Insight into the experiment and extraction mechanism for separating carbazole from anthracene oil with quaternary ammonium-based deep eutectic solvents

Carbazole is an irreplaceable basic organic chemical raw material and intermediate in industry.The separation of carbazole from anthracene oil by environmental benign solvents is important but still a challenge in chemical engineering.Deep eutectic solvents (DESs) as a sustainable green separation solvent have been proposed for the separation of carbazole from model anthracene oil.In this research,three quaternary ammonium-based DESs were prepared using ethylene glycol (EG) as hydrogen bond donor and tetrabutylammonium chloride (TBAC),tetrabutylammonium bromide or choline chloride as hydrogen bond acceptors.To explore their extraction performance of carbazole,the conductor-like screening model for real solvents (COSMO-RS) model was used to predict the activity coefficient at infinite dilution (γ^(∞)) of carbazole in DESs,and the result indicated TBAC:EG (1:2) had the stronger extraction ability for carbazole due to the higher capacity at infinite dilution (C^(∞)) value.Then,the separation performance of these three DESs was evaluated by experiments,and the experimental results were in good agreement with the COSMO-RS prediction results.The TBAC:EG (1:2) was determined as the most promising solvent.Additionally,the extraction conditions of TBAC:EG (1:2) were optimized,and the extraction efficiency,distribution coefficient and selectivity of carbazole could reach up to 85.74%,30.18 and 66.10%,respectively.Moreover,the TBAC:EG (1:2) could be recycled by using environmentally friendly water as antisolvent.In addition,the separation performance of TBAC:EG (1:2) was also evaluated by real crude anthracene,the carbazole was obtained with purity and yield of 85.32%,60.27%,respectively.Lastly,the extraction mechanism was elucidated byσ-profiles and interaction energy analysis.Theoretical calculation results showed that the main driving force for the extraction process was the hydrogen bonding ((N–H...Cl) and van der Waals interactions (C–H...O and C–H...π),which corresponding to the blue and green isosurfaces in IGMH analysis.This work presented a novel method for separating carbazole from crude anthracene oil,and will provide an important reference for the separation of other high value-added products from coal tar.

Determination of Benzo[a]pyrene in Edible Oil by High Performance Liquid Chromatography-Fluorescence Detector (HPLC-FL)

In this study, an optimized high performance liquid chromatography-fluorescence detector (HPLC-FL) method for the determination of benzo[a]pyrene in edible oil was established. HPLC was performed with Thermo Fisher Scientific C18 column (250 mm×4.6 mm, 5 μm) as the chromatographic column and acetonitrile and water as the mobile phase, and the excitation wavelength and emission wavelength of fluorescence detector were 286 and 430 nm, respectively. The response was high, and the linear range was 0.5-10.0 ng/ml. The lowest limit of detection was 0.11 ng/ml, and the average recovery was 92.5%. This method is suitable for quantitative analysis of benzo[a]pyrene content in edible oil.

Establishment and evaluation of a mouse model of skin squamous cell carcinoma induced by 7, 12-dimethyl-benzodiazepine [a] anthracene at high dose

Objective: To establish a mouse model of skin squamous cell carcinoma induced by high dose 7, 12-dimethyl-benzodiazepine [a] anthracene, and to provide experimental basis for the prevention and treatment of skin squamous cell carcinoma. Methods: 36 BALB/c mice were randomly divided into 3 groups, respectively is: the high dose group (group A), low dose group (group B), and blank control group (group c), recorded in the process of building body weight in mice, and skin changes back, local skin tissue for building after HE staining of PCNA immunohistochemical staining, and the expression of PCNA positive staining level as reflected in mice skin squamous cell carcinoma of the malignant indicators, using the Image Pro Plus calculation PCNA staining of integral optical density (IOD) and positive area, and the results were statistically analyzed. Results: at 12 weeks, there were statistically significant differences in tumor number and tumor volume between group A and group B (P<0.05), and the tumor bearing rate was up to 83.3% and 66.7%, and the incidence of SCC was up to 75% and 33.3%. Immunohistochemical analysis showed statistical differences in IOD and positive area of mice in groups A, B and C (P<0.01). Conclusion: compared with the mouse model of skin squamous cell carcinoma established by the traditional two-stage protocol, this model has the characteristics of shorter experimental period, higher tumor formation rate and easier operation, providing a reliable experimental basis for the study of skin squamous cell carcinoma.

