Intramolecular carbon isotope distribution of acetic acid in vinegar

Compound-specific carbon isotope analysis of acetic acid is useful for origin discrimination and quality control of vinegar. Intramolecular carbon isotope distributions, which are each carbon isotope ratios of the methyl and carboxyl carbons in the acetic acid molecule, may be required to obtain more detailed information to discriminate such origin. In this study, improved gas chromatography-pyrolysis-gas chromatography-combustion-isotope ratio mass spectrometry (GC-Py-GC-C-IRMS) combined with headspace solid-phase microextraction (HS-SPME) was used to measure the intramolecular carbon isotope distributions of acetic acid in 14 Japanese vinegars. The results demonstrated that the methyl carbons of acetic acid molecules in vinegars produced from plants were mostly isotopically depleted in (13)C relative to the carboxyl carbon. Moreover, isotopic differences (δ(13)C(carboxyl) - δ(13)C(methyl)) had a wide range from -0.3 to 18.2‰, and these values differed among botanical origins, C3, C4, and CAM plants.     

Isotopic fractionation by saprotrophic microfungi: Effects of species, temperature and the age of colonies

Saprotrophic fungi represent an important resource for a number of fungivorous and omnivorous soil animals, but little is known about the patterns of isotopic fractionation by soil fungi. We grew five common species of saprotrophic microfungi in laboratory cultures on simple artificial substrate based on carbohydrates derived either from C3 or C4 plants. Fungal cultures were kept at 15, 20 or 25 °C. Isotopic composition of carbon (¹³C/¹²C) and nitrogen (¹⁵N/¹⁴N) in bulk fungal tissue was determined after 11, 21 and 32 days. The fractionation of carbon and nitrogen stable isotopes was species-specific, but generally did not differ in C3- and C4-based growth media. The Zygomycete Mucor plumbeus did not differ in δ¹³C from the carbon source used, though Ascomycetes (Alternaria alternata, Cladosporium cladosporioides, Trichoderma harzianum and Ulocladium botrytis) were depleted in heavy carbon relative to the carbon source by 0.5-0.9‰. Three species were significantly depleted in ¹⁵N relative to the sodium nitrate that was used as a single source of nitrogen. In all species, δ¹⁵N but not δ¹³C tended to increase with the age of fungal colonies. The effect of temperature on δ¹⁵N was weak and inconsistent in different species. In contrast, all fungi except T. harzianum accumulated more ¹³С at 25 °C than at 15 °C. The overall variation in the isotopic signatures of saprotrophic fungi growing in identical conditions reached 8‰ for δ¹⁵N and 2.5‰ for δ¹³C due to species-specific differences in the isotopic fractionation and the age of individual fungal colonies. This variation should be incorporated into the interpretation of the isotopic composition of fungivorous soil animals.     

Carbon isotopes of water‐extractable organic carbon in a depth profile of forest soil imply a dynamic relationship with soil carbon

Although considerable research has been conducted on the importance of recent litter compared with older soil organic matter as sources of dissolved organic carbon (DOC) in forest soils, a more thorough evaluation of this mechanism is necessary. We studied water‐extractable organic carbon (WEOC) in a soil profile under a cool‐temperate beech forest by analysing the isotopic composition (¹³C and ¹⁴C) of WEOC and its fractions after separation on a DAX‐8 resin. With depth, WEOC became more enriched in ¹³C, which reflects the increasing proportion of the hydrophilic, isotopically heavier fraction. The ¹⁴C content in WEOC and its fractions decreased with depth, paralleling the ¹⁴C trend in soil organic matter (SOM). These results indicate a dynamic equilibrium of WEOC and soil organic carbon. The dominant process maintaining the WEOC pool in the mineral soil appears to be the microbial release of water‐soluble compounds from the SOM, which alters in time‐scales of decades to centuries.     

