• Title/Summary/Keyword: MS2

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Structural Analysis of Cu Binding Site in [Cu(I)·d(CpG)·d(CpG)-2H]-1 Complex

  • Im, Yu-Jin;Jung, Sang-Mi;Kang, Ye-Song;Kim, Ho-Tae
    • Bulletin of the Korean Chemical Society
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    • v.34 no.4
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    • pp.1232-1236
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    • 2013
  • The Cu cation binding sites of $[Cu(I){\cdot}d(CpG){\cdot}d(CpG)-2H]^{-1}$ complex have been investigated to explain the $[Cu{\cdot}DNA]$ biological activity caused by the Cu association to DNA. The structure of $[Cu(I){\cdot}d(CpG){\cdot}d(CpG)-2H]^{-1}$ complex was investigated by electrospray ionization mass spectrometry (ESI-MS). The fragmentation patterns of $[Cu(I){\cdot}d(CpG){\cdot}d(CpG)-2H]^{-1}$ complex were analyzed by MS/MS spectra. In the MS/MS spectra of $[Cu(I){\cdot}d(CpG){\cdot}d(CpG)-2H]^{-1}$ complex, three fragment ions were observed with the loss of d(CpG), {d(CpG) + Cyt}, and {d(CpG) + Cyt + dR}. The Cu cation binds to d(CpG) mainly by substituting the $H^+$ of phosphate group. Simultaneously, the Cu cation prefers to bind to a guanine base rather than a cytosine base. Five possible geometries were considered in the attempt to optimize the $[Cu(I){\cdot}d(CpG){\cdot}d(CpG)-2H]^{-1}$ complex structure. The ab initio calculations were performed at B3LYP/6-31G(d) level.

Comparative Proteomic Analyses of the Yeast Saccharomyces cerevisiae KNU5377 Strain Against Menadione-Induced Oxidative Stress

  • Kim, Il-Sup;Yun, Hae-Sun;Jin, In-Gnyol
    • Journal of Microbiology and Biotechnology
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    • v.17 no.2
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    • pp.207-217
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    • 2007
  • The Saccharomyces0 cerevisiae KNU5377 strain, which was isolated from spoilage in nature, has the ability to convert biomass to alcohol at high temperatures and it can resist against various stresses [18, 19]. In order to understand the defense mechanisms of the KNU5377 strain under menadione (MD) as oxidative stress, we used several techniques for study: peptide mass fingerprinting (PMF) by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry (MS) followed by two-dimensional (2D) gel electrophoresis, liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS), and surface-enhanced laser desorption ionization-time of flight (SELDI-TOF) technology. Among the 35 proteins identified by MALDI-TOF MS, 19 proteins including Sod1p, Sod2p, Tsa1p, and Ahp1p were induced under stress condition, while 16 proteins were augmented under normal condition. In particular, five proteins, Sod1p, Sod2p, Ahp1p, Rib3p, Yaf9p, and Mnt1p, were induced in only stressed cells. By LC-ESI-MS/MS analysis, 37 proteins were identified in normal cells and 49 proteins were confirmed in the stressed cells. Among the identified proteins, 32 proteins were found in both cells. Five proteins including Yel047cp and Met6p were only upregulated in the normal cells, whereas 17 proteins including Abp1P and Sam1p were elevated in the stressed cells. It was interesting that highly hypothetical proteins such as Ynl281wp, Ygr279cp, Ypl273wp, Ykl133cp, and Ykr074wp were only expressed in the stressed cells. SELDI-TOF analysis using the SAX2 and WCX2 chips showed that highly multiple-specific protein patterns were reproducibly detected in ranges from 2.9 to 27.0 kDa both under normal and stress conditions. Therefore, induction of antioxidant proteins, hypothetical proteins, and low molecular weight proteins were revealed by different proteomic techniques. These results suggest that comparative analyses using proteomics might contribute to elucidate the defense mechanisms of KNU5377 under MD stress.

