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Determination of specific volatile organic compounds synthesised during Tuber borchii fruit body development by solid-phase microextraction and gas chromatography/mass spectrometry.

Fruit body development is a particular phase of the Tuber life cycle, characterised by the aggregation of different types of hyphae, i.e., vegetative hyphal cells and highly specialised reproductive hyphae (asci). In order to identify the volatile organic compounds (VOCs) produced in different stages of the Tuber borchii ripening fruit body, solid-phase microextraction with gas chromatography and mass spectrometry was used. The volatile organic compounds were extracted using a DVB/CAR/PDMS 50/30 microm fiber placed for 10 min at room temperature in the truffle headspace. The results obtained reveal 49 compounds each of which was present only in a particular stage of maturation. 1-octen-3-ol, aromadendrene, alpha-farnesene and other terpenoid compounds were of particular interest, and their possible biological roles are discussed. The production of aromadendrene in the completely unripe fruit body suggests the existence of communication events in the early stage of ascomata formation between the fungus and the host plant. alpha-Farnesene could represent a chemotactic attractant to saprophytic organisms in order to disperse the fungal spores in the environment. The identification of the VOCs produced by truffles during their maturation could give information about the processes underlying this phase of Tuber life cycle.

Ascomycota↗

Determination of microbial volatile organic compounds from Staphylococcus pasteuri against Tuber borchii using solid-phase microextraction and gas chromatography/ion trap mass spectrometry.

The mycelium of Tuber borchii Vittad., a commercial truffle species, is used as a model system for in vitro ectomycorrhizal synthesis, infected seedling production and biotechnological applications. Our fungal cultures were accidentally contaminated with a Staphylococcus pasteuri strain, showing a strong antifungal activity against T. borchii mycelium. In order to identify the antifungal volatile agents produced by S. pasteuri, solid-phase microextraction (SPME) with gas chromatography and mass spectrometry (GC/MS) was used. Using this method 65 microbial volatile organic compounds (MVOCs), synthesized by this bacterium in either single or in fungal-bacterial dual culture, were identified. SPME combined with GC/MS may be a useful method for the determination of MVOCs involved in the antifungal activity. These results showed that bacteria with unusual biological activities could be a major problem during large-scale production of inoculum for truffle-infected seedling.

Fungi↗

Influence of volatile compounds produced by yeasts predominant during processing of Coffea arabica in East Africa on growth and ochratoxin A (OTA) production by Aspergillus ochraceus.

The effects of volatile compounds produced during coffee processing by Pichia anomala, P. kluyveri and Hanseniaspora uvarum on growth of Aspergillus ochraceus and production of ochratoxin A (OTA) were studied. On malt extract agar (MEA) and on coffee agar (CA), exposure of A. ochraceus to the gaseous phase of malt yeast glucose peptone (MYGP) plates inoculated with P. anomala, P. kluyveri and H. uvarum inhibited fungal growth, with the two Pichia spp. showing the strongest effect. The main esters and alcohols produced by the three yeasts were ethyl acetate, isobutyl acetate, 2-phenyl ethyl acetate, ethyl propionate and isoamyl alcohol. The individual esters and alcohols were found to affect fungal growth. The most effective compound in inhibiting fungal growth was 2-phenyl ethyl acetate; which at 48 microg/l headspace completely inhibited growth of A. ochraceus. Exposure of A. ochraceus to the gaseous phase of MYGP plates inoculated with P. anomala, P. kluyveri and H. uvarum prevented production of OTA. On CA medium, only the headspace of P. anomala and P. kluyveri prevented OTA production. Furthermore, when A. ochraceus was exposed to the headspace of the individual volatile compounds, 2-phenyl ethyl acetate was the most effective in preventing OTA production. Prevention of OTA seems to be due to reduction of fungal biomass.

Africa, Eastern↗

A new mass screening method for methylmercury poisoning using mercury-volatilizing bacteria from Minamata Bay.

