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ESI-MS Analysis (esi-m + analysis)
Selected AbstractsFluorescent, internally quenched, peptides for exploring the pH-dependent substrate specificity of cathepsin BJOURNAL OF PEPTIDE SCIENCE, Issue 7 2006Paolo Ruzza Abstract Cathepsin B is a cysteine protease that in tumor tissues is localized in both acidic lysosomes and extracellular spaces. It can catalyze the cleavage of peptide bonds by two mechanisms: endoproteolytic attack with a pH optimum around 7.4, and attack from the C -terminus with a pH optimum at 4.5,5.5. In this work, seven fluorescent, internally quenched, decapeptides have been synthesized using the prototypical cathepsin B selective substrate Z-Phe-Arg-AMC as a lead, and used to identify the structural factors determining the susceptibility of peptides to hydrolysis at acidic and neutral pH values. Each peptide differs from the others in one amino acid (residue 6) and contains a highly fluorescent Nma group linked to the ,-amino function of the N -terminal Orn residue and a Dnp group linked to the side chain of the Lys8 residue acting as a quencher. Proteolytic cleavage was monitored by measuring the increase of fluorescence at 440 nm upon excitation at 340 nm, and the cleavage sites were determined by HPLC followed by ESI-MS analysis. Peptides containing Ala or Phe at position 6 are good substrates for the enzyme at both pH 5.0 and 7.4. By contrast, those containing Glu, Asp, Lys or Val are not cleaved at all by cathepsin B at pH 7.4, and are poorly hydrolyzed at pH 5.0. These findings provide new information for the rational design of cathepsin B-activated peptide-containing anticancer drugs. Copyright © 2006 European Peptide Society and John Wiley & Sons, Ltd. [source] Secondary metabolites from Paronychia argenteaMAGNETIC RESONANCE IN CHEMISTRY, Issue 1 2008Alessandra Braca Abstract Two new oleanane saponins (1 and 2) and one new flavonol glycoside (3) together with six known flavonoids, were isolated from the aerial parts of Paronychia argentea. Their structures were elucidated by 1D and 2D NMR experiments including 1D-TOCSY, DQF-COSY, NOESY, HSQC, and HMBC spectroscopy, as well as ESI-MS analysis. Copyright © 2007 John Wiley & Sons, Ltd. [source] Electrospray ionization tandem mass spectral analysis of oxidation products of precursors of sulfur mustardsRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 16 2006Vijay Tak Electrospray ionization tandem mass spectral (ESI-MSn) analysis of thiodiglycol, bis(2-hydroxyethylthio)alkanes (BHETAs) and their mono-, di-, tri-, and tetraoxygenated compounds was carried out to obtain their characteristic spectra for ESI-MS analysis. These compounds are important markers of chemical warfare agents, namely sulfur mustards. ESI-MSn (n,,,3) analysis of a compound by collisionally induced dissociation in an ion trap gives rise to mass spectra that are somewhat similar to electron ionization mass spectra. These ESI-MSn spectra can be used for compound identification. Under ESI-MS and ESI-MS/MS the compounds mostly produced [M+NH4]+, [M+H]+ and [M+HH2O]+ ions. Fragmentations of these even-electron precursors in the ion trap gave rise to characteristic product ions via neutral loss of O2, H2O, C2H4, HCHO, C2H4O, C2H4S, HSC2H4OH and C2H4SO. Fragmentation routes of these compounds are proposed that rationalize the formation of product ions in ESI-MSn analysis. Copyright © 2006 John Wiley & Sons, Ltd. [source] Analysis of the composition of immunoconjugates using size-exclusion chromatography coupled to mass spectrometryRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 13 2005Alexandru C. Lazar Recombinant monoclonal antibody drug products play an increasingly important role in the treatment of various diseases. Antibodies are large, multi-chain proteins and antibody preparations often contain several molecular variants, which renders them heterogeneous. The heterogeneity is further increased in immunoconjugates prepared by covalently linking several drug molecules per antibody molecule. As part of the product characterization, the molecular weights of the antibodies or their drug conjugates need to be measured. Electrospray ionization mass spectrometry (ESI-MS) is well suited for the analysis of recombinant antibodies and immunoconjugates. Sample preparation is an important element of ESI-MS analysis, in particular samples need to be freed of interfering charged species, such as salts and buffer components. In this paper, Amicon centrifugal filters, reversed-phase high-performance liquid chromatography (HPLC), and size-exclusion HPLC were evaluated for sample desalting. Size-exclusion HPLC, using aqueous acetonitrile as the mobile phase, directly coupled to ESI-MS provided the best performance and was optimized for the study of immunoconjugates. The results showed that antibodies carrying covalently linked maytansinoid molecules generated charge envelope profiles that differ from those of the non-conjugated antibody. For the determination of the distribution of the various conjugate species in an immunoconjugate sample prepared by randomly linking in the average 3.6 drug molecules per antibody molecule, the experimental conditions needed to be carefully selected to allow acquisition of the whole spectrum containing the charge envelopes of all species. Copyright © 2005 John Wiley & Sons, Ltd. [source] An HPLC-MS method for simultaneous estimation of ,,, -arteether and its metabolite dihydroartemisinin, in rat plasma for application to pharmacokinetic studyBIOMEDICAL CHROMATOGRAPHY, Issue 7 2003M. Rajanikanth Abstract This manuscript reports, the development and validation of a sensitive and selective assay method for simultaneous determination of ,,, -arteether and its metabolite dihydroartemisinin (DHA) in rat plasma by liquid chromatography,mass spectrometry. Chromatographic separations were achieved by gradient elution of the analytes with an initial composition of methanol,potassium acetate buffer (pH 4; 73:27, v/v) to 100% methanol in 3 min and maintained for 5 min on a Spheri-10, RP18 (100 × 4.6 mm i.d.) column following an RP18 (30 × 4.6 mm i.d.) guard column. The total ef,uent from the column was split so that one-tenth was injected into the electrospray LC/MS interface. ESI-MS analysis was performed using a Micromass Quattro II Triple Quadrupole Mass Spectrometer equipped with an electrospray source. The MS analysis was carried out at cone voltage of 22 V with a scan range of 200,500 Da. The analytes were quanti,ed from the [M+ K]+ ion chromatograms of ,,, -arteether at m/z 352, DHA at m/z 323, artemisinin at m/z 321 and propyl ether analogue of arteether at m/z 365. Liquid,liquid extractions with a combination of n -hexane and hexane,ethyl acetate (8:2) were used to isolate ,,, -arteether and DHA from rat plasma. The method was validated and gave good accuracy and precision for the studied domain. Linearity in serum was observed over the range 4.375,70 ng/mL for a -arteether and 10,160 ng/mL for , -arteether and DHA. Percentage bias (accuracy) and within- and between-assay precision were well within the acceptable range. This method was applied to study the pharmacokinetics following oral administration of ,,, -arteether (30 mg/kg) in rats. Copyright © 2003 John Wiley & Sons, Ltd. [source] | ||||||||||