Surfactant-assisted hydrothermally synthesized MoS_2 samples with controllable morphologies and structures for anthracene hydrogenation

MoS_2 samples with controllable morphologies and structures were synthesized using surfactantassisted hydrothermal processes.The effects of surfactants（PEG,PVP,P123,SDS,AOT,and CTAB）on the morphologies and structures of MoS_2 samples were investigated.The results revealed that spherical,bulk-like,and flower-like MoS_2 particles assembled by NH4~＋-intercalated MoS_2 nano-sheets were synthesized.The morphologies of the MoS_2 samples and their structures（including the slab length and the number of stacked layers） of MoS_2 nano-sheets in these samples could be controlled by adjusting the surfactants.Mono-dispersed spherical MoS_2 particles could be synthesized with PEG via the creation of MoS_2 nano-sheets with slab lengths shorter than 15 nm and fewer than six stacked layers.Possible formation mechanisms of these MoS_2 samples created via surfactant-assisted hydrothermal processes are proposed.Further,the catalytic activities of MoS_2 samples for anthracene hydrogenation were evaluated in a slurry-bed reactor.The catalyst synthesized with the surfactant PEG exhibited the highest catalytic hydrogenation activity.Compared with the other catalysts,it had a smaller particle size,mono-dispersed spherical morphology,shorter slab length,and fewer stacked layers;these were all beneficial to exposing its active edges.This work provides an efficient approach to synthesize transition metal sulfides with controllable morphologies and structures.

基于β-糊精氧化石墨烯管尖固相萃取/高效液相色谱检测生活饮用水中苯并[a]芘

以自制的β-糊精氧化石墨烯(β-CD@GO)装填入10mL移液器吸头作为固相萃取剂,结合高效液相色谱法对生活饮用水中苯并[a]芘进行检测,并对固相萃取条件进行了优化。苯并[a]芘在0.1~50μg/L范围内线性关系良好(r=0.9996),检出限为3ng/L(S/N=3)。在加标水平为0.5、5、40μg/L时,方法回收率为89.2%~92.5%,相对标准偏差为3.7%~5.2%。该方法操作简便快速、选择性好、灵敏度高、有机溶剂消耗少,可用于生活饮用水中苯并[a]芘快速检测分析。

Inhibitory effects of RRR-α-tocopheryl succinate on benzo (a) pyrene (B (a) P)-induced forestomach carcinogenesis in female mice

AIM To study the inhibitory effects of VES( RRR-α-tocopheryl Succinate, VES ), aderivative of natural Vitamin E, on benzo (a)pyrene (B (a) P)-induced forestomach tumor infemale mice.METHODS The model of B (a)P-inducedforestomach tumor was established according tothe methods of Wattenberg with slightmodifications. One hundred and eighty femalemice (6 weeks old) were divided into six groupsequally; negative control (Succinic acid),vehicle control ( Succinate + B (a) P), positivecontrol(B(a) P), high VES(2.5g/kg. b. w + B(a)P), Iow VES(1 .25 g/kg. b. w + B(a) P) ig as wellas VES by ip (20 mg/kg, b. w + B(a) P). Exceptthe negative control group, the mice wereadministrated with B(a)P ig. and correspondingtreatments for 4 weeks to study the anti-carcinogenetic effect of VES during the initiationperiod. The experiment lasted 29 weeks, inwhich the inhibitory effects of VES both ontumor incidence and tumor size were tested.RESULTS The models of B (a)P-inducedforestomach tumor in female mice wereestablished successfully. Some werecauliflower-like, others looked like papilla, evena few were formed into the ulcer cavities. VES at1.25 g/kg. b. w, 2.5 g/kg. b.w. by ig and 20 mg/kg. b. w. via ip could decrease the number oftumors per mouse (1.7 ± 0. 41, 1.6 ± 0.34 and 1.1±0.43), being lower than that of B(a)P group(5.4 ± 0.32, P＜0.05). The tumor incidence wasinhibited by 18.2%, 23.1% and 50.0%. VES at1.25g/kg.b.w., 2.5 g/ kg.b.w. by ig and20 mg/kg. b.w. via ip reduced the total volumeof tumors per mouse (54.8 ± 8.84, 28.4 ± 8.32and 23.9± 16.05), being significantly lower thanthat of B(a)P group (150.2±20.93, P＜0.01).The inhibitory rates were 63.5%, 81.1% and84.1%, respectively.CONCLUSION VES has inhibitory effects on B(a) P-induced forestomach carcinogenesis infemale mice, especially by ip and it may be apotential anti-cancer agent in vivo.

