Peptides eluted from peripheral blood cells of HLA-B*2705 healthy donor were analyzed by LC MALDI MS/MS and LC ESI FTMS techniques. The sequences of 92 peptide ligands identified from one healthy blood donor by LC MALDI-TOF MS/MS were compared with those previously published from in vitro long-term cell cultures available in SYFPEITHI database and splenocytes. It was found that 18 sequences confirmed within 1ppm mass error by LC ESI FTMS were already described and 3 of them matched with those previously reported from HLA-B*2705 splenocytes. Another 38 sequences validated within the same mass error were not found in SYFPEITHI database and are identified here for the first time. Finally, 36 sequences (5 sequences already published in SYFPEITHI database) were evaluated by LC MALDI-TOF MS/MS but no matches in the list of monoisotopic masses obtained from LC ESI FTMS were found.
- MeSH
- Alcohol Oxidoreductases MeSH
- Spondylitis, Ankylosing genetics metabolism MeSH
- Autoimmunity genetics MeSH
- Databases, Protein MeSH
- Adult MeSH
- Endopeptidases metabolism MeSH
- Financing, Organized MeSH
- Genetic Predisposition to Disease MeSH
- Histones genetics metabolism MeSH
- HLA-B27 Antigen analysis immunology isolation & purification MeSH
- Spectrometry, Mass, Electrospray Ionization MeSH
- Protein Interaction Domains and Motifs MeSH
- Blood Cells immunology metabolism MeSH
- Humans MeSH
- Peptide Mapping MeSH
- Peptides analysis immunology isolation & purification MeSH
- HSC70 Heat-Shock Proteins genetics immunology blood MeSH
- GTP-Binding Protein alpha Subunits, Gs genetics immunology blood MeSH
- Sequence Alignment MeSH
- Software MeSH
- Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization MeSH
- Check Tag
- Adult MeSH
- Humans MeSH
- Male MeSH
- Publication type
- Comparative Study MeSH
This new analytical approach for high-throughput and comprehensive lipidomic analysis of biological samples using ultrahigh-performance supercritical fluid chromatography (UHPSFC) with electrospray ionization-mass spectrometry (ESI-MS) is based on lipid class separation using 1.7 μm particle bridged ethylene hybrid silica columns and a gradient of methanol-water-ammonium acetate mixture as a modifier. The method enables a fast separation of 30 nonpolar and polar lipid classes within 6-min analysis time covering six main lipid categories including fatty acyls, glycerolipids, glycerophospholipids, sphingolipids, sterols, and prenols. Individual lipid species within lipid classes are identified based on positive- and negative-ion full scan and tandem mass spectra measured with high mass accuracy and high resolving power. The method is used for the quantitative analysis of lipid species in biological tissues using internal standards for each lipid class. This high-throughput, comprehensive, and accurate UHPSFC/ESI-MS method is suitable for the lipidomic analysis of large sample sets in clinical research.
- MeSH
- Glycerophospholipids analysis MeSH
- Spectrometry, Mass, Electrospray Ionization methods MeSH
- Lipids analysis MeSH
- Metabolomics methods MeSH
- Pentanols analysis MeSH
- Sphingolipids analysis MeSH
- Sterols analysis MeSH
- Chromatography, Supercritical Fluid methods MeSH
- Tandem Mass Spectrometry MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Liquid chromatography-electrospray tandem mass spectrometry (LC/ESI-MS/MS) was used to analyze phospholipids from three species of the anaerobic beer-spoilage bacterial genus Pectinatus. Analysis of total lipids by HILIC (Hydrophilic Interaction Liquid Chromatography) column succeeded in separating diacyl- and plasmalogen phospholipids. Plasmalogens were then analyzed by means of the ESI-MS/MS and more than 220 molecular species of four classes of plasmalogens (PlsCho (choline plasmalogen), PlsEtn (ethanolamine plasmalogen), PlsGro (glycerol plasmalogen), and PlsSer (serine plasmalogen)) were identified. Major molecular species were c-p19:0/15:0 PlsEtn and PlsSer, which accounted for more than 4% of the total lipids.
