Effect of the Compositions of Sample and Polymer Sorbents on the Extraction of Volatile Compounds by Solid-Phase Microextraction

A comparative study was performed by solid phase microextraction and capillary gas chromatography to establish the ability of four polymer sorbents of different compositions to extract and concentrate volatile organic compounds from the gas phase above an aqueous solution. All polymer sorbents sorbed nonpolar monoterpene hydrocarbons via a cooperative mechanism with almost equal and high efficiency. Sorbents based on polymethyl disiloxane and its mixture with divinylbenzene were more effective in extracting acetates and sesquiterpenes. As the concentration of these compounds in the gas phase increased, their binding by sorbents decreased. It was found that the determination of polar compounds depended on the presence of a solvent in the system. Compounds that are highly soluble in water (alcohols, ketones, etc.) had low coefficients of distribution between gas and water phases. Consequently, their sorption to any of the polymer sorbents was negligible. In the absence of the solvent, the degree of their extraction from the gas phase above the sample was high. It was shown that the actual composition of compounds in the initial mixture of essential oils could significantly differ from their composition in the gas phase. This method is convenient and informative for the purpose of studying the composition of volatile compounds in the gas phase that determine the flavor of the product.     

The Influence of Oxygen on the Production of Volatile Compounds by Dipodascus aggregates

The production of volatile compounds by Dipodascus aggregatus was studied in relation to the oxygen concentration in the medium. Oxygen concentration was determined with a Clark oxygen electrode and volatile compounds in the atmosphere above the culture by a gas chromatographic technique. Shake cultures of the fungus in its stationary phase of growth were very sensitive to a decrease in oxygen concentration in the presence of residual glucose. Anaerobic conditions induced production of volatile compounds that continued for many hours. The pattern of production of volatile compounds observed under conditions of low oxygen concentration during the stationary phase of growth differed from that obtained under aerobic conditions during the exponential phase of growth.     

The influence of different nitrogen sources on the production of volatile compounds by Dipodascus aggregatus

Summary The yeast fungus Dipodascus aggregatus was grown aerobically on 9 different nitrogen sources and the production of volatile compounds determined by a gas chromatographic head-space technique. Excellent growth was supported by glutamine, aspartic acid, asparagine, (NH₄)₂-tartrate and NH₄H₂PO₄. Valine, leucine, and particularly isoleucine were utilized with a somewhat lower growth rate. Lysine was rapidly utilized after a prolonged lag phase.The highest production of volatile compounds was obtained from leucine and isoleucine. At least 20 volatile compounds were formed from each of them and many products were detected in high concentrations. Intermediate amounts of volatile compounds were produced from asparagine, the ammonium salts and valine, and low amounts from lysine, glutamine and aspartic acid.Ethyl acetate was a major product irrespective of the nitrogen source used. Regarding the pattern of volatile compounds produced, leucine, isoleucine and valine had much in common. Most of the volatile products formed from these amino acids contained a branched carbon chain and at least three high-boiling components eluted later than n-amyl acetate from the gas chromatographic column. The other six nitrogen sources could be grouped together. In general the same volatile compounds were formed from these sources, but the quantities of the individual compounds differed. Only one component eluted later than n-amyl acetate. No basic difference in production of volatile compounds was observed between the ammonium salts and -amino compounds like lysine and asparagine.     

Liquid chromatographic methods for the determination of endogenous nucleotides and nucleotide analogs used in cancer therapy: A review

Endogenous ribonucleotides and deoxyribonucleotides play a crucial role in cell function. The determination of their levels is of fundamental interest in numerous applications such as energy metabolism, biochemical processes, or in understanding the mechanism of nucleoside analog compounds. Nucleoside analogs are widely used in anticancer therapy. Their mechanisms of action are related to their structural similarity with natural nucleotides. Numerous assays have been described for the determination of endogenous nucleotides or anticancer nucleotide analogs in different matrices such as cellular cultures, tissue or peripheral blood mononuclear cells. The determination of these compounds is challenging due to the large difference of concentrations between ribonucleotides and deoxyribonucleotides, the presence of numerous endogenous interferences in complex matrices and the high polarity of the molecules due to the phosphate moiety. The extraction was generally performed at low temperature and was based on protein precipitation using acid or solvent mixture. This first phase could be coupled with extraction or cleaning step of the supernatant. Liquid chromatography coupled with UV detection and based on ion-exchange chromatography using non-volatile high salt concentrations was largely described for the quantification of nucleotides. However, the development of LC–MS and LC–MS/MS during the last ten years has constituted a sensitive and specific tool. In this case, analytical column was mostly constituted by graphite or C18 stationary phase. Mobile phase was usually based on a mixture of ammonium buffer and acetonitrile and in several assays included a volatile ion-pairing agent. Mass spectrometry detection was performed either with positive or negative electrospray mode according to compounds and mobile phase components. The purpose of the current review is to provide an overview of the most recent chromatographic assays (over the past ten years) developed for the determination of endogenous nucleotides and nucleotide analogs used in cancer therapy. We focused on sample preparation, chromatographic separation and quantitative considerations.     