The fat that matters: Soil food web analysis using fatty acids and their carbon stable isotope signature

Chemical taxonomy based upon the composition of lipids is widely applied to investigate microbial communities and fatty acids have recently been employed to connect soil microbial and faunal food webs as well as to elucidate functional groups at higher trophic levels. The additional use of compound-specific isotopic analysis of ¹³C/¹²C ratios in fatty acids allows assessing specific trophic links and belowground carbon fluxes. In this review systematic patterns and processes underlying variations in the composition of fatty acids and their ¹³C/¹²C ratio are described. The emphasis is on biomarker fatty acids, their incorporation and modification, effects of pool size, and analytical methods. Further development of the application of fatty acid profiling to soil ecology should include both advances in experimental research and growth of theory. Accordingly, areas in which future experimentation can lead to progress in soil food web analysis are identified. Overall, combining fatty acid biomarker and their isotopic ratios will allow detailed insight into belowground trophic interactions.     

Microbial phospholipid biomarkers and stable isotope methods help reveal soil functions

This review targets microbial phospholipid biomarkers, their isotope analysis and their ability to reveal soil functions. The amount and composition of phospholipid fatty acids (PLFAs) measured in environmental samples strongly depend on the methodology. To achieve comparable results the extraction, separation and methylation method must be kept constant. PLFAs patterns are sensitive to microbial community shifts even though the taxonomic resolution of PLFAs is low. The possibility to easily link lipid biomarkers with stable isotope techniques is identified as a major advantage when addressing soil functions. Measurement of PLFA isotopic ratios is sensitive and enables detecting isotopic fractionation. The difference between the carbon isotopic ratio of single PLFAs and their substrate (Δ¹³C) can vary between −6 and +11‰. This difference derives from the fractionation during biosynthesis and from substrate inhomogeneity. Consequently, natural abundance studies are restricted to quantifying substrate uptake of the total microbial biomass. In contrast, artificial labelling enables quantifying carbon uptake into single PLFAs, but labelling success depends on homogeneous and undisturbed label application. Current developments in microbial ecology (e.g. ¹³C and ¹⁵N proteomics) and isotope techniques (online monitoring of CO₂ isotope ratios) will likely improve soil functional interpretations in the future. ¹³C PLFA analysis will continue to contribute because it is affordable, sensitive and allows frequent sampling combined with the use of small amounts of ¹³C label.     

Stable carbon isotopic composition of the wine and CO2 bubbles of sparkling wines: detecting C4 sugar additions

Sparkling wines have become a popular beverage in recent years, and the production of these wines is subject to adulteration during fermentation. This study investigated the stable carbon isotopic composition (expressed as delta(13)C) of the wine and of the CO(2) bubbles produced during the second fermentation for a number of sparkling wines produced in different countries around the world. Carbon isotope ratio analyses were used to estimate the addition of sugar obtained from C(4) plants (sugar cane or corn). The average delta(13)C values of the Brazilian brut, demi-sec, and doux sparkling wines were -20.5 +/- 1.2 per thousand (n = 18), -18.1 +/- 1.3 per thousand (n = 9), and -15.8 per thousand (n = 1), respectively. These values were statistically heavier (more positive carbon isotope ratio values) than the average delta(13)C of sparkling wines produced in other parts of South America (Argentina and Chile, -26.1 +/- 1.6 per thousand, n = 5) and Europe (France, Germany, Italy, Portugal, and Spain, -25.5 +/- 1.2 per thousand, n = 12), but not statistically different from sparkling wines produced in the United States or Australia. The most likely explanation for differences in the carbon isotope ratios of wines from these different regions is the addition of C(4) sugar during the production of some sparkling wines from Australia, Brazil, and the United States. The isotopic composition of the CO(2) bubbles (delta(13)C-CO(2)) followed similar trends. The average delta(13)C-CO(2) of most of the Brazilian and Argentine sparkling wines was -10.8 +/- 1.2 per thousand (n = 23), indicating that the likely source of carbon for the second fermentation was sugar cane. Conversely, the average delta(13)C-CO(2) of most of the sparkling wines produced in Chile and Europe was -22.0 +/- 1.2 per thousand (n = 13), suggesting that a different sugar (most likely sugar beet) was most used in the second fermentation. It was concluded that in many cases, the carbon isotope ratios of sparkling wine and CO(2) bubbles can provide valuable information about the sugar sources.     

