• 한국환경보건학회지
• /
• 제37권6호
• /
• pp.474-481
• /
• 2011

This study investigated urinary metabolites of polycyclic aromatic hydrocarbons (PAHs) in the urine of smokers and non-smokers by liquid chromatography triple quordrupole tandem mass spectroscopy (LC/MS/MS). Compounds analyzed for urinary biomarkers of PAHs were five mono-hydroxylated PAHs metabolites; 1-naphthol, 2-naphthol, 1-hydroxypyrene(1-OHP), 3-phenanthrol, 2-fluorenol. Urine samples were pretreated by enzymatic hydrolysis and solid phase extraction method. Smokers were composed of 17 men and five women; non-smokers 17 men and 16 women. Smoking increased urinary concentrations of five PAHs metabolites significantly higher than those of nonsmokers. Statistically significant correlations among the five PAHs metabolites were shown. The results suggest that LC/MS/MS technology should be useful in the environmental health discipline.

• Biomolecules & Therapeutics
• /
• 제4권3호
• /
• pp.251-256
• /
• 1996

The metabolism of carbinoxamine, 2-[(4-chlorophenyl)-2-pyridinyl-methoxy]-N, N-dimethylethaneamine, was studied in adult male volunteers after an oral dose of 15 mg. Solvent extracts of urine obtained with or without enzyme hydrolysis were analyzed by gas chromatography-mass spectrometry after derivatization with MSTFA/TMSCl (N-methyl-N-trimethylsilyltrifluoroacetamide/trimethyl chlorosilane). The structures of metabolites were determined based on the electron impact (EI) and chemical ionization (CI) mass spectra. Nonconjugated metabolites identified in the urine were carbinoxamine, nor-carbinoxamine, and bits-nor-carbinoxamine. Parent drug, nor-carbinoxamine, and bits-nor-carbinoxamine were also detected as conjugated forms. These metabolites observed in human urine were different from those previously reported in the rat. Urinary excretions of carbinoxamine were reached to maxima in 4 hours after drug administration with 4.9%-8.1% and 2.5-4.2% of the dose excreted during 24 h as carbinoxamine and its glucuronide, respectively.

• Archives of Pharmacal Research
• /
• 제15권2호
• /
• pp.109-114
• /
• 1992

For the risk assessment of human exposure to volatile halogenated hydrocarbons, a dynamic purge trap/on-column cryofocusing method using capillary gas chromatograph-$^{63}Ni$ electron capture detector and thermal desorption unit was applied to analyze the free forms, metabolites of 1, 1, 2-trichloroethylene and 1, 1, 2, 2-tetrachloroethylene. The urine sample was diluted with distilled water, hydrolyzed and sealed. Then the inert gas was infused to purge out free 1, 1, 2-trichloroethylene, free 1, 1, 2, 2-tetrachloroethylene and urichloroethanol. These compounds were trapped to $Tenax^R$ / GC-gas trap device throughout clean up tube. Being undertectable to gas chromatograph directly, trichloroacetic acid was methyl esterificated and trapped in the manner above mentioned. The optimal incubation time to get best recovery of methyl ester was 4 hours at $60^circ$C. The concentrations of free volatile halogenated hydrocarbons and their metabolites in urine were obtained of free volatile halogenated hydrocarbons and their metabolites in urine were obtained from 5 healthy volunteers. This analytical method is expected to make the biological monitoring more precise and convenient.

• Bulletin of the Korean Chemical Society
• /
• 제19권2호
• /
• pp.194-196
• /
• 1998

The metabolism of dromostanolone (2α-methyl-5α- androstan-17β-ol-3-one) was studied in three adult volunteers after oral dose of 20 mg. Solvent extracts of urine obtained after enzyme hydrolysis were derivatized with MSTFA/TMCS and MSTFA/TMIS. The structures of intact drug and its metabolites were determined by gas chromatography/mass spectrometry (GC/MS) in electron impact (EI) mode. The major metabolite (2α-methyl-5α- androstan-3α-ol-17-one), its 3β-epimer, parent compound, and several hydroxylated metabolites including intact drug were detected by comparing total ion chromatograms of control urine with that of the administered sample. Two epimers of 2α-methyl-5α- androstan-3,17β-diol were detected using selected ion monitoring. The maximum excretion of dromostanolone and 2α-methyl-5α- androstan-3α-ol-17-one was reached in 6.2-15 hr. The half-life of intact dromostanolone was 5.3 hr. About 3.0% of the administered amount was found to be excreted within 95 hr as unchanged form.