Photochemical behavior of benzo[a]pyrene on soil surfaces under UV light irradiation

The rates of photodegradation and photocatalysis of benzo [a]pyrene （BaP） on soil surfaces under UV light have been studied. Different parameters such as temperature, soil particle sizes, and soil depth responsible for photodegradation, catalyst loads and wavelength of UV irradiation blamed for photocatalysis have been monitored. The results obtained indicated that BaP photodegradation follows pseudo-first-order kinetics. BaP photodegradation was the fastest at 30℃ . The rates of BaP photodegradation at different soil particle size followed the order： less than 1 mm〉less than 0.45 mm〉less than 0.25 mm. When the soil depth increased from 1 mm to 4 ram, the half-life increased from 13.23 d to 17.73 d. The additions of TiO2 or Fe2O3 accelerated the photodegradation of BaP, and the photocatalysis of BaP follows pseudo-first-order kinetics. Changes in catalyst loads of TiO2 （0.5%, 1%, 2%, and 3% （wt）） or Fe203 （2%, 5%, 7%, and 10% （wt）） did not significantly affect the degradation rates. Both BaP photocatalysis in the presence of TiO2 and Fe2O3 were the fastest at 254 nm UV irradiation.

Biodegradation of benzo[a]pyrene in soil by Mucor sp.SF06 and Bacillus sp.SB02 co-immobilized on vermiculite

Two indigenous microorganisms, Bacillus sp. SB02 and Mucor sp. SF06, capable of degrading polycyclic aromatic hydrocarbons （PAHs） were co-immobilized on vermiculite by physical adsorption and used to degrade benzo[a] pyrene （BaP）. The characteristics of BaP degradation by both free and co-immobilized microorganism were then investigated and compared. The removal rate using the immobilized bacterial-fungal mixed consortium was higher than that of the freely mobile mixed consortium. 95.3% of BaP was degraded using the co-immobilized system within 42 d, which was remarkably higher than the removal rate of that by the free strains. The optimal amount of inoculated co-immobilized system for BaP degradation was 2%. The immobilized bacterial-fungal mixed consortium also showed better water stability than the free strains. Kinetics of BaP biodegradation by co-immobilized SF06 and SB02 were also studied. The results demonstrated that BaP degradation could be well described by a zero-order reaction rate equation when the initial BaP concentration was in the range of 10--200 mg/kg. The scanning electronic microscope （SEM） analysis showed that the co-immobilized microstructure was suitable for the growth of SF06 and SB02. The mass transmission process of co-immobilized system in soil is discussed. The results demonstrate the potential for employing the bacterial-fungal mixed consortium, co-immobilized on vermiculite, for in situ bioremediation of BaP.

Antioxidant responses to benzo[a]pyrene,tributyltin and their mixture in the spleen of Sebasticus marmoratus

It has been reported that there is an interaction between Benzo[a]pyrene （BaP）, a widespread carcinogenic polycyclic aromatic hydrocarbon, and tributyltin （TBT）, an organometal used as an antifouling biocide. This study was therefore designed to examine the potential in vivo influence of BaP, TBT and their mixture on splenic antioxidant defense systems of Sebastiscus marmoratus. The fish were exposed to water containing environmentally relevant concentrations of BaP, TBT and their mixture. Spleens were collected for biochemical analysis after exposure for 7, 25, 50 d and after recovery for 7, 20 d. Cotreatment with BaP and TBT for 7 d potentiated the induction of glutathione peroxidase （GPx） activity by BaP or TBT alone. The cotreatment for 25 and 50 d resulted in inhibition of GPx activity, which was similar to the effect of TBT. Splenic glutathione S-transferase （GST） activities were significantly elevated in S. marmoratus exposed to BaP starting from 7 d and remained high up to 25 d. However, no further activity change was found with prolonged exposure. Cotreatment of BaP and TBT primarily inhibited the GST activity, which was similar to the effect of TBT. Cotreatment with BaP and TBT for 25 or 50 d potentiated the depletion of GSH （glutathione） by BaP or TBT alone. MDA （malondialdehyde） contents in spleen of S. marmoratus were not significantly altered compared with the control during the test period. Spleen, as an immune organ, is sensitive to exposure of BaP or TBT. It should have an effective mechanism to counteract oxidative damage. Antioxidative defense systems in spleen of S. marmoratus should be considered as potential biomarkers. Short-term exposure of BaP or TBT could result in induction of antioxidant defense system. A significant decrease of these indices, such as GSH, GST, GPx might indicate more severe contamination.

