulted in the detection of 10 of the 12 pharmaceuticals and all of the antiseptics sought. Ten of 15 surrogates were synthesized from pure analytes by a simple H-D exchange reaction employing D(2)O and D(2)SO(4). Measured recoveries were sensitive to matrix effects and varied substantially among analytes, indicative of the limitations associated with using a single surrogate standard.

Effects of chain length, chlorination degree, and structure on the octanol-water partition coefficients of polychlorinated n-alkanes.

Log octanol-water partition coefficients (log Kow) of 40 synthesized polychlorinated n-alkanes (PCAs) with different chlorination degrees were determined using reversed-phase high performance liquid chromatography (RP-HPLC). In addition, log Kow values of a technical mixture namely Cereclor 63L as well as 15 individual in house synthesized C10, C11, and C12 chloroalkanes with known chlorine positions were estimated. Based on these results, the effects of chain length, chlorination degree, and structure were explored. The estimated log Kow values ranged from 4.10 (polychlorinated n-decanes with 50.2% chlorine content) to 11.34 (polychlorinated n-octacosanes with 54.8% chlorine content) for PCAs and from 3.82 (1,2,5,6,9,10-hexachlorodecane) to 7.75 (1,1,1,3,9,11,11,11-octachlorododecane) for the individual chloroalkanes studied. The results showed that log Kow value was influenced linearly at a given chlorine content by chain length, while a polynominal effect was observed in dependence on the chlorination degree of an alkane chain. Chlorine substitution pattern influenced markedly the log Kow value of chloroalkanes.

Exposure to selected limonene oxidation products: 4-OPA, IPOH, 4-AMCH induces oxidative stress and inflammation in human lung epithelial cell lines.

Limonene oxidation products (LOPs) have gained interest on their harmful health effects over time. Recently, studies have shown that the selected LOPs: 4-oxopentanal (4-OPA), 3-isopropenyl-6-oxo-heptanal (IPOH) and 4-acetyl-1-methylcyclohexene (4-AMCH) have sensory irritation effects in mice and inflammatory effects in human lung cells. This study was therefore undertaken to investigate the potential capacity of 4-OPA, IPOH and 4-AMCH to cause cell membrane damage, oxidative stress and inflammation in human bronchial (16HBE14o-) and alveolar (A549) epithelial cell lines. Overall results suggest that 4-OPA, IPOH have cytotoxic effects on human lung cells that might be mediated by ROS: the highest concentration applied of IPOH [500 µM] enhanced ROS generation by 100-fold ± 7.7 (A549) and 230-fold ± 19.9 (16HBE14o-) compared to the baseline. 4-OPA [500 µM] increased ROS levels by 1.4-fold ± 0.3 (A549) and by 127-fold ± 10.5 (16HBE14o-), while treatment with 4-AMCH [500 µM] led to 0.9-fold ± 0.2 (A549) and 49-fold ± 12.8 (16HBE14o-) increase. IPOH [500 µM] caused a decrease in the thiol-state balance (e.g. after 2 h, GSH:GSSG was reduced by 37% compared to the untreated 16HBE14o-cells). 4-OPA [500 µM] decreased the GSH:GSSG by 1.3-fold change in A549 cells and 1.4-fold change in 16HBE14o-cells. No statistically significant decrease in the GSH:GSSG in A549 and 16HBE14o-cell lines was observed for 4-AMCH [500 µM]. In addition, IPOH and 4-OPA [31.2 µM] increased the amount of the inflammatory markers: RANTES, VEGF and EGF. On the other hand, 4-AMCH [31.2 µM] did not show inflammatory effects in A549 or 16HBE14o-cells. The 4-OPA, IPOH and 4-AMCH treatment concentration and time-dependently induce oxidative stress and/or alteration of inflammatory markers on human bronchial and alveolar cell lines.

Determination and levels of the biocide ortho-phenylphenol in canned beers from different countries.

A method was developed for the determination of the biocide ortho-phenylphenol (biphenyl-2-ol; OPP) in beer, using deuterated OPP as an internal standard. A new liquid-liquid extraction procedure, employing acetonitrile, diethyl ether, and n-pentane, afforded rapid phase separation. The evaporated extract was derivatized with pentafluorobenzyl bromide in a water-acetonitrile mixture that was buffered with potassium carbonate, followed by extraction of the derivative into cyclohexane and analysis by gas chromatography-mass spectrometry in electron ionization mode. The method enables the detection of OPP in 50 mL of beer at concentrations as low as 0.1 microg/L and provides a linear range of quantification of 0.5-40 microg/L. Samples from 61 beers canned over the past 12 years and sold in 27 countries were analyzed for OPP. In 40 of them, the target compound was present at concentrations of 1.2-40 microg/L. Our investigations indicate that the ends of the cans, which contain sealing material presumably treated with OPP, are responsible for this contamination.

