Controversy on health-based guidance values for bisphenol A-the need of criteria for studies that serve as a basis for risk assessment.

Since 2006, the responsible regulatory bodies have proposed five health-based guidance values (HBGV) for bisphenol A (BPA) that differ by a factor of 250,000. This range of HBGVs covers a considerable part of the range from highly toxic to relatively non-toxic substances. As such heterogeneity of regulatory opinions is a challenge not only for scientific risk assessment but also for all stakeholders, the Senate Commission on Food Safety (SKLM) of the German Research Foundation (DFG) analyzed the reasons for the current discrepancy and used this example to suggest improvements for the process of HBGV recommendations. A key aspect for deriving a HBGV is the selection of appropriate studies that allow the identification of a point of departure (PoD) for risk assessment. In the case of BPA, the HBGV derived in the 2023 EFSA assessment was based on a study that reported an increase of Th17 cells in mice with a benchmark dose lower bound (BMDL40) of 0.53 µg/kg bw/day. However, this study does not comply with several criteria that are important for scientific risk assessment: (1) the selected end-point, Th17 cell frequency in the spleen of mice, is insufficiently understood with respect to health outcomes. (2) It is unclear, by which mechanism BPA may cause an increase in Th17 cell frequency. (3) It is unknown, if an increase of Th17 cell frequency in rodents is comparably observed in humans. (4) Toxicokinetics were not addressed. (5) Neither the raw data nor the experimental protocols are available. A further particularly important criterion (6) is independent data confirmation which is not available in the present case. Previous studies using other readouts did not observe immune-related adverse effects such as inflammation, even at doses orders of magnitude higher than in the Th17 cell-based study. The SKLM not only provides here key criteria for the use of such studies, but also suggests that the use of such a "checklist" requires a careful and comprehensive scientific judgement of each item. It is concluded that the Th17 cell-based study data do not represent an adequate basis for risk assessment of BPA.

Commentary of the SKLM to the EFSA opinion on risk assessment of N-nitrosamines in food.

Dietary exposure to N-nitrosamines has recently been assessed by the European Food Safety Authority (EFSA) to result in margins of exposure that are conceived to indicate concern with respect to human health risk. However, evidence from more than half a century of international research shows that N-nitroso compounds (NOC) can also be formed endogenously. In this commentary of the Senate Commission on Food Safety (SKLM) of the German Research Foundation (DFG), the complex metabolic and physiological biokinetics network of nitrate, nitrite and reactive nitrogen species is discussed with emphasis on its influence on endogenous NOC formation. Pioneering approaches to monitor endogenous NOC have been based on steady-state levels of N-nitrosodimethylamine (NDMA) in human blood and on DNA adduct levels in blood cells. Further NOC have not been considered yet to a comparable extent, although their generation from endogenous or exogenous precursors is to be expected. The evidence available to date indicates that endogenous NDMA exposure could exceed dietary exposure by about 2-3 orders of magnitude. These findings require consolidation by refined toxicokinetics and DNA adduct monitoring data to achieve a credible and comprehensive human health risk assessment.

Quantification of odorous and potentially harmful substances in acrylic paint.

Within this study sixteen odour active substances (1-butanol, butyl acetate, 3-methyl-4-heptanone, butyl acrylate, styrene, ethylbenzene, propylbenzene, cumene, sec-butylbenzene, benzaldehyde, 2-ethylhexyl acrylate, naphthalene, 1-methylnaphthalene, 2-methylnaphthalene, 1,2-dimethylnaphthalene, 1,7-dimethylnaphthalene), that have previously been identified in acryl paints were quantified in ten black acrylic paint samples using GC-GC-MS (heart-cut two-dimensional gas chromatography-mass spectrometry) after prior extraction and purification via solvent assisted flavour evaporation. In this case, the preceding analysis of the most abundant odour active volatiles via GC-O (gas chromatography-olfactometry) provided indications on which smell properties might be linked with elevated and concerning levels of possibly harmful substances. Based on that work, four samples raised attention due to their mainly repelling or unpleasant odour with mainly aromatic, solvent-like, mothball-like or geranium-like smell impressions. The remaining samples either showed fruity/fermented, fruity/apple juice-like, mushroom-like or cocoa-like/earthy odour qualities. The repelling and unpleasant odour was generally linked to naphthalene or benzene derivatives that are of concern with regard to being potential carcinogens. With concentrations ranging as high as from 3.75 mg/kg to 143.41 mg/kg, n-butanol was found to be the odour active substance with the highest concentrations in all but one paint sample. The results showed that all paints contain different and specific patterns of the analysed odour active substances and volatiles. Whereas only two samples revealed no elevated concentration of any of the quantified substances, two samples (A9 and A10) showed elevated levels for all or nearly all substances. The concentrations for single substances were generally lower than the recommended exposure limit and therefore unlikely to cause any adverse effects with regards to toxicity and irritation as single constituents. However, as a group of substances these might exert adverse health effects due to combined or synergistic effects. Based on these observations, the occurrence of combinations of potentially carcinogenic substances in products which are occupationally used on a daily basis or are in direct skin contact should thus be regarded with care in the future.

