• Title/Summary/Keyword: electrospray ionization (ESI)

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Characterization of a Collagenase-1 Inhibitory Peptide Purified from Skate Dipturus chilensis Skin (홍어류(Dipturus chilensis) 껍질로부터 분리 정제된 collagenase-1 저해 펩타이드의 특성)

  • Park, Sung-Ha;Lee, Jung-Kwon;Jeon, Joong-Kyun;Byun, Hee-Guk
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.44 no.5
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    • pp.456-463
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    • 2011
  • We attempted to isolate a collagenase-1 inhibitory peptide from skate Dipturus chilensis skin protein. The protein from skate skin was digested by various enzymes (alcalase, ${\alpha}$-chymotrypsin, neutrase, papain, pepsin, and trypsin) to produce a collagenase-1 inhibitory peptide. The collagenase-1 inhibitory activity of the peptides obtained was measured by gelatin digestion assay. Among the six hydrolysates, pepsin hydrolysate exhibited the highest collagenase-1 inhibitory activity. The peptide showing strong collagenase-1 inhibitory activity was purified by Sephadex G-25 gel chromatography and HPLC using an octadecylsilyls (ODS) column. The amino acid sequence of purified collagenase-1 inhibitory peptide was identified to be Asn-Leu-Asp-Val -Leu-Glu-Val-Phe (961 Da) by quadrupole time of flight (Q-TOF) and electrospray ionization mass spectrometry (ESI-MS) mass spectroscopy. The $IC_{50}$ value of purified peptide was 87.0 ${\mu}M$. Moreover, the peptide did not exhibit cytotoxic effects on human dermal fibroblast cell lines.

LC-MS/MS Profiling-Based Secondary Metabolite Screening of Myxococcus xanthus

  • Kim, Ji-Young;Choi, Jung-Nam;Kim, Pil;Sok, Dai-Eun;Nam, Soo-Wan;Lee, Choong-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.19 no.1
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    • pp.51-54
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    • 2009
  • Myxobacteria, Gram-negative soil bacteria, are a well-known producer of bioactive secondary metabolites. Therefore, this study presents a methodological approach for the high-throughput screening of secondary metabolites from 4 wild-type Myxococcus xanthus strains. First, electrospray ionization mass spectrometry (ESI-MS) was performed using extracellular crude extracts. As a result, 22 metabolite peaks were detected, and the metabolite profiling was then conducted using the m/z value, retention time, and MS/MS fragmentation pattern analyses. Among the peaks, one unknown compound peak was identified as analogous to the myxalamid A, B, and C series. An analysis of the tandem mass spectrometric fragmentation patterns and HR-MS identified myxalamid K as a new compound derived from M. xanthus. In conclusion, LC-MS/MS-based chemical screening of diverse secondary metabolites would appear to be an effective approach for discovering unknown microbial secondary metabolites.

2'-Hydroxylation of Genistein Enhanced Antioxidant and Antiproliferative Activities in MCF-7 Human Breast Cancer Cells

  • Choi, Jung-Nam;Kim, Doc-Kyu;Choi, Hyung-Kyoon;Yoo, Kyung-Mi;Kim, Ji-Young;Lee, Choong-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.19 no.11
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    • pp.1348-1354
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    • 2009
  • Bioconversion of the isoflavonoid genistein to 2'-hydroxygenistein (2'-HG) was performed using isoflavone 2'-hydroxylase (CYP81E1) heterologously expressed in yeast. A monohydroxylated product was analyzed by liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) and NMR spectrometry and was identified as 2'-HG. An initial bioconversion rate of 6% was increased up to 14% under optimized conditions. After recovery, the biological activity of 2'-HG was evaluated. Bioconverted 2'-HG showed higher antioxidant activity against 1,1-diphenyl-2-picryl hydrazine (DPPH) and 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radicals than did genistein. Furthermore, 2'-HG exhibited greater antiproliferative effects in MCF-7 human breast cancer cells than did genistein. These results suggest that 2'-hydroxylation of genistein enhanced its antioxidant activity and cell cytotoxicity in MCF-7 human breast cancer cells.