- MeSH
- Aldehydes chemistry MeSH
- Phospholipids analysis MeSH
- Spectrometry, Mass, Electrospray Ionization methods MeSH
- Humans MeSH
- Fatty Acids chemistry MeSH
- Molecular Structure MeSH
- Pectinatus chemistry MeSH
- Beer microbiology MeSH
- Plasmalogens analysis MeSH
- Tandem Mass Spectrometry methods MeSH
- Chromatography, High Pressure Liquid methods MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Unusual glucose-substituted cardiolipins (Glcx-CLs) in three genera of thermophilic bacteria, having more than one glycosidically linked glucose to the hydroxyl of the central glycerol of Glcx-CLs were identified for the first time in thermophilic bacteria of the genera Geobacillus, Meiothermus, and Thermus. The number of glucoses reached up to five units. The structure of glycosidically linked oligosaccharides was determined based on shotgun analysis MS (electrospray high-resolution tandem mass spectrometry), partially methylated alditol acetates were identified by GC-MS, both electron ionization (EI) and positive chemical ionization (PCI), hydrophilic interaction liquid chromatography (HILIC) separation and identification of CLs glycosides by high resolution MS-ESI, and digestion by specific glycosidases.
After shining as the ultimate separation - sequencing technique used for the successful completion of the Human Genome Project, in the early 2000s CE experienced lowered popularity among separation scientists. The renewed interest in recent years relates to the separation needs, especially in proteomics, metabolomics, and glycomics, where CE complements liquid chromatography techniques. This interest is further boosted by the regulators requiring additional separation techniques for characterization of newly developed pharmaceuticals. This paper gives a short overview of recent developments in the on-line interfacing of CE separation techniques with electrospray ionization/mass spectrometric analysis. Both the instrumentation and selected CE/ESI/MS applications including analyses of peptides, proteins, and glycans are discussed with the stress on research published in the past 3 years. Techniques related to the proteomic and glycomic analyses such as sample preconcentration, on-line protein digestion, and analyte derivatization prior CE/ESI/MS analysis are also included.
- MeSH
- Electrophoresis, Capillary methods MeSH
- Spectrometry, Mass, Electrospray Ionization methods MeSH
- Humans MeSH
- Peptides analysis chemistry MeSH
- Polysaccharides analysis chemistry MeSH
- Proteins analysis chemistry MeSH
- Proteomics methods MeSH
- Systems Integration MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Review MeSH
Phospholipids and glycolipids from two recently described species belonging to the thermophilic genus Anoxybacillus were analyzed by liquid chromatography-electrospray tandem mass spectrometry (LC/ESI-MS/MS). Analysis of total lipids from the facultatively anaerobic A. bogrovensis on a HILIC (Hydrophilic Interaction LIquid Chromatography) column succeeded in separating diacyl- and plasmalogen phospholipids. The LC/ESI-MS/MS analysis of the strict aerobe A. rupiensis revealed the presence of different unique polar lipids, predominantly alanyl-, lysyl-, and glucosyl-phosphatidylglycerols and cardiolipins. Each of the classes of polar lipids was then analyzed by means of the ESI-MS/MS and more than 140 molecular species of six lipid classes from A. bogrovensis and nearly 200 molecular species of nine classes of polar lipids from A. rupiensis were identified. Five classes of unidentified polar lipids were detected in both strains. Plasmalogens were thus determined for the first time in a facultatively anaerobic bacterium, i.e. A. bogrovensis.
A rapid and precise method for the identification and quantification of cysteinyl leukotrienes (leukotriene C(4), leukotriene D(4) and leukotriene E(4)), essential markers of bronchial asthma, in exhaled breath condensate was developed. The protocol consists of immunoaffinity separation and a detection step, liquid chromatography combined with electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). In particular, the selected reaction monitoring mode was used for its extremely high degree of selectivity and the stable-isotope-dilution assay for its high precision of quantification. The developed method was characterized with a high precision (≤ 7.7%, determined as RSD), an acceptable accuracy (90.4-93.7%, determined as recovery), a low limit of detection (≤ 2 pg/ml EBC) and a low limit of quantification (≤ 10 pg/ml EBC). It was compared to other simple, clinically appropriate combinations of pre-treatment methods (solid phase extraction and lyophilization) with LC/MS. Finally, the method (a combination of immunoaffinity separation with LC-MS) was successfully tested in a clinical study where a significant difference was found in the concentration levels of cysteinyl leukotrienes between patients with occupational bronchial asthma and healthy subjects.