Influence of elongation and desaturation on chemosensory properties in acrylates and their corresponding 1-alken-3-ones.

Acrylates as well as 1-alken-3-ones are both known to be odour active substances but are generally identified in different materials. Nonetheless, butyl acrylate and 1-octen-3-one were both found to elicit a similar mushroom-like odour in previous studies. This led to the question of whether acrylates and enones with the same overall chain length generally elicited similar odours and whether they had similar odour thresholds. Overall, most of the investigated substances showed a mushroom-like, geranium-like or fruity odour. In contrast, short chained substances elicited garlic-like, lighter gas-like or glue-like, odour qualities, suggesting a correlation between the odour quality and the overall chain length. The results showed that only between the analogue structures butyl acrylate and 1-octen-3-one as well as hexyl acrylate and 1-decen-3-one could similar odour qualities be observed. All investigated substances showed low odour threshold values in air between 0.0032 ng/lair (1-hexen-3-one) and 55 ng/lair (1-dodecen-3-one). Overall, 1-alken-3-ones revealed a higher dependency on the chain length when compared to their respective acrylates. The introduction of a second terminal double bond led to a decrease of OT values in case of the acrylates and to an increase in case of the ketones that neither contained a second terminal double bond nor a double bond located close to the carbonyl group. Despite their structural similarities, the results suggest that both substance classes are perceived in a different manner and are therefore likely to be recognized by different types of receptors or are related to different activation patterns in multi-receptor stimulation processes.

Smells like new car or rather like an old carriage? - Resolution of the decay behavior of odorants in vehicle cabins during usage.

The typical new car smell is not only perceived directly after vehicle delivery. Vehicle interiors maintain their characteristic odors for a period of time during use even though the gas composition of the vehicle interior changes due to external influences. To obtain deeper insights into the odorant composition of a passenger cabin, this study aimed at characterizing the gas phase of two vehicle interiors at defined time intervals after vehicle delivery, and use by a customer in a controlled environment using a targeted odorant analysis. Thereby, the decrease in the general emissions in the cars did not coincide with the decay behavior of the odorants due to the chemical characteristics such as polarity of the odorants. Identification of the odorants in the vehicle interior during use revealed three groups of odor contributors exhibiting different decay behaviors: (i) odorants vaporizing rapidly via elevated temperature; (ii) odorants released by continued diffusion from materials; and (iii) fragrance chemicals of the customer. After 23 weeks of vehicle use, octanal, p-chloro-m-cresol, nonanal, p-tert-butylphenol, γ-nonalactone, and unsaturated aldehydes and ketones represented the most important odorants in the vehicle interior constituting the investigated car odor. The results of a descriptive sensory analysis corresponded with the identified odorants.

Follow your nose - Traveling the world of odorants in new cars.