Organochlorine levels in edible fish from the Marmara Sea, Turkey.

Samples of 12 edible fish species from the Marmara Sea were analyzed for organochlorines (PCBs, DDTs, HCB, HCHs, toxaphene, etc.). The results showed that the total concentrations ranged from 329.41 ng/g fat to 1453.87 ng/g fat. DDT group components made up almost half or more of organochlorine contamination. Levels in red mullet were compared with those from neighbor seas. The sum of DDTs as well as HCHs concentrations were markedly lower than in the Black Sea but higher in the Aegean Sea and Mediterranean. Thus, inflow from the Black Sea might be considered as a contamination source for DDT and HCH contamination. On the other hand, total PCB concentration (sum of congeners 28, 52, 101, 118, 138, 153 and 180) detected in this study was comparable or lower than those from the Aegean Sea and Mediterranean. Toxaphene was a minor contaminant. Measured values were below maximum residue levels for human consumption.

Inflammatory effects induced by selected limonene oxidation products: 4-OPA, IPOH, 4-AMCH in human bronchial (16HBE14o-) and alveolar (A549) epithelial cell lines.

Limonene, a monoterpene abundantly present in most of the consumer products (due to its pleasant citrus smell), easily undergoes ozonolysis leading to several limonene oxidation products (LOPs) such as 4-acetyl-1-methylcyclohexene (4-AMCH), 4-oxopentanal (4-OPA) and 3-isopropenyl-6-oxoheptanal (IPOH). Toxicological studies have indicated that human exposure to limonene and ozone can cause adverse airway effects. However, little attention has been paid to the potential health impact of specific LOPs, in particular of IPOH, 4-OPA and 4-AMCH. This study evaluates the cytotoxic effects of the selected LOPs on human bronchial epithelial (16HBE14o-) and alveolar epithelial (A549) cell lines by generating concentration-response curves using the neutral red uptake assay and analyzing the inflammatory response with a series of cytokines/chemokines. The cellular viability was mostly reduced by 4-OPA [IC50=1.6mM (A549) and 1.45mM (16HBE14o-)] when compared to IPOH [IC50=3.5mM (A549) and 3.4mM (16HBE14o-)] and 4-AMCH [IC50 could not be calculated]. As a result from the inflammatory response, IPOH [50µM] induced an increase of both IL-6 and IL-8 secretion in A549 (1.5-fold change) and in 16HBE14o- (2.8- and 7-fold change respectively). 4-OPA [50µM] treatment of A549 increased IL-6 (1.4-times) and IL-8 (1.3-times) levels, while in 16HBE14o- had an opposite effect. A549 treated with 4-AMCH [50µM] elevate both IL-6 and IL-8 levels by 1.2-times, while in 16HBE14o- had an opposite effect. Based on our results, lung cellular injury characterized by inflammatory cytokine release was observed for both cell lines treated with the selected chemicals at concentrations that did not affect their cellular viability.

Relative developmental toxicity of short-chain chlorinated paraffins in Zebrafish (Danio rerio) embryos.

Short-chain chlorinated paraffins (SCCPs) are ubiquitous in the environment and might cause adverse environmental and human health effects. Little is known about the relative toxicity of different SCCP compounds especially during development. The objective of this study was to characterize and compare effects of seven SCCP groups at environmentally relevant levels, using a zebrafish (Danio rerio) model. Observations on malformation, survival rates at 96 h post fertilization (hpf), and hatching rates at 72 hpf indicated that the C10- groups (C10H18Cl4, 1,2,5,6,9,10-C10H16Cl6 and C10H15Cl7) were more toxic than the C12- groups (C12H22Cl4, C12H19Cl7 and 1,1,1,3,10,12,12,12-C12H18Cl8) and Cereclor 63L. The C10- groups were also more potent than C12- groups and Cereclor 63L in decreasing thyroid hormone levels. Among the three compounds within the C10- group, the compounds with less chlorine content had stronger effects on sub-lethal malformations but less effects on triiodothyronine (T3) and tetraiodothyronine (T4). Only C10H18Cl4 significantly decreased the mRNA expression of tyr, ttr, dio2 and dio3 at a dose-dependent manner suggesting that the specific mode of actions differ with different congeners. The mechanisms of disruption of thyroid status by different SCCPs could be different. C10H18Cl4 might inhibit T3 production through the inhibition effect on dio2. These results indicate that SCCP exposure could alter gene expression in the hypothalamic-pituitary-thyroid (HPT) axis and thyroid hormone levels. The mechanisms of disruption of thyroid status by different SCCPs could be different. Our results on the relative developmental toxicities of SCCPs will be useful to reach a better understanding of SCCP toxicity supporting environmental risk evaluation and regulation and used as a guidance for environmental monitoring of SCCPs in the future.