• Toxicological Research
• /
• 제27권1호
• /
• pp.15-18
• /
• 2011

The toxicities of phenanthrene (PH) and pyrene (PY) are less than benzo(a)pyrene (BaP), but both compounds are found in higher concentrations in the air, feed, and food. Most PAHs are metabolized to hydroxylated compounds by the hepatic cytochrome P450 monooxigenases system. Metabolites are excreted into urine and feces. We determined concentrations of PH, PY and BaP in muscle and hydroxylated metabolites, 3-OH-PH, 1-OH-PY, and 3-OH-BaP, respectively, in urine from dairy cattle (n = 24). We also evaluated the relationship between parent compounds in muscle and their metabolites in urine. Concentrations of PH and PY in muscle ranged from 0.7~4.8 ng/g ($1.8{\pm}1.7$) and 0.4~4.1 ng/g ($1.2{\pm}1.2$), respectively. Concentrations of 3-OH-PH and 1-OH-PY in urine ranged from 0.1~5.9 ng/ml ($2.9{\pm}3.7$) and 0.5~3.6 ng/ml ($1.9{\pm}2.3$), respectively. Correlation coefficient for PY concentration in muscle versus 1-OH-PY in urine was 0.657 and for PH concentration in muscle versus 3-OH-PH in urine was 0.579. Coefficient determination for PY and PH concentrations in muscle was 0.886 and for 1-OH-PY and 3-OH-PH in urine was 0.834. This study suggests that 1-OH-PY and 3-OH-PH could be used as biomarkers for PAHs exposure in dairy cattle.

• Animal Bioscience
• /
• 제36권7호
• /
• pp.1130-1142
• /
• 2023

Objective: Cow urine possesses several bioactive properties but the responsible components behind these bioactivities are still far from identified. In our study, we tried to identify the possible components behind the antimicrobial activity of cow urine by exploring the peptidome and metabolome. Methods: We extracted peptides from the urine of Sahiwal cows belonging to three different physiological states viz heifer, lactation, and pregnant, each group consisting of 10 different animals. The peptides were extracted using the solid phase extraction technique followed by further extraction using ethyl acetate. The antimicrobial activity of the aqueous extract was evaluated against different pathogenic strains like Staphylococcus aureus, Escherichia coli, and Streptococcus agalactiae. The safety of urinary aqueous extract was evaluated by hemolysis and cytotoxicity assay on the BuMEC cell line. The urinary peptides were further fractionated using high-performance liquid chromatography (HPLC) to identify the fraction(s) containing the antimicrobial activity. The HPLC fractions and ethyl acetate extract were analyzed using nLC-MS/MS for the identification of the peptides and metabolites. Results: A total of three fractions were identified with antimicrobial activity, and nLC-MS/MS analysis of fractions resulted in the identification of 511 sequences. While 46 compounds were identified in the metabolite profiling of organic extract. The urinary aqueous extract showed significant activity against E. coli as compared to S. aureus and S. agalactiae and was relatively safe against mammalian cells. Conclusion: The antimicrobial activity of cow urine is a consequence of the feeding habit. The metabolites of plant origin with several bioactivities are eliminated through urine and are responsible for their antimicrobial nature. Secondly, the plethora of peptides generated from the activity of endogenous proteases on protein shed from different parts of tissues also find their way to urine. Some of these sequences possess antimicrobial activity due to their amino acid composition.

• 분석과학
• /
• 제15권3호
• /
• pp.185-189
• /
• 2002

Cyclofenil은 국제올림픽위원회(IOC)에서 금지약물로 규정하고 있다. 이에 따라 본 실험에서는 GC/MS를 사용하여 소변으로부터 cyclofenil의 복용여부를 검사할 수 있는 방법을 만들었다. 이를 위하여 cyclofenil 및 그 대사체들의 추출 회수율을 측정하였는데 pH 5-9 사이가 최적의 추출 조건이었으며, 소변으로는 모 약물인 cyclofenil은 배설되지 않아서 검출하지 못하였고 대신에 그 대사체들을 검출함으로써 cyclofenil의 복용여부를 검출할 수 있었다. 따라서 대사체들은 가수분해 후 pH 9.6에서 에테르로 추출하여 MSTFA로 유도체화시켜서 GC/MS로 분석하였다.