High-resolution gas chromatographic profiles of volatile organic compounds produced by microorganisms at refrigerated temperatures.

Three different strains of bacteria isolated from spoiled, uncooked chicken were grown in pure culture on Trypticase soy agar supplemented with yeast extract. The volatile organic compounds produced by each culture were concentrated on a porous polymer precolumn and analyzed by high-resolution gas chromatographic mass spectrometry. Twenty different compounds were identified. Both qualitative and quantitative differences in the chromatographic profiles from each culture were found.     

High-resolution gas chromatographic profiles of volatile organic compounds produced by microorganisms at refrigerated temperatures.

Three different strains of bacteria isolated from spoiled, uncooked chicken were grown in pure culture on Trypticase soy agar supplemented with yeast extract. The volatile organic compounds produced by each culture were concentrated on a porous polymer precolumn and analyzed by high-resolution gas chromatographic mass spectrometry. Twenty different compounds were identified. Both qualitative and quantitative differences in the chromatographic profiles from each culture were found.     

A gas chromatographic investigation of the influence of different carbon sources on the production of volatile compounds by Dipodascus aggregatus

Summary The yeast fungus Dipodascus aggregatus was cultivated aerobically on different carbon sources. The growth was measured turbidimetrically and related to the simultaneus production of volatile compounds, which was determined by a gas chromatographic head-space technique. The same culture could be analyzed many times and up to 15 components were detected in a chromatogram recorded in 18 min. The main peaks were identified and the chromatograms quantitatively evaluated by peak height measurements.All the carbon sources tested were utilized for growth even if the lag phase was prolonged on xylose and ethanol. The production of volatile compounds from the different carbon sources decreased as follows; ethanol>glucose>fructose>glycerol >xylose and succinic acid. A good carbon source for growth could be unsuitable for the formation of volatile products and vice versa.The time course production of volatile components was recorded. On each separate carbon source the formation of volatiles was correlated to growth until the end of the exponential phase of growth.An extraction procedure including the addition of an internal standard was used to determine the exact concentrations of 9 components at near optimum production. The amounts varied between 0.1–32 mg/l medium when the fungus was grown on 5% glucose.     

Supramolecular interactions of solid human serum albumin with binary mixtures of solvent vapors.

Sorption isotherms of organic compounds on solid human serum albumin (HSA) from binary vapor mixtures were determined by gas chromatographic headspace analysis. The shape of sorption isotherms depends on molecular structure of studied sorbates. The 'active' compounds capable to sorb effectively on dry HSA increase the sorption of 'passive' compounds unable to be sorbed by dry HSA in absence of the third component. The critical hydration of HSA is required for sorption activation of 'passive' sorbates if water is taken as 'active' component. Ethanol and acetonitrile exhibit such activation effect without threshold. 'Passive' sorbates are able to produce cooperative activation effect on the sorption of 'active' component. Hydration history effect is observed for sorption on prehydrated HSA and HSA hydrated in situ. Obtained results were interpreted in terms of clathrate formation by 'passive' sorbate (substrate) and 'active' component inside the HSA (receptor) binding centers.     

Sorption of components from a mixture of essential oils by cryotextured cornstarches

Sorption by cryotextured cornstarches of components of the aqueous phase of a mixture of essential oils was studied by capillary gas chromatography. The amount of cryotexture-sorbed substances depended linearly on their concentration in the initial gel. The sorption of components from the mixture by starch polysaccharides was mainly associated with hydrophobic cooperative interactions, which resulted in the formation of supramolecular structures and inclusion complexes. The structure of the compounds was a major factor determining the degree of sorption. Sorption of monoterpene carbohydrates was the most pronounced. We revealed a synergistic increase in the degree of sorption from the mixture as compared to binding of individual compounds.     

Enantiomer separation of 1,4-sulfanylalcohols by conventional and low-temperature gas chromatography

Since 1990, the family of organosulfur molecules has assumed increasing importance in pheromone, flavour, and fragrance chemistry. Depending on the constitution of the functional groups, various volatile sulfur-containing compounds are chiral. However, hitherto it has been impossible to study the chirality of 1,4-sulfanylalcohols, since no adequate enantioselective analytical technique has been available. Here we report on the enantiomer separation of ten volatile 1,4-sulfanylalcohol homologues by applying an heptakis(6-O-tert-butyldimethylsilyl-2,3-di-O-methyl)-beta-cyclodextrin phase, involving in some cases the use of a low-temperature gas chromatographic (ltGC) technique. The results are expected to open research potential on the asymmetry of volatile organosulfur molecules, particularly in the fields of pheromone, flavour, and fragrance research.     

