Differential transfer of organic micropollutants through intestinal barrier using Caco-2 cell line

Food seems to be one of the main ways of animal and human contamination with polycyclic aromatic hydrocarbons (PAHs) and dioxins. In vivo studies showed a blood absorption of these xenobiotics after their ingestion. Our work aimed at studying the in vitro transfer of PAHs and dioxins through intestinal barrier. Caco-2 cells were cultivated on permeable filters to measure transepithelial permeability of (14)C labeled 2,3,7,8-tetrachlorodibenzo-p-dioxin, benzo[a]pyrene, pyrene, and phenanthrene, which differed in their physicochemical properties. The results showed that the molecules were able to cross intestinal cell layers. All the molecules were detected associated with cells, even if the dioxin was the less uptaken compound. Phenanthrene appeared in basal media faster, and its level after a 6-h exposure was respectively 1.1, 2, and 7 times higher than pyrene, benzo[a]pyrene, and 2,3,7,8-tetrachlorodibenzo-p-dioxin levels. These findings suggest that intestinal epithelium plays a key role in selective permeability and then in bioavailibility of micropollutants.     

Effect of exposure to soil-bound polycyclic aromatic hydrocarbons on milk contaminations of parent compounds and their monohydroxylated metabolites

The aim of this study was to determine the transfer kinetics of soil-bound polycyclic aromatic hydrocarbons to milk in lactating cows. Soil (500 g/day) fortified with fluorene (104 microg/g dry soil), phenanthrene (82 microg/g), pyrene (78 microg/g), and benzo[a]pyrene (33 microg/g) was administered to three dairy cows via a rumen cannulas for 28 consecutive days. Parent compounds and their major metabolites in milk were measured using gas chromatography-mass spectrometry. Secretion of parent compounds in milk did not increase significantly (P > 0.05) over the control values measured before supply. Target monohydroxylated metabolites were not detected in control samples, but 2-hydroxy fluorene, 3-hydroxy phenanthrene, and 1-hydroxy pyrene were present in milk by the second day of dosing. The highest concentrations of metabolites in milk (31-39 ng/mL) were for 1-hydroxy pyrene at days 7 and 14 of dosing. The observed plateaus for 3-hydroxy phenanthrene and 2-hydroxy fluorene were lower (respectively, 0.69 and 2.79 ng/mL) but significantly increased in comparison to the control samples. Contrarily, 3-hydroxy benzo[a]pyrene was not detected in milk at any sampling time. These results suggested a notable metabolism of the parent compounds after their extraction from soil during the digestive transfer. Thus, the metabolization of fluorene and pyrene can lead to higher concentrations of metabolites than of parent compounds in milk. Despite the absence of a significant transfer of parent PAHs to milk, the appearance of metabolites raises the questions of their impact on human health.     

Characterization and Sources of PAHs and Potentially Toxic Metals in Urban Environments of Sevilla (Southern Spain)

The purpose of this study was to determine the degree of PAH contamination and the association of PAHs with metals in urban soil samples from Sevilla (Spain). Fifteen polycyclic aromatic hydrocarbons-PAHs (naphthalene, acenaphthene, fluorene, phenanthrene, anthracene, fluoranthene, pyrene, benzo[a]anthracene, chrysene, benzo[b]fluoranthene, benzo[k]fluoranthene, benzo[a]pyrene, dibenzo[a,h]anthracene, benzo[g,h,i]perylene, indeno[1,2,3-c,d]pyrene) and seven metals (Cd, Cr, Cu, Mn, Ni, Pb, Zn) have been evaluated in representative urban soil samples. Forty-one top soils (0–10 cm) under different land use (garden, roadside, riverbank and agricultural allotment) were selected. PAHs from soil samples were extracted by sonication using dichloromethane. The simultaneous quantification of 15 different PAH compounds were carried out by HPLC using multiple wavelength shift in the fluorescence detector. For qualitative analysis a photo diode-array detector was used. Metal (pseudo-total) analysis was carried out by digestion of the soils with aqua regia in microwave oven. The mean concentration of each PAH in urban soils of Sevilla showed a wide range, they are not considered highly contaminated. The results of the sum of 15 PAHs in Sevilla soils are in the range 89.5–4004.2 μg kg^?1, but there seems not to be a correlation between the concentration of PAHs and the land use. Of the 15 PAHs examined, phenanthrene, fluoranthene and pyrene were present at the highest concentrations, being the sum of these PAHs about 40% of the total content. Although metal content were not especially high in most soils, there are significant hints of moderate pollution in some particular spots. Such spots are mainly related with some gardens within the historic quarters of the city. The associations among metals and PAHs content in the soil samples was checked by principal components analysis (PCA). The largest values both for ‘urban’ metals (Pb, Cu and Zn) and for PAHs were mainly found in sites close to the historic quarters of the city in which a heavy traffic of motor vehicles is suffered from years.     

