Small Volatile Compounds Research Articles

Helichrysum microphyllum subsp. tyrrhenicum: a possible correlation between small volatile compounds and phloroglucinol content

The chemical composition of the flowered aerial parts of four samples of Helichrysum microphyllum subsp. tyrrhenicum collected in South-West Sardinia was investigated with a combined focus on volatile constituents and phloroglucinols to find a possible correlation with the presence of arzanol endowed with the major anti-inflammatory activity. The volatile constituents were analysed by GC-MS as EO-HD and with HS-SPME identifying a total of 95 compounds of which 70 and 77 by EO-HD and HS-SPME respectively. The profile of the non-volatile phloroglucinols was investigated by HPLC-MS/MS. Arzanol concentrations ranged from 2.79 to 21.87 mg/g, helipyrone showed the same trend but in lower concentration. Surprisingly, leaves and stems contain higher concentration of phloroglucinols than the flowers. The concentration of arzanol was positively correlated to the one of γ-curcumene and ethylpyrone in the EO, while a negative correlation was observed with the monoterpene limonene and linalool as well as with the sesquiterpene 5-eudesmen-11-ol.

Stomatal closure prevents xylem transport of green leaf volatiles and impairs their systemic function in plants.

Plants perceive environmental stresses as whole organisms via distant signals conveying danger messages through their vasculature. In parallel to vascular transport, airborne plant volatile compounds, including green leaf volatiles (GLVs), can bypass the lack of vascular connection. However, some small volatile compounds move through the vasculature; such vascular transport is little known about GLVs. Here we illustrate GLV alcohols as solutes move within xylem vessels in Zea mays. We describe GLV alcohols, including Z-3-hexen-ol and its isomer E-3-hexen-ol, which is not synthesized in maize, moving through the transpiration stream via xylem vessels. Since transpiration is mediated by the stomatal aperture, closing stomata by two independent methods diminishes the transport of GLV alcohol and its isomer. In addition, the lower transport of GLV alcohols impairs their function in inducing terpenoid biosynthesis, suggesting that xylem transport of GLV alcohols plays a significant role in their systemic function. Our study suggests that GLV alcohols, in addition to airborne signals, are transported through xylem vessels. Our findings can be critical in future studies about the perception and function of these compounds in plants.

Early Prediction of Shiraz Wine Quality Based on Small Volatile Compounds in Grapes

Wine producers perform early wine quality prediction based on berry morphology, the taste of the berry and the measurement of basic chemical parameters. Incorporating analysis on grape and wine volatiles could potentially achieve a more accurate prediction of wine quality, but forming these models requires careful selection of grapes, controlled fermentations, and standardised quality assessment. Here, we present 3 models for the prediction of quality in Shiraz wine. Modelling was performed by general regression analysis with 4-fold cross-validation: Model 1 (R2 = 99.97% and 4-foldR2 = 97.61%) for prediction of wine quality from wine volatiles, Model 2 (R2 = 99.89% and 4-foldR2 = 98.42%) for early prediction of wine quality from free-bound and glycosidically bound grape volatiles, and Model 3 (R2 = 91.62% and 4-foldR2 = 80.21%) for the prediction of wine quality from free grape volatiles only. The accuracy of these models presents an advancement in the early prediction of wine quality and provides a valuable tool to assist grape growers and winemakers to support the understanding of quality in the vineyard to better direct scarce resources.

Laser Ablation Secondary Electrospray Ionization for In Situ Mass Spectrometric Interrogation of Acoustically-Levitated Droplets.

The composition of acoustically levitated droplets was probed by a novel combination of mid-IR laser evaporation and subsequent postionization via secondary electrospray ionization. The combination of microliter samples and subnanoliter sampling provided time-resolved interrogation of droplets and allowed for a kinetic investigation of the laser-induced release of the analyte, which was found to strongly depend on the analytes. The observed substance-specific delayed release of the analytes permitted baseline-separated discrimination of the analytes, ideal for the study of complex samples. The additionally applied postionization scheme was found to enable efficient detection of small volatile compounds as well as peptides. The detection of small molecules and peptides occurred under very different sampling geometries, pointing to two distinct underlying ionization mechanisms. Overall, our results suggest that the experimental setup presented in this study can serve as a widely applicable platform to study chemical reactions in acoustically levitated droplets as model reactors.