Comparison of volatile leaf compounds and herbivorous insect communities on three willow species

 Plants produce volatile compounds known to influence insect preferences for oviposition and feeding. To examine whether volatile leaf compounds are correlated with the herbivorous insect community, we analyzed volatile compounds in leaves from three co-occurring willow species, Salix serissaefolia, S. eriocarpa, and S. integra, and investigated their associated insect communities in 3 months across different years. The gas chromatographic profiles of volatile compounds were highly specific to each willow species and remained constant in the study months. In a comparison between the chemical composition of the volatile compounds and the taxon composition of the insect communities, dissimilarity patterns in chemical composition among the three willow species were very close to those in herbivorous insect communities. These findings indicate that willow leaves produce specific volatiles that are highly correlated with the community structure of herbivorous insects associated with them. Received: October 10, 2002 / Accepted: March 17, 2003     

爱玉子花序挥发物成分以及对其传粉小蜂的吸引作用

利用动态顶空吸附法和GC-MS分析,研究接受期以及传粉或产卵5d后爱玉子(Ficus awkeotsang)雌、雄隐头花序挥发物成分。研究结果表明:爱玉子隐头花序挥发物的组成成分复杂,种类繁多,主要成分是对传粉小蜂具有显著吸引作用的芳樟醇。组成雌、雄花序的接受期挥发物信号的化合物种类与数量不同,但两者之间的共有化合物却占花序挥发物总量的70%以上,且接受期雌、雄花序挥发物组成比例相似,小蜂无法通过花序挥发物信号的差异,区别雌、雄花序,从而支持"榕树雌、雄花序接受期挥发物存在相互模拟现象"的假说。爱玉子传粉(或产卵)前后花序挥发物信号发生变化,表现在花序在传粉(或产卵)后,一些挥发性化合物在榕果中消失或相对含量减少(如芳樟醇),有些化合物的相对含量增加(如苯甲酸甲酯),并出现新的化合物(如2-乙基己醇);在化合物组成上,萜类化合物相对含量下降,芳香族化合物、脂肪族化合物相对含量上升。嗅觉仪实验表明爱玉子接受期雌、雄花序挥发物对其传粉榕小蜂有极显著的吸引作用,而传粉(或产卵)5d后的雌、雄花序挥发物对传粉榕小蜂有显著的趋避作用。传粉小蜂对雌、雄接受期花序挥发物的选择没有偏向性。榕果通过挥发物的释放量和成分的改变反映出花发育与被授粉(被产卵)状况,传粉小蜂通过接收榕果发出的化学信息,判断选择适合的寄主。研究对于爱玉子的高产栽培以及植物与昆虫专性共生化学生态学机制的理论研究具有重要的意义。     

Three mammal species distinction through the analysis of scats chemical composition provided by comprehensive two-dimensional gas chromatography

In the last years, new approaches use scats analysis in ecological studies, like gas chromatographic techniques, has been developed. The aim of this study is to use VOCs (volatile organic compounds) released from scats to distinguish three mammal species: the ungulates red and fallow deer and the carnivore Eurasian otter through the application of SPME/GC × GC–ToFMS. This chromatographic technique has never been used in ecological studies. Chromatogram contour plot was effective in distinguishing between the carnivore Eurasian otter and the two ungulates. Additionally, a subset of 21 selected compounds, including hydrocarbons, ketones, aldehydes, alcohols and terpenes, allowed a distinction between Eurasian otter, red deer and fallow deer. Eurasian otter scats were characterized mainly by the presence of aldehydes that may arise from oxidative degradation of unsaturated fatty acids present in fish species, included in their diet. Red deer volatile profile is characterized by terpenes compounds, presumably related with diet, while fallow deer achieved also chemical communication markers: 2-propanone and hexane. The scat volatile profiles provide information that explains their odour and also the type of diet (herbivorous or carnivorous) and intra-specific communication through chemical signals. Being able to distinguish species using the proposed methodology is an asset to wildlife ecology and management studies.     

Production of volatile organic compounds by the yeast fungus Dipodascus aggregatus

Summary The yeast fungus Dipodascus aggregatus was grown aerobically in a synthetic nutrient solution and the volatile compounds produced were concentrated. Identification of the volatiles was performed by combined gas chromatographymass spectrometry or by one of these methods. The compounds identified were 11 esters, 9 alcohols, 5 acids and 3 carbonyls.The time course production of volatile neutral compounds was followed. During the phase of no apparent growth only a few substances were formed (mostly alcohols). The rapid phase of growth was characterized by an intense synthesis of many compounds in relatively high concentrations and later a sudden decrease in the number and amounts of substances. A slow successive, decline in the number and amounts of volatile components was observed during the phase of no net growth.The volatiles emitted by the fungus were concentrated, when most of the compounds were most abundant and the relative amounts of the major volatile neutral compounds were determined. The main components were ethyl acetate, ethyl propionate and ethanol.     

