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Quantitation in gradient high performance liquid chromatography/inductively coupled mass spectrometry investigated
Catherine J Duckett1, Nigel J C Bailey, Heather Walker
1Biological Chemistry, Division of Biomedical Sciences, Faculty of Medicine, Imperial College of Science, Technology and Medicine, Sir Alexander Fleming Building, South Kensington, London SW7 2AZ, UK.
Abstract:
The use of directly coupled high performance liquid chromatography/inductively coupled plasma mass spectroscopy (HPLC/ICPMS) employing chlorine ((35)Cl/(37)Cl) detection has been investigated with respect to the detection and quantitation of the drugs diclofenac and chlorpromazine. By integration of peak areas in the 'chloratogram' (the chlorine specific HPLC chromatogram), a calibration curve was constructed, from which the concentrations could be determined. Chlorine detected HPLC/ICPMS is quantitative over a wide range of concentrations of pharmaceutical relevance for metabolite detection and the results reproducible (standard deviation +/- 0.43%) over multiple injections. Application of gradient chromatography and variation in the bulk mobile phase physicochemical properties has little effect on the ICPMS detection response for these compounds. This work indicates that the use of HPLC/ICPMS is likely to be quantitatively reliable for metabolism studies for a range of chlorinated xenobiotics.