Isolation of marine benzo[a]pyrene-degrading Ochrobactrum sp. BAP5 and proteins characterization

A bacterial strain BAP5 with a relatively high degradation ability of benzo[a]pyrene （BaP） was isolated from marine sediments of Xiamen Western Sea, China and identified as Ochrobactrum sp. according to 16S rRNA gene sequence as well as Biolog microbial identification system. Strain BAP5 could grow in mineral salt medium with 50 mg/L of BaP and degrade about 20% BaP after 30 d of incubation. Ochrobactrum sp. BAP5 was able to utilize other polycyclic aromatic hydrocarbons （PAHs） （such as phenanthrene, pyrene and fluoranthene） as the sole carbon source and energy source, suggesting its potential application in PAHs bioremediation. The profile of total soluble protein from Ochrobactrum sp. BAP5 was also investigated. Some over- and special-expressed proteins of strain BAP5 when incubated with the presence of BaP were detected by two-dimensional polyacrylamide gel electrophoresis, and found to be related with PAHs metabolism, DNA translation, and energy production based on peptide fingerprint analysis through matrix-assisted laser desorption/ionization-time of flight mass spectrometry.

In vitro Study on Role of Hsp70 Expression in DNA Damage of Human Embryonic Lung Cells Exposed to Benzo[a]pyrene

Objective Benzo[a]pyrene (B[a]P), a ubiquitous environmental pollutant, is a potent procarcinogen and mutagen that can elicit tumors, leading to malignancy. Heat shock proteins (Hsp) have been shown to protect cells against damages caused by various stresses including exposure to numerous chemicals. Whether Hsps, or more specifically Hsp70, are involved in repair of B[a]P-induced DNA damage is currently unknown. Methods We assessed the potential role of the inducible form of Hsp70 in B[a]P-induced DNA damage of human embryonic lung (HEL) cells using immunoblot and the comet assay (i.e., the single cell gel electrophoresis assay). Results Exposure to B[a]P induced a dose-dependent decrease in the level of Hsp70, but a dose-dependent +-increase in DNA damage both in untreated (control) HEL cells and in cells preconditioned by a heat treatment. Heat preconditioning prior to B[a]P exposure potentiated the effect of B[a]P at a low dose (10 μmol/L), but appeared to be protective at higher doses. There was a negative correlation between Hsp70 level and DNA damage in the non-preheated as well as in the preconditioned cells. Conclusion These data suggest that exposure of HEL cells to B[a]P may induce a dose-dependent reduction in the levels of the inducible Hsp70. The detailed mechanisms for the reduction of Hsp70 levels by B[a]P and the role of Hsp70 in DNA damage under different concentrations of B[a]P remains to be determined.

Interlayer Energy Transfer from Naphthalene to Anthracene Chromophores Organized in Langmuir-Blodgett Films

Interlayer energy transfer between 2, 3-naphtho-10-hexadecylaza-15-crown-5 (NC16) and N-[1-(9-methoxyanthryl)]decylaza-15-crown-5 (A10C) within multilayer Langmuir-Blodgett films has been studied by steady-state fluorescence spectra. The donor and acceptor could be separated precisely by inserting stearic acid (SA) spacers. The efficiency of the energy transfer increases with the decrease in the donor-acceptor distance by a quadratic manner, suggesting the donor excitations are delocalized in the layer.