Development and validation of a selective and sensitive LC-MS/MS method for determination of misoprostol acid in human plasma: Application to bioequivalence study

  • Park, Yong Sun;Kim, Keun Nam;Kim, Ye Na;Kim, Jung Hwan;Kim, Jin Young;Sim, Soo Ji;Lee, Heon Soo
    • Analytical Science and Technology
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    • v.28 no.1
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    • pp.17-25
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    • 2015
  • A rapid, sensitive and specific method was developed and validated using electrospray ionization (ESI) tandem mass spectrometry (LC-MS/MS) for determination of misoprostol acid in human plasma. Misoprostol $acid-d_5$ was used as in internal standard (IS). The analyte and IS were extracted by simple one step solid phase extraction (SPE). Linearity in plasma was obtained over the concentration range 10~3000 pg/mL and lower limit of quantification (LLOQ) was identifiable and reproducible at 10 pg/mL. The intra- and inter-day precision values were below 9% and the accuracy was ranged from 93.81% to 102.02% at all four quality control samples. The method was has been successfully applied for routine assay to support pharmacokinetic study of misoprostol acid in human plasma after an oral administration of 0.4 mg misoprostol.

Polymethoxylated Flavone Extracts from Citrus Peels for Use in the Functional Food and Nutraceutical Industry

  • Yao, Xiaolin;Pan, Siyi;Duan, Chunhong;Yang, Fang;Fan, Gang;Zhu, Xinrong;Yang, Shuzhen;Xu, Xiaoyun
    • Food Science and Biotechnology
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    • v.18 no.5
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    • pp.1237-1242
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    • 2009
  • Polymethoxylated flavones (PMFs) extracted from Citrus sinensis 'Jincheng' peel were characterized by chromatographic and spectroscopic techniques. Seven individual PMF were identified. 3, 3', 4', 5, 6, 7-hexamethoxyflavone (HEX), nobiletin (NOB), heptamethoxyflavone (HEP), 5-demethylnobiletin (DN), and tangeretin (TAN) were characterized through electrospray ionization mass spectrometry (ESI-MS) in positive mode of protonated molecular ions $[M+H]^+$, the diagnostic fragment ions, together with the UV-Vis spectra and high performance liquid chromatography (HPLC) elution order from literature data. Sinensetin (SIN) and tetramethyl-O-scutellarein (SCU) were isolated and identified through their MS, $^1H$ nuclear magnetic resonance (NMR) and UV-Vis spectral studies. The levels of PMFs in peels from different cultivars of citrus fruits grown in China were determined for the first time. The results showed that C. aurantium 'Bitter orange' peel was the most promising variety for HEP. C. sinensis peel was a good source for SIN and SCU.

Rapid Determination of L-carnitine in Infant and Toddler Formulas by Liquid Chromatography Tandem Mass Spectrometry

  • Ahn, Jang-Hyuk;Kwak, Byung-Man;Park, Jung-Min;Kim, Na-Kyeoung;Kim, Jin-Man
    • Food Science of Animal Resources
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    • v.34 no.6
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    • pp.749-756
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    • 2014
  • A rapid and simple analytical method for L-carnitine was developed for infant and toddler formulas by liquid chromatography tandem mass spectrometry (LC-MS/MS). A 0.3 g of infant formula and toddler formula sample was mixed in a 50 mL conical tube with 9 mL water and 1 mL 0.1 M hydrochloric acid (HCl) to chemical extraction. Then, chloroform was used for removing a lipid fraction. After centrifuged, L-carnitine was separated and quantified using LC-MS/MS with electrospray ionization (ESI) mode. The precursor ion for L-carnitine was m/z 162, and product ions were m/z 103 (quantitative) and m/z 85 (qualitative), respectively. The results for spiked recovery test were in the range of 93.18-95.64% and the result for certified reference material (SRM 1849a) was within the range of the certificated values. This method could be implemented in many laboratories that require time and labor saving.

