A COMPARISON OF HPLC WITH FLUORESCENCE DETECTION AND FIXED WAVELENGTH FLUORESCENCE METHODS FOR THE DETERMINATION OF POLYCYCLIC AROMATIC HYDROCARBON METABOLITES IN FISH BILE

Fish exposed to polycyclic aromatic hydrocarbons (PAHs) accumulate only a small amount of PAH in their tissues. Hydrophobic PAHs are readily transformed into hydrophilic metabolites and excreted in bile. Bile samples of five fish species were collected from the Baltic Sea and analyzed using two methods: high-performance liquid chromatography (HPLC) with fluorescence detection and a fixed wavelength fluorescence method. With the HPLC method PAH compounds were quantitated after hydrolysis, and with the fluorescence method without enzyme-assisted hydrolysis. In HPLC analysis the major metabolite in all fish species, 1-hydroxypyrene, was the only metabolite detected in perch and salmon samples. 1-Hydroxyphenanthrene was detected in a few flounder and in most eelpouts. The correlation between the two methods for pyrene-type metabolites was good (r² = 0.773).     

DISTRIBUTION OF PRIORITY POLYCYCLIC AROMATIC HYDROCARBONS IN THE SEDIMENT OF LAKE BALATON,HUNGARY

The determination of 13 PAH pollutants was carried out on sediment samples collected at 27 sites at the Lake Balaton, Hungary. The aim was to investigate the distribution patterns of PAHs and the correlation of source-sink relationship. Sediment samples were collected from the upper 10 cm and from 20 to 70 cm depth. The dry mass ratio of the fine grain-size fraction (<0.063 mm) and the coarse sand sediments (0.063–500 mm) were analyzed. Principal component analysis (PCA) was performed on the PAH compositional data for 110 samples to estimate the distribution of PAHs in different compartments. The average concentration of PAHs was found as 132 μg/kg dry weight (11–1734 μg/kg) for all sites and depth. Considering the harbors, at some sites, 930–950 μg/kg of total PAHs were obtained. The ratio of phenantrene/anthracene (PHE/AN) and fluoranthene/pyrene (FA/PY) indicated that most of the samples showed pyrogenic origin. It can be established that the upper 10 cm of the sediment is significantly more polluted than the deeper layers. The interim sediment quality guideline (ISQG) values and the probable effect level (PEL) were used to compare our findings with other data. No concentrations of PAHs were found higher than either ISQG and PEL values of samples collected inside of the lake, so the sediment has not been associated with adverse biological effects. However, the maximum concentrations of 7 out of 9 PAH compounds found in samples of harbors were higher than ISOG values but lower than PELs. Analysis of the harbor sediments revealed an elevated amount of contamination probably derived from the fuel of ships.     

Hepatotoxicity of the mycotoxin penicillic acid: A pharmacokinetics consideration

The hepatotoxicity of penicillic acid (PA), a carcinogenic mycotoxin, was substantiated by a variety of hepatic functional tests. Involvement of an active metabolite as the toxic species was proposed. The toxicity of PA was dependent on the route of administration with intraperitoneal (ip) being the most toxic followed by intravenous (iv) and oral. This difference in toxicity was explained by the kinetic data for PA if liver were assumed to be the site of activation. One-, 2- and 3-compartment open models were proposed to fit the plasma parent compound concentration after oral, ip, and iv administration of PA. Liver, kidneys, heart, lungs and spleen contained more radioactivity than brain, fat and muscle after [¹⁴C] - PA administration. Only a fraction of the radioactivity in the blood was detected as the parent compound. Most of the recovered radioactivity in the kidneys and liver was in the cytosol fraction. [¹⁴C]PA was readily metabolized in the liver. The metabolites were excreted in the bile and effectively cleared by the kidneys. Fecal and respiratory CO₂ were minor excretory routes. Over 90% of the urinary and 99% of the biliary metabolites were not extracted with polar organic solvents. Three water-soluble metabolites (derived from GSH or cysteine) were resolved by HPLC in urine and bile. About 10% of the urinary metabolites were detected as glucuronide conjugates. These data supported the hypothesis that an active metabolite which can be detoxified by GSH is involved in the toxicity of PA.     

