• Title/Summary/Keyword: Mass Spectrometry (MS)

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Enrichment Strategies for Identification and Characterization of Phosphoproteome

  • Lee, Sun Young;Kang, Dukjin;Hong, Jongki
    • Mass Spectrometry Letters
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    • v.6 no.2
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    • pp.31-37
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    • 2015
  • Phosphorylation upon protein is well known to a key regulator that implicates in modulating many cellular processes like growth, migration, and differentiation. Up to date, grafting of multidimensional separation techniques onto advanced mass spectrometry (MS) has emerged as a promising tool for figuring out the biological functions of phosphorylation in a cell. However, advanced MS-based phosphoproteomics is still challenging, due to its intrinsic issues, i.e., low stoichiometry, less susceptibility in positive ion mode, and low abundance in biological sample. To overcome these bottlenecks, diverse techniques (e.g., SCX, HILIC, ERLIC, IMAC, TiO2, etc.) are continuously developed for on-/off-line enrichment of phosphorylated protein (or peptide) from biological samples, thereby helping qualitative/quantitative determination of phosphorylated protein and its phosphorylated sites. In this review, we introduce to the overall views of enrichment tools that are universally used to selectively isolate targeted phosphorylated protein (or peptide) from ordinary ones before MS-based phospoproteomic analysis.

Superconducting Tunnel Junction Detectors for Mass Spectrometry

  • Ohkubo, M.;Zen, N.;Kitazume, T.;Ukibe, M.;Shiki, S.;Koike, M.
    • Progress in Superconductivity
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    • v.14 no.2
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    • pp.77-81
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    • 2012
  • With conventional mass spectrometry (MS), ions are separated according to mass/charge (m/z) ratios. We must speculate the z values to obtain the m values. Superconducting tunnel junction (STJ) detectors can solve this problem, and true mass spectrometry becomes possible instead of m/z spectrometry. The STJ detectors were installed in MS instruments with a variety of ion sources. As an example, we report fragmentation analysis of a non-covalent protein complex of hemoglobin.

Organic matrix-free imaging mass spectrometry

  • Kim, Eunjin;Kim, Jisu;Choi, Inseong;Lee, Jeongwook;Yeo, Woon-Seok
    • BMB Reports
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    • v.53 no.7
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    • pp.349-356
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    • 2020
  • Mass spectrometry (MS) is an ideal tool for analyzing multiple types of (bio)molecular information simultaneously in complex biological systems. In addition, MS provides structural information on targets, and can easily discriminate between true analytes and background. Therefore, imaging mass spectrometry (IMS) enables not only visualization of tissues to give positional information on targets but also allows for molecular analysis of targets by affording the molecular weights. Matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) MS is particularly effective and is generally used for IMS. However, the requirement for an organic matrix raises several limitations that get in the way of accurate and reliable images and hampers imaging of small molecules such as drugs and their metabolites. To overcome these problems, various organic matrix-free LDI IMS systems have been developed, mostly utilizing nanostructured surfaces and inorganic nanoparticles as an alternative to the organic matrix. This minireview highlights and focuses on the progress in organic matrix-free LDI IMS and briefly discusses the use of other IMS techniques such as desorption electrospray ionization, laser ablation electrospray ionization, and secondary ion mass spectrometry.

Comparison of Liquid Chromatography-Mass/Mass Spectrometry (MS) and Gas Chromatography-MS for Quantitative Analysis of Indole-3-acetic acid and Indole-3-butyric acid from the Concentrated Liquid Fertilizer (Liquid Chromatography-Mass/Mass Spectrometry (MS)와 Gas Chromatography-MS를 이용한 농축 액상 비료제품 중 Indole-3-acetic acid 및 Indole-3-butyric acid 정량분석능 비교)

  • Kim, Jin Hyo;Park, Jong Min;Choi, Geun-Hyoung;Park, Yun-Ki;Im, Geon-Jae;Kim, Doo-Ho;Kwon, Oh-Kyung
    • Journal of Applied Biological Chemistry
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    • v.56 no.1
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    • pp.53-57
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    • 2013
  • In here, we investigated the quantitative analysis method of indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) with liquid chromatography-mass/mass spectrometry (LC-MS/MS) or gas chromatography-MS. Two ways of clean-up process were investigated for LC-MS/MS instrumental analysis of IAA, but both a simple dilution and hydrophile-lipophile balance (HLB) solid phase extraction (SPE) were not met the optimal recovery rates for quantitative analysis. On the other hand, the clean-up method for GC-MS was finally optimized through HLB-SPE from 250-folds diluted sample and methylation with trimethylsilyl chloride in methanol for 4 h. The limit of detection for methyl ester of IAA and IBA were both 1.4 mg/L, and recovery rates showed 93-107% from the concentrated liquid fertilizer.