The turnover of carbohydrate carbon in a cultivated soil estimated by 13C natural abundances

Understanding the chemical composition of soil organic matter (SOM) requires the determination of the dynamics of each class of compounds. We measured the dynamics of carbon in neutral carbohydrates by use of natural ¹³C labelling in an experimental wheat and maize sequence extending over 23 years. The isotopic composition of individual neutral monosaccharides was determined in hydrolysed particle‐size fractions by gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS) of trimethylsilyl (TMS) derivatives. The sensitivity in terms of ¹³C/¹²C ratios ranged between 1 and 2‰ depending on the monosaccharide. The age distribution of neutral sugar carbon was very similar to that of total soil carbon. Particulate organic matter (POM) was characterized by the predominance of glucose and xylose of vegetal origin. In POM > 200 µm, the mean age of sugar‐C (5 years) was slightly less than that of total carbon (7 years). Xylose was younger than glucose. The fine fraction 0–50 µm contained mainly glucose, arabinose, galactose, xylose, fucose and mannose, which had predominantly microbial origins. The mean age of carbohydrate carbon in the fraction 0–50 µm was between 60 and 100 years and was similar to that of total organic carbon (OC). No difference in the age of carbon between the individual monosaccharides was found. The POM fraction 50–200 µm had an intermediate signature and turnover. Considering the typical lability of carbohydrates, the relatively great age of carbohydrate carbon may be explained by physical or chemical protection from degradation, as well as by recycling of soil organic matter carbon by soil microbes.     

稳定性碳同位素方法在稻田甲烷研究中的应用

稻田甲烷产生、氧化等过程不仅影响甲烷排放量,还影响其稳定性碳同位素组成;反之,稻田所排放甲烷的稳定性碳同位素组成也可用来定量研究甲烷的产生和氧化过程。20世纪80年代以来,国外已将稳定性碳同位素方法广泛应用在稻田甲烷的研究中。本文介绍了稳定性碳同位素方法的基本原理及其在稻田甲烷产生、氧化过程研究中的应用,指出了该方法在稻田甲烷研究中的不确定因素,为我国稻田甲烷的深入研究提供了新的有力手段。     

Prospects and problems of using the methods of geochemistry of stable carbon isotopes in soil studies

A review of world investigations into the geochemistry of stable isotopes—a new area for soil science—is presented. Studies of the behavior of stable isotopes in soils are being developed in two major directions: reconstruction of the environmental conditions and indication of the soil processes. Investigations into the reconstruction of the environmental conditions are based on the assumption that the isotopic composition of carbon in the soil humus is inherited from and reflects the isotopic composition of carbon in the corresponding vegetation (the source of soil humus). At the same time, studies into the indication of soil processes are based on the effects of fractionation of carbon isotopes in the course of transformation of carbon compounds. In this case, the isotopic composition of carbon in the soil humus should differ from that in the initial plant material. The analysis of available data suggests that the current state of research does not allow assessing the entire diversity of the effects of isotopic fractionation in soils. New data on the isotopic composition of carbon of soil organic matter and carbonates are being actively accumulated at present. However, it is obvious that the effect of isotopic fractionation should be taken into account in the paleoenvironmental and paleoclimatic reconstructions.     

Cadmium Aqueous Exposure and Uptake of the Estuarine Isopod Cyathura carinata

We studied the isotopic composition of organic matter in the sediments of eight mountain lakes located in the Tatra Mountains (Western Carpathians, Poland). The sediments of the lakes were fine and course detritus gyttja, mud, and sand. The total organic carbon content varied from 0.5 to 53 %. The C/N ratio indicated that in-lake primary production is the major source of the organic matter in the lakes located above the treeline, whereas terrestrial plant fragments are the major organic compounds in the sediments of dystrophic forest lakes. We also found that a clear trend of isotopic curves toward lower values of δ ¹³C and δ ¹⁵N (both ~3 ‰) began in the 1960s. This trend is a sign of the deposition of greater amounts of NO _x from the combustion of fossil fuels, mainly by vehicle engines. The combustion of fossil fuels in electric plants and other factories had a smaller influence on the isotopic composition. This trend has been weaker since the 1990s. Animal and human wastes from pastures and tourism had a surprisingly minor effect on lake environments. These data are contrary to previous data regarding lake biota and suggest the high sensitivity of living organisms to organic pollution.     