The Effect of Local Cooking Methods on Polycyclic Aromatic Hydrocarbons (PAHs) Contents in Beef,Goat Meat,and Pork as Potential Sources of Human Exposure in Kisumu City,Kenya

Roasted meat is known to be a major source of human exposure to PAHs. The contribution of direct-heat charcoal-roasted, electric- oven grilled, and shallow-pan fried meat to human exposure in Kisumu City was not known although the three modes of cooking meat are very prevalent. This study analyzed the concentrations of the PAHs in raw beef, goat meat, and pork, investigated the effect of direct-heat charcoal roasting, electric-oven grilling, and shallow-pan frying on these concentrations, and compared their concentration levels with international standards for foods in order to assess the potential risks to consumers. Samples were taken from three popular meat-roasting hotels within Kisumu City, Kenya. Extraction of PAHs was done using liquid-liquid partition after saponification with alcoholic potassium hydroxide followed by clean-up on a silica gel column and final analysis by gas chromatography-mass spectrometry (GC-MS). Roasting and shallow-pan frying introduced new PAHs and significantly (P ≤ 0.05) increased the concentrations of those existing in raw meat. Direct-heat charcoal roast beef had 5 new PAHs and a total mean PAH content of 17.88 μg/kg, compared with a mean of 1.39 μg/kg for raw beef, with the potent dibenz(a,h)anthracene also being detected. Direct-heat charcoal roasted goat meat had three new PAHs and a total mean PAH content of 4.77 μg/kg, compared with a mean of 2.13 μg/kg in raw meat, with the potent benzo(a)pyrene concentration being 8.84% of the total mean PAH. Fried pork had 7 new PAHs and a total mean PAH content of 3.47 μg/kg, compared with a mean total of 0.17 μg/kg, detected in the raw meat. Roast beef had the highest individual PAH concentration (5.03 μg/kg) and highest total PAHs concentration (17.88 μg/kg), both being higher than acceptable EU limits. The PAHs from local raw and cooked meat were characterized and quantified for the first time in Kisumu City and the study therefore provided the needed baseline data on PAHs in raw and cooked meat.     

Bioaccumulation of PAHs and Their Hydroxylated Metabolites in Common Carp (Cyprinus Carpio Linnaeus 1758) in Controlled Environment

Abstract

This study aimed to determine the incorporation of PAHs into muscle of Cyprinus carpio. Three model compounds (phenanthrene, pyrene and benzo[a]pyrene) were orally administered during 56 days, in a series of three increasing doses (0, 100, 500 μg of each PAHs per kg of fish). PAHs and their hydroxylated metabolites were analyzed by a HPLC-Fluorimetry method. Results of this study showed a significant increase of phenanthrene and pyrene in muscle. Benzo[a]pyrene incorporation to muscle was not effective. Concerning hydroxylated metabolites, they were not detected in muscle of carp. Steady states for phenanthrene and pyrene were obtained after 28 days of exposure. Low incorporation to muscle was observed at equilibrium for phenanthrene (6% and 3%) and pyrene (3% and 1%), depending of the dose tested.     

Evaluation of PAH Metabolites in Bile of Common Carp,Cyprinus carpio L. with Fixed Wavelength Fluorescence in a Field and Laboratory Study

The seasonal and inter-gender variability of polycyclic aromatic hydrocarbon (PAH) metabolites in the bile of common carp (Cyprinus carpio) was investigated with a field study in Karakaya Dam Lake (KDL) and a laboratory-conducted study with the fish samples of KDL and Sultansuyu Dam Lake (Turkey). The KDL has been polluted by urban, industrial, and agricultural activities for many years. A seasonal trend was seen in PAH metabolite levels with higher levels detected in May 2005 samples in KDL. Their levels in the bile of fish were not found to be related to sex which showed that gender is not a confounding factor on PAH uptake. The results showed significant decreases in bile metabolite levels in the KDL fish by approximately 75% versus increases in the SDL fish after acclimation at laboratory conditions. These results supported that PAH metabolites in bile were not stable and short-term differences in living conditions may lead to great differences in their levels. Moreover, these rapid changes in bile metabolites with acclimation may also indicate that dam lakes were not badly polluted with PAHs and the feeding status of fish should be taken into consideration when using bile metabolites as biomarkers.     