A simplified mass screening method for methylmercury exposure was developed using methylmercury-volatilizing bacteria from Minamata Bay. Some bacteria can transform methylmercury into mercury vapor. Most mercury in the hair is methylmercury, which is readily extracted with HCl solution. Black spots are formed on X-ray film due to the reduction of Ag(+) emulsion with mercury vapor produced by methylmercury-volatilizing bacteria. By exploiting these characteristics, a screening method was developed, whereby the fur of rats injected with methylmercury chloride formed clear black spots on X-ray film, whereas the fur of rats injected with saline did not. Subsequently, 50 human hair samples were examined using this mass screening method. The method identified people who had high mercury concentration, over 20 microg/g. A few thousand hair samples may be screened in a day using this method because it is rapid, simple, and economical. This method, therefore, enables screening of persons with methylmercury poisoning in mercury-polluted areas.

Animals↗

Determination of skin:air partition coefficients for volatile chemicals: experimental method and applications.

The partition coefficient (PC) of a chemical in skin is an indicator of the capacity for a chemical in the skin and may reflect the rate at which a chemical penetrates the skin and enters into systemic circulation. In this study we present a simple method to measure the skin:air PC for volatile organic chemicals. Important considerations in the development of this method for a skin:air PC were the effect of size and shape of the skin sample, initial chemical concentration, and time to equilibrium in the skin. Clipped, whole-thickness skin was obtained from the dorsal surface of 8- to 16-week-old male F-344 rats. After removal of the hypodermis, skin was cut into strips and placed on the side of a glass vial. An organic chemical vapor was introduced into a sealed sample vial (initial concentration before equilibration was 203 ppm) and a corresponding reference vial, which were equilibrated at 32 degrees C. Headspace concentrations at equilibrium were used to determine a skin:air PC value. After developing the technique using dibromomethane, a skin:air PC value was determined for perchloroethylene, trichloroethylene, benzene, hexane, toluene, m-xylene, styrene, methyl chloroform, methylene chloride, carbon tetrachloride, halothane, and isoflurane. The skin:air PC values correlated with previously determined vapor permeability constants but correlated poorly with octanol/water PC values. This method provides a screening technique for predicting skin penetration of volatile chemicals.

Air↗

A comparison of headspace entrainment on Tenax with solid phase microextraction for the analysis of the aroma volatiles of cooked beef.

Headspace solid phase microextraction (SPME), using a Carboxen-polydimethylsiloxane fiber, was compared with headspace entrainment on Tenax TA, as a means of examining the aroma constituents of cooked beef. Extracts were analyzed using gas chromatography-mass spectrometry. Headspace entrainment on Tenax extracted higher amounts of most volatiles, particularly low-boiling compounds. However, polar volatiles, such as hydroxyfuranones and hydroxyketones, were present at higher levels in the SPME extracts. Some components were present only in the SPME extracts, including norfuraneol, furaneol, and cyclotene, which are key Maillard reaction intermediates.

Animals↗

Volatile nitrosamines in the main stream smoke of black tobacco.

The cigarettes studied are smoked through an automatic device. The mainstream smoke is trapped in Citrate-Phosphate buffer pH 4.5 containing 20 mM ascorbic acid. The volatile nitrosamines are extracted in dichloromethane, purified on alumina column, then analysed by gas-chromatography with a thermal energy analyzer (TEA) detector. The two main volatile nitrosamines quantified are: Nitrosodimethylamine and Nitrosopyrrolidine.

Nitrosamines↗

Contamination of toiletries and cosmetic products with volatile and nonvolatile N-nitroso carcinogens.

Commercially available cosmetics and toiletries were analyzed for contamination with volatile and nonvolatile N-nitrosamines. Of a total of 145 samples analyzed 50 were found to contain N-nitrosodimethylamine (max. value found 24 micrograms/kg), 26 samples were contaminated with N-nitrosomorpholine (max. value found 640 micrograms/kg), and 25 samples contained N-nitrosodiethanolamine, a non-volatile carcinogen (max. value found 1400 micrograms/kg). These results are discussed and compared with other published data on NDE1A in cosmetics, with reference to potential human exposure and to possible preventive measures.

