• Title/Summary/Keyword: Alpha spectrometry

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Derivatization of a-ketoacid for the Analysis of Aminotransferase Reaction Using Mass Spectrometry (질량 분석기를 이용한 트렌스아미나제 반응 분석을 위한 케토산 화학 수식법 개발)

  • Lee Chang Soo;Kim Yun Gon;Kim Eun Mi;Kim Byung Gee
    • KSBB Journal
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    • v.19 no.5
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    • pp.394-398
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    • 2004
  • A derivatization method in mass spectrometry for small molecular analysis was developed to solve the problems of volatility of many analytes, difficult ionization of analytes, and undiscriminating isobaric analytes. This derivatization method, oximation of $\alpha$-ketoacid, in the transaminase reaction leads to change of mass difference between amine reactants and ketoacid products. In addition, regardless of the kinds of ketoacid, the linear relationship between the peak intensity and its concentration (from 1 mM to 10 mM) shows that quantitative analysis of the conversion of the reaction can be executed by the analysis of peak intensity of the corresponding oximated ketoacids. Furthermore, this method can be used for identifying transaminase as well as determining its substrate specificity

Enrichment of Peptides using Novel C8-functionalized Magnetic Nanoparticles for Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometric Analysis

  • Song, Sun-Mi;Yang, Hyo-Jik;Kim, Jin-Hee;Shin, Seong-Jae;Park, Eun-Hye;Kim, Jeong-Kwon
    • Mass Spectrometry Letters
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    • v.2 no.2
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    • pp.53-56
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    • 2011
  • [ $C_8$ ]functionalized magnetic nanoparticles were synthesized by coating magnetic $Fe_3O_4$ nanoparticles with silicaamine groups using 3-aminopropyltriethoxysilane and by subsequently modifying the amine groups with chloro(dimethyl)octylsilane to produce octyl groups on the surface of the MNPs. The $C_8$-functionalized MNPs were used to enrich peptides from tryptic protein digests of myoglobin and ${\alpha}$-casein. The enriched peptides were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). MALDI-MS was also used to investigate desalting of the $C_8$-functionalized MNPs. Sample solutions were prepared in 1.0 M NaCl, and the successful removal of salt was observed. Enrichment with $C_8$-functionalized MNPs was very effective for separating and concentrating tryptic peptides.

Simultaneous Determination of α-Amanitin and β-Amanitin in Mouse Plasma Using Liquid Chromatography-High Resolution Mass Spectrometry

  • Bang, Young Yoon;Lee, Min Seo;Lim, Chang Ho;Lee, Hye Suk
    • Mass Spectrometry Letters
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    • v.12 no.3
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    • pp.112-117
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    • 2021
  • α-Amanitin and β-amanitin are highly toxic bicyclic octapeptides responsible for the poisoning of poisonous mushrooms such as Amanita, Galerina, and Lepiota by inhibiting RNA polymerase II, DNA transcription, and protein synthesis. A sensitive, simple, and selective liquid chromatography-high resolution mass spectrometric method using parallel reaction monitoring mode was developed and validated for the simultaneous determination of α- and β-amanitin in mouse plasma to evaluate the toxicokinetics of α- and β-amanitin in mice. Protein precipitation of 5 μL mouse plasma sample with methanol as sample clean-up procedure and use of negative electrospray ionization resulted in better sensitivity and less matrix effect. The calibration curves for α- and β-amanitin in mouse plasma were linear over the range of 0.5-500 ng/mL. The intra- and inter-day coefficient of variations and accuracies for α- and β-amanitin at four quality control concentrations were 3.1-14.6% and 92.5-115.0%, respectively. The present method was successfully applied to the toxicokinetic study of α- and β-amanitin after an oral administration of α- and β-amanitin at 1.5 mg/kg dose to male ICR mice.