Polycyclic aromatic hydrocarbon emissions of non-road diesel engine treated with non-thermal plasma technology

Non-road diesel engines are important polycyclic aromatic hydrocarbon (PAH) sources in the environment due to their high emission concentration compared to on-road diesel engines. Particle- and gas-phase PAH concentrations of a non-road diesel engine were investigated. Non-thermal plasma (NTP) as an effective after-treatment technology was used to reduce PAH emissions. The results showed that particle-phase PAH concentrations were 329.7 µg/m³, 3,206.7 µg/m³, and 1,185.7 µg/m³ without the action of NTP at three different engine loads respectively. Relatively low concentrations were measured for gas-phase PAHs. Excellent linearity was shown for particle-phase with total PAH concentrations both with, and without, NTP. The gas-phase PAH concentrations linearly increased with engine load without NTP. The five most abundant compounds of PAHs were among low molecular weight (LMW) and medium molecular weight (MMW) compounds. Total PAH cleaning efficiency was beyond 50% when treated with NTP at the three different engine loads. We hypothesized that naphthalene (Nap) concentrations increased greatly at 60% and 80% engine loads because it was produced within the plasma zone by decomposition of high molecular weight (HMW) PAHs. The PAHs content of particulate matter (PM) aggregation at 60% load was approximately three times higher than at 40% and 80% loads. High correlation values were observed for MMW PAHs with total PAH concentrations. Correlations of PAH concentration reduction could be important to clarify the PAH reduction mechanism with NTP technology.     

Particle Associated Polycyclic Aromatic Hydrocarbons (PAHS) in Urban Air of Agra

Total Suspended Particulate Matter (TSPM) samples were collected at Nunhai Agra from April to September 2006. The concentrations of 16 PAHs in aerosols were quantified. The dominated predominant PAHs in TSPM include high molecular weight (HMW) congeners BghiP, DbA, IP and BaP. Nap and Acy were not detected in any of the samples. The sum of 14 priority PAHs ranged from 150 to 480 ng m^?3 with a mean value of 269 ± 121 ng m^?3. The Results indicated that PAH concentrations at Nunhai were higher than in other industrial sites, but are comparable to those measured in several urban Chinese cities, however, and less than the industrial locations of China. Higher HMW PAH concentration were attributable to higher rates of emissions as well as or greater scavenging and adsorption of vapor phase PAH on available TSPM. Potential sources of PAHs in aerosols were identified using the diagnostic ratios between PAHs. Vehicular emissions were the main contributors of particulate-associated PAHs, with minor contribution from stationary combustion sources may also contribute to the particulate PAHs. PAHs in aerosols were predominantly from gasoline and diesel engines.     

Urban Runoff Contribution to Surface Sediment Accumulation for Polycyclic Aromatic Hydrocarbons in the Cotonou Lagoon,Benin

The carcinogenic and mutagenic properties of some polycyclic aromatic hydrocarbons (PAHs) and their widespread presence in the environment have led to many scientific investigations during the past three decades. In this study, surface sediments from six sampling sites in the Cotonou lagoon (three sites in the channel and three in Lake Nokoue) during the highest stormy season (May, June, and July) were used to assess the urban runoff contribution of PAH accumulation in this ecosystem. This assessment showed relatively high PAH loads (140-625 ng/g) during the rainy season and high inputs of organic materials (total organic carbon contents between 2.5 and 9.5% dry weight). Generally known as originating from different processes, PAHs detected in most of the Cotonou lagoon sampling sites were characterized by the predominance of the high-molecular-weight PAH over the low-molecular-weight PAH, which is an indicator of pyrolytic origin PAH inputs. PAH contamination in the Cotonou lagoon surface sediments was mainly from heavy urban traffic linked to the use of diesel vehicles and to the motorbike taxis proliferating across and around the city. Regression analysis carried out between total PAH concentration and total organic carbon contents showed, excluding the rural sampling site, a positive linear relationship for most samples ( R 2 = 0.9996), and confirmed the pyrogenic input of PAHs in the Cotonou lagoon. Mitigation measures were suggested to ameliorate the PAH pollution in the lagoon.     