Determination of S- and R-Amlodipine in Rat Plasma using LC-MS/MS After Oral Administration of S-Amlodipine and Racemic Amlodipine

  • Yoo, Hye-Hyun;Kim, Tae-Kon;Lee, Bong-Yong;Kim, Dong-Hyun
    • Mass Spectrometry Letters
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    • v.2 no.4
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    • pp.88-91
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    • 2011
  • The pharmacokinetic properties of S-amlodipine were studied using racemic amlodipine and single S-enantiomer (SK310) administration to rats. Plasma levels of the drug were determined using chiral liquid chromatography coupled with tandem mass spectrometry following solid phase extraction. The stereospecific analysis of amlodipine was performed on an ${\alpha}$-acid glycoprotein (AGP) column using a mobile phase comprising 10 mM ammonium acetate (pH 4.0) and propanol at a flow rate of 0.2 mL/min. This method was used to perform a comparative study of the pharmacokinetics of amlodipine and SK310. The results revealed that the pharmacokinetic profile of S-amlodipine after the administration of SK310 was comparable to that following the administration of the racemic mixture.

Tertiary Matrices for the Analysis of Polyethylene Glycols Using MALDI-TOF MS

  • Hong, Jangmi;Kim, Taehee;Kim, Jeongkwon
    • Mass Spectrometry Letters
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    • v.5 no.2
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    • pp.49-51
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    • 2014
  • The effectiveness of tertiary matrices composed of the combination of three common matrices (dihydrobenzoic acid (DHB), ${\alpha}$-cyano-4-hydroxycinnamic acid (CHCA), and sinapinic acid (SA)) was compared with that of single or binary matrices in the analysis of polyethylene glycol (PEG) polymers ranging from 1400 to 10000 Da using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). A tertiary matrix of 2,5-DHB+CHCA+SA was the most effective in terms of S/N ratios. CHCA and CHCA+SA produced the highest S/N ratios among the single matrices and the binary matrices, respectively. The improvement observed when using a tertiary matrix in analyses of PEG polymers by MALDI-TOF MS is believed to be due to the uniform morphology of the MALDI sample spots and synergistic effects arising from the mixture of the three matrix materials.

LC-MS/MS-based Proteomic Analysis to Identify Protein Phosphorylation in Emiliania huxleyi

  • Duong, Van-An;Nam, Onyou;Jin, EonSeon;Seo, Jae-Min;Park, Jong-Moon;Lee, Hookeun
    • Mass Spectrometry Letters
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    • v.12 no.4
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    • pp.163-171
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    • 2021
  • Emiliania huxleyi is a marine phytoplankton that plays a critical role in global carbon and sulfur cycling. The genome of E. huxleyi has been sequenced, and an in-depth proteomic profile of this organism has been reported. This study analyzed the phosphoproteome of E. huxleyi and identified its changes under calcium-limited conditions. A TiO2 microcolumn was used for phosphopeptide enrichment, followed by liquid chromatography-tandem mass spectrometry analysis. Overall, we identified 7,010 phosphorylated sites on 3,355 phosphopeptides associated with 2,929 phosphoproteins in E. huxleyi. Quantitative analysis revealed changes in the phosphoproteome in E. huxleyi when ambient conditions changed to calcium-limited conditions, notably the phosphorylation of some transporters was altered. This study provides an overview of protein phosphorylation in E. huxleyi and paves the way for further investigations of its biological functions.

Chiral Derivatization of Hydroxycarboxylic Acids Using 2,4,6-Trichlorobenzoyl Chloride as a Highly Efficient Regioselective Esterification Reagent for Gas Chromatography-Mass Spectrometry

  • Park, Jeong Hyeok;Han, Sang Yun
    • Mass Spectrometry Letters
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    • v.11 no.4
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    • pp.103-107
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    • 2020
  • We report the application of 2,4,6-trichlorobenzoyl chloride (often referred to as Yamaguchi esterification reagent) for the selective derivatization of the carboxylic group for GC-MS with the sample preparation method optimized for GC-MS analysis. The reagent was shown to be capable of selectively turning the carboxylic group into a reaction center, i.e., anhydride, of which the further reaction was directed to a near complete formation of required esters by unique steric and electronic effects of the reagent. Using the developed method, the chiral separation of hydroxycarboxylic acids by GC-MS using non-chiral columns was successfully demonstrated.

A Low-Cost Open-Source Air Sampler for the Sorbent Tube Sampling for TD-GC/MS Analysis

  • Jang, Hwa-yong;Oh, Han Bin
    • Mass Spectrometry Letters
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    • v.11 no.4
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    • pp.95-102
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    • 2020
  • In this study, we provide full descriptions of how to make a low-cost and completely open-source laboratory-made air sampler that will be used for sample adsorption for thermal desorption-gas chromatography mass spectrometry (TD-GC/MS) analysis. It is well known that harmful gases cause bad effects on human bodies, so it is necessary to identify the types and amounts of gases in industrial sites. One of the most commonly used methods for gas sampling is to utilize a sorbent tube using an air sampler. Commercially available air samplers are expensive, typically priced between $1,000 and $2,000, and their design often cannot be modified to fit the experiment. To address these shortcomings, we have developed a do-it-yourself (DIY) air sampler that is not only cheap enough, but also completely open-source. Furthermore, the performance of the fabricated air sampler was validated in conjunction with TD-GC/MS for the analysis of volatile compounds.