Thermophilic bacteria of four genera in contrast to the commonly used production strains such as Bacillus subtilis, produce homologs other than menaquinone (MK) with seven isoprene units. The number of isoprene units and the configuration of double bonds are essential factors for their biological activity. The goal was to obtain a strain of bacteria that produces a wide range of MK homologs and only all-trans geometrical isomers, which was the strain G. kaustophilus. Using off-line two-dimensional LC-tandem MS in columns with the RP18 phase and the COSMOSIL cholester phase (separation according to the geometric configuration of double bonds) it was shown that thermophilic bacteria grown at different temperatures produce only all-trans isomers of menaquinones from MK-5 (menaquinone with five isoprenyl units) to MK-15 (fifteen isoprenyl units). Therefore, G. kaustophilus appears to be a biotechnologically important strain produces only trans isomers and additionally homologs from 5 to 15 isoprene units.

• Klíčová slova
• Cis/trans isomers, LC/ESI–tandem MS, Menaquinones, Shotgun mass spectrometry, Thermophilic bacteria,
• MeSH
• Bacteria * MeSH
• butadieny * MeSH
• hmotnostní spektrometrie MeSH
• vitamin K 2 chemie   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• butadieny * MeSH
• isoprene MeSH Prohlížeč
• vitamin K 2 MeSH

Very long chain fatty acids (VLCFAs) were identified in four strains of the green alga Botryococcus braunii (Trebouxiophyceae). The algae contained a series of monoenoic fatty acids up to triacontenoic acid and further VLCFAs in amounts around 1% of total fatty acids. The separation of lipid classes using hydrophilic interaction chromatography revealed that the most abundant VLCFAs (28:2, 28:1 and 28:0) were contained in neutral lipids (triacylglycerols and/or diacylglycerols) and in phospholipids (phosphatidic acid and/or phosphatidylcholine). Using non-aqueous reversed-phase liquid chromatography tandem mass spectrometry (NARP-LC/MS2) of the appropriate collected fractions, molecular species of triacylglycerols containing one or two VLCFAs were described and phosphatidylcholines containing VLCFAs were separated for the first time. Because the presence of Botryosphaerella sudetica (Chlorophyceae) as contaminant of Botryococcus braunii strain Droop 1950/807-1 placed some doubts on the results of previous studies, a strain of this green alga of was also analyzed. In contrast to Botryococcus, C16, a substantially lower proportion of C18 polyunsaturated fatty acids and no VLCFAs were detected in Botryosphaerella.

• Klíčová slova
• Botryococcus, Botryosphaerella, Green alga, LC/ESI-tandem MS, Lipidomics, Phosphatidylcholine, Triacylglycerols, Very-long chain fatty acids,
• MeSH
• Chlorophyta chemie   MeSH
• chromatografie kapalinová MeSH
• fosfolipidy analýza   MeSH
• mastné kyseliny analýza   MeSH
• nenasycené mastné kyseliny chemie   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• fosfolipidy MeSH
• mastné kyseliny MeSH
• nenasycené mastné kyseliny 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.

• MeSH
• Anoxybacillus chemie   MeSH
• chromatografie kapalinová metody   MeSH
• fosfolipidy analýza   chemie   MeSH
• lipidy analýza   chemie   MeSH
• tandemová hmotnostní spektrometrie metody   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• fosfolipidy MeSH
• lipidy 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
• aldehydy chemie   MeSH
• fosfolipidy analýza   MeSH
• hmotnostní spektrometrie s elektrosprejovou ionizací metody   MeSH
• lidé MeSH
• mastné kyseliny chemie   MeSH
• molekulární struktura MeSH
• Pectinatus chemie   MeSH
• pivo mikrobiologie   MeSH
• plasmalogeny analýza   MeSH
• tandemová hmotnostní spektrometrie metody   MeSH
• vysokoúčinná kapalinová chromatografie metody   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• aldehydy MeSH
• fosfolipidy MeSH
• mastné kyseliny MeSH
• plasmalogeny MeSH

Liquid chromatography-mass spectrometry (LC-MS) is the method of choice for the untargeted profiling of biological samples. A multiplatform LC-MS-based approach is needed to screen polar metabolites and lipids comprehensively. Different mobile phase modifiers were tested to improve the electrospray ionization process during metabolomic and lipidomic profiling. For polar metabolites, hydrophilic interaction LC using a mobile phase with 10 mM ammonium formate/0.125% formic acid provided the best performance for amino acids, biogenic amines, sugars, nucleotides, acylcarnitines, and sugar phosphate, while reversed-phase LC (RPLC) with 0.1% formic acid outperformed for organic acids. For lipids, RPLC using a mobile phase with 10 mM ammonium formate or 10 mM ammonium formate with 0.1% formic acid permitted the high signal intensity of various lipid classes ionized in ESI(+) and robust retention times. For ESI(-), the mobile phase with 10 mM ammonium acetate with 0.1% acetic acid represented a reasonable compromise regarding the signal intensity of the detected lipids and the stability of retention times compared to 10 mM ammonium acetate alone or 0.02% acetic acid. Collectively, we show that untargeted methods should be evaluated not only on the total number of features but also based on common metabolites detected by a specific platform along with the long-term stability of retention times.