Can wine quality be predicted by small volatile compounds? A study based on performance of wine show entries and their volatile profiles

AbstractIt is widely recognized that wine aroma critically influences wine quality, yet the extent to which volatile compound composition in wine determines wine quality has proven difficult to define, beyond the negative influence of taint compounds. While some relationships between concentration of volatile aroma compounds and wine quality have been made, this has been usually based on single molecules, in laboratory wines and without reference to experienced wine panels. Here, we collected and analysed 157 commercial Shiraz wines from a competitive show over 2 years. We found significant (P < .05) correlations between specific volatiles and the panel's wine quality scores. However, these correlations were not always consistent between years. We could not explain medals given to wine entries solely based on multivariate analysis of their composite volatile composition. Here, we suggest that aroma determination in judging wine quality does not completely explain the assessment of wine, and appearance (colour, clarity) and taste may be more influential factors. There are considerable practical difficulties in establishing a convincing wine quality prediction model based on analysis of volatile compounds alone.

Glucose-6-P/phosphate translocator2 mediates the phosphoglucose-isomerase1-independent response to microbial volatiles.

In Arabidopsis (Arabidopsis thaliana), the plastidial isoform of phosphoglucose isomerase (PGI1) mediates photosynthesis, metabolism, and development, probably due to its involvement in the synthesis of isoprenoid-derived signals in vascular tissues. Microbial volatile compounds (VCs) with molecular masses of <45 Da promote photosynthesis, growth, and starch overaccumulation in leaves through PGI1-independent mechanisms. Exposure to these compounds in leaves enhances the levels of GLUCOSE-6-PHOSPHATE/PHOSPHATE TRANSLOCATOR2 (GPT2) transcripts. We hypothesized that the PGI1-independent response to microbial volatile emissions involves GPT2 action. To test this hypothesis, we characterized the responses of wild-type (WT), GPT2-null gpt2-1, PGI1-null pgi1-2, and pgi1-2gpt2-1 plants to small fungal VCs. In addition, we characterized the responses of pgi1-2gpt2-1 plants expressing GPT2 under the control of a vascular tissue- and root tip-specific promoter to small fungal VCs. Fungal VCs promoted increases in growth, starch content, and photosynthesis in WT and gpt2-1 plants. These changes were substantially weaker in VC-exposed pgi1-2gpt2-1 plants but reverted to WT levels with vascular and root tip-specific GPT2 expression. Proteomic analyses did not detect enhanced levels of GPT2 protein in VC-exposed leaves and showed that knocking out GPT2 reduced the expression of photosynthesis-related proteins in pgi1-2 plants. Histochemical analyses of GUS activity in plants expressing GPT2-GUS under the control of the GPT2 promoter showed that GPT2 is mainly expressed in root tips and vascular tissues around hydathodes. Overall, the data indicated that the PGI1-independent response to microbial VCs involves resetting of the photosynthesis-related proteome in leaves through long-distance GPT2 action.

Template Imprinting Versus Porogen Imprinting of Small Molecules: A Review of Molecularly Imprinted Polymers in Gas Sensing.

The selective sensing of gaseous target molecules is a challenge to analytical chemistry. Selectivity may be achieved in liquids by several different methods, but many of these are not suitable for gas-phase analysis. In this review, we will focus on molecular imprinting and its application in selective binding of volatile organic compounds and atmospheric pollutants in the gas phase. The vast majority of indexed publications describing molecularly imprinted polymers for gas sensors and vapour monitors have been analysed and categorised. Specific attention was then given to sensitivity, selectivity, and the challenges of imprinting these small volatile compounds. A distinction was made between porogen (solvent) imprinting and template imprinting for the discussion of different synthetic techniques, and the suitability of each to different applications. We conclude that porogen imprinting, synthesis in an excess of template, has great potential in gas capture technology and possibly in tandem with more typical template imprinting, but that the latter generally remains preferable for selective and sensitive detection of gaseous molecules. More generally, it is concluded that gas-phase applications of MIPs are an established science, capable of great selectivity and parts-per-trillion sensitivity. Improvements in the fields are likely to emerge by deviating from standards developed for MIP in liquids, but original methodologies generating exceptional results are already present in the literature.

Mammalian derived lipocalin and secretoglobin respiratory allergens strongly bind ligands with potentially immune modulating properties.