- MeSH
- Asthma diagnosis MeSH
- Chromatography, Liquid methods MeSH
- Cysteine analysis MeSH
- Breath Tests methods MeSH
- Spectrometry, Mass, Electrospray Ionization methods MeSH
- Immunoassay methods MeSH
- Leukotrienes analysis MeSH
- Humans MeSH
- Adolescent MeSH
- Young Adult MeSH
- Exhalation MeSH
- Check Tag
- Humans MeSH
- Adolescent MeSH
- Young Adult MeSH
- Male MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- Evaluation Study MeSH
- Research Support, Non-U.S. Gov't MeSH
The triterpenoid plant hormones brassinosteroids (BRs) are believed to influence almost every aspect of plant growth and development. We have developed a sensitive mass spectrometry-based method for the simultaneous profiling of twenty-two naturally occurring brassinosteroids including biosynthetic precursors and the majority of biologically active metabolites. Using ultra-high performance liquid chromatographic (UHPLC) analysis, the run time was reduced up to three times (to 9 min) in comparison to standard HPLC BRs analyses, the retention time stability was improved to 0.1-0.2 % RSD and the injection accuracy was increased to 1.1-4.9 % RSD. The procedures for extraction and for two-step purification based on solid-phase extraction (SPE) were optimised in combination with subsequent UHPLC analysis coupled to electrospray ionisation tandem mass spectrometry (ESI-MS/MS) using Brassica flowers and Arabidopsis plant tissue extracts. In multiple reaction monitoring (MRM) mode, the average detection limit for BRs analysed was close to 7 pg, and the linear range covered up to 3 orders of magnitude. The low detection limits for this broad range of BR metabolites enabled as little as 50 mg of plant tissue to be used for quantitative analyses. The results of determinations exploiting internal standards showed that this approach provides a high level of practicality, reproducibility and recovery. The method we have established will enable researchers to gain a better understanding of the dynamics of the biosynthesis and metabolism of brassinosteroids and their modes of action in plant growth and development.
- MeSH
- Arabidopsis chemistry MeSH
- Brassica napus chemistry MeSH
- Brassinosteroids analysis MeSH
- Solid Phase Extraction methods MeSH
- Spectrometry, Mass, Electrospray Ionization methods MeSH
- Limit of Detection MeSH
- Reproducibility of Results MeSH
- Plant Extracts chemistry MeSH
- Tandem Mass Spectrometry methods MeSH
- Chromatography, High Pressure Liquid methods MeSH
- Publication type
- Journal Article MeSH
- Validation Study MeSH
Members of Hymenochaetaceae fungi are among well-known macromycetes with various medicinal properties. The aim of this study was to investigate the biological activities of Phellinus tuberculosus and Fuscoporia ferruginosa collected in Iran. The antimicrobial, antioxidant, and cytotoxic activities of the two species were examined, and their phenolic and polysaccharide contents were quantified. Compounds were characterized by HPLC-DAD chromatography and LC-ESI-MS/MS spectroscopy. According to our results, the antibacterial and antioxidant effects of P. tuberculosus extracts were stronger than F. ferruginosa. Also, the effect of hydroalcoholic extracts was higher than the aqueous extract. Gram-positive bacteria were more sensitive to all extracts, especially Streptococcus mutans with a MIC of 0.7 mg/mL and MBC of 6.25 mg/mL. HPLC-DAD analyses detected gallic acid, caffeic acid, and syringic acid in both fungi. The LC-ESI-MS/MS confirmed the detected compounds in HPLC-DAD and showed the presence of several phenolic compounds such as phellifuropyranone, phelligridin, and hispidin, besides others. This study showed that F. ferruginosa and P. tuberculosus are potent medicinal fungi with antibacterial and antioxidant properties, with no toxic effect on normal HDF cells, and possess various bioactive compounds including styrylpyrone-type phenols with well-known bioactivities.
- MeSH
- Anti-Bacterial Agents * chemistry isolation & purification pharmacology MeSH
- Antioxidants * chemistry isolation & purification pharmacology MeSH
- Basidiomycota * chemistry metabolism MeSH
- Chromatography, Liquid MeSH
- Gram-Positive Bacteria drug effects MeSH
- Phellinus chemistry MeSH
- Tandem Mass Spectrometry MeSH
- Chromatography, High Pressure Liquid MeSH
- Publication type
- Journal Article MeSH
- Geographicals
- Iran MeSH
