• Title/Summary/Keyword: Metabolites Analysis

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Effects of ethanol and phenobarbital on hemoglobin adducts formation in rats exposed to benzidine (흰쥐에서 에탄올과 phenobarbital이 벤지딘의 헤모글로빈 부가체 형성에 미치는 영향)

  • Kim, Chi Nyon;Lee, Se Hoon;Kim, Hyun-Soo;Youn, Young-Shik;Roh, Jaehoon
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.11 no.2
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    • pp.118-125
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    • 2001
  • Recently, biochemical analysis using hemoglobin adduct is frequently performed to evaluate the exposure to chemical carcinogens. However, data on the effect of co-exposure with other chemicals on hemoglobin adduct formation are seldom provided. The objective of this study is to evaluate the effects of pretreatment of ethanol(EtOH) and phenobarbital(PB), which are known to affect metabolism of xenobiotics, on the formation of hemoglobin adducts in the rats(Sprague-Dawley) administered benzidine(BZ). The experimental rats were divided into control, EtOH, and P8 groups. Rats were pretreated with EtOH or PB 24 hours before the oral administration of BZ. Blood sampling was taken before the administration of the chemicals and 0.5, 3, 6, 9, 12, 24, 48, 72, 96, and 144 hours after the administration of the BZ in 5 rats each. The blood was separated into hemoglobin and plasma immediately after taking the blood samples, and the adducts were undergone basic hydrolysis to convert them into aromatic amines. Hydrolyzed BZ, monoacetylbenzidine (MABZ), and 4-aminobiphenyl(4ABP) were separated by reversed-phase liquid chromatography without derivatization, and quantitative analyses of them were performed by a highperformance liquid chromatograph equipped with electrochemical detector. The quantitative amount of the metabolites was expressed by hemoglobin binding index(HBI), BZ-, MABZ-, and 4ABP-HBI of EtOH and PB groups were increased more than those of control group. These results are attributable to the fact that EtOH and PB induced N-hydroxylation related to the hemoglobin adduct formation. The ratio of N-acetylation (viz, MABZ-HBI/BZ-HBI) showed no significant difference between EtOH group and control group. It means that EtOH increased N-hydroxylation and N-acetylation in a similar degree. The N-acetylation ratio of PB group was relatively lower than control group because the PB increased N-hydroxylation induction. The N-acetylation ratios of all groups were higher than 1 during the entire experimental period. This result suggests that the effects of EtOH or PB need to be considered in the biochemical monitoring for the assessment of intermittent exposure of benzidine.

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Treatment Effect of Green Manure Crops on Content of γ-Oryzanols from Korean Rice Variety, Unkwangbyeo (녹비 작물 처리에 따른 운광벼 중 감마오리자놀 변화)

  • Kim, Heon-Woong;Shin, Jae-Hyeong;Lee, Min-Ki;Lee, Sung-Hyeon;Jang, Hwan-Hee;Cho, Hyun-Suk;Lee, Jeong-Tae;Jeon, Weon-Tai;Kim, Jung-Bong
    • Korean Journal of Environmental Agriculture
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    • v.34 no.2
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    • pp.98-104
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    • 2015
  • BACKGROUND: ${\gamma}$-Oryzanol, a mixture of ferulic acid esters of triterpene alcohols and sterols, are a nutritionally important group of rice secondary metabolites. The ${\gamma}$-Oryzanol content and composition were found to vary with enviromental factors such as growth temperature, varietal origin, and cultivation method. Therefore, the effect of green manure treatments will be also be an important factor in their content. METHODS AND RESULTS: The ${\gamma}$-Oryzanols extracted using dichloromethane/methanol were analyzed equipped liquid chromatography with a diode array detector. A total of ten components of ${\gamma}$-oryzanol including ${\Delta}7$-stigmastenyl ferulate were isolated of which, cycloartenyl ferulate, 24-methylenecycloartanyl ferulate, campesteryl ferulate and sitosteryl ferulate were identified as the major components in Korean rice variety, Unkwang. The ${\gamma}$-oryzanol content in rice, in the presence of nitrogen fertilization using green manure was similar to conventional nitrogen fertilization, but was higher than the control(no fertilizations). The ${\gamma}$-oryzanol ontent in rice, in the presence of hairy vetch as green manure crop was the highest compared to other crops (opium-poppy, crimson clover, cornflower). As a result of PLS-DA using SIMCA 11.0 ver. as multivariate analysis program on the basis of total data, in all samples, the specific pattern and cluster of ${\gamma}$-oryzanol scores according to green manure crops and conditions were confirmed with possible distinguishing nitrogen effects. CONCLUSION: The nitrogen contained in the green manure crops is considered to play a major role in the formation of ${\gamma}$-oryzanol. Hairy vetch which contains higher nitrogen increased the concentration of ${\gamma}$-oryzanol in rice.