Polycyclic aromatic hydrocarbons in soils of Vasilievsky Island (St. Petersburg)

The composition and accumulation patterns of priority polycyclic aromatic hydrocarbons (PAHs) in soils of Vasilievsky Island in Saint Petersburg were studied. Concentrations of benzo[a]pyrene were found to exceed maximum permissible concentrations in all the samples, and the maximum recorded concentration exceeded the MPC by 50 times. Concentrations of other PAHs also exceeded the background values. The main soil pollutants were found to be fluoranthene, pyrene, benzo[b]fluoranthene, benzo[a]pyrene, and benzo[g, h, i] perylene, the part of which in the total content of PAHs was 65–80%.     

Formation of polycyclic aromatic hydrocarbons in northern and middle taiga soils

An integrated study of the qualitative and quantitative composition of polycyclic aromatic hydrocarbons (PAHs) in the atmospheric precipitation-soil-lysimetric water system was performed using high performance liquid chromatography. It was shown that the accumulation of low-molecular PAHs (phenanthrene, anthracene, fluoranthene, pyrene, benz[a]anthracene, and chrysene) in soils is due to the transformation of organic matter and the regional transport and deposition of PAHs with atmospheric precipitation on the underlying surface. High-molecular polyarenes (benz[b]fluoranthene, benz[k]fluoranthene, benz[a]pyrene, dibenz[a,h]anthracene, benzo[ghi]perylene, and indeno[1,2,3-cd]pyrene) mainly result from the decomposition of soil organic matter.     

多环芳烃微生物降解机理研究进展

主要阐述了微生物降解PAHs的机理,比较了典型微生物种类芽孢杆菌(Bacillus)和分支杆菌(M.vanbaalenii PYR-1)对同种PAHs菲的代谢过程不同之处,并分析了分支杆菌(M.vanbaalenii PYR-1)对菲和芘的代谢机理,指出微生物氧化降解PAHs主要从其K区和湾区开始,而K区氧化是有毒PAHs降解的主要可能途径,资料显示真菌比细菌对苯并[a]芘的降解能力更强,同时给出了微生物代谢菲、芘和苯并[a]芘的降解图,以便做进一步研究。     

多环芳烃高效降解菌的筛选

以多环芳烃(PAHs)菲、蒽、芘、■和苯并(a)芘为供试物,对土著混合菌和引进菌同时进行筛选实验。结果表明,引进菌和土著混合菌经过驯化后对菲、蒽、芘、■和苯并(a)芘均具有一定的降解能力。其中,在pH=6时,混合菌U03在48h内对5种PAHs的降解率均相对较高,分别为:72.38%;64.46%;65.77%;66.49%和64.77%,并且其的降解速率在各菌剂中同样最快,通过SPSS数理统计分析软件对数据进行处理后得出,混合菌U03可在较短时间内达到较好的降解目的。室内模拟试验证明混合菌U03具有较强的降解PAHs的能力,混合菌的协同作用有利于污染土壤中PAHs的降解。     

