• Journal of Life Science
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• v.8 no.4
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• pp.400-409
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• 1998

Formation of adduct was studied in benzo(a)pyrene(BP)- and doxorubicin(Dx)-treated human NC-37 cells and isolated nuclei. Major adducts formed were determined by fluorescence absorption spectrophotometery and DNA-lin-ked protein assay. When isolated nuclei were exposed to carcinogens BP and DMBA, and anticancer drugs m-AMSA, ellipticine and Dx, varying degrees of adduct formation occured between DNA-protein complex and these drugs. When the mixture was centrifuged 1.7 M sucrose solution, binding BP and DMBA appeared to be similar between the sediment and the supernatant. When the sediment was centrifuged again with 0.35% polymin-P, the amount of BP bound was 2-fold greater in the protein(1077$\pm$55cpm) than in DNA fraction (470$\pm$20cpm), whereas that of DMBA was 1.6-fold greater in the DNA than in protein fraction. In the case of m-AMSA, ellipticine and Dx, the amount of binding was slightly greater in supernatant than in sediment in centrifugation with 1.7 M sucrose, and more than 3 times greater in the DNA- than in protein- fraction in centrifugation with 0.35% polymin P. DNA fractions which associated with a subset of nonhistone chromosomal protein were isolated from NC-37 cells exposed to $^{3}$H-BP and $^{14}$C-Dx. They were separated into two distince components DNA-S and DNA-P by centrifugation with 2M Nacl chromatin extraction. The results indicated that the amount of $^{3}$H-BP bound was 6.0-fold greater in DNA-P as compared with DNA-S, while that of $^{14}$C-Dx binding appreaed to be 6.2-fold greater in DNA-S than in DNA-P fraction. When $^{3}$H-BP binding wasdetermined in the presence of cold Dx, the amount of binding was reduced only in the DNA-P fraction, indicating that the interaction between DNA and protein is decreased. Gene expression by these drugs, BP treated cells were increased to compare with nomal cells but reduced by treatment with BP-Dx. These results suggest that the protein moiety which tightly bound to DNA-P fraction may play an important role in the regulation of gene expression.

• Korean journal of food and cookery science
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• v.11 no.4
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• pp.351-355
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• 1995

The inhibitory effects of methanol, ethanol, chloroform/ methanol and water extracts from natural and cultural yams on the mutagenicity in the cooked pork (broiled and panfried) and the chemically induced mutagen, sodium azide, benzo(a)pyrene and 2-aminofluorene were investigated using salmolla typhimurium TA 100. In the presence of the S9 mixture, ethanol extract from natural yam showed high inhibitory effect on the mutagenicity of the cooked pork. But benzo(a)pyrene, supposed to be produced in mutagen during cooking, did not show high inhibitory effect in same extract. Besids, the yam extract on the mutagenicity of the sodium azide without S9 mixture showed low inhibitory effect. However 2-aminoflourene with S9 mixture showed high inhibitory effect, 91.5％.

• Journal of Environmental Science International
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• v.13 no.4
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• pp.413-420
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• 2004

The objectives of present study were to investigate the effects of benzo[a]pyrene(BaP) on cytotoxicity, lipid peroxidation and antioxidant enzymes in rat hepatocyte primary culture. Primary cultures of rat hepatocytes were incubated for 24 hr, 48 hr or 72 hr in the presence of various concentrations (0, 10, 20, 30, 50 or 100 $\mu.$ M) of BaP. Cytotoxicity and cell viability were determined by measuring glutamic oxaloacetic transaminase(GOT) activity, lactate dehydrogenase(LDH) activity and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide(MIT) value. Lipid peroxidation was evaluated using thiobarbituric acid reactive substances(TBARS) assay. Effects on antioxidant system were determined by measuring glutathione peroxidase(GPx) activity, glutathione reductase(GR) activity and glutathione concentration. Activities of GOT and LDH, MTT value as well as TBARS concentration were not affected by up to 100 $\muM$ of BaP for 24 hr incubation. However, BaP at the concentration of 50 $\muM$ for 48 hr incubation or at the concentration of 30 $\muM$ for 72 hr incubation began to increase LDH activity and TBARS concentration but decrease MTT value, representing that BaP caused cytotoxicity and decreased cell viability in dose- and time-dependent manners. GPx activity began to be decreased by BaP at the concentration of 50 $\muM$ for 72 hr incubation. Whereas, GR activity began to be decreased by BaP at the concentration of 20 $\muM$ for 72 hr incubation. Glutathione concentration began to be decreased by BaP at the concentration of 20 $\muM$ for 72 hr incubation and was further reduced to 90% by 100 $\muM$ of BaP. These results demonstrate that BaP caused cytoctoxicity and decreased cell viability by increasing lipid peroxidation and decreasing glutathione concentration as well as activities of GPx and GR.