Carcinogens↗

Comparison of the effects of volatile anesthetics in varying concentrations on brain energy metabolism with brain ischemia in rats.

The effects of varying concentrations and types of volatile anesthetics on neurochemical sequelae of brain ischemia were evaluated in the rat. Rats were assigned to treatment defined by a 3 x 3 design (anesthetic type and dose) with 5 rats/cell. Each group received halothane, enflurane, or isoflurane 0.5, 1.0, or 2.0 MAC (minimal alveolar concentration). This was followed by preischemic plasma glucose sampling, 5 min hypotension (30 mmHg) and 5 min decapitation cerebral ischemia. Preischemia plasma glucose increased with increasing anesthetic concentration and was highest in the isoflurane groups, varying from a low (+/- SD) of 7.19 +/- 1.79 mumol/ml in the 0.5 MAC halothane group to a high of 12.68 +/- 3.65 mumol/ml in the 2.0 MAC isoflurane group. End-ischemic brain lactate correlated with preischemic plasma glucose (r = 0.5, alpha = 0.5). We conclude that increasing concentration of volatile anesthesia with iv phenylephrine blood pressure support produces higher levels of plasma glucose and brain lactate with cerebral ischemia.

Anesthetics↗

Effect of volatile anesthetics on the circular dichroism of bilirubin bound to human serum albumin.

The characteristic circular dichroism of bilirubin bound to human serum albumin undergoes a remarkable sign inversion on addition of halothane, chloroform and other volatile anesthetics. This sign inversion, which is completely reversed by removal of the anesthetic, reflects a pronounced conformational change of the bound ligand; probably a complete inversion of chirality. The observation suggests that association of volatile anesthetics with proteins can markedly alter the internal topography of receptor sites and potentially influence the stereoselectivity of ligand binding.

Anesthetics↗

Structural relationships between the endogenous volatile urinary metabolites of experimentally diabetic rats and certain neurotoxins (l).

High resolution glass capillary gas chromatography and GC/MS were utilized to examine qualitative and quantitative variations from normal of urinary volatile metabolites of long-term alloxan and streptozotocin diabetic rats. Volatile metabolites were structurally compared with known neurotoxins to examine any possible relationship between these metabolites and the development of the diabetic polyneuropathy.

Animals↗

Pyrrolizidine alkaloids, flavonoids and volatile compounds in the genus Eupatorium. Eupatorium cannabinum L., an ancient drug with new perspectives.

Within the scope of a study of antitumour compounds in higher plants a survey is given concerning the presence of pyrrolizidine alkaloids, flavonoids and volatile compounds in Eupatorium species. Preliminary results of a phytochemical study of these compounds in E. cannabinum are also presented. From the results of a GC-MS analysis of an alkaloid extract from aerial parts of E. cannabinum the conclusion can be drawn, that the composition of pyrrolizidine alkaloids is more complicated than reported in literature. This is caused by the fact that different stereoisomers exist. The presence of at least two alkaloids with a molecular weight of 283 (supinine or isomers) and four alkaloids with a molecular weight of 299 (echinatine or isomers) could be shown. In subterranean plant material also other pyrrolizidine alkaloids are present. A great number of flavonoids, also as glycosides, have been shown in Eupatorium species, often in low quantities. Rutin, present in many Eupatorium species, could not be detected in subterranean parts of E. cannabinum. Relatively little attention has been paid to the analysis of volatile compounds (essential oils) of Eupatorium species. Thymol derivatives are often reported to be present in Eupatorium species. Thirty-five compounds could be detected by means of a GC-MS analysis in the essential oil of E. cannabinum about which no literature data were available.

Chemical Phenomena↗

Effect of volatile and gaseous metabolites of germinating pea seeds on Micromycetes.

