Targeted proteomics recently proved to be a technique for the detection and absolute quantification of proteins not easily accessible to classical bottom-up approaches. Due to this, it has been considered as a high fidelity tool to detect potential warfare agents in wide spread kinds of biological and environmental matrices. Clostridium perfringens toxins are considered to be potential biological weapons, especially the epsilon toxin which belongs to a group of the most powerful bacterial toxins. Here, the development of a target mass spectrometry method for the detection of C. perfringens protein toxins (alpha, beta, beta2, epsilon, iota) is described. A high-resolution mass spectrometer with a quadrupole-Orbitrap system operating in target acquisition mode (parallel reaction monitoring) was utilized. Because of the lack of commercial protein toxin standards recombinant toxins were prepared within Escherichia coli. The analysis was performed using proteotypic peptides as the target compounds together with their isotopically labeled synthetic analogues as internal standards. Calibration curves were calculated for each peptide in concentrations ranging from 0.635 to 1101 fmol/μL. Limits of detection and quantification were determined for each peptide in blank matrices.

Faculty of Military Health Sciences University of Defense in Brno Třebešská 1575 CZ 500 01 Hradec Králové Czech Republic

Faculty of Military Health Sciences University of Defense in Brno Třebešská 1575 CZ 500 01 Hradec Králové Czech Republic Department of Biology Faculty of Science University of Hradec Kralove Hradecká 1285 CZ 500 03 Hradec Kralove Czech Republic

Military Health Institute Military Medical Agency Tychonova 1 CZ 160 00 Prague 6 Czech Republic

Citace poskytuje Crossref.org