LC−ESI−MS Dotaz Zobrazit nápovědu
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
- alkoholoxidoreduktasy MeSH
- ankylózující spondylitida genetika metabolismus MeSH
- autoimunita genetika MeSH
- databáze proteinů MeSH
- dospělí MeSH
- endopeptidasy metabolismus MeSH
- financování organizované MeSH
- genetická predispozice k nemoci MeSH
- histony genetika metabolismus MeSH
- HLA-B27 antigen analýza imunologie izolace a purifikace MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací MeSH
- interakční proteinové domény a motivy MeSH
- krevní buňky imunologie metabolismus MeSH
- lidé MeSH
- peptidové mapování MeSH
- peptidy analýza imunologie izolace a purifikace MeSH
- proteiny tepelného šoku HSC70 genetika imunologie krev MeSH
- proteiny vázající GTP - alfa-podjednotky Gs genetika imunologie krev MeSH
- sekvenční seřazení MeSH
- software MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- srovnávací studie MeSH
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
- bronchiální astma diagnóza MeSH
- chromatografie kapalinová metody MeSH
- cystein analýza MeSH
- dechové testy metody MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací metody MeSH
- imunoanalýza metody MeSH
- leukotrieny analýza MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- vydechnutí MeSH
- Check Tag
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem 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.
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
- antibakteriální látky * chemie izolace a purifikace farmakologie MeSH
- antioxidancia * chemie izolace a purifikace farmakologie MeSH
- Basidiomycota * chemie metabolismus MeSH
- chromatografie kapalinová MeSH
- grampozitivní bakterie účinky léků MeSH
- Phellinus chemie MeSH
- tandemová hmotnostní spektrometrie MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Írán MeSH
The great research interest in the quantification of reactive carbonyl compounds (RCCs), such as methylglyoxal (MGO) in biological and environmental samples, is reflected by the fact that several publications have described specific strategies to perform this task. Thus, many reagents have also been reported for the derivatization of RCCs to effectively detect and quantify the resulting compounds using sensitive techniques such as liquid chromatography coupled with mass spectrometry (LC-MS). However, the choice of the derivatization protocol is not always clear, and a comparative evaluation is not feasible because detection limits from separate reports and determined with different instruments are hardly comparable. Consequently, for a systematic comparison, we tested 21 agents in one experimental setup for derivatization of RCCs prior to LC-MS analysis. This consisted of seven commonly employed reagents and 14 similar reagents, three of which were designed and synthesized by us. All reagents were probed for analytical responsiveness of the derivatives and stability of the reaction mixtures. The results showed that derivatives of 4-methoxyphenylenediamine and 3-methoxyphenylhydrazine-reported here for the first time for derivatization of RCCs-provided a particularly high responsiveness with ESI-MS detection. We applied the protocol to investigate MGO contamination of laboratory water and show successful quantification in a lipoxidation experiment. In summary, our results provide valuable information for scientists in establishing accurate analysis of RCCs.
The main analytical benefit of this study is the development of methods enabling a rapid determination of total lipids of algae by lipidomic analysis and detailed identification and quantification of a complex mixture of natural TAGs by silver-LC/APCI-MS and NARP-LC/APCI-MS. Both types of chromatography can readily identify, both qualitatively and semiquantitatively, triacylglycerols containing 16:3 and 16:4 acids in the molecule. We conclude that the genus Chloromonas is a major producer of C16 PUFAs mostly contained in TAGs. Since more detailed studies in this field have been stymied by the shortage of 16:3 and 16:4 FAs, we decided to study the alga Chloromonas as a potential biotechnological source of C16 PUFAs.
Rapid and precise method for the determination of 8-iso-prostaglandin F(2alpha), an essential marker of the oxidative stress, in exhaled breath condensate (EBC) was developed. The protocol consisted of stable isotope dilution, immunoseparation combined with selective and sensitive LC-ESI-MS/MS operated in multiple reaction monitoring (MRM) mode. The imprecision of the developed method was below 8.8%, the parameter of mean inaccuracy was determined as <9.6% (0-250pg of 8-iso-prostaglandin F(2alpha)/ml EBC). The limit of detection (LOD) was 1 pg/ml EBC and limit of quantification (LOQ) 5 pg/ml EBC. A significant difference in 8-iso-prostaglandin F(2alpha) content between the group of asbestosis patients and healthy volunteers was found.
- MeSH
- dechové testy MeSH
- dinoprost analogy a deriváty analýza MeSH
- financování organizované MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací metody MeSH
- kalibrace MeSH
- lidé MeSH
- reprodukovatelnost výsledků MeSH
- senzitivita a specificita MeSH
- tandemová hmotnostní spektrometrie metody MeSH
- vysokoúčinná kapalinová chromatografie metody MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- validační studie MeSH
The sensitive assay method was developed for a parallel, rapid and precise determination of the most prominent oxidative stress biomarkers: 8-iso-prostaglandin F(2α) a lipid oxidation biomarker, o-tyrosine an amino acid oxidation biomarker and 8-hydroxy-2'-deoxy-guanosine a nucleic acid oxidation biomarker. The method consisted of a pre-treatment part, freeze drying (lyophilization), serving the purpose of biomarkers concentration from the exhaled breath condensate and detection method LC-ESI-MS/MS, where 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 is characterized by the following parameters: the precision was higher than 84.3% and the mean accuracy (relative error) was determined lower than 11.6%. The method was tested on samples obtained from patients diagnosed with asbestosis and silicosis, occupational diseases induced by oxidative stress, and then compared with samples from healthy subjects. The difference in biomarkers' concentration levels found between the two groups was statistically significant.
- MeSH
- azbestóza metabolismus MeSH
- biologické markery analýza MeSH
- chromatografie kapalinová MeSH
- dechové testy MeSH
- dospělí MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací metody MeSH
- lidé MeSH
- lyofilizace MeSH
- mladý dospělý MeSH
- oxidační stres * MeSH
- silikóza metabolismus MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem 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.
- MeSH
- Bacteria MeSH
- chromatografie kapalinová metody MeSH
- glukosa MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací metody MeSH
- kardiolipiny * analýza MeSH
- plynová chromatografie s hmotnostně spektrometrickou detekcí MeSH
- tandemová hmotnostní spektrometrie * metody MeSH
- Publikační typ
- časopisecké články MeSH
Glucuronidation of the non-steroidal anti-inflammatory chiral drug flobufen and its major metabolite M17203 has been implicated as an important mechanism of flobufen elimination. To characterize flobufen metabolism by O-glucuronidation, new liquid chromatographic method (LC) coupled with ESI-MS was developed to detect the conjugates of flobufen and its metabolites formed in vitro in rat liver microsomes. Discovery DSC-18 LT cartridge columns were utilized for solid phase extraction (SPE) and Discovery C18 column (150 mm x 2.1 mm, 5 microm particle size) was used for LC separation. Chiral inversion of flobufen and its metabolites enantiomers was checked by special 1-allyl-(5R,8S,10R)-terguride column (150 mm x 4.6 mm). O-Glucuronidation of the S-enantiomer displayed a typical Michaelis-Menten kinetics, whereas the R-enantiomer exhibited a substrate inhibition type of kinetics. The study of glucuronidation of M17203 led to kinetics with sigmoidal characteristics.
- MeSH
- butyráty analýza MeSH
- financování organizované MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací metody MeSH
- jaterní mikrozomy metabolismus MeSH
- krysa rodu rattus MeSH
- potkani Wistar MeSH
- vysokoúčinná kapalinová chromatografie metody MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
