Novel Techniques for Identification and Characterization of Proteins Loaded on Gels in Femtomole Amounts

A combination of techniques is presented allowing gel-purified protein identification in the femtomole range using matrix-assisted-laser-desorption-ionization mass spectrometry. The proteins are detected in the primary gel by a sensitive negative staining procedure, transferred, and concentrated in a secondary gel matrix. There, they are digested in the presence of H₂ ¹⁸O and their sequences are predicted (1) by peptide mass fingerprinting, (2) by comparing the post-source-decay (PSD) spectra with theoretical spectra of candidate isobaric peptides using a computer algorithm called MassFrag, and (3) by a manual readout of the ¹⁸O/¹⁶O-labeled fragmentation ions in the PSD spectra.     

How to Use Secondary Data on Seafood Contamination for Probabilistic Exposure Assessment Purposes? Main Problems and Potential Solutions

Seafood consumption is related to both favorable health benefits of nutrients and to potential adverse health impacts of chemical contamination. To quantify the magnitude of this dilemma, probabilistic intake assessments can be performed. One step in such a procedure involves the determination of nutrient and contaminant concentrations in seafood for which data need to be collected. This article describes the process of building up a database containing previously published contaminant concentrations in seafood, and defining input distributions characterizing the variability. During the constitution of the database, several problems influencing the comparability of the data were encountered related to (1) sampling plans of the published data; (2) sample handling prior to analysis; (3) analytical methodologies; (4) the format of reporting results; and (5) missing data. Different solutions for these methodological problems have been developed. Contaminant concentrations ranges per gram fresh weight of 2.4–4390.0 ng for mercury, 0.1–5736.6 ng for the sum of indicator PCB, 0.002–115.000 pg TEQ for the sum of all PCBs, 0.002–34.400 pg TEQ for dioxins, and 0.006–126.000 pg TEQ for total of dioxin-like compounds were found. This work confirms that more guidelines are needed to standardize the analytical methodologies to be used and the format for result reporting in order to improve the comparability of data critical to conduct a human intake and risk-benefit assessment.