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  • Analytical Chemistry and Spectroscopy  (2,287)
  • 1995-1999  (2,287)
  • 1
    Electronic Resource
    Electronic Resource
    Enantiomeric separation and sensitive determination of D, L-amino acids derivatized with fluorogenic benzofurazan reagents on pirkle type stationary phases (1995)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 9 (1995), S. 10-17 
    Details
    ISSN: 0269-3879
    Keywords: 4-fluoro-7-nitro-2,1,3-benzoxadiazole ; NBD-F ; 4-(N,N-dimethylaminosulphonyl)-7-fluoro-2,1,3-benzoxadiazole ; BDD-F ; enantimoeric separation ; D,L-amino acid ; biological sample ; Pirkle type stationary phase ; HPLC ; Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The enantiomeric separations of D, L-amino acids derivatized with fluorogenic reagents, 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F), 4-(N,N-dimethylaminosulphonyl)-7-fluro-2,1-3,-benzoxadiazole (DBD-F) and 4-aminosulphonbly-7-fluoro-2,1,3- benzoxadiazole (ABD-F) by high-performance liquid chromatography (HPLC) ON various Pirkle type chiral stationary phases (CSPs, Sumichiral OA series) with citric acid in methanol as a mobile phase were studied. Since the least retention and no separation was observed for the derivatives of racemic phenylalanine methyl-ester,-amide and a drug without an α-carboxyl group, the carboxylic acid group of the amino acid derivatives seemed to contribute to the enantioselective fixation of the derivatives through hydrogen bonding on the N-acyl-amino acid amide moiety of the CSP. The enantioselective retention of the derivatives was attained through the (S) or (R) configuration of valine, phenylglycine, naphthylglycine, naphthylethylamine or the tert-leucine moiety in the CSP. The 2, 1, 3-benzoxadazole (benzofurazan) moiety in the derivatives helps the effective fixation of the derivatives through a π-πinteraction with an aromatic moiety such as a 3, 5-dinitrophenyl group in the Pirkle type chiral stationary phases.D-Amino acids in biological samples were easily determined utilizing the present derivatization with NBD-F, enantiomeric separation and fluorometric detection (530 nm em/470nm ex) following deproteinization of biological samples (serum or brain homogenate) with methanol and centrifugation. The applications of the method were clearly demonstrated by the following results; D-Ala was detected in sera of healthy volunteers at a level of 0.48-3.10μM. D-Lys was found in the serum of a patient with myeloma and requiring renal dialysis, and D-Ser was found in rat and bovine cerebrum. Peak identification was performed by use of different types of stationary phases especially those bearing the opposite configuration to that of the chiral centre.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bmc.1130090103
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  • 2
    Electronic Resource
    Electronic Resource
    Mannitol prevents methionine sulphoxidation mediated electrophoretic heterogeneity of apolipoprotein A-I (1995)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 9 (1995), S. 28-31 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Hybrid isoelectic focusing of apolipoprotein A-I in polyacrylanide gels with immobilized pH-gradients under non-denaturing conditions resulted in the occurrence of additional bands which could prevent the specific and sensitive detection of genetic variants. Hybrid isoelectric focusing of tow chromatographically distinguishable apolipoprotien A-I isoforms that differ by sulphoxidaton of methionine residues, apo A-I(Met) and apo A-I(MetSO), revealed that the additional bands were caused by this post-translational modification. Several antioxidative additives and conditions were compared for their ability to prevent methionine sulphoxidation in apoliporotein A-I In the presence of 200 g/L mannitol in the gel, apolipoprotein A/I focused as a single band. Since methionine sulphoxidation in proteins is a general phenomenon either taking place in vivo or in vitro by isoelectric focusing, we conclude that isoelectric focusing in the presence of mannitol will improve the quality of resolution of many proteins in gels with immobilized pH-gradients.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bmc.1130090106
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  • 3
    Electronic Resource
    Electronic Resource
    A highly sensitive method to quantify triazolam and its metabolites with liquid chromatography - mass spectrometry (1995)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 9 (1995), S. 48-51 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: We established a highly sensitive method to determine triazolam and its major metabolites, α-hydroxytriazolam and 4-hydroxytriazolam, in human plasma and urine with a liquid chromatography-mass spectrometry system which incorporates an atmospheric chemical ionization interface. A plasma sample and a urine sample after solvent extraction were injected into an ODS column of reversed phase with a mobile phase in a linear solvent gradient of initially 50 mM ammonium acetate (pH 4.0) 50%: methanol 50% and 15 min later methanol 100%; quantification limits of as low as 20 pg/mL at an SNR of 3 were obtained for each compound. With diazepam as an internal standard, recovery yields of 84, 81, and 77% were obtained for triazolam, α-hydroxytriazolam and 4-hydroxytriazolam, respectively.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bmc.1130090110
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  • 4
    Electronic Resource
    Electronic Resource
    Sensitive determination of salmon calcitonin, by means of pre-column derivatization, HPLC and fluorometric determination (1995)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 9 (1995), S. 52-55 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A high sensitive analytical method for salmon calcitonin (sCT) was established by means of derivatization with a fluorescence labelling agent, DBD-F, followed by HPLC separation and fluorescence detection at 558 nm with excitation at 430 nm.Under optimized conditions (DBD-F, 0.1 M borate buffer (pH 8.5): acetonitrile (70:30, v/v)), three molecules of DBD-F reacted with one sCT molecule in the presence of 1 mM sodium dodecyl sulphate (SDS) at 50 oC for 3 h. The detection limit for the sCT derivative was 20 fmol. A linear relationship was obtained between the amount of sCT on column and the peak heights in the range of 35-8000 fmol.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bmc.1130090111
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  • 5
    Electronic Resource
    Electronic Resource