Concentrations and Source Identification of Polycyclic Aromatic Hydrocarbons (PAHs) in Mangrove Sediments from North of Persian Gulf

The concentrations of 16 Environmental Protection Agency (EPA) priority polyaromatic hydrocarbons (PAHs) were quantified in surface sediments from Hormozgan Province mangroves, south of Iran in dry and wet seasons. Sampling stations were selected in Laft and Khamir mangroves with international importance. Polyaromatic hydrocarbons varied from 75.24 ± 11.24 to 581.94 ± 637.39 ng/g dry weight basis. Pollution sources and their contribution for polyaromatic hydrocarbons pollution in sediments of Hormozgan mangroves were appointed based on molecular ratios and statistical methods, including principal components analysis (PCA) and multiple linear regression/principal components analysis (PCA/MLR) tests. Both of pyrogenic and petrogenic sources contributed in detected concentrations of PAHs. Contribution percentages of pyrogenic and petrogenic origins were estimated at 73.20 and 26.79%, respectively. Temporal variations showed that sediments contained higher levels of ∑PAHs in wet season than dry time. However, the mean detected ∑PAHs was lower than international quality guidelines; the high concentration of PAHs was found in Laft mangrove, suggesting the presence of PAH polluted localized area.     

THE FATE AND BEHAVIOR OF POLYCYCLIC AROMATIC HYDROCARBONS (PAHS) THROUGH FEEDING AND EXCRETION OF ANNELIDS

The concentration of polycyclic aromatic hydrocarbons (PAHs) in sediment and excrements of annelids was measured by gas chromatography-mass spectrometry (GC/MS) and the movement of PAHs in the tidal flat was investigated. The PAH concentrations ranged 4.4–80.4 and 61.2–286.9 μg/kg-dry for the excrement of Arenicola basiliensis and of Marphysa sanguinea, respectively; the former contained about 10 times as much PAHs as the sediment and the latter about 100 times. The difference in the PAH concentration between the two organisms was attributed to their feeding behavior; M. sanguinea intakes much more detritus containing PAHs than A. basiliensis. Moreover, after the excrement of M. sanguinea had stood on the sediment for 2 hours, the PAH concentrations decreased to half. The reduction of the PAHs may arise from chemical changes owing to microorganisms and/or enzymes in the excrement of M. sanguinea.     

Occurrence of Polycyclic Aromatic Hydrocarbons in Smoke Flavourings

The presence of Polycyclic Aromatic Hydrocarbons (PAHs) in four liquid smoke flavourings of different origins has been studied, in order to test whether a relation between their PAH content and their composition could be established, and to evaluate whether the use of these flavourings could involve a risk to human health. The samples were subjected to an alkaline treatment, extracted with cyclohexane, cleaned up, and the PAHs determined by gas chromatography-mass spectrometry. The results reveal a wide range of PAHs, especially in the commercial flavourings studied, which present higher concentrations of both total and carcinogenic PAHs than those obtained in the laboratory. Benzo(a)pyrene has been detected in three of the samples studied, but its levels do not exceed, in any of the cases, the 10 μg/kg value fixed by the FAO/WHO.     

POLYCYCLIC AROMATIC HYDROCARBONS IN BALTIC SEA BIVALVES

Concentrations of parent polycyclic aromatic hydrocarbons (PAHs) in the bivalves Macoma balthica and Astarte borealis were studied as an indication of the state of the Baltic Sea. Samples were collected between 1999 and 2001 from the Gulf of Finland, the Eastern Gotland Basin, and the Southern Baltic Proper. PAHs were quantified by liquid chromatography with fluorescence detection. The sum of 12 PAHs varied between 44 and 298 ng/g (ww), with the most contaminated bivalves found in Hanö Bight (the Bornholm basin). High-molecular-weight PAHs predominated among the PAHs. The PAH profiles of M. balthica differed from those determined in sediment from the same area. Both pyrolytic and petrogenic origins were indicated. Toxic equivalency factors evaluated as benzo[a]pyrene equivalents (BaPEs) were used to assess the PAH contamination. BaPEs were higher than values for M. balthica found in the southern part of the Gulf of Finland in 1995 and much lower than values measured in 1987 after the most serious oil spill in the Gulf of Finland in the past 25 years.     

