Computerized Interpretation of H P L C Chromatogramms by Means of Absorbance Ratio Method and Derivative Spectroscopy

  • U. Wellner
  • H. K. Biesalski
Part of the Lecture Notes in Medical Informatics book series (LNMED, volume 25)


As in other chromatographic processes it is the aim of high pressure liquid chromatography (HPLC) to identify the components of a mixture, whereby there may be some information on the class of substances in question. The analytical procedure can shortly be desribed as follows. The mixture is injected in a mobile phase (solvent) and passes a column where the separation takes place. The separated components leave the column at different retention times. A following detection device generates a signal as a function of concentration (chromatographic peak). Usually the components are identified by relating retention times of external standards to the times of occurence of chromatographic peaks. A System for the computer aided evaluation of such chromatograms was developed by the authors (1). That system was designed in such a way that all components of the mixture to be analyzed are supposed to separate completely, e.g. only chromatograms with distinct peaks were allowed, while overlapping peaks are rejected by a checking algorithm. An inevitable shortcome of this algorithm is, that total overlapping of peaks — which result in a single peak (with undetectable shoulders) — cannot be recognized, so that it was the responsibility of the HPLC- operator to ensure the absence of such envents.


High Pressure Liquid Chromatography Chromatographic Peak Pure Substance Relate Retention Time Absorbance Ratio 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    BIESALSKI HK, WELLNER U Computer aided evaluation of HPLC (High Pressure Liquid Chromatography) with fluorometric detection. Lecture Notes in Medical Informatics 16:64–69 (1982)CrossRefGoogle Scholar
  2. 2.
    ROSS AC Separation of long chain fatty acid esters of; retinol by high performance liquid chromatography. Anal. Biochem.115:324–330 (1981)PubMedCrossRefGoogle Scholar
  3. 3.
    PAANAKER JE, GROENENDIJK GWT Separation of geometric isomers of retinyl ester, retinal and retinol, pertaining to the visual cycle, by high performance liquid chromatography. J Chrom 168:125–132 (1979)CrossRefGoogle Scholar
  4. 4.
    BIESALSKI HK, HAFNER G, BÄSSLER KH Separation of long chain fatty acid esters of retinol in biological samples using isocratic adsorption chromatography (HPLC). To be publishedGoogle Scholar
  5. 5.
    O’HAVER TC Potential clinical applications of derivative and wavelength modulation spectrometry. Clin Chem 25:1548–53 (1079)Google Scholar
  6. 6.
    BIESALSKI HK, WELLNER U Qualitative analysis of chromatographic peaks by derivative spectroscopy. To be publishedGoogle Scholar
  7. 7.
    JOST R, MACLEAN W, STOVEKEN J Confirmatory identification of HPLC peaks using absorbance ratios at several wavelength. Chrom Newsletter 4:1–4 (1976)Google Scholar
  8. 8.
    HEIN H, JÖSTER R Die Leistungsfähigkeit des photometrischen Detektors LC55 mit variabler Wellenlänge beim Einsatz in der Hochdruck-Flüssigchromatographie. Appl. Chrom.28 (1977)Google Scholar
  9. 9.
    MALCZEWSKI ML, GRUSHKA E Multiple peak recognition in high per — formance liquid chromatography by fast fourier transfromation. J Chrom Sci 19:187–194 (1981)Google Scholar
  10. 10.
    METZGER HD True peak area separation of overlapping peaks in gas chromatograms by means of a process computer. Chromatographia 3:64–70 (1970)CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1985

Authors and Affiliations

  • U. Wellner
    • 1
  • H. K. Biesalski
    • 1
  1. 1.Institute of Physiological Chemistry IIUniversity of MainzMainzGermany

Personalised recommendations