Synthesis of two major toxaphene components and their photostabilities.

The synthesis of 2-endo,3-exo,5-endo,6-exo,8,9,10-heptachlorobornane (B7-1001, Hp-Sed) and 2-endo,3-exo,5-endo,6-exo,8,9,9,10-octachlorobornane (B8-1412) is described. Both compounds are components of toxaphene, an insecticide that has been widely used in the past. B7-1001 is an important toxaphene congener, comprising up to 99% of total toxaphene concentrations found in fish and sediment samples from treated lakes. B8-1412 is also a significant component of toxaphene contamination in samples from biota. In synthesizing the compounds, 2-exo,3-endo,6-endo,8,9,10-hexachlorobornane (B6-913) was obtained by reduction of the well-known toxaphene component P 32 (B7-515, 2,2,5-endo,6-exo,8,9,10-heptachlorobornane), which was itself isolated from the chlorination products of (+)-camphene. Chlorination of B6-913 provided B7-1001 in 49.5% yield, and P 32 and four other heptachlorobornanes were also detected in the reaction mixture. Structures of two of the heptachlorobornanes were elucidated by MS and NMR as 2-exo,3-endo,6-endo,8,9,9,10-heptachlorobornane (B7-1461) and 2-exo,3,3,6-endo,8,9,10-heptachlorobornane (B7-1303). B8-1412 was isolated from the product mixture obtained by chlorination of 2-exo,3-endo,6-endo,8,9,9,10-heptachlorobornane. Photolysis experiments at lambda = 254 nm revealed that B6-913 is photochemicaly the most stable compound of the seven toxaphene compounds studied, with a t(1/2) of 213 h. B7-1001, having a t(1/2) of 82 h, was the second most stable compound. B8-1412 was degraded more rapidly, with a t(1/2) of 28.8 h, than B7-1001, but was still much more stable than P 50 (B9-1679, 2-endo,3-exo,5-endo,6-exo,8,8,9,10,10-nonachlorobornane), which had a t(1/2) of 9.4 h, despite its reputation as a very persistant compound. Under the same experimental conditions hexachlorobenzene (HLB) and octachlorodibenzodioxine (OCDD) were consumed very quickly with t(1/2) = 0.0025 and 0.0015 h, respectively.

Analysis of short chain chlorinated paraffins in sediment samples from the Czech Republic by short-column GC/ECNI-MS.

Top layer sediment samples from the Czech Republic were analyzed to obtain the preliminary information about contamination of the region by chlorinated paraffins. Sediment samples from three locations with different industrial discharges were taken over the period of 2 years. The analysis was performed by short-column gas chromatography-electron capture negative ion mass spectrometry (SCGC/ECNI-MS). Short chain chlorinated paraffin sediment concentrations varied between 24.00 and 45.78 ng g(-1) (dry weight, d.w.) in the Kosetice area, 16.30-180.75 ng g(-1) (dry weight) in the Zlín area and 4.58-21.57 ng g(-1) dry weight in the Beroun area. Highly chlorinated undecanes prevailed in the samples. On the basis of these results, more detailed studies should be conducted to determine the magnitude and extent of chlorinated paraffin contamination in these regions.

Determination of volatile compounds in different hop varieties by headspace-trap GC/MS--in comparison with conventional hop essential oil analysis.

A headspace (HS)-trap method in combination with gas chromatography (GC)-mass spectrometry (MS) was developed for the determination of volatile constituents in hops. The highly sensitive HS-trap system reduces the detection limit by using up to four trap enrichment cycles. Seventy hop samples of different varieties and cultivation regions, from the 2008 harvest, were examined using the HS-trap-GC-MS method and the established "European Brewery Convention" (EBC) method for hop essential oil analysis. Twenty-one different volatiles were quantified for each hop sample. For all compounds, except caryophyllene oxide, a strong correlation was found between the results of the HS-trap method and the EBC method. Experiments have revealed that the EBC method using steam distillation is not appropriate for thermolabile compounds, such as caryophyllene oxide, due to decomposition during boiling. The HS-trap method is fast and sensitive, requires small sample amounts and minimal sample preparation, and is easy to apply.