Volatile organic compounds of the vehicle interior are well investigated, but only limited information is available on the odorants of the passenger cabin. To close this gap, we aimed at specifically elucidating the odor, as a general proof of principle, of two new cars with different seat upholstery in a controlled environment using a targeted odorant analysis. In a first step, odor profiles were evaluated by a descriptive sensory analysis. Then, potent odorants of the passenger cabins were characterized by gas chromatography-olfactometry and ranked according to their odor potency via odor extract dilution analysis. Using this approach, 41 potent odorants were detected, and 39 odorants were successfully identified by two-dimensional gas chromatography-mass spectrometry/olfactometry. In a third step, important odorants of the vehicle interior were quantified by means of internal standard addition. The most dominant odorants could be assigned to several specific substance classes comprising esters, saturated and unsaturated aldehydes, unsaturated ketones, rose ketones, phenolic and benzene derivatives, and pyrazines, occurring in a concentration range between 0.05 and 219 ng/L in air. Of these potent odorants, the aldehydes 2-butylhept-2-enal, 2-propyloct-2-enal, and (Z)-2-butyloct-2-enal are reported here for the first time as odorants in the environment of a passenger cabin.

Odorant Metabolism in Humans.

Odorants are relatively small molecules which are easily taken up and distributed in the human body. Despite their relevance in everyday life, however, only a limited amount of evidence about their metabolism, pathways, and bioactivities in the human body exists. With this Review, we aim to encourage future interdisciplinary research on the function and mechanisms of the biotransformation of odorants, involving different disciplines such as nutrition, medicine, biochemistry, chemistry, and sensory sciences. Starting with a general overview of the different ways of odorant uptake and enzymes involved in the metabolism of odorants, a more precise description of biotransformation processes and their function in the oral cavity, the nose, the lower respiratory tract (LRT), and the gastrointestinal tract (GIT) is given together with an overview of the different routes of odorant excretion. Finally, perspectives for future research are discussed.

Sesquiterpenes and sesquiterpenoids harbor modulatory allosteric potential and affect inhibitory GABAA receptor function in vitro.

Naturally occurring compounds such as sesquiterpenes and sesquiterpenoids (SQTs) have been shown to modulate GABAA receptors (GABAA Rs). In this study, the modulatory potential of 11 SQTs at GABAA Rs was analyzed to characterize their potential neurotropic activity. Transfected HEK293 cells and primary hippocampal neurons were functionally investigated using electrophysiological whole-cell recordings. Significantly different effects of ß-caryophyllene and α-humulene, as well as their respective derivatives ß-caryolanol and humulol, were observed in the HEK293 cell system. In neurons, the concomitant presence of phasic and tonic GABAA R configurations accounts for differences in receptor modulation by SQTs. The in vivo presence of the γ2 and δ subunits is important for SQT modulation. While phasic GABAA receptors in hippocampal neurons exhibited significantly altered GABA-evoked current amplitudes in the presence of humulol and guaiol, negative allosteric potential at recombinantly expressed α1 ß2 γ2 receptors was only verified for humolol. Modeling and docking studies provided support for the binding of SQTs to the neurosteroid-binding site of the GABAA R localized between transmembrane segments 1 and 3 at the (+ α)-(- α) interface. In sum, differences in the modulation of GABAA R isoforms between SQTs were identified. Another finding is that our results provide an indication that nutritional digestion affects the neurotropic potential of natural compounds.

Isolation of sesquiterpenoids from Matricaria chamomilla by means of solvent assisted flavor evaporation and centrifugal partition chromatography.

The (semi)volatile fraction of Matricaria chamomilla L., an annual herbal plant from the family of Asteraceae, contains high quantities of sesquiterpenes and sesquiterpenoids. A method was developed to achieve isolation and separation of these compounds, using a combination of solvent assisted flavor evaporation (SAFE) and solid support-free liquid-liquid chromatography. The biphasic liquid solvent system n-heptane/ethyl acetate/methanol/water, 5/2/5/2 v/v/v/v (Arizona S) was elaborated as a suitable solvent system for the simultaneous separation of the target compounds. The lab-scale liquid-liquid chromatography separation performed in a countercurrent chromatography (CCC) column was successfully transferred to a semi-preparative centrifugal partition chromatography (CPC) column, which enabled the isolation of artemisia ketone, artemisia alcohol, α-bisabolone oxide A, and (E)-en-yn-dicycloether. α-Bisabolol oxide A and (Z)-en-yn-dicycloether co-eluted, but were successfully separated by subsequent size-exclusion chromatography (SEC). Similarly, spathulenol and α-bisabolol oxide B were obtained as a mixture, and were separated by means of column chromatography using silica gel as stationary phase. The isolated compounds were characterized by means of nuclear magnetic resonance spectroscopy (NMR) and gas chromatography-mass spectrometry (GC-MS).