Combined effect of tributyltin and benzo[a] pyrene on the levels of sex hormone and vitellogenin in female Sebastiscus marmoratu

Tributyltin（TBT）, an organometal used as an antifouling biocide, has been reported to induce masculinization of fish. Benzo [a]pyrene （BaP）, a widespread carcinogenic polycyclic aromatic hydrocarbon, has been reported that its microsomal metabolites can produce an estrogenic response when tested in vitro. This study was therefore designed to examine the potential in vivo influence of TBT, BaP and their mixture on sex hormone levels in serum of Sebastiscus marmoratus, which were given 2 separate intraperitoneally （ip） injections（a single injection every 7 d） of TBT（0.5, 1, 5 and 10 mg/kg）, BaP（0.5, 1, 5 and 10 mg/kg）, or both in combination（0.5, 1, 5 and 10 mg/kg）; control fish received olive oil vehicle only. Six days after the 2nd injection, serum samples were collected and analyzed for sex hormone levels and alkali labile protein phosphorus （ALPP）, which is related to the yolk precursor protein vitellogenin. The pollutants at all doses significantly reduced serum testosterone, estradiol and ALPP content after 2 injections compared with the corresponding controls. The reduction of the estradiol levels should be response for the decrease of the vitellogenin levels. The results in the present study suggested that aromatase seems not the major target acted by TBT and BaP in fish. This study demonstrated that TBT or BaP exposure both inhibit the reproductive potential in female Sebastiscus marmoratus. Combined effect of TBT and BaP on the serum testosterone, estradiol and ALPP was not antagonism from the anticipation.

Synthesis,Bioactivity and Crystal Structure Analysis of Novel Benzo[d]isothiazol-3(2H)-ones

Two compounds,3-oxo-N-o-tolylbenzo[d]isothiazole-2（3H）-carboxamide （1） and N-（2-methoxyphenyl）-3-oxobenzo[d]isothiazole-2（3H）-carboxamide （2）,were synthesized from the initial compound benzo[d]isothiazol-3（2H）-one （BIT） and characterized by 1 H NMR,IR and elemental analysis,respectively.The single crystals of compounds 1 and 2 were obtained and determined by X-ray diffraction analysis.The preliminary results of biological activity experiment show that some of the title compounds exhibited a favorable antimicrobial activity.

Isolation and purification of carbazole contained in anthracene slag by extraction combined with medium pressure liquid chromatography

Ultrasonic-assisted extraction(UAE)combined with medium pressure liquid chromatography(MPLC)was designed for carbazole separation from anthracene slag(AS).The effects of liquid/solid ratio,temperature,and extraction times on carbazole separation were investigated.When using CC14 and ethyl acetate as extraction solvents and combining with MPLC,carbazole recovery and purity are 75.1%and 95.4%,respectively.The mechanism for carbazole separation were presumed by examining intermolecular interactions such as N-H…π,π-π,and C-Cl…πinteractions.These results demonstrate that UAE/MPLC has a considerable potential as a green and promising strategy for separating and purifying carbazole and other chemicals from AS.

Synthesis and Crystal Structure of 2-Ethoxy-3-n-butyl-benzofuro [2,3-d]pyrimidin-4(3H)-one

The crystal structure of the new title compound 2-ethoxy-3-n-butyl- benzofuro[2,3d]pyrimidin-4（3H）-one （C16H18N2O3, Mr = 286.32） has been prepared and determined by singlecrystal X-ray diffraction. The crystal is of monoclinic, space group P21/c with a = 13.7167（14）, b = 13.113（1）, c = 8.378（1） A, β = 98.992（2）^o, V = 1488.4（3） A^3, Z = 4, Dc = 1.278, F（000） = 608, μ = 0.089 mm^-1, MoKa radiation （2 = 0.71073）, R = 0.0498, wR = 0.1238 for 2336 observed reflections with I 〉 2σ（I）. X-ray diffraction analysis reveals that all ring atoms in the benzo[4, 5]furo [2,3-d] pyrimi- dinone moieties are almost coplanar.

Modulation of Behavior and Glutamate Receptor mRNA Expression in Rats after Sub-chronic Administration of Benzo(a)pyrene

Objective The present study aimed to test whether exposure to benzo（a）pyrene [B（a）P] affects spatial learning and short-term memory by modulating the expression of the Gria1 and Grin2a glutamate receptor subunit genes in the hippocampus.Methods Thirty-six 21-24-day-old,male rats were randomly assigned into high-,medium-,and low-dose toxin exposure groups （6.25,2.5,and 1 mg/kg,respectively） and a control group,each containing nine rats.The behavioral performance of adult rats exposed to sub-chronic administration of B（a）P was monitored by learning and memory tests （Morris water maze）.Real-time PCR assays were used to quantify Gria1 and Grin2a gene expression in the hippocampus.Results At medium and high doses,B（a）P impaired spatial learning performance.The crossing-platform-location frequency and the time spent swimming in the platform area,which both relate to short-term memory,were significantly decreased in B（a）P-treated rats compared with controls.The level of Gria1 mRNA increased 2.6-5.9-fold,and the level of Grin2a mRNA increased 10-14.5-fold,with a greater fold increase associated with higher doses of B（a）P.Conclusion We demonstrated that sub-chronic administration of B（a）P inhibits spatial learning and short-term memory,and increases Gria1 and Grin2a expression in the hippocampus.This suggests a relationship of B（a）P exposure levels with Gria1 and Grin2a expression and impairment of short-term and spatial memory.