Trace determination of pharmaceuticals and other wastewater-derived micropollutants by solid phase extraction and gas chromatography/mass spectrometry.

The presence of pharmaceuticals and other wastewater-derived micropollutants in surface and groundwaters is receiving intense public and scientific attention. Yet simple GC/MS methods that would enable measurement of a wide range of such compounds are scarce. This paper describes a GC/MS method for the simultaneous determination of 13 pharmaceuticals (acetaminophen, albuterol, allopurinol, amitriptyline, brompheniramine, carbamazepine, carisoprodol, ciclopirox, diazepam, fenofibrate, metoprolol, primidone, and terbinafine) and 5 wastewater-derived contaminants (caffeine, diethyltoluamide, n-butylbenzene sulfonamide, n-nonylphenol, and n-octylphenol) by solid phase extraction (SPE) and derivatization with BSTFA. The method was applied to the analysis of raw and treated sewage samples obtained from a wastewater treatment plant located in the mid-Atlantic United States. All analytes were detected in untreated sewage, and 14 of the 18 analytes were detected in treated sewage.

Aerobic biotransformation of polybrominated diphenyl ethers (PBDEs) by bacterial isolates.

Polybrominated diphenyl ethers (PBDEs) are flame retardants that have been used in consumer products and furniture for three decades. Currently, very little is known about their fate in the environment and specifically about their susceptibility to aerobic biotransformation. Here, we investigated the ability of the polychlorinated biphenyl (PCB) degrading bacteria Rhodococcus jostii RHA1 and Burkholderia xenovorans LB400 to transform mono- through hexa-BDEs at ppb levels. We also tested the PBDE transforming abilities of the related strain Rhodococcus sp. RR1 and the ether-degrading Pseudonocardia dioxanivorans CB1190. The two PCB-degrading strains transformed all of the mono- through penta-BDEs and strain LB400 transformed one of the hexa-BDEs. The extent of transformation was inversely proportional to the degree of bromination. Strains RR1 and CB1190 were only able to transform the less brominated mono- and di-BDE congeners. RHA1 released stoichiometric quantities of bromide while transforming mono- and tetra-BDE congeners. LB400 instead converted most of a mono-BDE to a hydroxylated mono-BDE. This is the first report of aerobic transformation of tetra-, penta,- and hexa-BDEs as well as the first report of stoichiometric release of bromide during PBDE transformation.

Structure of the toxaphene compound 2,5-endo,6-exo,8,9,9,10,10-octachlorobornene-2: a temperature-dependent formation of two rotamers.

The irradiation of 2,2,3-exo,5-endo,6-exo,8,9,9,10,10-decachlorobornane in n-hexane at 254 nm leads to a spontaneous Cl2 elimination as the major reaction pathway. This results finally in the main product 2,5-endo,6-exo,8,9,9,10,10-octachlorobornene-2, of which the structure could be elucidated with the help of X-ray, 1H and 13C NMR, IR, and MS. Temperature-dependent 1H NMR spectroscopic investigations have shown that the -CHCl2 groups located at C1 and C7 are able to rotate slowly under normal circumstances. If such measurements, however, are exerted at low temperatures (-10 to -60 degrees C), so can be seen that two rotamers are formed due to the hindrance of the free rotation about the bonds C1-C10, C7-C8, and C7-C9, which for the first time could be revealed for a toxaphene compound. Furthermore, as all 1H NMR chlorobornane spectra known so far show only sharp and clear signals, it can be assumed that chlorobornane compounds as main toxaphene components have fixed bonds, which requires to indicate chlorine atoms within the tentacles such as "a", "b", and "c" for characterizing the correct position. Those fixed tentacles are probably the reason that many toxaphene congeners remain very stable in environmental compartments, and particularly the biotic and abiotic transformation may strongly be hindered by the inflexibility of the tentacles.

Enrichment of the glycoalkaloids alpha-solanine and alpha-chaconine from potato juice by adsorptive bubble separation using a pH gradient.

For the first time, the solanidine alkaloids alpha-solanine and alpha-chaconine could be quantitatively enriched from potato juice by Adsorptive Bubble Separation (ABS) with a pH gradient. The enrichment into the foam was influenced by the pH value, bubble size, and gas flow rate. The efficiency was highest on using diluted samples with a concentration between 2 and 6 mg L(-1) of the alkaloids at pH 6.0. The experiments with a standard solution of each alkaloid confirmed that these substances can be quantitatively enriched into the 'spumat' without surface active potato proteins. The transfer into the foam fraction under these conditions was similar to that from the aqueous potato extract.