Analysis for Compatibility of Daehwangmokdan-tang and Its Pharmacological Activities (대황목란탕(大黃牧丹湯)의 배오(配伍)분석과 그 약리활성)

  • Kim, Do Hoy;Yoon, Michung;Shin, Soon Shik
    • Herbal Formula Science
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    • v.26 no.1
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    • pp.81-102
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    • 2018
  • Objectives : I analysed daehwangmokdan-tang's compatibility principle by the system of chief, deputy, assistant, and envoy and investigated pharmacological activities by categorizing with chemical components, molecular level, cellular level, animal level and human level based on Korean and Chinese studies for this formula. Methods : Daehwangmokdan-tang's compatibiltity principle was examined by the system of chief, deputy, assistant, and envoy. I looked into studies that presented in Korea from 1956 to 2016 about daehwangmokdan-tang through KOREA INSTITUTE OF ORIENTAL MEDICINE, Korean medicine information system (OASIS) and in Chinese for 20 years about daehwangmokdan-tang through China National Knowledge Infrastructure, CNKI. Then classify into chemical components, molecular level, cellular level, animal level and human level to analyse. Results : According to the system of chief, deputy, assistant, and envoy, chief herb is Rhei Radix et Rhizoma and Moutan Cortex, deputy herb is Natrii Sulfas and Persicae Semen, assistant and envoy herbs are Trichosanthis Semen. The amount of extraction of paeonol, total anthraquinone, and conjugated anthraquinone from daehwangmokdan-tang with the formulation of the system of chief, deputy, assistant, and envoy was the highest, and in the formulation of chief herb and deputy herb, the extraction amount of paeonol and conjugated anthraquinone was the lowest. With other formulations, the amount of extraction of total anthraquinone and conjugated anthraquinone was improved, although the degree was different. In particular, when it is blended with Persicae Semen as a deputy herb, the extraction amount of total anthraquinone and conjugated anthraquinone of Rhei Radix et Rhizoma as a chief herb is greatly increased, and the extraction amount of paeonol is rather different, but it is lowered. It was found that the amount of Mg Ca K Na in daehwangmokdan-tang was the highest. Pharmacological activities can be detected in inflammatory mediators and enzymes for molecular level. For cellular level, it can be determined in lipopolysaccharide (LPS)-stimulated RAW 264.7 cell line. In mouse and rats for animal level and human level, in inflammatory diseases (acute appendicitis, acute pancreatitis, acute cholecystitis, acute abdominal disease, ect.), pharmacological activities was caught. Conclusions : From the above results, daehwangmokdan-tang is composed in line with the system of chief, deputy, assistant, and envoy, suggesting that there is certain rationality and scientific. Pharmacological activities of daehwangmokdan-tang are effective to anti-inflammation, improvement of sepsis, analgesic, muscle relaxation, and improvement of intestinal flora and its metabolites. Daehwangmokdan-tang is consistent with the action of diuresis and anti-inflammation in terms of the content of elements, suggesting that there is action of purging fire and removing blood stasis, defecation detoxification, cooling blood and clearing heat and activating blood and dispersing stasis.

The Possible Mechanisms Involved in Citrinin Elimination by Cryptococcus podzolicus Y3 and the Effects of Extrinsic Factors on the Degradation of Citrinin