Studying the Effects of Two Various Methods of Composting on the Degradation Levels of Polycyclic Aromatic Hydrocarbons (PAHs) in Sewage Sludge

The research comprised of studying the effect composting sewage sludge with sawdust and vermicomposting with earthworm Eisenia fetida has on the degradation of 16 polycyclic aromatic hydrocarbons (PAHs). Raw rural sewage sludge prior composting was more contaminated with PAHs than urban sewage sludge, in both cases exceeding EU cutoff limits of 6 mg/kg established for land application. Dibenzo[a,h]anthracene (DBahAnt), acenaphtylene (Acy) and indeno[1,2,3-c,d]pyrene (IPyr) were predominant in rural sewage sludge, whilst the urban sewage sludge contained the highest concentrations of benzo[b]fluoranthene (BbFl), benzo[k]fluoranthene (BkFl) and indeno[1,2,3-c,d]pyrene (IPyr). Thirty days of composting with sawdust has caused a significant reduction of 16 PAHs on average from 26.07 to 4.01 mg/kg (84.6%). During vermicomposting, total PAH concentration decreased on average from 15.5 to 2.37 mg/kg (84.7%). Vermicomposting caused full degradation of hydrocarbons containing 2 and 6 rings and significant reduction of PAHs with 3 aromatic rings (94.4%) as well as with 5 aromatic rings (83.2%). The lowest rate of degradation (64.4%) was observed for hydrocarbons with 4 aromatic rings such as fluoranthene, benzo(a)anthracene, chrysene and pyrene. On the other hand, the highest level of degradation was determined for PAHs with 2 rings (100%), 3 rings (88%) and 6 aromatic rings in the molecule (86.9%) after composting with sawdust. Acenaphthene and pyrene were found to be the most resistant to biodegradation during both composting methods.     

1-Hydroxypyrene in milk and urine as a bioindicator of polycyclic aromatic hydrocarbon exposure of ruminants

Urinary 1-hydroxypyrene (1-OH-pyrene) is now largely considered to be a valuable biomarker of exposure of man and animals to pyrene and other polycyclic aromatic hydrocarbons (PAHs). However, from a practical and agronomic standpoint, the question remains whether such biomarking capability still holds when 1-OH-pyrene is analyzed in milk produced by ruminants. To assess this hypothesis, four goats were daily submitted to three different amounts of pyrene oral ingestion, together with phenanthrene and benzo(a)pyrene (1, 7, and 49 mg/day during 1 week each). An HPLC-fluorometric analysis of 1-OH-pyrene in milk revealed a perfect correlation between pyrene doses and 1-OH-pyrene detected in milk, thus fully confirming the biomarking capability of 1-OH-pyrene and providing information on its transfer coefficient toward milk. Transfer equations such as the ones found in the present study could be used as a valuable and practical risk assessment tool in (i) the accurate monitoring of exposure of ruminants to pyrene and (ii) the evaluation of occupational and environmental exposure of ruminants to PAH mixtures.     

Rapid, semimicro method for determination of polycyclic aromatic hydrocarbons in shellfish by automated gel permeation/liquid chromatography

A simple, rapid, easily automated method is described for the determination of polycyclic aromatic hydrocarbons (PAHs) in shellfish such as American lobster (Homarus americanus) and blue mussel (Mytilus edulis). PAHs are extracted from small amounts (1-8 g) of tissue by saponification in 1N ethanolic potassium hydroxide followed by partitioning into 2,2,4-trimethylpentane. This solution is evaporated just to dryness by rotary evaporation and the residue is dissolved in cyclohexane-dichloromethane (1 + 1) for gel permeation chromatography (GPC) on Bio-Beads SX-3. The GPC procedure is ideal as a screening method in the range 25-18 000 ng PAHs/g tissue. If individual PAH measurements are required, the appropriate GPC fraction is collected and PAHs are separated by reverse phase liquid chromatography (LC) with fluorometric detection. Individual PAHs at concentrations as low as 0.25-10 ng/g can be determined. Recoveries of added fluoranthene, pyrene, benz[a]anthracene, chrysene, benzo[e]pyrene, benzo[b]fluoranthene, benzo[k]fluoranthene, benzo[a]pyrene, dibenz[a,h]anthracene, benzo[ghi]perylene, and indeno[1,2,3-cd]pyrene were quantitative, with relative standard deviations ranging from 0.0 to 16.9%.     

