Probabilistic limit of detection for ricin identification using a shotgun proteomics assay

Journal Article · Tue Oct 01 00:00:00 EDT 2019 · Analytical Chemistry

DOI:https://doi.org/10.1021/acs.analchem.9b02721· OSTI ID:1569006

Heller, Natalie C.  ^[1]; Garrett, Alaine M. ^[2]; Merkley, Eric D.  ^[1]; Cendrowski, Stephen ^[3]; Melville, Angela M. ^[1]; Arce, Jennifer S. ^[1]; Jenson, Sarah C.  ^[1]; Wahl, Karen L. ^[1]; Jarman, Kristin H. ^[1]

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1. BATTELLE (PACIFIC NW LAB)
2. OTHER GOVERNMENT AGENCIES
3. National Bioforensic Analysis Center

Robust and highly specific methods for the detection of the protein toxin ricin are of interest to the law enforcement community. In previous studies, methods based on liquid chromatography-tandem mass spectrometry shotgun proteomics have been proposed. The successful implementation of this approach relies on specific data evaluation criteria addressing (1) the quality of the mass spectrometric data, (2) the confidence of peptide identifications (peptide-spectrum matches), and (3) the number and sequence specificity of peptides detected. We present such data evaluation criteria, and use a novel approach to establish the limit of detection for this ricin assay. Specifically, we use logistic regression to determine the probability of detection for individual ricin peptides at different concentrations. We then apply basic rules from probability theory, combining these individual peptide probabilities into an overall assay limit of detection. This procedure yields an assay limit of detection for ricin at 42.5 ng on column or 21.25 ng/µL for a 2-µL injection. We also show that, in spite of the conventional wisdom that detergents are deleterious to mass spectrometric analyses, the presence of Tween-20 did not prevent detection of ricin peptides, and indeed assays performed in buffers that included Tween-20 gave better results than assays performed using other buffer formulations with or without detergent removal.

• Research Organization: Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
• Sponsoring Organization: USDOE
• DOE Contract Number: AC05-76RL01830
• OSTI ID: 1569006
• Report Number(s): PNNL-SA-144376
• Journal Information: Analytical Chemistry, Vol. 91, Issue 19
• Country of Publication: United States
• Language: English