Nosy confirmation: reconstitution of the characteristic odor of softwood via quantitative analysis and human sensory evaluation.

The significance of wood odors beyond flavoring effects for barbecues and wine aged in barrels has recently been under discussion. Wood has an immense influence on our physical and mental condition through our palate, such as positive health and mood-stimulating effects. As a result of increased public interest, the key odorants from various natural wood species were recently decoded. To gain profound insights into the contribution of single odorants to the overall scent of distinct wood types, this study compares several softwood species (Scots pine, incense cedar, European larch, Norway spruce, and white fir) by odor profile analysis. Nine odor-active constituents, which were previously detected in those woods, were quantified using stable isotope dilution analysis (SIDA). Odor activity values (OAVs) calculated on the basis of odor thresholds (OTs) determined in cellulose revealed hexanal, octanal, (E)-non-2-enal, p-cresol, vanillin, and thymoquinone as dominant odorants for wood odor. Recombination experiments by mixing the odorants in their naturally occurring concentrations in a cellulose matrix confirmed the successful characterization of the key odorants for Scots pine and incense cedar wood.

Structure-Odor Activity Studies on Derivatives of Aromatic and Oxygenated Monoterpenoids Synthesized by Modifying p-Cymene.

Thymoquinone was recently reported as having a unique pencil-like odor and being the impact compound for the cedar-like and cedar wood-based product smell such as pencils. The compounds thymol and carvacrol are structurally related odorants commonly found in plants and foods such as thyme and oregano, also having a significant contribution to their overall aroma. However, a systematic elucidation of the sensory properties in this class of oxygenated, aromatic monoterpenoids has not been carried out. To close this gap and gain new insights into structure-odor relationships leading to pencil-like and woody odors, 19 structurally related derivatives of p-cymene starting from thymol and carvacrol were synthesized and characterized. The compounds had odor thresholds ranging from 2.0 ng/L air to 388.8 ng/L air, being lowest for thymol and carvacrol and highest for thymohydroquinone. The compounds smelled mostly thyme-like, oregano-like, and pencil-like with phenolic, earthy, and medicinal variations in their odor character, which could be successfully linked to structural motifs.

Identification and characterisation of odorants in a squishy toy using gas chromatography-mass spectrometry/olfactometry after thermal extraction.

Soft, squashable toys known as squishies have become increasingly popular amongst children. In this study, one such toy was evaluated sensorially by a trained panel and analytically using gas chromatography-mass spectrometry/olfactometry (GC-MS/O) after thermal extraction of the sample. Sensory analysis revealed the presence of an intense and unpleasant odour exhibited by the sample. The smell was dominated by almond- and inflatable swimming aid-like, as well as malty and glue-like notes, but also pleasant odours that were described as caramel-like and coconut-like. GC-MS/O analysis identified 2-butoxyethanol, cyclohexanone, γ-nonalactone, and ethyl maltol as being the main causative substances for the overall odour of the product. The data additionally indicated that the pleasant smelling substances γ-nonalactone (coconut-like smell) and ethyl maltol (caramel-like smell) were intentionally added by the manufacturer to mask the unpleasant odour of the solvents.

Correlations between odour activity and the structural modifications of acrylates.

Acrylates (acrylic esters) are versatile monomers that are widely used in polymer formulations because of their highly reactive α,ß-unsaturated carboxyl structure. Commonly used acrylates such as butyl acrylate are known to emit a strong unpleasant odour, and the monomers are therefore potential off-odorants in acrylic polymers. However, up to now, the odour properties of structurally related acrylic esters have not been characterised in detail. To obtain deeper insights into the smell properties of different acrylates, we investigated the relationship between the molecular structure and odour thresholds as well as the odour qualities of 20 acrylic esters, nine of these synthesised here for the first time. The OT values of 16 acrylates fell within the range from 0.73 to 20 ng/Lair, corresponding to a high-odour activity. Moreover, sec-butyl acrylate and 2-methoxyphenyl acrylate showed even lower OT values of 0.073 and 0.068, respectively. On the other hand, the OT values of the hydroxylated acrylates 2-hydroxyethyl acrylate and 2-hydroxypropyl acrylate were 5-244 times higher than those of the other compounds, demonstrating that the presence of a hydroxyl group obviously favours odour inactivity.