Input and behavior of polycyclic aromatic hydrocarbons in arable,fallow, and forest soils of the taiga zone (Tver oblast)

Contents of 11 most prevalent polycyclic aromatic hydrocarbons (PAHs) in snow and soils of arable, fallow, and forest areas significantly remote from impact technogenic sources of polyarenes have been examined in the Torzhok district of Tver oblast. From the analysis of snow samples, the volumes and composition of PAHs coming from the atmosphere onto the areas of different land use have been determined. Light hydrocarbons prevail in PAHs. They make up 65–70% of total PAHs; their share in soils reaches almost 95%. An increase in the content of PAHs is revealed in fallow soils compared to arable and afforested areas. A direct relationship is revealed between the lateral distribution of total PAHs and the content of organic carbon. The distribution of total PAHs is surface-accumulative in forest soils, mainly uniform in arable soils, and deepaccumulative in fallow soils. PAH groups characterized by similar radial distributions and ratios between their reserves in snow and soils are distinguished: (1) fluorene and phenanthrene, (2) biphenyl and naphthalene, (3) benzo(a)anthracene, chrysene, perylene, and benzo[a]pyrene, and (4) anthracene and benzo[ghi]pyrene.     

In vitro bioaccessibility of soil-bound polycyclic aromatic hydrocarbons in successive digestive compartments in cows

Ruminants, which have a central place in the food chain, ingest soil that may contain pollutants. The bioaccessibility of three different polycyclic aromatic hydrocarbon compounds from soil was studied using an in vitro model based on the digestive tract of cows. For this purpose, pasture soil was spiked with (14)C radio-labeled compounds, aged, and then exposed to conditions which simulated the digestive compartments of the rumen, abomasum, and intestines. Our results show that aging generally reduced the bioaccessibility of all the compounds tested. Total bioaccessibilty in the first digestive compartment, i.e., the rumen, depended on the considered compound: elevated for phenanthrene (17-24%), moderate for pyrene (6.6-8.1%), and low for benzo[ a]pyrene (2.3-3.6%). Bioaccessibility was very low in abomasal acidity (generally <2%) and intestinal colloids (<8%). The liquid phases of intestinal medium successfully extracted compounds from freshly contaminated soil (25-28%), but the bioaccessibilty dropped markedly after aging (17% for phenanthrene and <9% for the more lipophylic compounds). Total bioaccessibilty in this in vitro model ranged from 11% for benzo[ a]pyrene in aged soil to 58% for phenanthrene in freshly contaminated soil, and the bioaccessibility of this latter compound was always higher compared to pyrene or benzo[ a]pyrene. Residual soil contained around half of the initial load, the highest residual levels being of benzo[ a]pyrene, which confirms the observed bioaccessibility.     

Effect of roasting conditions on the polycyclic aromatic hydrocarbon content in ground Arabica coffee and coffee brew