Metallurgical Study of Bronze Relics Excavated from Sanoesa Temple, Chongju (청주(淸州) 사뇌사지(思惱寺址) 출토 청동유물의 금속학적 조사)

  • Kwon, H.N.;Yu, H.S.;Ahn, B.C.
    • Journal of Conservation Science
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    • v.9 no.1
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    • pp.1-10
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    • 2000
  • In 1993, many bronze artifacts were excavated from the Sanoesa Temple(思惱寺), Chongju, Chungbuk. Twelve items were selected and chemically analyzed with AA Spectrometry and ICP-Atomic Emission Spectrometry. They were also observed under the optical microscopy and SEM. According to the results from chemical analysis, production method and use, these artifacts were classified into four groups: casting, wrought and welding products, and bells. Cast products, probably used for ritual, were alloy of 70% Cu, 10% Sn and 20% Pb. They showed ${\alpha}+{\beta}$ phase as a typical microstructure of casting. The ${\delta}$ phase was rarely observed due to the small amount of Sn. These artifacts included more lead than other alloys. They showed segregation like island-shape on the lead part. Wrought products used for daily too1s. were alloy of 80% Cu and 20% Sn. Since they were consist of ${\alpha}$ phase and martensite ${\beta}$ phase, it could be presumed that they were heat-treated. The production method could be identified from twinned grains in ${\alpha}$ phase. Lead was not included in because it had a bad effect to alloy. The bells were alloyed with 85% Cu, 10% Sn, 5% Pb or 90% Cu and 10% Sn. They show the dendrite structure because they were cast and alloyed with many tin. Weldinged were alloyed with 83% Cu, 12% Sn and 5% Pb. lt showed the fine dendrite structure because of fast cooling in air.

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Antioxidant Effect of Korean Red Ginseng Extract on Aqueous Linoleic Acid and LDL (수용성계의 Linoleic Acid와 LDL에 대한 한국산 홍삼의 산화방지효과)

  • Lee, J.W.;Lee, S.K.;Do, J.H.;Sung, H.S.;Lee, H.O.
    • Applied Biological Chemistry
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    • v.40 no.4
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    • pp.283-288
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    • 1997
  • The antioxidant activities of Korean Red Ginseng Extract(RGE) and ${\alpha}-tocopherol$, as reference compound, were tested with HPLC and fluorometry which measure the MDA after reacting an aqueous 1% linoleic acid buffer solution, and LDL(1 mg protein/ml) buffer solution with $H_2O_2{\;}and{\;}FeCl_2$. The generation of conjugated-diene in LDL(0.25 mg protein/ml) was also measured by spectrometry. MDA determination showed the antioxidant effect on linoleic acid oxidation with oxidation inhibition ratio of 71.8% and 76.1%, respectively, by addition of 1000 ppm RGE and 100 ppm ${\alpha}-tocopherol$. LDL(1 mg protein/ml) oxidation was inhibited by 25.2% and 21.2%, respectively, by addition of 200 ppm REG and 100 ppm ${\alpha}-tocopherol$. The generation of conjugated diene in LDL(0.25 mg protein/ml) was also inhibited by 44.2%, by addition of 50 ppm RGE.

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In Vitro Steroidogenesis on Oocyte Development in the Starry Flounder, Platichthys stellatus

