Calibration based on the "single-point calibration method", a simple exponential transformation of the response function of an evaporative light scattering detector was improved and applied to analysis of selected saccharides under hydrophilic interaction chromatography mode (a polar phase LiChrospher100 DIOL, mobile phase acetonitrile/water). The improved approach to the calibration procedure yielded a calibration curve with an excellent linearity (quality coefficient <5%). This quantitative evaluation of chromatograms of D-galactose suggested that not only anomers but even pyranose and furanose forms of the anomers could be resolved--the resulting calculations of abundance of the anomeric form strongly correlated with data from the literature obtained mostly by NMR studies (analogous results were also obtained for D-arabinose, D-glucose, and D-mannose). Because of the rapid separation (retention time less than 10 min), the observed correlation enabled to monitor anomeric conversion (mutarotation) of monosaccharides.

• MeSH
• kalibrace MeSH
• monosacharidy analýza   MeSH
• vysokoúčinná kapalinová chromatografie metody   MeSH
• Publikační typ
• časopisecké články MeSH

Saccharides and their derivatives are typical polar analytes without a suitable UV-chromophore that are nowadays analyzed by HPLC (high-performance liquid chromatography) under HILIC (hydrophilic interaction liquid chromatography) mode. Usually an evaporative light scattering detector (ELSD) is utilized which, however, gives a nonlinear response. A procedure to overcome the problem of mutarotating (time-varying) analytes recorded with such a nonlinear response detector is described. The procedure was applied for determination of glucosamine in two commercially available pharmaceutical formulations containing the common inorganic ions that the detector gives a response to. Under optimized conditions, both the anomers of glucosamine were separated and could be determined separately. Owing to the short retention time of the analyte (a run time <4 min) and relatively slow kinetics of the anomeric conversion (equilibration time 2.5 h), mutarotation could be monitored and corresponding rate constants calculated.

• MeSH
• chromatografie kapalinová metody   MeSH
• glukosamin analýza   chemie   MeSH
• hydrofobní a hydrofilní interakce MeSH
• isomerie MeSH
• lineární modely MeSH
• radiační rozptyl * MeSH
• reprodukovatelnost výsledků MeSH
• světlo MeSH
• Publikační typ
• časopisecké články MeSH

• MeSH
• biochemie MeSH

• Konspekt
• Lékařské vědy. Lékařství
• Učební osnovy. Vyučovací předměty. Učebnice
• NLK Obory
• biochemie
• NLK Publikační typ
• učebnice vysokých škol
• laboratorní cvičení

• MeSH
• organická chemie MeSH

• Konspekt
• Chemie. Mineralogické vědy
• NLK Obory
• chemie, klinická chemie