  • Zhang, Xiaoyun;Lin, Zhen;Apaliya, Maurice Tibiru;Gu, Xiangyu;Zheng, Xiangfeng;Zhao, Lina;Abdelhai, Mandour Haydar;Zhang, Hongyin;Hu, Weicheng
    • Journal of Microbiology and Biotechnology
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    • v.27 no.12
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    • pp.2119-2128
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    • 2017
  • Citrinin (CIT) is a toxic secondary metabolite produced by fungi belonging to the Penicillium, Aspergillus, and Monascus spp. This toxin has been detected in many agricultural products. In this study, a strain Y3 with the ability to eliminate CIT was screened and identified as Cryptococcus podzolicus, based on the sequence analysis of the internal transcribed spacer region. Neither uptake of CIT by cells nor adsorption by cell wall was involved in CIT elimination by Cryptococcus podzolicus Y3. The extracellular metabolites of Cryptococcus podzolicus Y3 stimulated by CIT or not showed no degradation for CIT. It indicated that CIT elimination was attributed to the degradation of intracellular enzyme(s). The degradation of CIT by C. podzolicus Y3 was dependent on the type of media, yeast concentration, temperature, pH, and initial concentration of CIT. Most of the CIT was degraded by C. podzolicus Y3 in NYDB medium at 42 h but not in PDB medium. The degradation rate of CIT was the highest (94%) when the concentration of C. podzolicus Y3 was $1{\times}10^8cells/ml$. The quantity of CIT degradation was highest at $28^{\circ}C$, and there was no degradation observed at 3$5^{\circ}C$. The study also showed that acidic condition (pH 4.0) was the most favorable for CIT degradation by C. podzolicus Y3. The degradation rate of CIT increased to 98% as the concentration of CIT was increased to $20{\mu}g/ml$. The toxicity of CIT degradation product(s) toward HEK293 was much lower than that of CIT.

Antibacterial Activity of Pharbitin, Isolated from the Seeds of Pharbitis nil, against Various Plant Pathogenic Bacteria

  • Nguyen, Hoa Thi;Yu, Nan Hee;Park, Ae Ran;Park, Hae Woong;Kim, In Seon;Kim, Jin-Cheol
    • Journal of Microbiology and Biotechnology
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    • v.27 no.10
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    • pp.1763-1772
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    • 2017
  • This study aimed to isolate and characterize antibacterial metabolites from Pharbitis nil seeds and investigate their antibacterial activity against various plant pathogenic bacteria. The methanol extract of P. nil seeds showed the strongest activity against Xanthomonas arboricola pv. pruni (Xap) with a minimum inhibition concentration (MIC) value of $250{\mu}g/ml$. Among the three solvent layers obtained from the methanol extract of P. nil seeds, only the butanol layer displayed the activity with an MIC value of $125{\mu}g/ml$ against Xap. An antibacterial fraction was obtained from P. nil seeds by repeated column chromatography and identified as pharbitin, a crude resin glycoside, by instrumental analysis. The antibacterial activity of pharbitin was tested in vitro against 14 phytopathogenic bacteria, and it was found to inhibit Ralstonia solanacearum and four Xanthomonas species. The minimum inhibitory concentration values against the five bacteria were $125-500{\mu}g/ml$ for the n-butanol layer and $31.25-125{\mu}g/ml$ for pharbitin. In a detached peach leaf assay, it effectively suppressed the development of bacterial leaf spot, with a control value of 87.5% at $500{\mu}g/ml$. In addition, pharbitin strongly reduced the development of bacterial wilt on tomato seedlings by 97.4% at $250{\mu}g/ml$, 7 days after inoculation. These findings suggest that the crude extract of P. nil seeds can be used as an alternative biopesticide for the control of plant diseases caused by R. solanacearum and Xanthomonas spp. This is the first report on the antibacterial activity of pharbitin against phytopathogenic bacteria.

Characterization and screening of pyrrolizidine alkaloids by UPLC-MS/MS: Application to honey (UPLC-MS/MS를 이용한 벌꿀제품의 피롤리지딘 알칼로이드 잔류실태 및 분석법 선진화)

  • Ryu, Hoejin;Kim, Oukhee;Lee, Eunsoon;Kim, Misun;Kim, Jeong-gon;Yun, Eunsun;Kim, Hyunjeong;Kim, Musang
    • Analytical Science and Technology
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    • v.32 no.6
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    • pp.252-261
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    • 2019
  • Pyrrolizidine alkaloids (PAs) are a group of secondary metabolites that are produced by plants all over the world as a defense mechanism against herbivores. To date, over 660 PAs have been identified from more than 6,000 plant species that have been reported to be widely present in plants belonging to Asteraceae, Boraginaceae, and Fabaceae. This study describes an analytical method based on UPLC-MS/MS for the quantitation of 7 pyrrolizidine alkaloids (Lycopsamine, Echimidine, Retrorsine, Retrorsine N-oxide, Senecionine, Heliotrine, and Trichodesmine) in honey, and was applied to 84 honey samples for validation. Quantitation was performed based on a matrix-matched calibration to compensate for the matrix effect on the electrospray ionization. Good linear calibrations were obtained for all 7 PAs in the spiked honey samples (2.575-202.14 ㎍/kg; R2 ≥ 0.998). The extraction recoveries for most of the PAs in the honey samples were in the range of 81 %-108 %. The analysis showed that 6 of the 84 honey samples were contaminated by the PAs with the mean total sum of PAs being 47.19 ㎍/kg, and the concentrations of the PAs were observed to be in the range of 1.76-202.1 ㎍/kg. The retronecine type compounds (Echimidine, Lycopsamine) were the most frequently found PAs in honey. These data provide useful information for the assessment of human risk posed by the consumption of honey contaminated PAs.