The distribution of the stable carbon isotope (13C/12C) in fractions of soil organic matter

The carbon-isotopic composition of fulvic and humic acid from the A horizons of eight soil types, developed under a wide variety of climatological conditions, was measured. The fulvic acid is always enriched in ¹³C as compared with the humic acid from the same soil by a rather constant factor of 0.9?. The fulvic acids are isotopically closer to the plant source of the organic matter and thus represent an intermediate stage in the formation of humic substances. Depth sections of peat soil showed that carbon isotopes can be used to evaluate the dynamic nature of the fulvic-acid fraction. With depth, a transfer of carbon groups from polysaccharides to fulvic acid is seen. Based on isotopic evidence it is shown that in addition to formation of β-humus, part of the fulvic acid is condensed with depth to a stable humic fraction — humin.     

Natural abundance δN and δC of DNA extracted from soil

We report the first simultaneous measurements of δ¹⁵N and δ¹³C of DNA extracted from surface soils. The isotopic composition of DNA differed significantly among nine different soils. The δ¹³C and δ¹⁵N of DNA was correlated with δ¹³C and δ¹⁵N of soil, respectively, suggesting that the isotopic composition of DNA is strongly influenced by the isotopic composition of soil organic matter. However, in all samples DNA was enriched in ¹³C relative to soil, indicating microorganisms fractionated C during assimilation or preferentially used ¹³C enriched substrates. Enrichment of DNA in ¹⁵N relative to soil was not consistently observed, but there were significant differences between δ¹⁵N of DNA and δ¹⁵N of soil for three different sites, suggesting microorganisms are fractionating N or preferentially using N substrates at different rates across these contrasting ecosystems. There was a strong linear correlation between δ¹⁵N of DNA and δ¹⁵N of the microbial biomass, which indicated DNA was depleted in ¹⁵N relative to the microbial biomass by approximately 3.4‰. Our results show that accurate and precise isotopic measurements of C and N in DNA extracted from the soil are feasible, and that these analyses may provide powerful tools for elucidating C and N cycling processes through soil microorganisms.     

Short-term C4 plant Spartina alterniflora invasions change the soil carbon in C3 plant-dominated tidal wetlands on a growing estuarine Island

Spartina alterniflora is an invasive C₄ perennial grass, native to North America, and has spread rapidly along the east coast of China since its introduction in 1979. Since its intentional introduction to the Jiuduansha Island in the Yangtze River estuary, Spartina alterniflora community has become one of the dominant vegetation types. We investigated the soil carbon in the Spartina alterniflora community and compared it with that of the native C₃Scirpus mariqueter community by measuring total soil carbon (TC), soil organic carbon (SOC), total soil nitrogen (TN), and the stable carbon isotope composition (δ¹³C) of various fractions. TC and SOC were significantly higher in Spartina alterniflora in the top 60 cm of soil. However, there was no significant difference in soil inorganic carbon (IC) between the two communities. Stable carbon isotopic analysis suggests that the fraction of SOC pool contributed by Spartina alterniflora varied from 0.90% to 10.64% at a soil depth of 0-100 cm with a greater percentage between 20 and 40 cm deep soils. The δ¹³C decreased with increasing soil depth in both communities, but the difference in δ¹³C among layers of the top 60 cm soil was significant (p<0.05), while that for the deeper soil layers (>60 cm) was not detected statistically. The changes in δ¹³C with depth appeared to be associated with the small contribution of residues from Spartina alterniflora at greater soil depth that was directly related to the vertical root distribution of the species.     

