High-performance liquid chromatography-mass spectrometry of porphyrins by using an atmospheric pressure interface.

A method is described for the high-performance liquid chromatography (HPLC) mass spectrometry analysis of porphyrin mixtures by using an atmospheric pressure interface, which can operate in two modes: pneumatically assisted elecrrospray and atmospheric pressure chemical ionization (APCI). Optimization of the conditions and evaluation of spectral information has been carried out by using direct injection of free-base and metallo porphyrin standards. The most effective results were obtained using APCI. HPLC-APCI mass spectrometry analysis of the demetallated vanadyl porphyrin fraction from the Triassic Serpiano oil shale has allowed rapid characterization of the distribution; more than 50 significant components are present. The presence of trace amounts of high molecular weight (>C33) cycloalkano porphyrins indicates the occurrence of photic zone anoxia in the ancient water column. This example illustrates the potential of the approach for studies of porphyrin mixtures of environmental or biological significance, which should be applicable to other types of metallo and free-base components that can be separated by HPLC under normal or reversed-phase conditions.