Differences in the effect of volatile and gaseous metabolites of germinating pea seeds on the germination of spores of Mucor racemosus and macroconidia of Fusarium oxysporum are described. Germination of spores of M. racemosus was inhibited by seed metabolites whereas germination of macroconidia of F. oxysporum was stimulated during the first two days and inhibition occurred only after further two days of germination of the seeds. A pronounced inhibition of germination of spores of both micromycetes took place due to absorption of CO2 from volatile and gaseous metabolites. Absorption of some components of seed metabolites in a KMnO4 solution led to a decrease of the inhibitory effect on germination of spores of M. racemosus and stimulatory effect on germination of macroconidia of F. oxysporum.

Absorption↗

Volatile hydrocarbons in the atmosphere of Athens, Greece.

This work presents the results of one-year monitoring study of Volatile Hydrocarbons, VHCs, in the atmosphere of Athens. It is the first systematic attempt to determine the VHC levels in the Athens' atmosphere with the very well known photochemical pollution problems. The purpose of this work was to create a database concerning VHCs in order to evaluate the photochemical pollution in this area (ozone creation, case studies and meteorology). Totally, 308 samples were collected at three different sites used in the state-monitoring programme involving the criteria pollutants. Air samples were collected on Tenax TA tubes and analysed by thermal desorption and dual column GC dual FID. Fifteen selected compounds were studied; 6 alkanes and 9 benzenoid compounds. The measured values of individual alkanes ranged from 0.39 pg m(-3) to 33 pg m(-3), and those of aromatics from 0.20 pg m(-3) to 616 pg m(-3). The sum of all 15 VHC concentrations ranged between 16 and 1697 pg m(-3). The time and spatial variations in the concentration of these compounds were assessed. Volatile hydrocarbons exhibited a clear seasonal and time cycle, showing higher concentrations during winter and early morning hours. Study of the spatial variations of VHC levels showed higher concentrations at the center of the city. The variation of toluene/benzene ratio and the correlation between VHCs, criteria pollutants (CO, NOx and O3) and meteorological parameters were also assessed. It was demonstrated that a trip-line of the VHCs concentration at the city center doubles the ozone concentration at peripheral areas under favourable meteorological conditions.

Air Pollutants↗

Leakage of volatile anaesthetics from agent-specific keyed vapourizer filling devices.

Agent-specific keyed vapourizer filling devices were designed to ensure that an anaesthetic vapourizer is filled with the correct agent. Since there appear to be no reports of possible loss of volatile agent or operating room pollution resulting from either the design or patterns of use of these devices, measurements were made with three anaesthetic agents and two methods of use. First, two bottles each of methoxyflurane, enflurane and halothane were fitted with a suitable filling device and the weight of agent lost from each bottle over six weeks was measured. Bottle #1 of each agent remained without agitation between weighings; bottle #2 was tipped to mimic filling of a vapourizer. Weight loss over the six week period was 2.76 and 3.15 per cent of the halothane, 2.22 and 2.43 per cent of the enflurane, and 0.58 and 0.96 per cent of the methoxyflurane, for bottles #1 and #2, respectively. Second, pollution was measured with an infra-red analyser for halothane, using bottles #1 and #2, as described above, and a third bottle on which the filling device was replaced by the screw-on cap after each filling of the vapourizer. Vapour loss was undetectable for bottle #1, between 25 and 30 ppm for bottle #2, and between 350 and 400 ppm for bottle #3. Thus, although the design of the filling devices results in loss of the anaesthetic agent, this loss represents potential pollution only when the device is replaced by the screw-on cap between use. Therefore, when using filling devices, these should be left on the bottle of volatile agent between fillings to decrease operating room pollution.

Air Pollution↗

Volatilization of mercury by resting mercury-resistant bacterial cells.