Allergens from furry animals frequently cause sensitization and respiratory allergic diseases. Most relevant mammalian respiratory allergens belong either to the protein family of lipocalins or secretoglobins. Their mechanism of sensitization remains largely unresolved. Mammalian lipocalin and secretoglobin allergens are associated with a function in chemical communication that involves abundant secretion into the environment, high stability and the ability to transport small volatile compounds. These properties are likely to contribute concomitantly to their allergenic potential. In this study, we aim to further elucidate the physiological function of lipocalin and secretoglobin allergens and link it to their sensitizing capacity, by analyzing their ligand-binding characteristics. We produced eight major mammalian respiratory allergens from four pet species in E.coli and compared their ligand-binding affinities to forty-nine ligands of different chemical classes by using a fluorescence-quenching assay. Furthermore, we solved the crystal-structure of the major guinea pig allergen Cav p 1, a typical lipocalin. Recombinant lipocalin and secretoglobin allergens are of high thermal stability with melting temperatures ranging from 65 to 90°C and strongly bind ligands with dissociation constants in the low micromolar range, particularly fatty acids, fatty alcohols and the terpene alcohol farnesol, that are associated with potential semiochemical and/or immune-modulating functions. Through the systematic screening of respiratory mammalian lipocalin and secretoglobin allergens with a large panel of potential ligands, we observed that total amino acid composition, as well as cavity shape and volume direct affinities to ligands of different chemical classes. Therefore, we were able to categorize lipocalin allergens over their ligand-binding profile into three sub-groups of a lipocalin clade that is associated with functions in chemical communication, thus strengthening the function of major mammalian respiratory allergens as semiochemical carriers. The promiscuous binding capability of hydrophobic ligands from environmental sources warrants further investigation regarding their impact on a molecule's allergenicity.

Analysis of genotoxic N-nitrosamines in active pharmaceutical ingredients and market authorized products in low abundance by means of liquid chromatography – tandem mass spectrometry

In 2018, high levels of the IARC class IIA carcinogen N-nitrosodimethylamine (NDMA) were analytically verified in the active pharmaceutical ingredient (API) valsartan, resulting in extensive regulatory action on angiotensin-II-receptor antagonists and recall of finished drug products by the pharmaceutical industry to ensure patient safety. The root cause of contamination was the unintended reaction of common reagents utilized during drug synthesis. This lead to serious effects on drug quality and immediate regulatory action. Thus, routine analysis of drug product contents are inevitable and necessitate thoroughly performed work up procedures of the product as well as adequate validated analytical methods. The nature of N-nitrosamines (NA), ranging from small, semi-volatile compounds up to highly polar molecules, effort sophisticated requirements in terms of instrumental analysis. Up today, gas as well as liquid chromatographic devices coupled to mass spectrometers are the most widespread systems for analysis. Gas chromatographic – mass spectrometric (GC-MS) systems, obviously superior towards liquid chromatography – mass spectrometry (LC-MS) for detecting small volatile compounds like NDMA, reach their limits for broadly designed studies including polar or acidic NA. In this study, a complementary and highly sensitive approach by means of liquid chromatography – tandem mass spectrometry (LC-MS/MS) is presented, including detection of 13 NA deduced from major classes of secondary amines. Thereby, the fully validated approach was performed in accordance to ICH and European Medicines Agency (EMA) guidelines. Quantitative proof-of-concept measurements with various APIs and market authorized tablets as representative drug formulations conclude applicability for further presumably contaminated substances. The approach employs organic or inorganic extraction steps with solid phase extraction (SPE). The limit of detection for the most prominent NA, NDMA and N-diethylnitrosamine (NDEA), were both 0.025 parts-per-billion (ppb) per matrix, respectively.

Action mechanisms of small microbial volatile compounds in plants.

Microorganisms communicate with plants by exchanging chemical signals throughout the phytosphere. Before direct contact with plants occurs, beneficial microorganisms emit a plethora of volatile compounds that promote plant growth and photosynthesis as well as developmental, metabolic, transcriptional, and proteomic changes in plants. These compounds can also induce systemic drought tolerance and improve water and nutrient acquisition. Recent studies have shown that this capacity is not restricted to beneficial microbes; it also extends to phytopathogens. Plant responses to microbial volatile compounds have frequently been associated with volatile organic compounds with molecular masses ranging between ~ 45Da and 300Da. However, microorganisms also release a limited number of volatile compounds with molecular masses of less than ~45Da that react with proteins and/or act as signaling molecules. Some of these compounds promote photosynthesis and growth when exogenously applied in low concentrations. Recently, evidence has shown that small volatile compounds are important determinants of plant responses to microbial volatile emissions. However, the regulatory mechanisms involved in these responses remain poorly understood. This review summarizes current knowledge of biochemical and molecular mechanisms involved in plant growth, development, and metabolic responses to small microbial volatile compounds.

