Identification of Anticoagulant Adulterants in Seized Material and Biological Samples

Warfarin and superwarfarins are anticoagulant agents widely used in commercial rodenticides and in the treatment of bleeding disorders. The major compounds of the class include brodifacoum, bromodiolone, and difenacoum, all of which have a history of involvement in many types of forensic casework such as suicides, homicides, and accidental and deliberate poisonings. They have also been identified as potential chemical terror agents. Recently these substances have emerged as toxic adulterants in synthetic cannabinoid, cocaine, and marijuana casework, and have led to hundreds of hospitalizations and at least five deaths in the US. Toxic clinical effects of anticoagulant exposure include spontaneous internal and external bleeding, and can lead to death. The detection of these compounds in forensic casework is challenging due to the small amounts present in samples and their chemical properties such as their large mass, low volatility, polarity, and thermal instability. The goal of the proposed project is to systematically assess the effectiveness of current methodologies for the detection of anticoagulant adulterants and develop a variety of validated methods and workflow options, suited to different crime and toxicology laboratory capabilities and resources. Once complete, the validated methods will be used to examine 300 synthetic cannabinoid casework samples from current authentic seized drug cases and 68 authentic de-identified blood and serum samples from cases of suspected anticoagulant poisoning from the 2018 outbreak. The workflow proposed will include both nominal mass and high-resolution mass spectrometry methods using GC-MS, HPLC-UV, LC-MS/MS and LC-TRAP. The deliverables and outcomes for this project, in addition to the traditional conference presentations and peer reviewed publications, will include a critical assessment of existing analytical approaches addressing their limitations, a portfolio of validated methods on multiple platforms, comparative databases of anticoagulant products including digital and electronic spectral data, and instructional recorded presentations describing the principles and performance of the methods. Dissemination will be accomplished through an online toolkit of practical analytical approaches and resources for the analysis of anticoagulant adulterants in forensic casework in forensic laboratories with a range of resources and technology. Note: This project contains a research and/or development component, as defined in applicable law, and complies with Part 200 Uniform Requirements - 2 CFR 200.210(a)(14). CA/NCF