• Archives of Pharmacal Research
• /
• 제7권1호
• /
• pp.23-31
• /
• 1984

An improved mass spectrometric method for multicomponent analysis of metabolites in urine, well-suited for clinical biochemistry, is described. The method involves solvent elution of the metabolites from an adsorbent and the concentration of the eluate on a microadsorption column. This is administered by a direct inlet probe into the ionizing source of field ionization mass spectrometry (FIMS), which yield a molecular weight profile of the metabolites. The procedure provides rapidly (within one hour) reproducible profiles from a small volume of urine. The optimization of the sampling technique and the reproducibility are discussed.

• 대한의생명과학회지
• /
• 제11권4호
• /
• pp.509-515
• /
• 2005

This study was conducted to determine the kinetics of cyclohexane metabolites (the biomarker on cyclohexane exposure), the changes of hepatic cyclohexane metabolizing enzyme activities and the metabolites of cyclohexane in urine or serum. The rats were sacrificed at 2, 4, 8, 12 and 24 hr after administration of one dose of cyclohexane (1.56 g/kg body weight, i.p.). The metabolites of cyclohexane in urine were identified as cyclohexanol, cyclohexanone, trans-l,2-cyclohexanediol and 1,4-cyclohexanediol with cyclohexane metabolite being 124.00, 0.78, 23.28 and 2.75 (g/g of creatinine, $1\times10^{-3}$). Most of the cyclohexanol and trans-l,2-cyclohexanediol were determined to be in the form of $\beta-glucuronide$ conjugates, whereas cyclohexanone and 1 ,4-cyclohexanediol were found as free forms. In toxicokinetics of serum cyclohexane metabolites, cyclohexanol showed a rapid increase, reaching the plateau at 4 hr, after this time rapidly decreased throughout 24 hr. Changes of cyclohexanone also showed the similar pattern with cyclohexanol except somewhat lower concentration. Trans-l,2-cyclohexanediol, however, showed a gradual increase until 12 hr with the continued same levels throughout 24 hr. On the other hand, 1,4-cyclohexanediol was detected as trace levels at 4 and 12 hr, respectively. The administration of cyclohexane led to a significant increase of hepatic aniline hydroxylase activity from 2 to 8 hr. The activity of hepatic alcohol dehydrogenase showed a significant increase at 4 hr and then were recovered to the level of the control at 24 hr. On the other hand, there were no differences in liver weightlbody weight between the control and cyclohexane-treated animals. However, there were the changes of aniline hydroxylase and alcohol dehydrogenase activities on time-dependent pattern after cyclohexane treatment, which influence on the degree of cyclohexane metabolites both in blood and urine. These results suggest that differential determination of cyclohexane metabolites in urine and serum may be able to be as a biomarker of cyclohexane-exposure in the body. But in this fields further study is needed.

• 분석과학
• /
• 제23권2호
• /
• pp.179-186
• /
• 2010

본 연구는 건강한 성인남자로부터 ibuprofen의 인체 내 대사물질의 규명 및 배설에 대한 연구를 gas chromatography-mass spectrometry (GC/MS) 법으로 수행하였다. Ibuprofen 복용 후 3시간 단위로 15시간까지 소변을 채취하여 실험하였다. 이 중 glucuronide가 포합된 대사체로부터 포합체를 떼어내기 위해 6 M HCl을 통해 $100^{\circ}C$에서 30분 동안 가열하여 산 가수분해 시켰다. 가수분해 된 뇨 중 ibuprofen과 그 대사체를 추출하기 위하여 액체-액체 추출법을 적용하였다. 모약물과 대사체의 추출 최적조건을 찾기 위하여 pH 3, 5 및 8에서 수행하였으며 그 중 pH 3에서 가장 좋은 회수율을 보여 주었다. 또한 미량의 대사체 검출을 위하여 trimethylsilylation (TMS) 유도체 반응을 적용시킨 후 GC-MS를 이용하여 분석하였다. 본 연구에서는 모약물을 포함한 총 5개의 대사체를 검출하였으며, 이들 대사물질들은 주로 산화과정에 의해 약물 모핵에 hydroxylation 또는 carboxylation된 화합물이었다. 또한 시간대별로 채취한 시료에서 각각의 대사체의 배설율의 변화를 관찰함으로써 시간대별 배설율 및 축적률을 구할 수 있었다.