The mercuric ion reduction system encoded by the Hg2+ inducible mer operon confers bacterial resistance to mercuric ion. The mer A gene product which is a FAD-containing enzyme catalyzes the reduction of Hg2+ to volatile elemental mercury with the help of intracellular thiols and NADPH as a cofactor (Schottel 1974; Summers and Silver 1978; Fox and Walsh 1982; Misra 1992). Our earlier studies have shown that growing cells of different mercury-resistant bacteria reduce Hg2+ compounds to Hg(O) (Ray et al. 1989; Pahan et al. 1990a; Gachhui et al. 1989). We have also shown the effect of thiol compounds and flavins on mercury-degrading enzyme activities in mercury-resistant bacteria (Pahan et al. 1990b). Here we report that resting cells of mercury-resistant bacteria survive in a buffer system for several hours, synthesize inducible mercury-degrading enzymes and volatilize mercury from a mercury-containing buffer system. We know of no information regarding studies of mercury-degrading enzymes in resting mercury-resistant bacterial cells.

Azotobacter↗

Mutagenicity of N-nitrosodiethylamine in the Ames test with S. typhimurium TA1535 is due to volatile metabolites and is not dependent on cytochrome P4502E1 induction.

N-Nitrosodiethylamine (NDEA) is carcinogenic in all investigated animal species at relatively low dosages. No threshold has been detected for these carcinogenic effects. The substance has been extensively investigated in various in vitro systems, revealing only weak mutagenicity at relatively high dosages. We reinvestigated NDEA in the Ames test with Salmonella typhimurium TA1535 to establish appropriate modifications of the standard Ames test protocol, to achieve a dose-dependent mutagenic response at a reasonably low dose range. Two main modifications were evaluated. Since the metabolism of dialkylnitrosamines is postulated to be mainly dependent on cytochrome P4502E1, a pyrazole-induced rat liver S9 was applied. The second modification involved a gastight preincubation, since metabolites of NDEA might evaporate from the incubation mixture. Cytochrome P4502E1 induction in Wistar rats was achieved by pyrazole treatment. For comparison, a rat liver S9-fraction produced by beta-naphtoflavone/phenobarbital induction was used. N-Nitrosopyrrolidine served as positive control for pyrazole-induced S9-mix with TA1535. NDEA showed no mutagenic response under all test conditions in the presence of pyrazole-induced S9-mix. A strong mutagenic response, exceeding the base rate up to 15-fold at a dose range of 25-1000 microg/plate, was observed using beta-naphtoflavone/phenobarbital-induced S9-mix, gastight preincubation and TA1535. In conclusion the Ames test with gastight preincubation can be useful for the testing of volatile compounds or substances leading to gaseous metabolites. The weak response of NDEA in the Ames test observed previously seems mainly to be due to the volatile character of its mutagenic metabolites. Our results do not support the hypothesis that cytochrome P4502E1 is a major toxifying enzyme for the formation of Ames-test-positive metabolites from NDEA.

Animals↗

SPME analysis of volatile compounds from unfermented olives subjected to thermal treatment.

The effect of different types of thermal treatment, designed to increase the product's shelf-life, on the volatile composition of "Campo Real" unfermented table olives, has been studied by headspace solid-phase microextraction (HS-SPME) and GC-MS analysis. Different SPME fibres were evaluated to determine their selectivity for a mixture of the main components of the different spices used in "Campo Real" olive dressing. Of the different fibres investigated, the polydimethylsiloxane/divinylbenzene (PDMS/DVB) fibre was selected for analysis of the olive brines, which contained nine main aroma components. The types of thermal treatment were sterilisation (121 degrees C, 15 min) and four pasteurisation conditions (60 degrees C or 80 degrees C each for 5 or 9 min). Pasteurisation did not lead to significant changes in the amounts of these nine volatile compounds; the 2-butanol signal was reduced by treatment at 80 degrees C. On the other hand, sterilisation of the brine resulted in an decrease in the signals from these compounds and the appearance of a new, high signal for benzaldehyde; the origin of this has not yet been determined. Results suggest that the selected pasteurisation conditions do not significantly modify the typical, and valued, aroma characteristics of "Campo Real".

Benzenesulfonates↗