• Title/Summary/Keyword: Liquid Chromatography-tandem Mass Spectrometry

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Determination of Phthalate Metabolites in Human Serum and Urine as Biomarkers for Phthalate Exposure Using Column-Switching LC-MS/MS

  • Jeong, Jee-Yeon;Lee, Ji-Hyun;Kim, Eun-Young;Kim, Pan-Gyi;Kho, Young-Lim
    • Safety and Health at Work
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    • v.2 no.1
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    • pp.57-64
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    • 2011
  • Objectives: Although phthalates like dibutyl phthalate (DBP) and di-2-ethylhexyl phthalate (DEHP) are commonly used as plasticizers and their metabolites are especially suspected of reproductive toxicity, little is known about occupational exposure to those phthalates. The aim of this study was to assess the utility of measuring the metabolite concentrations of DBP and DEHP in serum and urine samples as an indicator of occupational exposure to those phthalates. Methods: Phthalate metabolites were analyzed by using column-switching high-performance liquid chromatography tandem mass spectrometry (LC-MS/MS). Results: We detected phthalate metabolites in serum and urine matrices at approximately 10-fold lower than the limit of detection of those metabolites in the same matrix by LC-MS/MS without column switching, which was sufficient to evaluate concentrations of phthalate metabolites for industrial workers and the general population. Conclusion: The accuracy and precision of the analytical method indicate that urinary metabolite determination can be a more acceptable biomarker for studying phthalate exposure and adverse health outcomes.

Simulateous Determination for the Contents of Caffeine and Chlorogenic Acid in Coffee Beans (원두커피내 카페인 및 클로로겐산 함량 평가를 위한 동시분석법)

  • Shin, Jeoung Hwa;Ahn, Yun Gyong
    • The Korean Journal of Community Living Science
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    • v.24 no.1
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    • pp.5-12
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    • 2013
  • Caffeine is an alkaloid of the methylxanthine family known as a central nervous system stimulant, temporarily warding off drowsiness and restoring alertness in humans. There is a recommended upper limits of caffeine for health because a high dose can cause negative effects. Chlorogenic acid is a natural polyphenol compound known to have an antioxidant activity. In this study, the contents of caffeine and chlorogenic acid in coffee beans from different origins(Costa Rica, Indonesia, Vietnam) were determined by using liquid chromatography-tandem mass spectrometry(LC-MS/MS). The experiment offers more selectivity and sensitivity for those compounds compared with conventional methods such as UV/VIS spectrophotometry. The average concentrations of caffeine and chlorogenic acid in coffee beans origined in Costa Rica were 15.05 mg/g and 5.33 mg/g respectively. In the case of coffee beans origined in Indonesia, the average concentrations were 13.10 mg/g for caffeine and 3.75 mg/g for chlorogenic acid. Vietnamese coffee showed that the average concentrations were 17.79 mg/g for caffeine and 1.12 mg/g for chlorogenic acid. This study can contribute to a better understanding of the contents of caffeine and chlorogenic acid in various coffee beans in order to evaluate dietary intake.

Maternal 3-methylcrotonyl-coenzyme A carboxylase deficiency with elevated 3-hydroxyisovalerylcarnitine in breast milk

  • Cho, Kyung Lae;Kim, Yeo Jin;Yang, Song Hyun;Kim, Gu-Hwan;Lee, Jun Hwa
    • Clinical and Experimental Pediatrics
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    • v.59 no.sup1
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    • pp.41-44
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    • 2016
  • We report here a case of maternal 3-methylcrotonyl-coenzyme A carboxylase (3-MCC) deficiency in a Korean woman. Her 2 infants had elevated 3-hydroxyisovalerylcarnitine (C5-OH) on a neonatal screening test by liquid chromatography-tandem mass spectrometry (LC-MS/MS), but normal results were found on urine organic acid analysis. The patient was subjected to serial testing and we confirmed a maternal 3-MCC deficiency by blood spot and breast milk spot test by LC-MS/MS, serum amino acid analysis, urine organic acid and molecular genetic analysis that found c.838G>T (p.Asp280Tyr) homozygous mutation within exon 9 of the MCCB gene. Especially, we confirmed marked higher levels of C5-OH on breast milk spot by LC-MS/MS, in the case of maternal 3-MCC deficiency vs. controls.