Roasting is a critical process in coffee production as it enables the development of flavor and aroma. At the same time, roasting may lead to the formation of nondesirable compounds, such as polycyclic aromatic hydrocarbons (PAHs). In this study, Arabica green coffee beans from Cuba were roasted under controlled conditions to monitor PAH formation during the roasting process. Roasting was performed in a pilot spouted bed roaster, with the inlet air temperature varying from 180 to 260 degrees C, using both dark (20 min) and light (5 min) roasting conditions. Several PAHs were determined in both roasted coffee samples and green coffee samples. Also, coffee brews, obtained using an electric coffee maker, were analyzed for final estimation of PAH transfer coefficients to the infusion. Formation of phenanthrene, anthracene, and benzo[a]anthracene in coffee beans was observed at temperatures above 220 degrees C, whereas formation of pyrene and chrysene required 260 degrees C. Low levels of benzo[g,h,i]perylene were also noted for dark roasting under 260 degrees C, with simultaneous partial degradation of three-cycle PAHs, suggesting that transformation of low molecular PAHs to high molecular PAHs occurs as the roasting degree is increased. The PAH transfer to the infusion was quite moderate (<35%), with a slightly lower extractability for dark-roasted coffee as compared to light-roasted coffee.     

某氮肥厂场地土壤PAHs污染特征研究

采用现场采样及室内测试方法对广州某氮肥厂原料车间和油库区土壤中16种优控多环芳烃（PAHs）的含量进行调查研究,分析了EPAHs含量及其组成特征和垂直分布特征,并在此基础上进行了源解析。结果表明,分析样品中∑PAHs范围在10-7795μg·kg,原料车间土壤中的∑PAHs小于油库区土壤中的,菲、芘、荧蒽、并（b）荧蒽、苯并（a）芘为主要污染物;油库土壤0-40cm的样品中16种PAHs均有检出,∑PAHs和单体分布基本一致;原料车间土壤∑PAHs和单体浓度随着地面深度的增加而减少。通过对单组分比值（菲／蒽,荧蒽／芘）的分析可以看出油库区土壤中PAHs来源于石油和燃烧源,而原料车间污染源主要为燃烧源。     

Rapid magnetic solid-phase extraction based on magnetic multiwalled carbon nanotubes for the determination of polycyclic aromatic hydrocarbons in edible oils

In this study, magnetic multiwalled carbon nanotubes were fabricated by a simple method and applied to magnetic solid-phase extraction (MSPE) of eight heavy molecular weight polycyclic aromatic hydrocarbons (PAHs) including chrysene, benzo[a]anthracene, benzo[b]fluoranthene, benzo[k]fluoranthene, benzo[a]pyrene, indeno[1,2,3-cd]pyrene, dibenzo[a,h]anthracene and benzo[g,h,i]perylene from edible oil samples. Several parameters affecting the extraction efficiency were investigated, including the type and volume of desorption solvent, extraction and desorption time, washing solution and the amount of sorbent. Under the optimized conditions, a simple and effective method for the determination of PAHs in edible oils was developed by coupling with gas chromatography-mass spectrometry (GC-MS). The whole pretreatment process was rapid, and it can be accomplished within 10 min. The limits of quantitation for the target PAHs were found to be 0.34-2.9 ng/g. The recoveries in oil sample were in the range 87.8-122.3% with the RSDs less than 6.8% (intraday) and 9.6% (interday). This method was successfully applied to the analysis of PAHs in seven kinds of edible oils from local markets.     

混合菌降解土壤中多环芳烃的试验研究

PAHs生物降解程度受多种因素影响。通过筛选驯化PAHs降解菌,研究混合菌对土壤中菲、芘、苯并（a）蒽、苯并（b）荧蒽、苯并（k）荧蒽、茚并（1,2,3-cd）芘的生物降解性能,并考察污染时间对土壤中PAHs降解效果的影响。结果表明,筛选的混合菌具有很强的PAHs降解能力,缩短了PAHs生物降解的半衰期,且PAHs起始降解速率较快,之后趋于平缓。27d内土壤中的菲、芘、苯并（a）蒽、苯并（b）荧蒽、苯并（k）荧蒽、茚并（1,2,3-cd）芘的平均降解率分别为98.14%、89.97%、88.47%、63.55%、65.24%、60.49%,其中菲在5d之内的降解率高于93%。污染210d的土壤中各PAHs的起始降解速率高于污染50d的土壤,因此污染时间越长,PAHs生物降解的停滞期越短。     