Analysis of human breath samples with a multi-bed sorption trap and comprehensive two-dimensional gas chromatography (GCxGC)

A multi-bed sorption trap designed to quantitatively collect volatile organic compounds from large-volume vapor samples and inject them into a gas chromatograph is combined with a comprehensive two-dimensional gas chromatograph (GCxGC) for the analysis of organic compounds in human breath samples. The first-column effluent of the GCxGC is modulated by a single-stage, resistively-heated and air-cooled segment of 0.18-mm i.d. stainless steel column using the same stationary phase as the first column. Cooling gas is provided by a two-stage conventional refrigeration system, and thus no consumables other than carrier gas and electric power are required. The sorption trap uses four discreet beds, three containing different grades of graphitized carbon and one containing a carbon molecular sieve. The ordering of the beds in the trap tube is from the weakest to strongest adsorbent during sample collection. Breath samples are collected in gas sampling bags, and samples are passed through the trap at a flow rate of about 50 cm3/min. After sample collection, hydrogen carrier gas flow is initiated in the direction opposite to the sample collection flow, and the metal trap tube is resistively heated to inject a sample plug into the GCxGC. Performance data for the combined GCxGC/sorption-trap instrument is described, and human breath-sample chromatograms are presented.     

Diurnal Changes in Uric Acid Metabolism of Rats Fed a Casein Diet

Direct gas chromatographic determination of tobacco smoke was developed. Tobacco smoke condensate was collected on a glass fiber filter and the components were converted into their trimethylsilyl derivatives and then subjected to glass capillary column gas chromatography. By this method, all tobacco smoke components, including unstable phenolic substances and water-soluble polyhydroxy compounds, were simultaneously determined. Compositional differences between lamina and midrib smoke of flue-cured tobacco leaves were also clarified by the method. The results indicate that there are quantitative differences in nicotine, phenols, levoglucosan, quinic acid γ-lactone and the other major components between lamina and midrib smoke.     

Sorption of Components from a Mixture of Essential Oils by Cryotextured Cornstarches

Sorption by cryotextured cornstarches of components of the aqueous phase of a mixture of essential oils was studied by capillary gas chromatography. The amount of cryotexture-sorbed substances depended linearly on their concentration in the initial gel. The sorption of components from the mixture by starch polysaccharides was mainly associated with hydrophobic cooperative interactions, which resulted in the formation of supramolecular structures and inclusion complexes. The structure of the compounds was a major factor determining the degree of sorption. Sorption of monoterpene hydrocarbons was the most pronounced. We revealed a synergistic increase in the degree of sorption from the mixture as compared to binding of individual compounds.__________Translated from Prikladnaya Biokhimiya i Mikrobiologiya, Vol. 41, No. 4, 2005, pp. 463–469.Original Russian Text Copyright © 2005 by Terenina, Misharina.     

Biomimetic cooperative interactions of dried cross-linked poly(N-6-aminohexylacrylamide) with binary mixtures of solvent vapors

Biomimetic cooperativity of hydration effect and effect of ethanol favorable for binding of bad organic sorbates were observed for their vapor sorption by cross-linked poly(N-6-aminohexylacrylamide) (PNAHAA) in the absence of liquid phase. The vapor sorption isotherms were determined for these systems by the static method of gas chromatographic headspace analysis at 298 K. The hydration above 0.09-0.13 g of H(2)O/(g of polymer) gives a cooperative increase in the PNAHAA binding affinity for benzene, cyclohexane, dioxane, and propanols up to a level which does not change by further hydration, indicating the polymer antiplasticization. Bad sorbates (dioxane, benzene) were observed to have a biomimetic cooperative influence on the binding of ethanol by the dried PNAHAA. This cooperativity does not occur in ternary systems with good nonhydroxylic sorbate acetonitrile.     

Modelling alterations in the partition coefficient in in vitro biological systems using headspace gas chromatography

Headspace gas chromatography was used to determine the physiological media–air partition coefficient (K) of four volatile organic solvents of industrial importance. The experimental conditions were those likely to be used in in vitro metabolic and toxicological studies on volatile compounds. The addition of solvent to the liquid phase from a stock solution in ethanol, or the presence of organic material at concentrations normally seen in in vitro studies, did not significantly alter the K value. Binary solvent addition resulted in a dose-dependent decrease in K for each solvent that was also influenced by the solvent solubility and the constituents of the liquid matrix. The aromatic solvents exerted the greatest effect and showed the greatest change in K value.