Different Patterns of Cyclin D1/CDK4-E2F-1/4 Pathways in Human Embryo Lung Fibroblasts Treated by Benzo[a]pyrene at Different Doses

Objective To investigate the roles of the cyclin D1/CDK4 and E2F-1/4 pathways and compare their work patterns in cell cycle changes induced by different doses of B[a]E Methods Human embryo lung fibroblasts （HELFs） were treated with 2 μmol/L or 100 μmol/L B[a]P which were provided with some characteristics of transformed cells （T-HELFs）. Cyclin D l, CDK4 and E2F-1/4 expressions were determined by Western blotting. Flow cytometry was used to detect the distribution of cell cycle. Results After B[a]P treatment, the proportion of the first gap （G 1） phase cells decreased. CDK4 and E2F-4 expression did not change significantly. In 2 μmol/L treated cells, a marked overexpression of cyclin D1 and E2F-1 was observed. However, in T-HELFs overexpression was limited to cyclin D1 only, and no overexpression of E2F-1 was observed. The decreases of G1 phase in response to B[a]P treatment were blocked in antisense cyclin D1 and antisense CDK4 transfected HELFs （A-D1 and A-K4） and T-HELFs （T-A-D1 and T-A-K4）. After 2 μmol/L B[a]P treatment, overexpression of E2F-1 was attenuated in A-D1, and E2F-4 expression was decreased significantly in A-K4. In T-A-D1 and T-A-K4, E2F-4 expression was increased significantly, compared with T-HELFs. The E2F-1 expression remained unchanged in T-A-D1 and T-A-K4. Condusions Cyclin DI/CDK4-E2F-1/4 pathways work in different patterns in response to low dose and high dose B[a]P treatment. In HELFs treated with 2 μmol/L B[a]P, cyclin D1 positively regulates the E2F-1 expression while CDK4 negatively regulates the E2F-4 expression; however, in HELFs treated with 100 μmol/L B[a]P, both cyclin D1 and CDK4 negatively regulate the E2F-4 expression.

Fluorescence Quenching of Anthracene by N,N-Diethylaniline and Phenothiazine in Triton X-100/n-C_(10)H_(21) OH/H_2O Microemulsion

The photo-induced electron transfer reactions of anthracene with N,N -diethylaniline(DEA) and phenothiazine(PTZ) occur in the membrane phase of a Triton X-100/ n -C 10 H 21 OH(1-decanol)/H 2O microemulsion. DEA and PTZ exist in the membrane phase of the microemulsion. Anthracene exists in the oil continuous phase of the W/O microemulsion and in the oil core and membrane phase of the O/W microemulsion.

Effects of benzo(a)pyrene exposure on the antioxidant enzyme activity of scallop Chlamys farreri

Scallop Chlamys farreri was exposed to different concentrations of benzo(a)pyrene (BaP) (0.5 μg/L, 1.0 μg/L, 10.0 μg/L and 50.0 μg/L) for 30 days in seawater. The 7-ethoxyresorufin O-deethylase (EROD) activity was significantly induced, and increased with the increasing BaP concentration. The glutathione-S-transferase (GST), superoxide dismutase (SOD), catalase (CAT), Glutathione peroxidase (GPx) activities increased in short time at low concentration of BaP, and was significantly depressed at high concentrations. Scallop gill was more sensitive to BaP than the digestive gland, and the digestive gland was the main tissue to deal with oxyradicals. The contents of malondialdehyde (MDA) increased with the exposure time and there was a positive correlation (concentration-effect) between the MDA content and the concentration of BaP. The biomarkers determined in this experiment had important roles in detoxification, and showed great potential as biomarkers for oxidative stress. Controlled laboratory experiments designed to simulate field exposure scenarios are particularly useful in ascertaining biomarkers suitable for use with complex contaminant mixtures in the marine environment.