秸秆施用对稻田CH4排放、溶解浓度及13C变化特征的影响

CH4 emission and the concentration of dissolved CH4 in soil solution and floodwater in a rice field and their stable carbon isotopic signatures as affected by straw application were investigated in 2009 in a field experiment at Jurong, Jiangsu Province, China. Straw application increased CH4 emission and CH4 concentration in the soil solution and floodwater. A positive seasonal correlation was also observed in the variation between CH4 flux and CH4 concentration in soil solution. The seasonal total CH4 emission (51.6 g CH4 m-2) in Treatment WS (straw applied) was about 168% higher than that in Treatment CK (without straw). The emitted CH4 and CH4 in soil solution were initially relatively enriched, then depleted and finally enriched again in 13C in both treatments, while CH4 in floodwater became isotopically heavier. The carbon isotopic signature of emitted CH4 and CH4 in floodwater averaged around -62‰ and -45‰ for both treatments, respectively, and was not significantly influenced by the application of straw. However, straw application caused the CH4 in soil solution to be significantly depleted in 13C during the middle of the rice season, and the mean δ13C value was lower in WS (-57.4‰) than in CK (-49.9‰). Calculation from the isotopic data showed that straw application increased the fraction of CH4 oxidized, causing no significant difference in the δ13C value of the emitted CH4 between the two treatments.     

Authentication of the origin of vanillin using quantitative natural abundance 13C NMR

The use of 13C isotopic distribution as an efficient means to determine the origin of vanillin has been substantiated. Using quantitative 13C NMR, the 13C/12C ratios at all eight carbon positions can be exploited. On a set of 21 samples of vanillin from five different origins, complete discrimination can be achieved. It is shown that, for many purposes, a rapid analysis in which only five sites are used is sufficient. However, improved discrimination using all eight sites is preferable to differentiate between different methods of production from natural ferulic acid or between natural and lignin-derived vanillin on the basis of the 13C/12C ratios characteristic of different origins. The C1 and C8 positions are demonstrated to be the most significant sites for discrimination using principle component analysis. However, aromatic carbon positions make an essential contribution, notably in differentiating between natural and lignin-derived vanillin.     

明胶骨原料碳、氮稳定同位素的地域性差异研究

为探讨不同地域来源的明胶骨原料中碳、氮稳定同位素的分布特征及其相关性,本试验采用元素分析-稳定同位素比率质谱仪测定了广西、内蒙古和山东3个主产区的牛骨样品及当地牧草饲料样品中的¹³C和¹⁵N同位素组成,并结合线性回归讨论了牛骨样品中δ¹³C和δ¹⁵N值的相关性与牛食谱组成的关系。结果表明,广西地区骨样品中δ¹³C和δ¹⁵N值的相关性较好(R²=0.401 8),而内蒙古和山东地区骨样品中δ¹³C和δ¹⁵N值的相关性较差(R²<0.1),反映了3地牛食物的来源稳定性及主要成分存在明显差异,对各地牛食谱结构推测的结果与实际牧草饲料样品中δ¹³C和δ¹⁵N值的地区分布规律也较为一致。本研究初步揭示了明胶骨原料中碳、氮稳定同位素产生地域性差异的原因,不仅为动物生态学研究提供了有益信息,更为动物制品产地溯源研究提供了新思路。     

泰国东部土地由森林转化为玉米地引起土壤有机碳流失及其储量的降低

Soil organic carbon(SOC) content and its stable carbon isotopic composition(within the upper 1 m) were measured to determine the e?ect of land-use changes from dry evergreen forest to maize fields in eastern Thailand.Digital land cover maps,derived from aerial photography and satellite images for years 1989,1996,and 2002 were used in association with field surveys and farmer interviews to derive land-use history and to assist in study site selection.Conversion from forest to maize cultivation for the duration of 12 years reduced SOC stocks at the rate of 6.97 Mg C ha-1 year-1.Reduction was most pronounced in the top 10 cm soil layer,which was 47% after 12 years of cultivation.Stable carbon isotope data revealed that the main fraction lost was forest-derived C.Generally low input rates of maize-derived C were not sufficient to maintain SOC at the level prior to forest conversion.After 12 years of continuous maize cultivation,the maize-derived C fraction made up about 20% of total SOC(5 Mg ha-1 of the total 25.31 Mg ha-1).     