Synergism of Green Tea Polyphenols and Phenanthrene Promotes the Death of Human Lung Adenocarcinoma Cell Line SPC-A-1

Green tea polyphenols (GTP), an anticarcinogen in tea leaves, can provide protection against various cancer types induced by polycyclic aromatic hydrocarbons (PAHs). However, current research about mutual effects has hardly focused on phenanthrene (PA), which is a dominant PAH with low toxicity detected in environment and many commercial tea samples. GTP and PA may be absorbed concurrently by the human body when people drink tea. In order to reveal the relation between intake of smoked tea and enhancement of cancer risk, in this study, growth inhibition, apoptosis, and cell cycle were assayed in human lung adenocarcinoma cell line SPC-A-1, and the interaction effects of GTP and PA was statistically analyzed. The results showed that GTP and PA inhibited cell growth in a concentration-dependent manner; their interaction significantly reduced the inhibition and affected the cell cycle; the final combination of PA and GTP exhibited a synergistic inhibition which caused cell death more acutely, and the cell cycle was arrested at S-phase, which means GTA could reduce the cancer risk even with drinking smoked tea in low concentration of PA.     

THE LOADS OF PAHS IN WASTEWATER AND SEWAGE SLUDGE OF MUNICIPAL TREATMENT PLANT

Abstract

Sewage and sewage sludge from a municipal wastewater treatment plant were analyzed for 16 EPA-PAHs. This plant is a classic mechanical-biological treatment plant, consisting of activated sludge technology with additional chemical treatment for the removal of phosphorus compounds. The process of sewage sludge treatment is carried out in closed as well as open sludge digesters. Primary and mechanically thickened sludge are passed through the fermentation process. Digested sludge is dewatered on filter-press through addition of flocculent. The measurements were obtained to investigate the effect of different treatment stages on PAH content in wastewater and sewage sludge. The following wastewater samples were collected: crude ones and those after sand trap, primary sedimentation, biological treatment and secondary sedimentation. Sewage sludge samples were collected from: primary sludge, digested sludge and dewatered sludge. PAH load in influent, mechanically and biologically treated sewage, as well as in raw digested and dewatered sludge, were calculated. PAHs were extracted from wastewater and sludge samples, with cyclohexane, dichloromethane using an ultrasonic method. Gas chromatograph equipped with mass spectrometry was used for qualitative and quantitative determination of PAHs. Mechanical and biological treatment proved the removal of 83–85% of PAHs from the influent. Despite this its daily PAH load introduced into the environment was high and reached 27–37% of PAH load in influent. In sewage sludge it was 46–70 g/d of PAHs (carcinogenic PAHs content 4–12%). In waste sludge (filter pressed sludge and sand from detrirer) PAH total load reached 42–68 g/d with (10–17% of carcinogenic PAHs).     

Vapour-phase and particulate-bound PAHs profile generated by a (SI/HCCI) engine from a winter grade commercial gasoline fuel

The vapour-phase and particulate-bound Aromatic Hydrocarbons, PAHs, generated by a V6 gasoline engine working in spark-ignition (SI) and homogeneous charge compression ignition (HCCI) modes were collected and analysed. All data were obtained during steady-state, fully warmed-up operation at different engine power levels (low and medium loads and mid-speed), and two different engine operation modes (SI and HCCI). The fuel used in this study was winter grade commercial gasoline fuel.The vapour-phase exhaust gases were passed through stainless-steel cartridges containing XAD-2 resin to capture PAHs. The PAHs were extracted from the resin with dichloromethane in an ultrasonic bath, the obtained extracts were later analysed qualitatively and quantitatively by GC-MS. The vapour-phase PAHs compounds observed from HCCI mode operated in low load were Naphthalene, Acenaphthylene and Acenaphthene only, while that obtained from SI mode under low load were Naphthalene, Acenaphthylene, Acenaphthene, Fluorene, Anthracene, Phenanthrene, Fluoranthene and Pyrene.The PAHs bound to particulates were trapped by using a complex of dilution tunnels with filter papers. The soluble organic fractions (SOF) of the trapped particulates were separated from the insoluble fraction (ISF) with the help of ultrasonic elution, and analysed by GC-MS method. The most abundant PAHs detected under selected operation condition for HCCI mode was Benzo[a]anthracene, followed by Chrysene, then Pyrene and pursued by Benzo[b]fluoranthene, in SI mode under same operation condition the highest PAH detected was Benzo[a]anthracene followed by Pyrene, Benzo[b]fluoranthene and Chrysene. Probable mechanisms for the production of some of the pyrosynthetic PAH were discussed.     