Identification of 1-Furan-2-yl-3-pyridin-2-yl-propenone, an Anti-inflammatory Agent, and Its Metabolites in Rat Liver Subcellular Fractions

  • Lee, Sang-Kyu;Jeon, Tae-Won;Basnet, Arjun;Jeong, Hye-Gwang;Lee, Eung-Seok;Jeong, Tae-Cheon
    • Archives of Pharmacal Research
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    • v.29 no.11
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    • pp.984-989
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    • 2006
  • 1-Furan-2-yl-3-pyridin-2-yl-propenone (FPP-3) has been characterized to have an anti-inflammatory activity through the inhibition of the production of nitric oxide and tumor necrosis $factor-{\alpha}$. In the present studies, the phase 1 metabolism of FPP-3 was investigated in rat liver microsomes and cytosols. When FPP-3 was incubated with rat liver microsomes and cytosols in the presence of NADPH. 2 major peaks were detected on a liquid chromatography/electrospray ionization-mass spectrometry. Two metabolites (i.e., M1 and M2) were characterized as reduced forms on propenone: M1 (1-furan-2-yl-3-pyridin-2-yl-propan-1-one) was the initial metabolite and M2 (1-furan-2-yl-3-pyridin-2-yl-propan-1-ol) was a secondary alcohol believed to be formed from M1.

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.

Investigation of the Effect of Acidity and Polyethylene Glycol on Electrochemical Deposition of Trivalent Chromium Ions

  • Phuong, N.V.;Kwon, S.C.;Lee, J.Y.;Kim, M.;Lee, Y.I.
    • Proceedings of the Korean Institute of Surface Engineering Conference
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    • 2011.05a
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    • pp.47-48
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    • 2011
  • The effect of solution acidity and organic additives, polyethylene glycol (PEG), on the trivalent chromium electroplating was systematically investigated in the view point of solution stability, electroreduction of trivalent chromium ions and characterization of deposition layer. It was found that, the concentration of fraction chromium complexes in the trivalent chromium bath containing formic acid is strongly depended on pH value. PEG molecules were stable in trivalent chromium bath containing formic acid via studies on electrospray ionization mass spectrometry (ESI-MS) and UV-Vis. However, the presence of PEG molecules decreased the reductive current of hydrogen evolution, increasing of current efficiency higher about 10 % compared with solutions without PEG. Moreover, PEG additives developed the nodular morphology during electroreduction of trivalent chromium ions with the increase of solution acidity and enhanced its current efficiency by maintaining the consumption of complexant, formic acid, at a low speed. In this study, the effect of solution acidity was emphasized important, there, it controlled the formation of complexes in the solution, cathodic film (CF) during deposition, and properties of deposited layer. By electrochemical quartz crystal microbalance (EQCM), studies show that chromium electrodeposition occurs via the formation of intermediate complexes and adsorption on the cathode surface, which hinder the penetration of ions from bulk solution to the cathode surface.

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Identification of Anti-Oxidant and Anti-Tyrosinase Activity of Phenolic Components Isolated from Betula schmidtii (박달나무로부터 분리된 페놀성 화합물의 항산화 및 Tyrosinase 저해 활성 연구)

  • Wang, Da-Hye;Chung, Ha Sook
    • The Korean Journal of Food And Nutrition
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    • v.34 no.5
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    • pp.553-559
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    • 2021
  • The aim of study to investigate the phytochemicals and biological activities the bark of Betula schmidtii. The studies consisted of the solvent extraction, followed by the isolation of phenolic components 1~3 from ethyl acetate-soluble fraction of Betula schmidtii Bark. Their chemical structures were identified as arbutin (1), ρ-coumaric acid (2) and ferulic acid (3) using Ultraviolet-Visible (UV-Vis) Spectrophotometer, Electrospray Ionization Mass Spectrometry (ESI-MS) (negative ion mode), 1H-Nuclear Magnetic Resonance (NMR), 13C-NMR, 1H-1H Correlation Spectroscopy (COSY) and 1H-13C Hetero Nuclear Multiple Quantum Correlation (HMQC) spectral data. Compounds 1~3 shows the anti-oxidant effect with IC50 values of 29.74±1.52, 21.32±1.07 and 34.41±1.24 in 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity, respectively. Also, compounds 1~3 exhibited mushroom tyrosinase inhibitory activity with IC50 values of 31.14±1.07, 42.54±1.46 and 69.22±1.43 µM, respectively.