Comparison of Dioxin Analytical Results Between SP-2331 and DB-5MS Capillary Columns in the HRGC/HRMS Analysis (HRGC/HRMS에 SP-2331 및 DB-5MS 컬럼 사용시 다이옥신 결과의 비교)

  • Hong, Jongki;Yang, Jeong-Soo;Shin, Jung-Hwa;Ahn, Yun-Gyong;Lee, Dai-Woon
    • Analytical Science and Technology
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    • v.17 no.2
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    • pp.184-191
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    • 2004
  • In this study, the separation of toxic dioxin 2,3,7,8-congeners by DB-5MS and SP-2331 GC columns which are widely used in HRGC/HRMS analysis was examined. Through the dioxin analysis of column performance check standard solution and fly ash sample, the isomer specific separation of 2,3,7,8-substituted dioxins from tetra to hexa-isomers on DB-5MS and SP-2331 columns were studied. The effect of I-TEQ value by these columns was also studied. The total concentrations of toxic dioxins for the column performance check standard solution were 508.4 ng/mL analyzed by DB-5MS and 515.8 ng/mL analyzed by SP-2331, respectively. The I-TEQ value obtained by both columns was shown to be almost equivalent for the column performance check standard solution and fly ash sample.

Effect of Water Temperature, Fish Age, and MS-222 Concentration on the Anesthetization of River Pufferfish, Takifugu obscurus (황복의 마취에 미치는 수온 및 연령과 MS-222 농도의 영향)

  • Seungyeon Lee;Phuong Thi Nguyen;Ho-Kyung Song;Sung Pyo Hur;Jin-Hyoung Kim
    • Korean Journal of Ichthyology
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    • v.35 no.2
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    • pp.67-74
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    • 2023
  • The river pufferfish (Takifugu obscurus) is a valuable species in aquaculture and genetic studies. Usage of fish anesthetics aids in the easier handling of fish during aquaculture. However, there are no studies on appropriate conditions required for effective anesthetization of pufferfish. This study aims to determine the optimal conditions (fish age, water temperature, anesthetic concentration) needed for the most common fish anesthesia, MS-222, to anesthetize T. obscurus. We tested three different water temperatures (20℃, 24℃, and 28℃), three different anesthetic concentrations (125 mg/L, 150 mg/L, and 175 mg/L), and two different fish ages (one- and two-year-old). Appropriate anesthetization conditions for T. obscurus ranged from 150 mg/L to 175 mg/L of MS-222 at 24℃ to 28℃ for one-year-old fish. For two-year-old fish, the appropriate conditions ranged from 150 mg/L to 175 mg/L of MS-222 at 28℃. However, to minimize side effects and risks, 150 mg/L of MS-222 at 24℃ for one-year-old fish and 175 mg/L of MS-222 at 28℃ for two-year-old fish are recommended for effective anesthetization.

Characteristic Effects of Dangnyo-hwan for Diabetes Control Studied Using LC-MS/MS and ICP (LC-MS/MS 및 ICP를 이용한 당뇨환의 화학적 특성 분석)

  • In, Jeong-do;Im, Dai-sig;Moon, Seung-ho;Ki, Won-ill
    • The Journal of Internal Korean Medicine
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    • v.36 no.3
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    • pp.217-227
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    • 2015
  • Objectives: The primary aim of this study was to identify bioactive compounds in Dangnyo-hwan, a Korean herbal medicine, through instrumental analysis using LC-MS/MS and ICP, and investigate its potential use in diabetes treatment. Methods: The extract of Dangnyo-hwan has 12 medicinal herbs; these were compared with 18 marker substances selected from literature survey. Results: LC-MS/MS analysis could detect 9 of the 18 bioactive compounds: citruline, catalpol, berberine, ginsenoside Rb1, ginsenoside Rg1, oleanolic acid, β-sitosterol, mangiferin, and schizandrin. While harmful heavy metals such as As, Pb, Cd, Hg, Ni, and Cu were not present in high concentrations, Zn concentration was 4.2 mg in 100 g Dangnyo-hwan. Conclusions: Instrumental analysis such as LC-MS/MS and ICP was successfully used to identify bioactive compounds in Dangnyo-hwan. Detection of 9 bioactive substances and Zn from the herb medicine is a valuable finding, and suggests that Dangnyo-hwan is a candidate medicine for diabetes. Further investigations like in vitro assay, percent GPR 119 activity, and percent human DGAT-1 inhibition are underway.