Proteostatic Regulation of MEP and Shikimate Pathways by Redox-Activated Photosynthesis Signaling in Plants Exposed to Small Fungal Volatiles.

Microorganisms produce volatile compounds (VCs) with molecular masses of less than 300 Da that promote plant growth and photosynthesis. Recently, we have shown that small VCs of less than 45 Da other than CO2 are major determinants of plant responses to fungal volatile emissions. However, the regulatory mechanisms involved in the plants’ responses to small microbial VCs remain unclear. In Arabidopsis thaliana plants exposed to small fungal VCs, growth promotion is accompanied by reduction of the thiol redox of Calvin-Benson cycle (CBC) enzymes and changes in the levels of shikimate and 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway-related compounds. We hypothesized that plants’ responses to small microbial VCs involve post-translational modulation of enzymes of the MEP and shikimate pathways via mechanisms involving redox-activated photosynthesis signaling. To test this hypothesis, we compared the responses of wild-type (WT) plants and a cfbp1 mutant defective in a redox-regulated isoform of the CBC enzyme fructose-1,6-bisphosphatase to small VCs emitted by the fungal phytopathogen Alternaria alternata. Fungal VC-promoted growth and photosynthesis, as well as metabolic and proteomic changes, were substantially weaker in cfbp1 plants than in WT plants. In WT plants, but not in cfbp1 plants, small fungal VCs reduced the levels of both transcripts and proteins of the stromal Clp protease system and enhanced those of plastidial chaperonins and co-chaperonins. Consistently, small fungal VCs promoted the accumulation of putative Clp protease clients including MEP and shikimate pathway enzymes. clpr1-2 and clpc1 mutants with disrupted plastidial protein homeostasis responded weakly to small fungal VCs, strongly indicating that plant responses to microbial volatile emissions require a finely regulated plastidial protein quality control system. Our findings provide strong evidence that plant responses to fungal VCs involve chloroplast-to-nucleus retrograde signaling of redox-activated photosynthesis leading to proteostatic regulation of the MEP and shikimate pathways.

Exploring the substrate specificity of Cytochrome P450cin

Cytochromes P450 are enzymes that catalyse the oxidation of a wide variety of compounds that range from small volatile compounds, such as monoterpenes to larger compounds like steroids. These enzymes can be modified to selectively oxidise substrates of interest, thereby making them attractive for applications in the biotechnology industry. In this study, we screened a small library of terpenes and terpenoid compounds against P450cin and two P450cin mutants, N242A and N242T, that have previously been shown to affect selectivity. Initial screening indicated that P450cin could catalyse the oxidation of most of the monoterpenes tested; however, sesquiterpenes were not substrates for this enzyme or the N242A mutant. Additionally, both P450cin mutants were found to be able to oxidise other bicyclic monoterpenes. For example, the oxidation of (R)- and (S)-camphor by N242T favoured the production of 5-endo-hydroxycamphor (65–77% of the total products, dependent on the enantiomer), which was similar to that previously observed for (R)-camphor with N242A (73%). Selectivity was also observed for both (R)- and (S)-limonene where N242A predominantly produced the cis-limonene 1,2-epoxide (80% of the products following (R)-limonene oxidation) as compared to P450cin (23% of the total products with (R)-limonene). Of the three enzymes screened, only P450cin was observed to catalyse the oxidation of the aromatic terpene p-cymene. All six possible hydroxylation products were generated from an in vivo expression system catalysing the oxidation of p-cymene and were assigned based on 1H NMR and GC-MS fragmentation patterns. Overall, these results have provided the foundation for pursuing new P450cin mutants that can selectively oxidise various monoterpenes for biocatalytic applications.

Health Effects of Small Volatile Compounds from East Asian Medicinal Mushrooms

Medicinal fungi, taken whole or as various forms of extracts, have been used to alleviate, cure or prevent human ailments since pre-historic times. In particular, Asian cultures have incorporated a variety of mushrooms into their medical practices. Chemically pure, bioactive metabolites from fungi have been a mainstay of modern pharmacological research and in addition to antibiotics, include anticancer agents, immunosuppressants, enzyme inhibitors, antagonist and agonists of hormones, and a variety of psychotropic substances. However, to date not many studies have focused on the possible health benefits of odorant volatile organic compounds (i.e., gas phase compounds). An analysis of these compounds for their health related effects will expand the range of compounds available for the treatment of chronic and acute diseases. This review highlights phenolic acids and monoterpenes from Asian medicinal mushrooms (AMMs), which not only produce pleasant odors but also have antioxidant and antibacterial effects. Odorant bioactive volatile phase compounds from medicinal mushrooms remain an essentially untapped source for future medicines, and AMMs remain a promising resource for future pharmacological research.