Degradation of $^{14}C-bifenox$ in Soils under Anaerobic Conditions (혐기적 조건의 토양에서 제초제 $^{14}C-bifenox$의 분해)

  • Kwak, Hyung-Ryul;Lee, Kang-Bong;Kim, Kil-Yong;Kim, Yong-Woong;Suh, Yong-Tack
    • Korean Journal of Environmental Agriculture
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    • v.19 no.4
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    • pp.300-308
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    • 2000
  • The degradation of herbicide $^{14}C-bifenox$ was studied in soils under anaerobic conditions. $^{14}C-bifenox$ was treated in silty loam and sandy loam soils, respectively at a rate of 2.1 mg/kg, and the soil was incubated under anaerobic conditions at $25^{\circ}C$ for 180 days. The mineralization, solvent extractable and non-extractable residues, degradation products of bifenox were investigated during the experiments. The relative amounts of $^{14}CO_2$ were 1.97 and 0.9% of applied $^{14}C$ in silty loam and sandy loam soils, respectively. The non-extractable residues of sandy loam soil increased dramatically up to 79.12% of applied $^{14}C$, and were higher than those of silt loam soil, suggesting physico-chemical properties and especially organic matter contributed to the difference of $^{14}C$ between two soils. The non-extractable residues were formed mainly humin fraction and increased with time. The major metabolites were nitrofen, 5-(2,4-dichlorophenoxy)-2-Nitrobenzoate, 2,4-dichlorophenoxy aniline and methyl 5-(2,4-dichlorophenoxy) anthranilate by GC/MS analysis. From the results of volatilization, mineralization and degradation of bifenox, bifenox was stable chemically and biologically in soil.

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Comparison of Chemical Constituents in Mung bean (Vigna radiata L.) Flour between Cultivation Regions and Seeding Dates

  • An, Yeon Ju;Kim, Mi Jung;Han, Sang Ik;Chi, Hee-Youn;Kwon, Chang;Kim, So Yeon;Yang, Yu Jin;Kim, Yun Ju;Moon, Hee Sung;Kim, Seung-Hyun;Chung, Ill-Min
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.65 no.4
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    • pp.457-467
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    • 2020
  • Legumes are one of the largest families of crop plants and are widely consumed and produced for their nutritional and commercial benefits. Mung bean (Vigna radiata L.) is a legume crop that contains various functional compounds ; moreover, it has strong antioxidant properties and is becoming an increasingly important food crop. However, most previous studies on mung beans have focused on their primary metabolites. In this study, we investigated the composition and contents of phenolic compounds, fatty acids, soyasapogenol and tocopherol in mung beans cultivated in different regions and cultivated at different seeding dates. Material analysis was conducted using the following methods: LC-MS/MS, GC-FID and HPLC-ELSD. In total, 57 different samples were analyzed. Thirteen phenolic compounds were detected in mung beans. Of these, vitexin and isovitexin were the most abundant compounds, accounting for approximately 99% of phenolic compounds. The difference in phenol compounds according to the seeding dates of mung bean was not statistically significant. The total fatty acid content in beans was the highest in Pyeongchang. Significant differences in total fatty acid content were found according to the cultivation regions. Crops grown in Sohyeon and Dahyeon showed the highest soyasapogenol B content in the Suwon region, and these were the lowest in Jeonju. The total tocopherol content of beans cultivated in Dahyeon and Sohyeon was the lowest and highest in Pyeongchang. Soyasapogenol B and total tocopherol content were not significantly different according to seeding dates. This study was conducted to obtain basic data for the cultivation of mung beans with a high content of various functional materials in terms of regional specialization and optimal seeding time.