Reconstruction of Late Glacial and Holocene landscape-climatic changes in the central Selenga Middle Mountains based on the isotopic composition of organic matter

The results of investigation into the composition of stable carbon and nitrogen isotopes of organic matter in the soils developed within soil–sedimentary sequences in the central part of the Selenga Middle Mountains in the Late Glacial and Holocene are presented. In the past 15000 years, the organic matter of the investigated soils has only been formed from the biomass of C3 plants (without the participation of C4 plants). This is confirmed by the of δ¹³С values from–27.00 to–23.35‰. A combined analysis of the parameters of the organic matter (С_org, N_total, C/N, δ¹³С, and δ¹⁵N) of soils formed in different periods makes it possible to assume that the isotopic composition of carbon and nitrogen reflects changes in the climate humidity during the Late Glacial and Holocene periods. The specified intervals of soil formation correspond to the climate humidification and stabilization of the surface owing to the development of dense vegetation. Aridization periods were characterized by the accumulation of sediments that buried soil horizons. The most pronounced stages of climate aridization occurred at the transition from the Late Glacial to the Holocene, from the Boreal to the Atlantic, and from the Atlantic to the Subboreal periods. The optimum soil-forming conditions existed in the periods of 11700–11000, 8800–6900, and 4700–1000 years ago, which is confirmed by the published data on the landscape-climatic changes in the adjacent areas in the past 15000 years.     

Soil organic matter in soil depth profiles: Distinct carbon preferences of microbial groups during carbon transformation

This study investigates how carbon sources of soil microbial communities vary with soil depth. Microbial phospholipid fatty acids (PLFA) were extracted from 0–20, 20–40 and 40–60 cm depth intervals from agricultural soils and analysed for their stable carbon isotopes (δ¹³C values). The soils had been subjected to a vegetation change from C3 (δ¹³C≈?29.3‰) to C4 plants (δ¹³C≈?12.5‰) 40 years previously, which allowed us to trace the carbon flow from plant-derived input (litter, roots, and root exudates) into microbial PLFA. While bulk soil organic matter (SOM) reflected ≈12% of the C4-derived carbon in top soil (0–20 cm) and 3% in deeper soil (40–60 cm), the PLFA had a much higher contribution of C4 carbon of about 64% in 0–20 cm and 34% in 40–60 cm. This implies a much faster turnover time of carbon in the microbial biomass compared to bulk SOM. The isotopic signature of bulk SOM and PLFA from C4 cultivated soil decreases with increasing soil depth (?23.7‰ to ?25.0‰ for bulk SOM and ?18.3‰ to ?23.3‰ for PLFA), which demonstrates decreasing influence of the isotopic signature of the new C4 vegetation with soil depth. In terms of soil microbial carbon sources this clearly shows a high percentage of C4 labelled and thus young plant carbon as microbial carbon source in topsoils. With increasing soil depth this percentage decreases and SOM is increasingly used as microbial carbon source. Among all PLFA that were associated to different microbial groups it could be observed that (a) depended on availability, Gram-negative and Gram-positive bacteria prefer plant-derived carbon as carbon source, however, (b) Gram-positive bacteria use more SOM-derived carbon sources while Gram-negative bacteria use more plant biomass. This tendency was observed in all three-depth intervals. However, our results also show that microorganisms maintain their preferred carbon sources independent on soil depth with an isotopic shift of 3–4‰ from 0–20 to 40–60 cm soil depth.     

陕西关中地区肉牛产地同位素溯源技术初探

为应用牛组织中C、N同位素组成的区域分布和同位素指标溯源肉牛产地,本文利用同位素比率质谱仪(IRMS)对陕西关中不同区县来源的牛尾毛样品的1δ3C和1δ5N值进行检测,通过聚类分析,研究陕西关中地区肉牛组织中同位素组成的区域分布情况。结果表明,杨凌区、眉县牛尾毛样品聚为一类;乾县、永寿县样品聚为一类;麟游县样品聚为一类;扶风县、岐山县和凤翔县分类不十分明显,分散在上述3类中。说明陕西关中不同地区肉牛组织中同位素组成存在差异,利用它们可进行肉牛地域的小范围溯源。