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Routine toxicological analysis is mostly focusedon the identification of non-organic and organic, chemicallydifferent compounds, but generally with low mass, usually notgreater than 500–600 Da. Peptide compounds with atomic masshigher than 900 Da are a specific analytical group. Several dozenof them are highly-toxic substances well known in toxicologicalpractice, for example mushroom toxin and animal venoms.In the paper the authors present an example of alpha-amanitinto explain the analytical problems and different original solutionsin identifying peptides in urine samples with the use ofthe universal LC MS/MS procedure.Materials and methodsThe analyzed material was urinesamples collected from patients with potential mushroomintoxication, routinely diagnosed for amanitin determination.Ultra filtration with centrifuge filter tubes (limited mass cutoff3 kDa) was used. Filtrate fluid was directly injected on thechromatographic column and analyzed with a mass detector(MS/MS).ResultsThe separation of peptides as organic, amphoteric compoundsfrom biological material with the use of the SPE techniqueis well known but requires dedicated, specific columns.The presented paper proved that with the fast and simple ultrafiltration technique amanitin can be effectively isolated fromurine, and the procedure offers satisfactory sensitivity of detectionand eliminates the influence of the biological matrix on analyticalresults. Another problem which had to be solved wasthe non-characteristic fragmentation of peptides in the MS/MSprocedure providing non-selective chromatograms. It is possibleto use higher collision energies in the analytical procedure,which results in more characteristic mass spectres, althoughit offers lower sensitivity.ConclusionsThe ultra filtration technique as a procedure ofsample preparation is effective for the isolation of amanitinfrom the biological matrix. The monitoring of selected masscorresponding to transition with the loss of water moleculeoffers satisfactory sensitivity of determination.

Analiza toksykologiczna peptydów w materiale biologicznym z wykorzystaniem spektrometrii mas na przykładzie oznaczania alfa-amanityny