甜菜与牧草间作对多环芳烃污染土壤的修复作用

通过盆栽试验方法,选择经济作物甜菜和牧草类黑麦草、苏丹草、香根草为供试植物,研究了甜菜与3种牧草分别间作及各自单作对多环芳烃（PAHs）菲、荧蒽、芘和苯并[a]芘污染土壤修复作用。结果显示：经6个月连续两茬种植试验后,所有种植植物的处理中土壤PAHs的去除率均高于无植物种植组,间作种植土壤PAHs的去除率高于单作,黑麦草、苏丹草、香根草与甜菜间作对土壤PAHs的去除率分别达到84.85%、79.96%、84.11%;在土壤污染条件下,间作模式更有利于甜菜生长;种植植物增强了土壤中多酚氧化酶和过氧化氢酶的活性,间作模式下二者活性高于单作4.37%~43.07%,过氧化氢酶较多酚氧化酶对PAHs土壤污染更敏感,可作为关键酶用于评价土壤PAHs污染状况。在不影响农业生产的前提下,修复植物牧草和经济作物甜菜间作种植模式显著提高了土壤PAHs的降解率。     

Determination of polycyclic aromatic hydrocarbons in commercial liquid smoke flavorings of different compositions by gas chromatography-mass spectrometry

The presence of polycyclic aromatic hydrocarbons (PAHs) in five commercial liquid smoke flavorings, used in the European food industry, was studied. The samples were subjected to an alkaline treatment, extracted with cyclohexane, cleaned up by means of solid-phase extraction tubes, and analyzed by gas chromatography-mass spectrometry. Three different procedures for the cleanup were tested. The results revealed the presence of 34 PAHs, some of them with methyl substituents. In all cases, the concentrations of compounds of low molecular weight were much higher than those of high molecular weight. Relationships between smoke flavoring compositions and PAH levels were also studied. Three of the samples contained high levels of both total and carcinogenic PAHs. Benzo[a]pyrene was also detected in these three samples, but its concentration did not exceed the 10 microg/kg level fixed by the FAO/WHO. Finally, a relation was found, first between the concentrations of total carcinogenic PAHs and benzo[a]pyrene and also between the concentrations of pyrene and benzo[a]pyrene. The latter ratio reveals that pyrene concentration could be very useful in predicting the level of benzo[a]pyrene and, consequently, in estimating the carcinogenicity arising from the presence of benzo[a]pyrene and other carcinogenic PAHs.     

湿热灭菌和氯化汞灭菌对双液相体系中PAHs降解的影响

A two-liquid-phase(TLP) soil slurry system was employed to quantify the efficiencies of autoclaving and mercuric chloride sterilization in the dissipation of polycyclic aromatic hydrocarbons(PAHs).The fates of 11 PAHs(naphthalene,fluorene,phenanthrene,anthracene,fluoranthene,pyrene,benzo(a)anthracene,benzo(a)pyrene,benzo(b)fluoranthene,benzo(k)fluoranthene,dibenzo(a,h)anthracene) were recorded over 113 days of incubation.No microorganisms were detected in the HgCl 2-sterilized soil slurries during the whole incubation period,indicating very effective sterilization.However,about 2%-36% losses of PAHs were observed in the HgCl 2 sterilized slurry.In contrast to the HgCl 2-sterilized soil slurry,some microorganisms survived in the autoclaved soil slurries.Moreover,significant biodegradation of 6 PAHs(naphthalene,fluorene,phenanthrene,anthracene,fluoranthene and pyrene) was observed in the autoclaved soil slurries.This indicated that biodegradation results of PAHs in the soil slurries,calculated on basis of the autoclaved control,would be underestimated.It could be concluded that the sterilization efficiency and effectiveness of HgCl 2 on soil slurry was much higher than those of autoclaving at 121℃ for 45 min.