• Klíčová slova
• LC-MS, additives, lipidomics, liquid chromatography, mass spectrometry, metabolomics, mobile phase, modifiers, optimization,
• MeSH
• chromatografie kapalinová metody   MeSH
• formiáty MeSH
• hmotnostní spektrometrie s elektrosprejovou ionizací metody   MeSH
• kyselina octová MeSH
• lipidomika * MeSH
• metabolomika metody   MeSH
• tandemová hmotnostní spektrometrie * metody   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• ammonium acetate MeSH Prohlížeč
• formiáty MeSH
• formic acid MeSH Prohlížeč
• kyselina octová 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
• Názvy látek
• cystein MeSH
• cysteinyl-leukotriene MeSH Prohlížeč
• leukotrieny MeSH

Galloyl esters of quercetin and taxifolin have been recently prepared semisynthetically as part of work towards modifying the solubility and modulating the biological activity of these natural flavonoids. In this paper we focused on the liquid chromatography-mass spectrometry (LC-MS) profiling of metabolites of 3-O-galloylquercetin and 7-O-galloyltaxifolin using human hepatocytes as the in vitro cell model. A subtoxic concentration (50μM) was used for both compounds and the formation of metabolites was monitored for 2h in hepatocytes and cultivation medium separately. Using negative electrospray ionization-quadrupole time-of-flight mass spectrometry (ESI-QqTOF MS), we identified different biotransformation patterns for the studied compounds. 3-O-Galloylquercetin is metabolized directly to glucuronides and methyl derivatives. In contrast, 7-O-galloyltaxifolin is oxidized to 7-O-galloylquercetin or cleaved to taxifolin, and consequently the products formed are sulfated or glucuronidated. The oxidative biotransformation of 3-O-galloylquercetin and 7-O-galloyltaxifolin is also accompanied by ester bond cleavage presumably by cellular enzymes (esterases) in a nonspecific manner. Our results provide fundamental insights into the biotransformation of monogalloyl esters of flavonoids and can be applied in investigations of the pharmaceutical potential of other galloylated polyphenolic substances.

• Klíčová slova
• 3-O-galloylquercetin, 3-O-galloylquercetin glucuronide, 3-O-galloylquercetin methyl derivative, 3GQ, 3GQG, 3GQM, 7-O-galloylquercetin, 7-O-galloyltaxifolin, 7GQ, 7GT, Cytotoxicity, ESI-QqTOF MS, GA, HPLC, LC–MS, Mass spectrometry, Metabolic transformation, PHE, Quercetin-3-O-gallate, SIM, TG, TS, Taxifolin-7-O-gallate, electrospray ionization-quadrupole time-of-flight mass spectrometry, gallic acid (gallate), high-performance liquid chromatography, liquid chromatography–mass spectrometry, phenyl, selected-ion monitoring, taxifolin glucuronide, taxifolin sulfate,
• MeSH
• biotransformace MeSH
• buňky NIH 3T3 MeSH
• chromatografie kapalinová metody   MeSH
• estery MeSH
• hepatocyty účinky léků   metabolismus   MeSH
• hmotnostní spektrometrie metody   MeSH
• kultivované buňky MeSH
• kyselina gallová chemie   farmakokinetika   toxicita   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• quercetin analogy a deriváty   chemie   farmakokinetika   toxicita   MeSH
• tandemová hmotnostní spektrometrie metody   MeSH
• zvířata MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• estery MeSH
• kyselina gallová MeSH
• quercetin MeSH
• quercitrin MeSH Prohlížeč
• taxifolin MeSH Prohlížeč

We have separated 2b myosin heavy chain (MyHC) isoform from the rat extensor digitorum longus muscle by SDS-PAGE and analyzed it by two subsequent mass spectrometry techniques. After tryptic digestion, the obtained peptides were identified by Matrix-Assisted Laser Desorption/Ionisation reflectron Time of Flight mass spectrometry (MALDI-TOF MS) and sequenced by liquid chromatography tandem mass spectrometry (ESI LC/MS/MS). The analyzed peptides proportionally covered 30 % of the 2b MyHC isoform sequence. The results suggest that the primary structure is identical with the highest probability to a NCBI database record ref|NP_062198.1|, representing the last updated record of rat 2b isoform. Nonetheless, four peptides carrying amino acid substitution(s) in comparison with the NCBI database record were identified.