The Lewis acidic and basic character of the internal HKUST-1 surface determined by inverse gas chromatography

Metal–organic frameworks (MOFs) have demonstrated great utilizability in separation applications, as in the separation of small volatile compounds via gas chromatography (GC). In the present work, HKUST-1 (Hong Kong University of Science and Technology), one of the best investigated MOFs, is used as a stationary phase for the gas chromatographic separation of various analytes possessing different modes of interaction due to their differences in polarity and the presence of free electron pairs. The system was investigated by inverse gas chromatography (IGC) to demonstrate in general how MOF materials can be quantitatively and qualitatively characterized in respect to their Lewis basic and acidic properties. Applying IGC theory, the investigation of the separation problem of benzene and its completely hydrogenated analogue cyclohexane was used to the determine the donor properties of the MOF linker benzene-1,3,5-tricarboxylic acid and the separation of diethyl ether, diisopropyl ether, tetrahydrofuran, and di-n-propyl ether to determine the acceptor properties of the coordinatively unsaturated sites of the copper(II)-secondary building unit (SBU), i.e. the nodal points of the MOF lattice.

Nanogram-Scale Preparation and NMR Analysis for Mass-Limited Small Volatile Compounds

Semiochemicals are often produced in infinitesimally small quantities, so their isolation requires large amounts of starting material, not only requiring significant effort in sample preparation, but also resulting in a complex mixture of compounds from which the bioactive compound needs to be purified and identified. Often, compounds cannot be unambiguously identified by their mass spectra alone, and NMR analysis is required for absolute chemical identification, further exacerbating the situation because NMR is relatively insensitive and requires large amounts of pure analyte, generally more than several micrograms. We developed an integrated approach for purification and NMR analysis of <1 µg of material. Collections from high performance preparative gas-chromatography are directly eluted with minimal NMR solvent into capillary NMR tubes. With this technique, 1H-NMR spectra were obtained on 50 ng of geranyl acetate, which served as a model compound, and reasonable H-H COSY NMR spectra were obtained from 250 ng of geranyl acetate. This simple off-line integration of preparative GC and NMR will facilitate the purification and chemical identification of novel volatile compounds, such as insect pheromones and other semiochemicals, which occur in minute (sub-nanogram), and often limited, quantities.

Characterisation by solid phase microextraction–gas chromatography–mass spectrometry of matrix changes of poly( l-lactide) exposed to outdoor soil environment

Films of poly( l-lactide), PLLA, were buried in outdoor environment in south Finland during two years. The changes in polylactide matrix were characterised by size exclusion chromatography and differential scanning calorimetry, and formation of volatile oligomers was monitored by solid phase microextraction (SPME)–gas chromatography–mass spectrometry. SPME sample preparation was found suitable for analysing small volatile compounds in solid PLLA samples. The oligomers in polylactide were readily extracted with polar CW/DVB fibre. The oligomers were analysed without any complicated sample handling procedure and no solvent was needed. Lactic acid, lactide and lactoyl lactic acid were extracted from the unaged and aged films. Lactide was observed in all films and its content did not change during the two years of soil burial. No lactic acid was observed in any of the polylactide films during the first year. After 20 months lactic acid and lactoyl lactic acid appeared as a result of hydrolysis. After 24 months the amount of lactoyl lactic acid decreased due to biotic hydrolysis of the film. The molecular weight M̄ n of polylactide increased and the polydispersity decreased during the first year. A slow chemical hydrolysis combined with the migration of small oligomers in soil caused the molecular weight changes for this period. During the second year of soil burial both M̄ n and M̄ w decreased and the polydispersity decreased towards 2. This indicates that a faster hydrolysis took place in the film. It was confirmed by the differential scanning calorimetry analysis of PLLA, which showed that during the second year in soil the thermal properties of the films ( T g and T m) were affected. PLLA was thus degraded by a combination of hydrolysis and biotic activity. In a first stage, after an induction period of 1 year, the film of PLLA undergoes hydrolysis. In a second stage microorganisms assimilate the small products of degradation while hydrolysis takes place.