Collision-induced dissociation as "mass spectrometric filter" for rapid screening of tetrapyrrole derivatives and their chelated metal species in complex biological and environmental samples.

ABSTRACT

RATIONALE Cyclic tetrapyrroles, such as chlorophylls and their diagenetic derivatives, are structurally diverse and often chelated with certain metal species in the natural environment. A high throughput analytical method enabling quick tetrapyrrole screening in complex samples will promote the study of tetrapyrrole biogeochemistry and probably discoveries of new tetrapyrroles. METHODS: Total lipid extracts of biological and environmental samples were injected onto a C18 column to separate compounds with a reverse-phase gradient. Collision-induced dissociation (CID) was performed at different energy levels, from 40 to 200 eV, on a quadrupole time-of-flight mass spectrometry (QTOF-MS) to identify cyclic tetrapyrroles in complex matrices. RESULTS: Under 200 eV CID cyclic tetrapyrroles exhibit a unique fragmentation behavior, the production of fragments larger than 300 Da. Utilizing such feature as a filter to extract product ions in the range of 300-500 Da, various cyclic tetrapyrrole derivatives are readily recognized in all tested biological and environmental samples. The 200 eV CID setup also dissociates chelated to porphyrin metals, including Cu, Fe, Mn, Ni, and V, as single-charged ions for direct MS detection. CONCLUSIONS: The 200 eV CID setup provides an efficient approach for the identification of cyclic tetrapyrroles, such as chlorophylls and fossil metalloporphyrins, in complex environmental samples. The direct detection of chelated to porphyrin metal ions with QTOF-MS shows the potential for compound-specific metal isotope analysis.