Quantitative Analysis of the Ten Phytochmicals in Acer tegmentosum Maxim by UPLC-MS/MS (UPLC-MS/MS를 이용한 산천목 중 10종 성분의 함량 분석)

  • Hwang, Youn-Hwan;Li, Wei;Yang, Hye Jin;Ma, Jin Yeul
    • Korean Journal of Pharmacognosy
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    • v.49 no.1
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    • pp.70-75
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    • 2018
  • Acer tegmentosum Maxim (ATM) has been used to treat hepatic disorders in traditional oriental medicine. However, there is little information about phytochemical constituents for quality control of ATM. In this study, we developed and established a simultaneous analytical method of the 10 marker compounds (three coumarins, 3 flavonoids, 1 lignan, 3 phenolics) in ATM using ultra-performance liquid chromatography-mass spectrometry (UPLC-MS/MS). Chromatographic separation of ten target analytes was achieved with a Waters Acquity UPLC BEH $C_{18}$ analytical column ($2.1{\times}100mm$, $1.7{\mu}m$), using a mobile phase of 0.1% (v/v) formic acid in water and acetonitrile with gradient elution. Identifications and quantitation of all analytes were performed using a Q-Exactive UPLC-MS/MS system. Correlation coefficients of the calibration curve for all analytes were ${\geq}0.9986$. The values of limits of detection and quantification of all analytes were 0.5-10.0 and 5.0-50.0 ng/mL, respectively. The established UPLC-MS/MS method successfully identified all target analytes in ATM, and the phytochemicals were 0.01-67.98 mg/g in its lyophilized water extract.

Component Analysis of Suaeda asparagoides Extracts (나문재 추출물의 성분 분석)

  • Yang, Hee-Jung;Park, Soo-Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.34 no.3
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    • pp.157-165
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    • 2008
  • In the previous study, the anti-oxidant activity of oxtract/fraction of Sueada aspparagoides(SA) and the stability test for the cream containing SA extract were investigated respectively[1,2]. In this study, the components of SA extract were analyzed by TLC, HPLC, and LC/ESI-MS/MS, $^1H$-NMR. TLC chromatogram of ethyl acetate fraction of SA extract revealed 5 bands $(SA1{\sim}SA5)$. HPLC chromatogram of aglycone fractions obtained from deglycoylation reaction of ethyl acetate fraction showed 2 bands (SAA 2 and SAA 1), which were identified as quercetin (composition ratio, 16.88%) and kaempferol (83.12%) in the order of elution time. Among 5 bands of TLC chromatogram, 4 bands $(SA2{\sim}SA5)$ also were Identified as kaempferol-3-O-glucoside (SA 2), quercetin-3-O-glucoside (SA3), kaempferol-3-O-rutinoside (SA 4), quercetin-3-O-rutinoside (SA 5) by LC/ESI-MS/MSMS/MS. respectively. The spectrum generated for SAA 1 by LC/ESI-MS/MS in the negative ion mode also gave the ion corresponding to the deprotonated aglycone $[M-H]^-$ (285m/z), the $^1H$-NMR spectrum contained signals [${\delta}$ 6.19 (1H, d, J=1.8Hz, H-6), ${\delta}$ 6.44 (1H, d, J=1.8Hz, H-8), ${\delta}$ 6.92 (2H, d, J=9.0Hz, H-3', 5'), ${\delta}$ 8.04 (2H, d, J=9.0Hz, H-2', 6', thus SAA 1 was identified as kaempferol. SAA 2 yielded the deprotonated agycone ion $[M-H]^-$ (301m/z), $^1H$-NMR spectrum showed signals [${\delta}$ 6.20 (1H, d, J=2.0Hz, H-6), ${\delta}$ 6.42 (1H, d, J=2.0Hz, H-8), ${\delta}$ 6.90 (1H, d, J=8.6Hz, H-5'), ${\delta}$ 7.55 (1H, dd, J=8.6, 2.2Hz, H-6'), ${\delta}$ 7.69 (1H, d, J=2.2Hz, H-2', thus SAA 2 was Identified as quercetin. In conclusion, with the anti-oxidant activity and the stability test reported previously, component analysis of SA extracts could be applicable to new cosmeceuticals.

