• Title/Summary/Keyword: Matrix-assisted laser desorption/ionization (MALDI)

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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.

Proteome Analysis of Vernalization-Treated Arabidopsis thaliana by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

  • Cho, Mi-Ran;Lee, Kyung-Hyeon;Hyun, You-Bong;Lee, Il-Ha;Kim, Hie-Joon
    • Bulletin of the Korean Chemical Society
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    • v.28 no.3
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    • pp.427-431
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    • 2007
  • In order to gain insight into the molecular changes at the protein level in plants exposed to low temperature for a long period of time (vernalization), proteome analyses of vernalization-treated Arabidopsis thaliana have been carried out by two-dimensional gel electrophoresis followed by matrix-assisted laser desorption/ ionization time-of-flight mass spectrometry. Fourteen proteins including ATP binding/GTP binding/translation elongation factor and glycine-rich RNA-binding protein 7 (GRP7) showed differential expression in vernalization-treated Arabidopsis thaliana. GRP7 showed the most dramatic increase in expression suggesting its involvement in response to vernalization treatment.

Selective or Class-wide Mass Fingerprinting of Phosphatidylcholines and Cerebrosides from Lipid Mixtures by MALDI Mass Spectrometry

  • Lee, Gwangbin;Son, Jeongjin;Cha, Sangwon
    • Bulletin of the Korean Chemical Society
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    • v.34 no.7
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    • pp.2143-2147
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    • 2013
  • Matrix assisted laser desorption/ionization (MALDI) mass spectrometry (MS) is a very effective method for lipid mass fingerprinting. However, MALDI MS suffered from spectral complexities, differential ionization efficiencies, and poor reproducibility when analyzing complex lipid mixtures without prior separation steps. Here, we aimed to find optimal MALDI sample preparation methods which enable selective or class-wide mass fingerprinting of two totally different lipid classes. In order to achieve this, various matrices with additives were tested against the mixture of phosphatidylcholine (PC) and cerebrosides (Cers) which are abundant in animal brain tissues and also of great interests in disease biology. Our results showed that, from complex lipid mixtures, 2,4,6-trihydroxyacetophenone (THAP) with $NaNO_3$ was a useful MALDI matrix for the class-wide fingerprinting of PC and Cers. In contrast, THAP efficiently generated PC-focused profiles and graphene oxide (GO) with $NaNO_3$ provided Cer-only profiles with reduced spectral complexity.

Enhancement of Analyte Ionization in Desoprtion/Ionization on Porous Silicon (DIOS)-Mass Spectrometry(MS)

  • Lee Chang-Soo;Kim Eun-Mi;Lee Sang-Ho;KIm Min-Soo;Kim Yong-Kweon;Kim Byug-Gee
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.3
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    • pp.212-217
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    • 2005
  • Desorption/ionization on silicon mass spectrometry (DIOS-MS) is a relatively new laser desorption/ionization technique for mass spectrometry without employing an organic matrix. This present study was carried to survey the experimental factors to improve the efficiency of DIOS-MS through electrochemical etching condition in structure and morphological properties of the porous silicon. The porous structure of silicon structure and its properties are crucial for the better performance of DIOS-MS and they can be controlled by the suitable selection of electrochemical conditions. The fabrication of porous silicon and ion signals on DIOS-MS were examined as a function of silicon orientation, etching time, etchant, current flux, irradiation, pore size, and pore depth. We have also examined the effect of pre- and post-etching conditions for their effect on DIOS-MS. Finally, we could optimize the electrochemical conditions for the efficient performance of DIOS-MS in the analysis of small molecule such as amino acid, drug and peptides without any unknown noise or fragmentation.

Free Radical Initiated Peptide Sequencing Using MALDI-TOF/TOF Mass Spectrometry

  • Song, Insu;Lee, Jae-ung;Baek, Jaehyeon;Cha, Sangwon;Han, Sang Yun;Oh, Han Bin
    • Mass Spectrometry Letters
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    • v.9 no.2
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    • pp.56-60
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    • 2018
  • In this study, matrix-assisted laser desorption/ionization (MALDI) was applied to the TEMPO-assisted FRIPS for the first time. We found that 3-HPA is the optimal matrix for the analysis of p-TEMPO-Bz-Sc-peptides, which gives minimal precursor fragmentations. MALDI-TOF/TOF experiments on p-TEMPO-Bz-Sc-peptides yielded mainly $[a_n+H]^+$, $[z_n+H]^+$, and $[y_n]^+-type$ products, indicating that radical-driven peptide fragmentation occurs in MALDI-TOF/TOF-MS.