Metagenomic and Proteomic Analyses of a Mangrove Microbial Community Following Green Macroalgae Enteromorpha prolifera Degradation

  • Wu, Yijing;Zhao, Chao;Xiao, Zheng;Lin, Hetong;Ruan, Lingwei;Liu, Bin
    • Journal of Microbiology and Biotechnology
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    • v.26 no.12
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    • pp.2127-2137
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    • 2016
  • A mangrove microbial community was analyzed at the gene and protein levels using metagenomic and proteomic methods with the green macroalgae Enteromorpha prolifera as the substrate. Total DNA was sequenced on the Illumina HiSeq 2000 PE-100 platform. Two-dimensional gel electrophoresis in combination with liquid chromatography tandem mass spectrometry was used for proteomic analysis. The metagenomic data revealed that the orders Pseudomonadales, Rhizobiales, and Sphingomonadales were the most prevalent in the mangrove microbial community. By monitoring changes at the functional level, proteomic analyses detected ATP synthase and transporter proteins, which were expressed mainly by members of the phyla Proteobacteria and Bacteroidetes. Members of the phylum Proteobacteria expressed a high number of sugar transporters and demonstrated specialized and efficient digestion of various glycans. A few glycoside hydrolases were detected in members of the phylum Firmicutes, which appeared to be the main cellulose-degrading bacteria. This is the first report of multiple "omics" analysis of E. prolifera degradation. These results support the fact that key enzymes of glycoside hydrolase family were expressed in large quantities, indicating the high metabolic activity of the community.

Establishment of Withdrawal Time of Erythromycin for the Cultured Black Rockfish (Sebastes schlegeli) after Oral Administration

  • Park, Mi-Jung;Choi, Kyu-Duck;Lee, Tae-Seek;Choi, Hye-Sung;Shim, Kil-Bo;Myeong, Jeong-In;Park, Mi-Seon;Shin, Il-Shik
    • Food Science and Biotechnology
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    • v.18 no.2
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    • pp.533-536
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    • 2009
  • Withdrawal time of erythromycin in the cultured black rockfish (Sebastes schlegeli) was investigated to provide regulatory authority and fishery industry with full information needed to secure food safety of the fish treated with erythromycin. Medication was carried out using the experimental diet containing erythromycin of which concentration was 200 mg/kg diet and its daily dose was about 40 mg/kg body weight. The withdrawal time needed to reduce antibiotic contents around 10.0 mg/kg accumulated during 9 days medication below 0.2 mg/kg was identified about 10 days. The antibiotic with 13.7 mg/kg of fish on the $9^{th}$ days of medication was completely depleted after 30 days from stop of medication.

Hepatotoxic Effect of 1-Bromopropane and Its Conjugation with Glutathione in Male ICR Mice

  • Lee Sang Kyu;Jo Sang Wook;Jeon Tae Won;Jun In Hye;Jin Chun Hua;Kim Ghee Hwan;Lee Dong Ju;Kim Tae-Oh;Lee Eung-Seok;Jeong Tae Cheon
    • Archives of Pharmacal Research
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    • v.28 no.10
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    • pp.1177-1182
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    • 2005
  • The hepatotoxic effects of 1-bromopropane (1-BP) and its conjugation with glutathione were investigated in male ICR mice. A single dose (1000 mg/kg, po) of 1-BP in corn oil to mice significantly increased serum activities of alanine aminotransferase and aspartate aminotransferase. Glutathione (GSH) content was dose-dependently reduced in liver homogenates 12 h after 1-BP treatment. In addition, 1-BP treatment dose-dependently increased levels of S-pro-pyl GSH conjugate at 12 h after treatment, as measured by liquid chromatography-electro-spray ionization tandem mass spectrometry. The GSH conjugate was maximally increased in liver at 6 h after 1-BP treatment (1000 mg/kg), with a parallel depletion of hepatic GSH content. Finally, 1-BP induced the production of malondialdehyde in liver. The present results suggest that 1-BP might cause hepatotoxicity, including lipid peroxidation via the depletion of GSH, due to the formation of GSH conjugates in male ICR mice.

Rapid Screening of Phospholipid Biomarker Candidates from Prostate Cancer Urine Samples by Multiple Reaction Monitoring of UPLC-ESI-MS/MS and Statistical Approaches

  • Lim, Sangsoo;Bang, Dae Young;Rha, Koon Ho;Moon, Myeong Hee
    • Bulletin of the Korean Chemical Society
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    • v.35 no.4
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    • pp.1133-1138
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    • 2014
  • Ultrahigh performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UPLC-ESI- MS/MS) provides a high-speed method to screen a large number of samples for small molecules with specific properties. In this study, UPLC-ESI-MS/MS with multiple reaction monitoring (MRM) was employed to screen urinary phospholipid (PL) content for biomarkers of prostate cancer. From lists of urinary PLs structurally identified using nanoflow LC-ESI-MS/MS, 52 PL species were selected for quantitative analysis in urine samples between 22 cancer-free urologic patients as controls and 45 prostate cancer patients. Statistical treatment of data by receiver operating characteristic (ROC) analysis yielded 14 PL species that differed significantly in relative concentrations (area under curve (AUC) > 0.8) between the two groups. Among PLs present at higher levels in prostate cancer urine, phosphatidylcholines (PCs) and phosphatidylinositols (PIs) constituted the major head group PLs (3 PCs and 7 PIs). For technical reasons, PL species of low abundance may be underrepresented in data from UPLC-ESI-MS/MS performed in MRM mode. However, the proposed method enables the rapid screening of large numbers of plasma or urine samples in the search for biomarkers of human disease.