  • Baek, Hea Ja;Kim, Dea Geun;Kim, Hyung Bae
    • Development and Reproduction
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    • v.17 no.4
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    • pp.421-426
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    • 2013
  • In this study, oocyte steroidogenesis are investigated in relation to oocyte development in the starry flounder, Platichthys stellatus, a marine multiple spawner. Vitellogenic (0.52 and 0.55 mm oocyte diameter) and mature oocytes (0.63, 0.66 and 0.71 mm oocyte diameter) were incubated in vitro in the presence of $[^3H]17{\alpha}$-hydroxyprogesterone ($[^3H]17{\alpha}$-OHP) as a precursor. Steroid metabolites were extracted from the incubated media and oocytes, the extracts were separated and identified by thin-layer chromatography (TLC), high performance liquid chromatography (HPLC) and gas chromatographymass spectrometry (GC-MS). The major metabolites produced from $[^3H]17{\alpha}$-OHP were androgens [androstenedione ($A_4$) and testosterone (T)] and estrogens [$17{\beta}$-estradiol ($E_2$) and estrone ($E_1$)] and progestins [$17{\alpha},20{\alpha}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\alpha}P$) and $17{\alpha},20{\beta}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}P$)] in vitellogenic and mature oocytes. The results from this study suggest the potential roles of $E_1$ in the oocytes with diameter 0.52-0.71 mm, $17{\alpha}20{\alpha}P$ and $17{\alpha}20{\beta}P$ at the oocytes of 0.63, 0.66 and 0.71 mm.

Production and Characterization of a New ${\alpha}$-Glucosidase Inhibitory Peptide from Aspergillus oryzae N159-1

  • Kang, Min-Gu;Yi, Sung-Hun;Lee, Jong-Soo
    • Mycobiology
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    • v.41 no.3
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    • pp.149-154
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    • 2013
  • An ${\alpha}$-glucosidase inhibitor was developed from Aspergillus oryzae N159-1, which was screened from traditional fermented Korean foods. The intracellular concentration of the inhibitor reached its highest level when the fungus was cultured in tryptic soy broth medium at $27^{\circ}C$ for five days. The inhibitor was purified using a series of purification steps involving ultrafiltration, Sephadex G-25 gel permeation chromatography, strong cation exchange solid phase extraction, reverse-phase high performance liquid chromatography, and size exclusion chromatography. The final yield of the purification was 1.9%. Results of the liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis indicated that the purified ${\alpha}$-glucosidase inhibitor was a tri-peptide, Pro-Phe-Pro, with the molecular weight of 360.1 Da. The IC50 value of the peptide against ${\alpha}$-glucosidase activity was 3.1 mg/mL. Using Lineweaver-Burk plot analysis, the inhibition pattern indicated that the inhibitor acts as a mixed type inhibitor.

Steroid Metabolism in the Blackfin Flounder Glyptocephalus stelleri during Oocyte Maturation (기름가자미(Glyptocephalus stelleri) 성숙기 난모세포에서의 성스테로이드 호르몬 대사물질 분석)

  • Lee, Hae Won;Baek, Hea Ja
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.48 no.4
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    • pp.483-488
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    • 2015
  • We studied oocyte steroidogenesis in the blackfin flounder Glyptocephalus stelleri as a region-specific species, in the East Sea of Korea during the spawning season. Maturing oocytes (0.76, 0.82, 0.88, and 0.91 mm in oocyte diameter) were incubated in vitro in the presence of [$^3H$] $17{\alpha}$-hydroxyprogesterone ($[^3H]17{\alpha}$-OHP) as a precursor. Steroid metabolites were extracted from the incubated medium and oocytes, and the extracts were separated and identified by thin-layer chromatography (TLC), high-performance liquid chromatography (HPLC) and gas chromatographymass spectrometry (GC/MS). The major metabolites produced from $[^3H]17{\alpha}$-OHP were androgens [androstenedione (A4) and testosterone (T)] and estrogens [$17{\beta}$-estradiol (E2) and estrone (E1)] and progestins [$17{\alpha},20{\alpha}$-dihydroxy-4-pregen-3-one ($17{\alpha}20{\alpha}P$) and $17{\alpha}20{\beta}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}P$)] in maturing oocytes. The metabolic rate of $17{\alpha}20{\beta}$ was elevated (29.04%) in oocytes measuring 0.88 mm (nucleus migration stage following the induction of germinal vesicle breakdown), but was very low in oocytes measuring 0.76, 0.82, and 0.91 mm (0.42, 0.67, and 2.62%, respectively). From these results, we suggest that $17{\alpha}20{\beta}P$ acts as a maturation-inducing steroid in the blackfin flounder.