    N-glycosylation site mapping of recombinant tissue plasminogen activator by micellar electrokinetic capillary chromatography (1995)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 9 (1995), S. 59-67 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: This report describes the N-glycosylation mapping of recombinant tissue plasminogen activator (rt-PA) using micellar electrokinetic capillary chromatography. The carbohydrate structures were tentatively assigned by comparison with the anion-exchange fractionated oligosaccharides and by a comparison with previously reported data. The separation was shown to rely mainly on the degree of sialylation of the oligosaccharides, allowing a quantitative determination of the proportion of neutral and mono- to tetrasialylated structures. Significant differences in the oligosaccharide distribution of the two variants of rt-PA, which differ by the presence (type I) or the absence (type II) of oligosaccharides at the Asn-184 site, were observed. The distribution of the oligosaccharides at each of the rt-PA glycosylation sites was then determined. Glycopeptides were prepared by tryptic digestion of rt-PA and isolated using two consecutive chromatographic procedures. The glycopeptides were finally treated with N-glycanase, and the resulting oligosaccharides were analysed by capillary electrophoresis. Oligosaccharide mapping revealed that the Asn-448 and Asn-184 sites carry the same population of complex-type oligosaccharides but that the relative amounts of each oligosaccharide vary markedly. High-pH anion-exchange chromatography performed on the desialylated oligosaccharides at each glycosylation site showed that the degree of microheterogeneity was related not only to the degree of sialylation but also to structural differences in the oligosaccharide sequences. From the results as a whole, we concluded that the Asn-448 site contains a greater proportion of heavily sialylated structures and has a higher degree of microheterogeneity. The Asn-117 site was demonstrated to contain a significant number of monosialylated structures (18.4%) in addition to the high-mannose-type oligosaccharides.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bmc.1130090202
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  • 6
    Electronic Resource
    Electronic Resource
    Determination of glutathione and related aminothiols by gas chromatography with flame photometric detection (1995)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 9 (1995), S. 85-89 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A selective and sensitive method for the determination of glutathione (GSH) and related aminothiols such as cysteine (Cys), cysteinylglycine (CysGly) and γ-glutamylcysteine (γ-GluCys) by gas chromatography (GC) has been developed. GSH and related aminothiols were converted into their N, S-isopropoxycarbonyl methyl ester derivatives and measured by GC with flame photometric detection using a short capillary column (5 m × 0.53 mm i.d.) of cross-linked DB-1. The calibration curves were linear in the range 1-25 nmol for GSH and in the range 0.2-5 nmol for other aminothiols, and the detection limits of GSH, Cys, CysGly and γ-GluCys were approximately 5, 0.2, 0.3 and 0.5 pmol per injection respectively. This method was successfully applied to blood samples without prior clean-up, and GSH and related aminothiols in these samples could be analysed without any influence from coexisting substances. Overall recoveries of GSH and other aminothiols added to blood samples were 88-107%. The analytical results of free and total blood GSH and related aminothiols in normal subjects are presented.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bmc.1130090206
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  • 7
    Electronic Resource
    Electronic Resource
    Masthead (1995)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 9 (1995) 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bmc.1130090301
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  • 8
    Electronic Resource
    Electronic Resource
    Determination of guanase activity in normal and pathological sera by high-performance liquid chromatography (1995)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 9 (1995), S. 130-134 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A simple HPLC assay for serum guanase based on the direct determination of enzymatically formed xanthine was applied to normal and pathological sera. The procedure is sensitive, precise (CV below 5%) and suitable for routine purposes, and the method requires only 50 μL of sample. Using this method the reference range as determined from the sera of 40 healthy adult controls is 0-1.1 U/L. In patients with various liver diseases serum guanase activities were found to be increased 5- to 50-fold compared with the normal mean value.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bmc.1130090304
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  • 9
    Electronic Resource
    Electronic Resource
    Stereoselective analysis of nadolol in human plasma (1995)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 9 (1995), S. 226-228 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A steroeselective method, involving a single liquid-liquid extraction step, was developed and validated for the analysis of nadolol in human plasma. The assay involved extraction of nadolol and desmethyl-nadolol as internal standard (IS) from alkalinized plasma into dichloromethane. The organic solvent was separated and evaporated under nitrogen at 40°C. A chiral derivatization scheme with (R)-(-)-napthylethylisocyanate (50 μL of 0.1% solution in dichloromethane for 60 min) was employed to convert the enantiomers of nadolol into the corresponding diastereomeric derivatives. The residue was reconstitutd in the mobile phase and injected onto a C-18 column. The mobile phase was a mixture of methanol: tetrahdrofuran: water (52:7:41 by vol) containing about 0.001% v/v of both phosphoric acid and tetramethylethylenediamine. Fluorimetric detection was performed at excitation 230 and emission 330 nm. The assay was specific for the enantiomers of nadolol and the lower limit of quantitation was 2 ng/mL for of the enantiomers. Analysis of quality control samples resulted in precision estimates of 7% RSD for inter-assay and 10.1% RSD for intra-assay and the predicted concentrations deviated less than 9.4% of the nominal values for the four enantiomers. The extraction recoveries of the individual enantiomer was about 70%. Stability of nadolol enantiomers were established for four freeze/thaw cycle periods and in the autosampler at 5°C for at least 116 h.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bmc.1130090507
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  • 10
    Electronic Resource
    Electronic Resource
    Announcement (1995)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 9 (1995), S. i 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bmc.1130090513
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