Urinary PAH Metabolites as Biomarkers of Environmental PAHs Exposure (환경성 PAHs 노출과 생체지표 연구)

  • Lee, Kyoung-Ho;Li, Zhung-Min;Cho, Soo-Hun;Kwon, Ho-Jang;Kang, Dae-Hee
    • Toxicological Research
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    • v.23 no.1
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    • pp.39-46
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    • 2007
  • The aim of the study was to see if there is any differences in urinary 1-hydroxypyrene glucuronide (1-OHPG) and 2-naphthol levels in children ($8{\sim}14$ years old) and their mothers ($30{\sim}46$ years old) living three cities in South Korea (Seoul, Incheon and Pohang) and three in China (Changchun, Datong and Kunming), where the levels of air pollution varies. The factors related with urinary biomarkers levels were also evaluated. The study subjects consisted of 118 Korean (60 children and 58 their mothers) and 120 Chinese (60 children and 60 their mothers). Urinary 1-OHPG was measured by synchronous fluorescence spectroscopy after immuno-affinity purification using monoclonal antibody 8E11 and urinary 2-naphthol concentrations were determined by HPLC with fluorescence detector. Information on recent consumption of diet containing high PAHs, environmental tobacco smoke (ETS), type of cooking and heating fuels, and other life-style characteristics were collected by self-administered questionnaire. The arithmetic mean of urinary 1-OHPG levels (n = 120, $mean{\pm}SD$, $6.77{\pm}7.96{\mu}mol/mol$ creatinine) in Chinese were 10 fold higher than those in Korean (n = 118, $0.62{\pm}0.61{\mu}mol/mol$ creatinine) (P < 0.01). Urinary 2-naphthol levels in Chinese (n = 119, $59.50{\pm}82.29{\mu}g/g$ creatinine) were significantly higher than those in Korean (n = 117, $25.09{\pm}46.56{\mu}g/g$ creatinine) (P < 0.01). Urinary 1-OHPG and 2-naphthol levels were significantly higher in children living the polluted cities in China (Datong and Chanchun, respectively). Multiple linear regression analysis indicated that living in factory area (vs. residential area) and use of coal stove as heating fuel were significant predictors for urinary 1-OHPG (overall model $R^2$= 0.46, n = 204). And ETS was predictor for urinary 2-naphthol levels in Korean ($R^2$ = 0.36, n = 46). These results indicated that urinary 1-OHPG and 2-naphthol levels were related with different ambient particulate air pollution, type of heating fuels and ETS.

Structure Analysis of pmcABCDEFT Gene Cluster for Degradation of Protocatechuate from Comamonas sp. Strain DJ-12 (Comamonas sp. Strain DJ-12로부터 Protocatechuate의 분해에 관여하는 pmcABCDEFT 유전자군의 구조 분석)

  • Kang Cheol-Hee;Lee Sang-Mhan;Lee Kyoung;Lee Dong-Hun;Kim Chi-Kyung
    • Korean Journal of Microbiology
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    • v.41 no.3
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    • pp.195-200
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    • 2005
  • Comamonas sp. strain DJ-12 is a bacterial isolate capable of degrading of 4-chlorobiphenyl (4CB) as a carbon and energy source. The degradation pathway was characterized as being conducted by consecutive reactions of the meta-degradation of 4CB, hydrolytic dechlorination of 4-chlorobenzoate (4CBA), hydroxylation of 4-hydroxybenzoate, and meta-degradation of protocatechuate to product TCA metabolites. The 6.8 kb fragment from the chromosomal DNA of Comamonas sp. strain DJ-12 included the genes encoding for the meta-degradation of PCA; the genes of protocatechuate 4,5-dioxygenase alpha and beta subunits (pmcA and pmcB), 4-carboxy-2-hydroxymuconate-6-semialdehyde dehydrogenase (pmcC), 2-pyrone-4,6-dicarboxylate hydrolase (pmcD), 4-oxalomesaconate (OMA) hydratase(pmcE), 4-oxalocitramalate (OCM) aldolase (pmcF), and transporter gene (pmcT). They were organized in the order of pmcT-pmcE-pmcF-pmcD-pmcA-pmcB-pmcC. The amino acid sequences deduced from the nucleotide sequences of pmcABCDEFT genes from Comamonas sp. strain DJ-12 exhibited 94 to $98\%$ homologies with those of Comamonas testosteroni BR6020 and Pseudomonas ochraceae NGJ1, but only 52 to $74\%$ with homologies Sphingomonas paucimobilis SYK-6, Sphingomonas sp. LB126, and Arthrobacter keyseri 12B.