• Analytical Science and Technology
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• v.36 no.6
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• pp.291-299
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• 2023

The benzo[a]pyrene in edible oils is extracted using methods such as Liquid-liquid, soxhlet and ultrasound-assisted extraction. However these extraction methods have significant drawbacks, such as long extraction time and large amount of solvent usage. To overcome these drawbacks, this study attempted to improve the current complex benzo[a]pyrene analysis method by applying the QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) method that can be analyzed in a simple and short time. The QuEChERS method applied in this study includes extraction of benzo[a]pyrene into n-hexane saturated acetonitrile and n-hexane. After extraction and distribution using magnesium sulfate and sodium chloride, benzo[a]pyrene is analyzed by liquid chromatography with fluorescence detector (LC/FLR). As a result of method validation of the new method, the limit of detection (LOD) and quantification (LOQ) were 0.02 ㎍/kg and 0.05 ㎍/kg, respectively. The calibration curves were constructed using five levels (0.1~10 ㎍/kg) and coefficient (R²) was above 0.99. Mean recovery ratio was ranged from 74.5 to 79.3 % with a relative standard deviation (RSD) between 0.52 to 1.58 %. The accuracy and precision were 72.6~79.4 % and 0.14~7.20 %, respectively. All results satisfied the criteria ranges requested in the Food Safety Evaluation Department guidelines (2016) and AOAC official method of analysis (2023). Therefore, the analysis method presented in this study was a relatively simple pretreatment method compared to the existing analysis method, which reduced the analysis time and solvent use to 92 % and 96 %, respectively.

• The Korean Journal of Zoology
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• v.31 no.2
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• pp.142-146
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• 1988

Metabolism of benzo(a)pyrene, the most thoroughly studied PAH, was studied in mouse placental microsomes incubated with $^3$H-labeled B(a)P. B(a)P metabolites were separated using HPLC fitted with a C18- $\mu$ Bondapak column. The single major metabolite by mouse placental microsomes induced by B(a)P was 7, 8-diol B(a)P, while 4, 5-diol B(a)P, 3-OH and quinones constituted minor metabolites. Treatment with 3-methyl-cholanthrene to mice resulted in indudion of hydroxy B(a)P and quinone compounds. Phenobarbital treated mouse placental microsomes also showed elevated level of B(a)P metabolism with 7, 8-diol B(a)P as a major metabolite.

• Journal of Microbiology and Biotechnology
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• v.13 no.4
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• pp.624-627
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• 2003

Benzo[a]pyrene (B[a]P) is an environmental pollutant that has been implicated in carcinogenesis. Saccharomyces cerevisiae was treated with B[a]P, and the responses of its cytochrome P450 (CYP) enzyme and DNA-damage checkpoint genes were examined through gene expression profiles using a reverse transcription polymerase chain reaction (RT-PCR). The DNA-damage checkpoint genes tested were the chk1 and pds1 genes, involved in a metaphase arrest, the swi6 gene targeted by G1 arrest, the pol2 gene related to S phase arrest, and the cln2 gene encoding a cyclin protein, all of which are based on rad9 and rad24. Among these genes, no noticeable effect was found when the cells were exposed to various concentrations of B[a]P. However, the transcriptional activity of CYP51 was significantly different when the cells were exposed to B[a]P. Accordingly, the present results indicate that cytochrome P450 plays a more significant role than DNA-damage checkpoint genes in the response of S. cerevisiae to B[a]P.