Revealing the Origin of Polycyclic Aromatic Hydrocarbons in an Urban Snowpack

Polyciclic aromatic hydrocarbons (PAHs) are ubiquitous pollutants in urban atmosphere. Several PAHs are known carcinogens or are the precursors to carcinogenic daughter compounds. Understanding the contributions of various PAH emission sources is critical to appropriately managing PAH levels in the environment. The sources of PAHs to urban snowpack in Shelekhov city, Eastern Siberia, characterized by extremely high levels of PAH accumulation in snow were determined by using end-member mixing approach. The best potential to distinguish PAH emission sources is exhibited by ratios of PAH pairs of the principal mass 228, 252, and 276. The ratios of PAH pairs were used as tracers of end-member PAH sources. The contributions of sources were calculated using systems of linear equations. The results obtained using ratios of PAH pairs were compared with those obtained using molecular diagnostic ratios. It was shown that the results obtained using diagnostic ratios as tracers are less reliable than the results obtained using the ratios of the sums of PAHs.     

Distribution and Removal of Polycyclic Aromatic Hydrocarbons in Two Italian Municipal Wastewater Treatment Plants in 2011–2013

PAH (Polycyclic Aromatic Hydrocarbons) analyses were carried out on samples from two Wastewater Treatment Plants (WWTPs) in Lombardy, similar for treatment sequences but fed on different influents: industrial component accounts for 70% at Alto Seveso plant while it is absent in Nosedo plant. Sampling concerned the influent and the effluent from activated sludge reactor and the final effluent after disinfection (ozonation for Alto Seveso and peracetic acid treatment for Nosedo). The concentrations of total PAHs were 5.3 ± 4.0 μg L^?1 and 2.4 ± 1.3 μg L^?1 in Alto Seveso and Nosedo influent, respectively. The lowest molecular weight PAHs had the highest concentrations in both plants; acenaphthene and naphthalene were the most important components in the influent to Alto Seveso and Nosedo WWTPs, respectively. The higher molecular weight compounds had the lowest concentrations and benzo(g,h,i)perylene and dibenzo(a,h)anthracene were never detected. Most of the PAH load entered biological treatment in dissolved form. For both plants PAHs were mostly removed in the biological section (96.5% and 89.5% for Alto Seveso and Nosedo, respectively), while disinfection had a minor role. Peracetic acid (Nosedo) seemed more efficient than ozone (Alto Seveso) in the removal of PAHs (4.18% and 0.89%, respectively). It is now necessary to confirm this result by using the same effluent for the two disinfection treatments.     

Analysis of Polycyclic Aromatic Hydrocarbons in Egyptian Petroleum Condensate Oils

The polycyclic aromatic hydrocarbons (PAH) in Egyptian condensates are analyzed for the first. A solid phase extraction (SPE) followed by gas chromatography-mass selective detection was used for their analysis. The method was calibrated for optimal extraction conditions. Excellent recoveries were found (78–114%) for the PAHs that were identified using a variety of standards and GC-MS spectra. The solid-phase extracted PAH fraction was further separated by HPLC on a Ag(I) mercaptopropanosilica gel to reduce the complexity of the sample by separating the PAHs based on the number of aromatic rings. The analytes were quantified using GC with a flame ionization detector. For this kind of sample SPE is a more convenient separation technique than an open column. PAHs containing two to four rings in the concentration range 0.6–11 μg/L were measured. Some preliminary geochemical hypotheses based on the analyzed PAHs and the previously analyzed S-containing aromatic compounds were formed as to the depositional environment and source rock type.