caa a22 4500 378917455 CHVBK 20180305123557.0 cr unu---uuuuu 161128e20041101xx s 000 0 eng 10.1515/CCLM.2004.251 doi (NATIONALLICENCE)gruyter-10.1515/CCLM.2004.251 Systematic toxicological analysis by high-performance liquid chromatography with diode array detection (HPLC-DAD) [Elektronische Daten] [Fritz Pragst, Matthias Herzler, Björn-Thoralf Erxleben] In recent years, photodiode array detectors (DADs) have been much improved with respect to wavelength accuracy and resolution, sensitivity, linearity and software operation. UV spectra of drugs measured with up-to-date DADs from different manufacturers are in excellent agreement, have the same quality as measured by a conventional UV spectrometer and are highly reproducible. The calculation of similarity parameters by the DAD software includes the entire range of the spectra compared and allows recognition of very small differences. It was shown in a systematic study of more than 2500 toxicologically relevant substances that UV spectra have a very high specificity with respect to substance structure. Therefore, HPLC-DAD in combination with a comprehensive database of UV spectra and retention parameters is one of the most efficient techniques used in systematic toxicological analysis (STA). Furthermore, the method is advantageous for the identification of metabolites, since in many cases they have the same or very similar UV spectra compared with their respective parent substances and their retention times on reversed-phase columns are shifted in a manner typical of the particular biotransformation reaction. Beside these general aspects, practical applications of HPLC-DAD for STA are reviewed with respect to sample preparation and chromatographic conditions. The efficiency of the method is demonstrated for an example of a routine procedure using liquid-liquid extraction with CH2Cl2 and protein precipitation for sample preparation, a system of three isocratic mobile phases with different acetonitrile/phosphate buffer ratios and RP8 columns for chromatography and a database of 2682 UV spectra and relative retention times for substance identification. © Walter de Gruyter Medical equipment & techniques nationallicence Medical diagnosis nationallicence Diseases & disorders nationallicence high-performance liquid chromatography nationallicence metabolites of drugs nationallicence photodiode array detector nationallicence systematic toxicological analysis nationallicence UV spectral library nationallicence Pragst Fritz Institute of Legal Medicine, University Hospital Charité, Berlin, Germany aut Herzler Matthias Federal Institute for Risk Assessment (BfR), Berlin, Germany aut Erxleben Björn-Thoralf Shimadzu Europe GmbH, Duisburg, Germany aut Clinical Chemistry and Laboratory Medicine Walter de Gruyter 42/11(2004-11-01), 1325-1340 1434-6621 42:11<1325 2004 42 cclm https://doi.org/10.1515/CCLM.2004.251 text/html Onlinezugriff via DOI 1 research article jats NATIONALLICENCE 856 40 https://doi.org/10.1515/CCLM.2004.251 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Pragst Fritz Institute of Legal Medicine, University Hospital Charité, Berlin, Germany aut NATIONALLICENCE 700 1- Herzler Matthias Federal Institute for Risk Assessment (BfR), Berlin, Germany aut NATIONALLICENCE 700 1- Erxleben Björn-Thoralf Shimadzu Europe GmbH, Duisburg, Germany aut NATIONALLICENCE 773 0- Clinical Chemistry and Laboratory Medicine Walter de Gruyter 42/11(2004-11-01), 1325-1340 1434-6621 42:11<1325 2004 42 cclm CC0 http://creativecommons.org/publicdomain/zero/1.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-gruyter