Effect of explant parts and medium components on in vitro regeneration in Osmunda cinnamomea var. forkiensis (꿩고비(Osmunda cinnamomea var. forkiensis) 기내 포자체 재생에 영향을 미치는 배양부위와 배지구성물질)

  • Kwon, Hyuk Joon;Shin, So Lim;Lee, Cheol Hee;Kim, Soo-Young
    • Journal of Plant Biotechnology
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    • v.44 no.4
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    • pp.448-453
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    • 2017
  • This study was carried out to find culture materials (explant parts) and medium components (medium type, sucrose and $NaH_2PO_4$ concentration) for in vitro propagation of Osmunda cinnamomea var. forkiensis sporophyte. The results of study: chopped segments of leaf blades, stipes, rhizomes and roots were cultured on a 1/2MS medium supplemented with 0.1% activated charcoal. Among these explant types, only the rhizome segments produced young sporophyte, regenerating vigorously on a 1/8MS medium. Adjusting the sucrose concentration to 2% and supplement to $50mg{\cdot}L^{-1}\;NaH_2PO_4$ in the 1/8MS medium proved to be more efficient for plant regeneration. Consequently, the addition of 0.1% activated charcoal to a modified 1/8MS medium (2% sucrose, $50mg{\cdot}L^{-1}\;NaH_2PO_4$, pH 5.8 and 0.8% agar) yielded the highest sporophyte regeneration.

Serial Flow Microwave Thermal Process System for Liquid Foods

  • Kim, Young-Jin;Lim, Seok-Won;Chun, Jae-Kun
    • Food Science and Biotechnology
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    • v.14 no.4
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    • pp.446-449
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    • 2005
  • Two single-magnetron heating systems (SM-HS), each with a helical glass heat exchanger and a cylindrical cavity, were combined to make a two-magnetron-in-series heating system (2MS-HS) in order to increase the heating capacity. A comparison using water showed that the heating performance of the 2MS-HS was increased by two-fold as compared to that of the SM-HS, resulting in energy saving of 7.0% in 2MS-HS. Pasteurization test of 2MS-HS conducted with model food (LB broth contaminated with Bacillus subtilis) showed two-fold higher treatment capacity compared to SM-HS. Relationships between outlet temperature of the processed food, flow rate, and residence time in the 2MS-HS were established for water. Optimum pasteurization capacity was 17 s, $73^{\circ}C$, at flow rate of 280 ml/min. The 2MS-HS could be applied to the small-scale pasteurization of liquid food.

A New Analytical Method for Erythromycin in Fish by Liquid Chromatography/Tandem Mass Spectrometry

  • Park, Mi-Jung;Park, Mi-Seon;Lee, Tae-Seek;Shin, Il-Shik
    • Food Science and Biotechnology
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    • v.17 no.3
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    • pp.508-513
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    • 2008
  • Erythromycin has been used to treat Streptococosis, Edwardsiel1osis, Vibriosis, Bacterial enteritis in the cultured fish. In this study, a rapid and effective erythromycin analysis method with new sample treatment protocol and liquid chromatography/tandem mass spectrometry (LC/MS/MS) system for fish products was developed. For the erythromycin extraction from fish muscle, the solvent mixture composed of 0.2% meta-phosphoric acid and methanol (6:4) showed good recovery rate, and the optimum extraction solvent volume was 20 mL. Erythromycin detection using LC/MS/MS were carried out under electro spray ionization (ESI) positive condition and erythromycin mass value 576.2 and 157.9. And the detection limit of the established method was 0.005 mg/kg in fish products. The recovery rate of the developed method applied to the fish species were as following, olive flounder, $87.6{\pm}5.0%$; black rockfish, $87.2{\pm}6.4%$; eel, $85.2{\pm}4.8%$; and rainbow trout, $86.0{\pm}6.2%$. In the established method in this study, the correlation of coefficient values ($R^2$) of erythromycin calibration curve (n=11) was 0.9998.