Structural changes at the myrtenol backbone reverse its positive allosteric potential into inhibitory GABAA receptor modulation.

GABAA receptors are ligand-gated anion channels that form pentameric arrangements of various subunits. Positive allosteric modulators of GABAA receptors have been reported as being isolated either from plants or synthesized analogs of known GABAA receptor targeting drugs. Recently, we identified monoterpenes, e.g. myrtenol as a positive allosteric modulator at α1ß2 GABAA receptors. Here, along with pharmacophore-based virtual screening studies, we demonstrate that scaffold modifications of myrtenol resulted in the loss of modulatory activity. Two independent approaches, fluorescence-based compound analysis and electrophysiological recordings in whole-cell configurations were used for analysis of transfected cells. C-atoms 1 and 2 of the myrtenol backbone were identified as crucial to preserve positive allosteric potential. A modification at C-atom 2 and lack of the hydroxyl group at C-atom 1 exhibited significantly reduced GABAergic currents at α1ß2, α1ß2γ, α2ß3, α2ß3γ and α4ß3δ receptors. This effect was independent of the γ2 subunit. A sub-screen with side chain length and volume differences at the C-atom 1 identified two compounds that inhibited GABAergic responses but without receptor subtype specificity. Our combined approach of pharmacophore-based virtual screening and functional readouts reveals that side chain modifications of the bridged six-membered ring structure of myrtenol are crucial for its modulatory potential at GABAA receptors.

Quantification of organic solvents in aquatic toys and swimming learning devices and evaluation of their influence on the smell properties of the corresponding products.

Based on the observation that the characteristic odour of inflatable aquatic toys for children is predominantly caused by residues of hazardous organic solvents, the concentrations of cyclohexanone, isophorone and phenol were determined in a selection of 20 products obtained from online suppliers located in Germany. Analytes were extracted with dichloromethane after the addition of non-labelled internal standards, and the volatile fraction was isolated using solvent-assisted flavour evaporation (SAFE). Extracts were then concentrated by Vigreux distillation and analysed by means of gas chromatography with mass spectrometric detection (GC-MS). Furthermore, each sample was evaluated regarding its specific olfactory properties by an expert sensory panel. While some samples did not contain significant amounts of solvents, cyclohexanone concentrations above the lower limit of quantification (LLOQ) were determined in nine samples with six samples containing high concentrations ranging from about 1 to 7 g/kg cyclohexanone. Isophorone concentrations above the LLOQ were observed in eight samples. Thereby, six products contained between 0.3 and 1.6 g/kg isophorone and the remaining two samples contained even about 5 g/kg isophorone, each. Likewise, phenol concentrations exceeded the LLOQ in 14 cases, with four samples containing elevated amounts ranging from about 140 to 280 mg/kg phenol.

Identification and quantification of glue-like off-odors in elastic therapeutic tapes.

Elastic therapeutic tapes are an important tool in the field of physical therapy and medicine. These tapes contain types of adhesive. However, sensory evaluations revealed the release of pronounced and irritating odors of the tapes. Negative odors were, amongst others, reported in elastic therapeutic tapes containing acrylic adhesives. In this study, the odor of four different tape samples was evaluated applying a descriptive analysis approach carried out by a trained sensory panel. Afterwards, the volatile compounds were recovered from the samples by solvent extraction and isolated by solvent-assisted flavor evaporation (SAFE). The obtained distillates were subsequently analyzed by gas chromatography-olfactometry (GC-O) and two-dimensional GC-O coupled with mass spectrometry (2D-GC-MS/O). To determine the most potent odorants in the distillates, odor extract dilution analyses (OEDA) were carried out. Thirty-one odorants were successfully identified using this approach, which were all described for the first time as odorants in tapes. Amongst the set of volatiles, unsaturated and saturated aldehydes were present, eliciting fatty, soapy, and citrus-like odor impressions, as well as a range of glue-like, moldy, and fruity smelling odor-active volatiles, such as 2-ethyl-1-hexanol, butyl benzoate, and 3-phenyltoluene. Based on their relative intensities, the concentrations of the glue-like smelling substances were determined: 2-ethyl-1-hexanol, present in all samples, was determined with concentrations ranging from 10 to 200 mg/kg in the investigated tapes.