• MeSH
• chromatografie kapalinová MeSH
• databáze proteinů MeSH
• elektroforéza v polyakrylamidovém gelu MeSH
• hmotnostní spektrometrie s elektrosprejovou ionizací * MeSH
• konformace proteinů MeSH
• kosterní svaly chemie   MeSH
• krysa rodu Rattus MeSH
• molekulární sekvence - údaje MeSH
• nesvalový myosin typu IIB chemie   izolace a purifikace   MeSH
• peptidové mapování MeSH
• sekvence aminokyselin MeSH
• spektrometrie hmotnostní - ionizace laserem za účasti matrice * MeSH
• tandemová hmotnostní spektrometrie * MeSH
• těžké řetězce myosinu chemie   izolace a purifikace   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• nesvalový myosin typu IIB MeSH
• nonmuscle myosin type IIB heavy chain MeSH Prohlížeč
• těžké řetězce myosinu MeSH

Lipid-like compounds containing a dimethylarsinoyl group, i.e. Me2As(O)-, have been identified by liquid chromatography/inductively coupled plasma mass spectrometry (LC/ICP-MS) and non-aqueous reversed-phase high-performance liquid chromatography (positive and/or negative high-resolution tandem electrospray ionization mass spectrometry (NARP-HPLC/HR-ESI+(-)-MS/MS) from three strains of green algae of the genus Coccomyxa (Trebouxiophyceae, Chlorophyta). The algae were cultivated in a medium containing 10 g arsenic/L, i.e. 133.5 mmol/L of Na2HAsO4.7H2O. After extraction by methyl-tert-butyl ether (MTBE), total lipids were analyzed by ICP-MS or ESI-MS without any further separation or fractionation. A total of 39 molecular species of arsenic triacylglycerols (AsTAG), 15 arsenic phosphatidylcholines (AsPC), 8 arsenic phosphatidylethanolamines (AsPE), 6 arsenic phosphatidylinositols (AsPI), 2 arsenic phosphatidylglycerols (AsPG) and 5 unknown lipids (probably ceramides) were identified. The structures of all molecular species were confirmed by tandem MS. Dry matter of the individual strains contained different amounts of total arsenolipids, i.e. C. elongata CCALA 427 (0.32 mg/g), C. onubensis (1.48 mg/g), C. elongata S3 (2.13 mg/g). On the other hand, there were only slight differences between strains in the relative abundances of individual molecular species. Possible biosynthesis of long-chain lipids with the end group Me2As(O) has also been suggested.

• Klíčová slova
• Arsenolipids, Coccomyxa elongata, Coccomyxa onubensis, Dimethylarsinoyl long chains, Green alga, Inductively coupled plasma mass spectrometry (ICP-MS), Non-aqueous reversed-phase high-performance liquid chromatography (NARP-HPLC), Positive/negative high-resolution tandem electrospray ionization mass spectrometry (HR-ESI-MS/MS), Trebouxiophyceae,
• MeSH
• arsenikové přípravky chemie   izolace a purifikace   metabolismus   MeSH
• Chlorella chemie   metabolismus   MeSH
• lipidy chemie   izolace a purifikace   MeSH
• molekulární struktura MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• arsenikové přípravky MeSH
• lipidy MeSH

Many compounds related to L-tryptophan (L-TRP) have interesting biological or pharmacological activity, and their abnormal neurotransmission seems to be linked to a wide range of neurodegenerative and psychiatric diseases. A high-throughput method based on ultra-high performance liquid chromatography connected to electrospray tandem mass spectrometry (UHPLC-ESI-MS/MS) was developed for the quantitative analysis of L-TRP and 16 of its metabolites in human serum and cerebrospinal fluid (CSF), representing both major and minor routes of L-TRP catabolism. The combination of a fast LC gradient with selective tandem mass spectrometry enabled accurate analysis of almost 100 samples in 24h. The standard isotope dilution method was used for quantitative determination. The method's lower limits of quantification for serum and cerebrospinal fluid ranged from 0.05 to 15nmol/L and 0.3 to 45nmol/L, respectively. Analytical recoveries ranged from 10.4 to 218.1% for serum and 22.1 to 370.0% for CSF. The method's accuracy ranged from 82.4 to 128.5% for serum matrix and 90.7 to 127.7% for CSF matrix. All intra- and inter-day coefficients of variation were below 15%. These results demonstrate that the new method is capable of quantifying endogenous serum and CSF levels of a heterogeneous group of compounds spanning a wide range of concentrations. The method was used to determine the physiological levels of target analytes in serum and CSF samples from 18 individuals, demonstrating its reliability and potential usefulness in large-scale epidemiological studies.

• Klíčová slova
• Cerebrospinal fluid, Neurochemicals, Serum, Tryptophan metabolism, UHPLC–MS/MS,
• MeSH
• biochemická analýza krve metody   MeSH
• chemické techniky analytické metody   MeSH
• indikátorové diluční techniky MeSH
• izotopy MeSH
• lidé MeSH
• reprodukovatelnost výsledků MeSH
• tandemová hmotnostní spektrometrie * MeSH
• tryptofan krev   mozkomíšní mok   MeSH
• vysokoúčinná kapalinová chromatografie * MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• izotopy MeSH
• tryptofan MeSH