Effect of Bisphenol A on Ovarian Steroidogenesis in Longchin Goby (Chasmichthys dolichognathus) (Bisphenol A가 점 망둑 (Chasmichthys dolichognothus)의 난소 스테로이드 호르몬 대사에 미치는 영향)

  • BAEK Hea-Ja;PARK Myoung-Hee;LEE Young-Don;KIM Hyung-Bae;KIM Jae-Won;YOO Myoung-Suk
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.37 no.3
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    • pp.192-196
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    • 2004
  • The in vitro effect of bisphenol A (BPA) on ovarian steroidogenesis of the longchin goby (Chasmichthys dolichognathus) was investigated. Oocytes taken during the maturing phase (vitellogenic, fully vitellogenic or germinal vesicle breakdown stage) were incubated with BPA (100 ng/mL) in the presence of exogenous precursor $^{3}H-17\alpha\;hydroxyprogesterone\;(^{3}H-17\alphaOHP).$ Steroids were extracted from the media and the isolated oocytes, and the extracts were separated and identified by thin layer chromatography and gas chromatography-mass spectrometry. The identities of the major metabolites were progestogens $[17{\alpha}-hydroxy,20{\alpha}-dihydroprogesterone\;(17{\alpha}20{\alpha}OHP)\;and\;17{\alpha}-hydrxy,20{\beta}-dihydroprogesterone\;(17{\alpha}20{\beta}OHP),$ androgens [androstenedione (A4) and testosterone (T)] and estrogens [estrone $(E_1)\;and\;estradiol-17{\beta}(E_2)].$ BPA treatment inhibited production of estrogens in all the maturing phases and progestogens in the germinal vesicle migrating stage. Percentage yield of estrogens was decreased with increased yield of androgens. In conclusion, BPA had an inhibitory effect on the conversion of $^3H-17\alphaOHP$ to estrogens and progestogens. These results demonstrate that BPA can act either estrogenic or anti-estrogenic effects.

Comparative Proteomic Analysis of Human Amniotic Fluid Supernatants with Down Syndrome Using Mass Spectrometry

  • Park, Ji-Sook;Cha, Dong-Hyun;Jung, Jin-Woo;Kim, Young-Hwan;Lee, Sook-Hwan;Kim, Young-Jun;Kim, Kwang-Pyo
    • Journal of Microbiology and Biotechnology
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    • v.20 no.6
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    • pp.959-967
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    • 2010
  • Down syndrome (DS) is an abnormality of the 21st chromosome that commonly occurs in children born to older women. Thus, amniotic fluid (AF) is usually collected from such women for prenatal diagnosis. This study analyzed human AF supernatants (AFS) using a mass spectrometric (MS) approach to search for candidate biomarkers of a DS pregnancy. The AFS were collected from older pregnant women at weeks 16-18 of their gestation by amniocentesis for cytogenetic analysis. The AFS from the pregnancies carrying DS (n=4) or chromosomally normal (n=6) fetuses, as revealed by the cytogenetic analysis, were then subjected to global protein profiling based on liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS). Affinity chromatography was also applied prior to the LC-ESI-MS/MS to minimize the masking effect of highly abundant albumin and immunoglobulin and thereby increase the diversity of the identified proteins. As a result, at least 30 new AFS proteins were identified and 44 AFS proteins were found to be differentially expressed between the DS and normal cases, where 6 of the proteins were unique to the DS cases and 11 were unique to the chromosomally normal cases. In addition, in the DS cases, 19 AFS proteins were downregulated and 8 were upregulated to varying degrees. A Western blot analysis confirmed the LC-ESI-MS/MS data, indicating that the combined detection of apolipoprotein A-II (apoA-II) and alpha-fetoprotein (AFP) could be a potential tool for diagnosing DS cases.