Applications of high-pressure liquid chromatography and field desorption mass spectrometry in studies of natural porphyrins and chlorophyll derivatives.
Mixtures of porphyrins derived from natural sources can be readily separated by high-pressure liquid chromatography both analytically and on a preparative scale. A variety of procedures have been developed not only for the esters but also for free acids, and on the analytical scale quantitation is easily achieved by visible absorption. The retention times are largely characteristic of the number of carboxylic acid side chains (or other polar groups) but further information can be obtained by mass spectrometric studies of the various fractions. Field desorption mass spectrometry is particularly useful for this purpose because the emitter wire can be dipped directly into the eluates. The field desorption spectra of porphyrin free acids and esters as well as their metal complexes give essentially molecular ions with little or no fragmentation in most cases, whereas electron-impact mass spectrometry, particularly of free acids, is impeded by the low volatility of porphyrins. Mixtures can also be analysed by field desorption mass spectrometry, and this provides not only a rapid qualitative assessment of the components of a mixture, but also a check on the subsequent chromatographic separations.
D000042:Absorption; D002734:Chlorophyll / Q000031:analogs & derivatives* / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid; D004583:Electrons; D005243:Feces / Q000032:analysis; D006801:Humans; D013058:Mass Spectrometry; D008745:Methylation; D011166:Porphyrins / Q000302:isolation & purification* / Q000652:urine
D011166:Porphyrins; D002734:Chlorophyll