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.

Rapid Identification of Staphylococcus Species Isolated from Food Samples by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

  • Kim, Eiseul;Kim, Hyun-Joong;Yang, Seung-Min;Kim, Chang-Gyeom;Choo, Dong-Won;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.29 no.4
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    • pp.548-557
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    • 2019
  • Staphylococcus species have a ubiquitous habitat in a wide range of foods, thus the ability to identify staphylococci at the species level is critical in the food industry. In this study, we performed rapid identification of Staphylococcus species using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight mass spectrometry (MALDI-TOF MS). MALDI-TOF MS was evaluated for the identification of Staphylococcus reference strains (n = 19) and isolates (n = 96) from various foods with consideration for the impact of sample preparation methods and incubation period. Additionally, the spectra of isolated Staphylococcus strains were analyzed using principal component analysis (PCA) and a main spectra profile (MSP)-based dendrogram. MALDI-TOF MS accurately identified Staphylococcus reference strains and isolated strains: the highest performance was by the EX method (83.3~89.5% accuracy) at species level identification (EDT, 70.3~78.9% accuracy; DT, less than 46.3~63.2% accuracy) of 24-h cultured colonies. Identification results at the genus level were 100% accurate at EDT, EX sample preparation and 24-h incubation time. On the other hand, the DT method showed relatively low identification accuracy in all extraction methods and incubation times. The analyzed spectra and MSP-based dendrogram showed that the isolated Staphylococcus strains were characterized at the species level. The performance analysis of MALDI-TOF MS shows the method has the potential ability to discriminate between Staphylococcus species from foods in Korea. This study provides valuable information that MALDI-TOF MS can be applied to monitor microbial populations and pathogenic bacteria in the food industry thereby contributing to food safety.

Effect of Cationization Agent Concentration on Glycan Detection Using MALDI TOF-MS

  • Kim, Inyoung;Shin, Dongwon;Paek, Jihyun;Kim, Jeongkwon
    • Mass Spectrometry Letters
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    • v.8 no.1
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    • pp.14-17
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    • 2017
  • The effect of cationization agent concentration on glycan detection via matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was investigated using $Na^+$ ions in the form of NaCl as the cationization agent. NaCl solution concentrations ranging from 1 mM to 1 M were investigated. Glycans from ovalbumin were mixed with the cationization agent solution and the 2,5-dihydroxybenzoic acid (2,5-DHB) matrix solution in a volume ratio of 1:1:1. The resulting mixture was loaded onto the MALDI plate. Two MALDI-TOF MS instruments (Voyager DE-STR MALDI-TOF MS and Tinkerbell RT MALDI-TOF MS) were used for detection of glycans. The best detection, in terms of the number of identified glycans, the peak intensity, and the signal-to-noise (S/N) ratio, was obtained with NaCl concentrations of 0.01-0.1 M for both MALDI-TOF MS instruments.

MALDI Mass Spectrometric Analysis of Nonderivatized Steroids Using Cyclodextrin-supported 2,5-Dihydroxybenzoic Acid as Matrix

  • Son, Jeongjin;Cha, Sangwon
    • Bulletin of the Korean Chemical Society
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    • v.35 no.5
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    • pp.1409-1412
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    • 2014
  • Sex hormones are important metabolites in vertebrates' development and reproduction. For rapid screening sex hormones, matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) is one of the promising analytical platforms, but MALDI MS faces many challenges in detecting steroids such as low ionization efficiency and matrix background interference. One potential strategy to overcome matrix interference in the low m/z region is using a cyclodextrin (CD)-supported matrix for steroid analysis since CD-supported matrixes are known to effectively suppress matrix-related ion signals. In this study, we aimed to find the optimal CD-supported matrix for the analysis of the nonderivatized sex steroids. Our results showed that the ${\alpha}CD$-supported 2,5-dihydroxybenzoic acid (DHB) matrix efficiently ionized all three major classes of sex hormones, estrogens, androgens, and progestagens, with low or no matrix background and also with high sensitivity. In addition, the ${\alpha}CD$-supported DHB matrix mainly generated molecular ions or protonated ions of sex hormones, and this enabled us to obtain information-rich tandem mass spectra which potentially lead to unambiguous identification of steroid species from complex metabolite mixtures.

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.