Identification of odorous compounds in oak wood using odor extract dilution analysis and two-dimensional gas chromatography-mass spectrometry/olfactometry.

Over the centuries, oak wood has been used in the maturation process of alcoholic beverages imparting aroma and flavor notes. Whereas several studies have dealt with the impact of oak wood on the chemical composition of, for example, wine aroma, only limited information is available on the odorant composition of unmodified and raw oak wood itself. To close this gap, a combination of human sensory and chemo-analytical techniques was applied for the elucidation of the chemical composition of oak odor, comprising extraction of the volatile fraction of oak wood by means of solvent-assisted flavor evaporation (SAFE) and subsequent mild concentration of the distillate. Odor extract dilution analysis (OEDA), which is based on gas chromatography-olfactometry (GC-O), was then applied for the targeted characterization of the odor-active compounds. Overall, a total of 97 odorants was identified via gas chromatography-mass spectrometry/olfactometry (GC-MS/O) and heart-cut two-dimensional gas chromatography-mass spectrometry/olfactometry (2D-GC-MS/O). The majority of these odorants comprised a series of terpenes, mainly mono- and sesquiterpenes, aldehydes, acids, and lactones, as well as a number of odorants containing a phenolic core moiety. Several odorants are reported here for the first time as volatile organic compounds in oak wood. Identification of the molecular composition of oak wood odor helps to establish a better understanding of the distinctive smell of oak wood, and offers the basis for unveiling its potential effects on humans when being exposed to oak wood smell in daily life. Graphical abstract ᅟ.

Development and Validation of a Food-Associated Olfactory Test (FAOT).

Olfactory tests are an important tool in human nutritional research for studying food preferences, yet comprehensive tests dedicated solely to food odors are currently lacking. Therefore, within this study, an innovative food-associated olfactory test (FAOT) system was developed. The FAOT comprises 16 odorant pens that contain representative food odors relating to different macronutrient classes. The test underwent a sensory validation based on identification rate, intensity, hedonic value, and food association scores. The accuracy of the test was further compared to the accuracy of the established Sniffin' Sticks identification test. The identification rates and intensities of this new FAOT were found to be comparable to the Sniffin' Sticks olfactory identification test. The odorant pens were also assessed chemo-analytically and were found to be chemically stable for at least 24 weeks. Overall, this new identification test for use in assessing olfaction in a food-associated context is valid both in terms of its use in sensory perception studies and its chemical stability. The FOAT is particularly suited to examinations of the sense of smell regarding food odors.

Resolving the chemical structures of off-odorants and potentially harmful substances in toys-example of children's swords.

Most children's toys on the market are primarily made out of plastic and other complex composite materials. Consumer complaints about offensive odors or irritating effects associated with toy products have increased in recent years. One example is the strongly perceivable negative odor reported for a particular series of toy swords. Characterizing the presence of contaminants, including those that have the potential to be deleterious to health, in such products is a significant analytical challenge due to the high baseline abundance of chemical constituents of the materials used in the products. In the present study, the nature of offensive odorants associated with toy sword products was examined by gas chromatography (GC). After initial sensory evaluations, the volatile compounds from the toy products were recovered using solvent extraction and solvent-assisted flavor evaporation. The extracts were analyzed using GC-olfactometry (GC-O) and two-dimensional GC-O coupled with mass spectrometry (GC-GC-MS/O). A total of 26 odor-active compounds, including aromatic hydrocarbons and phenols, were identified among numerous non-odorous volatile by-products. These substances also included polycyclic aromatic hydrocarbons, which were analyzed by GC-MS. Representative substances were naphthalene and 1,2-dihydronaphthalene that exhibited moldy, mothball-like odor impressions, and phenol derivatives with leather-like, phenolic, horse-stable-like smells. The odorants detected correlated with the assigned attributes from the sensory analyses. This study clearly shows that the detection and identification of such odorous contaminants can provide key indications of potentially harmful yet unknown substances in everyday products such as toys. Graphical abstract ᅟ.