Surface interactions between two of the main periodontal pathogens: Porphyromonas gingivalis and Tannerella forsythia

  • Zhu, Weidong;Lee, Seok-Woo
    • Journal of Periodontal and Implant Science
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    • v.46 no.1
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    • pp.2-9
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    • 2016
  • Purpose: Porphyromonas gingivalis and Tannerella forsythia have been implicated as the major etiologic agents of periodontal disease. These two bacteria are frequently isolated together from the periodontal lesion, and it has been suggested that their interaction may increase each one's virulence potential. The purpose of this study was to identify proteins on the surface of these organisms that are involved in interbacterial binding. Methods: Biotin labeling of surface proteins of P. gingivalis and T. forsythia and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis was performed to identify surface proteins involved in the coaggregating activity between P. gingivalis and T. forsythia. Results: It was found that three major T. forsythia proteins sized 161, 100, and 62 kDa were involved in binding to P. gingivalis, and P. gingivalis proteins sized 35, 32, and 26 kDa were involved in binding to T. forsythia cells. Conclusions: LC-MS/MS analysis identified one T. forsythia surface protein (TonB-linked outer membrane protein) involved in interbacterial binding to P. gingivalis. However, the nature of other T. forsythia and P. gingivalis surface proteins identified by biotin labeling could not be determined. Further analysis of these proteins will help elucidate the molecular mechanisms that mediate coaggregation between P. gingivalis and T. forsythia.

The Physiological Role of CPR1 in Saccharomyces cerevisiae KNU5377 against Menadione Stress by Proteomics

  • Kim, Il-Sup;Yun, Hae-Sun;Kwak, Sun-Hye;Jin, Ing-Nyol
    • Journal of Microbiology
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    • v.45 no.4
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    • pp.326-332
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    • 2007
  • In order to understand the functional role of CPRl in Saccharomyces cerevisiae KNU5377 with regard to its multi-tolerance characteristics against high temperatures, inorganic acids, and oxidative stress conditions, whole cellular proteins were analyzed via liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). This procedure was followed by two-dimensional (2-D) gel electrophoresis. Under menadione stress conditions, the 23 upregulated proteins were clearly identified only in the wild- type strain of KNU5377. Among the proteins, Sodl1p Tsa1p, Ahp1, Cpr1p, Cpr3, Ssb2p, and Hsp12p were identified as components of antioxidant systems or protein-folding related systems. The CPR1 protein could not be completely detected in the $cpr1{\Delta}$ mutant of KNU5377 and the other upregulated proteins in the wild-type strain evidenced a clear correlation with the results of immunoblot analysis. Moreover, a reduction in growth patterns (about 50%) could be observed in the $cpr1{\Delta}$ mutant, as compared with that of the wild-type strain under mild MD stress conditions. These results indicate that the upregulation of CPR1 may contribute to tolerance against MD as an inducer of oxidative stress.

Antioxidant and Anti-Inflammatory Effects of NCW Peptide from Clam Worm (Marphysa sanguinea)

  • Park, Young Ran;Park, Chan-Il;Soh, Yunjo
    • Journal of Microbiology and Biotechnology
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    • v.30 no.9
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    • pp.1387-1394
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    • 2020
  • Clam worms (Marphysa sanguinea) are a rich source of bioactive components such as the antibacterial peptide, perinerin. In the present study, we explored the physiological activities of a novel NCWPFQGVPLGFQAPP peptide (NCW peptide), which was purified from clam worm extract through high-performance liquid chromatography. Tandem mass spectrometry (MS/MS) revealed that NCW was a new peptide with a molecular weight of 1757.86 kDa. Moreover, NCW peptide exhibited significant antioxidant effects, causing a 50% inhibition of DPPH radical at a concentration of 20 μM without showing any cytotoxicity. These were associated with a reduction in the activity of catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and malondialdehyde (MDA) in LPS-stimulated RAW264. 7 cells. Furthermore, NCW peptide exhibited anti-inflammatory effects in LPS-stimulated RAW264.7 macrophages via inhibition of the abnormal production of pro-inflammatory cytokines including nitric oxide (NO), inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2). These anti-inflammatory effects of NCW peptide were associated with the inhibition of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α). Our results therefore suggest that this novel NCW peptide with antioxidant and anti-inflammatory effects could be a good therapeutic agent against inflammation-related diseases.