• Fisheries and Aquatic Sciences
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• v.9 no.2
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• pp.64-69
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• 2006

Cytochrome P450 (CYP1A) gene expression in the liver and sex steroid levels in plasma were investigated in olive flounder (Paralichthys olivaceus) exposed to tributyltin (TBT) and benzo[a]pyrene (BaP). We constructed a cDNA library and cloned a 230-base sequence encoding partial CYP1A DNA. The CYP1A gene expression level was estimated using northern blotting. Hepatic CYP1A mRNA levels in fish injected with BaP at 10 mg/kg body weight (b.w.) increased for 48 h after injection. However, fish injected with both BaP and TBT at 10 mg/kg b.w. showed no significant changes in CYP1A mRNA level after 48 h. Plasma concentrations of testosterone and $17{\beta}$-estradiol were not significantly different in males and females injected with BaP and TBT. We suggest that TBT-induced suppression of BaP bioactivity should be interpreted with caution in biomonitoring field studies.

• Fisheries and Aquatic Sciences
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• v.5 no.1
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• pp.28-31
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• 2002

Effects of cimetidine, a cytochrome P450 inhibitor, on the benzo[a]pyrene (BaP)-mediated cytochrome P450 induction and lipid peroxidation of liver in olive flounder, Paralichthys olivaceus, were investigated. Fish were fed either a cimetidine-supplemented diet or a cimetidine-free control diet once daily to satiation for 3 days. After 6 hrs of last feeding, the fish received intraperitoneal (i.p.) injection of BaP (20 mg/kg of body weight) dissolved in sterile corn oil $(100 \mu L)$ or received only a corn oil i.p. injection. At 1, 2, 3, and 7 days after the injection, hepatic cytochrome P450 and thiobarbituric acid reactive substances (TBARS), an indicator of lipid peroxidation, were analyzed. BaP injection resulted in an increase of hepatic cytochrome P450, and the fish fed the cimetidine-supplemented diet before injection of BaP showed delayed increase of hepatic cytochrome P450 compared to the fish fed a cimetidine-free diet and BaP injected. Injection of BaP clearly induced hepatic lipid peroxidation, and consistently higher TBAR values were shown in the fish fed a cimetidine­supplemented diet before injection of BaP than the fish injected with BaP alone.

• Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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• 2006.04a
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• pp.216-219
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• 2006

Water retention curve (WRC), one of soil hydraulic properties, is often approximated by property-transfer models (PTMs). Using the PTMs, we can estimate the WRCs from other physical properties such as particle-size distribution (PSD). The objective of this work was to investigate the performance of two PTMs with different origins for numerical simulations on transport of Benzo[a]pyrene in a soil. To do this, we chose both PTMs with different origins, i.e., (1) the lognormal distribution model (L anti NL models), and (2) the modified $Kov\'{a}cs$ model (MK model). The MK model showed tile worse performance in estimation of the WRCs. When transport of B[a]P was simulated, the MK model predicted to move farther than the L and NL models did, indicating that transport of B[a]P in a soil can be greatly influenced by the choice of PTMs.

• Journal of Food Hygiene and Safety
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• v.29 no.2
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• pp.141-145
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• 2014

To assess the health risk for benzo(a)pyrene by the intake of edible oils, 288 cases of edible oils collected from food markets were analysed using the high performance liquid chromatography with fluorescence detector. The levels of benzo(a)pyrene were from non-detection to $4.78{\mu}g/kg$, and the average was $0.11{\mu}g/kg$. The chronic daily exposures of benzo(a)pyrene for total population group and consumer-only group were estimated using the food consumption data in the fifth Korea National Health and Nutrition Examination Survey in 2011. The estimated daily intake of benzo(a)pyrene was $4.26{\times}10^{-3}ng/kg$ b.w./day for total population group and $7.64{\times}10^{-3}ng/kg$ b.w./day for consumer-only group. The MOE (margin of exposure) of benzo(a)pyrene for total population group and consumer-only group was $7.28{\times}10^7{\sim}1.74{\times}10^8$ and $3.95{\times}10^7{\sim}9.42{\times}10^7$, respectively. Accordingly, the health risk from benzo(a)pyrene caused by the intake of edible oils was considered as a very low level.