Development and Validation of Two Innovative Quantitative Liquid Chromatography Tandem Mass Spectrometry Methods for Forensic Toxicology Laboratories

The proliferation of liquid chromatography tandem mass spectrometry (LC-MS-MS) in forensic toxicology makes the development and validation of methods using generally accepted guidelines essential. The Virginia Department of Forensic Science (VADFS) will develop, validate, and disseminate two quantitative methods for whole blood and other biological matrices using LC-MS-MS that exceed the validation guidelines set forth by the Scientific Working Group for Forensic Toxicology (SWGTOX). An increased submission of designer drugs in VADFS Controlled Substances Section indicates a need for an accurate and robust quantitative method in the Toxicology Section. Limited literature references pertaining to concentrations associated with the effects of designer drugs makes interpretation or driving under the influence of drugs (DUID) and medicolegal death investigation challenging. A validated method for common designer drugs in whole blood will provide valuable concentration information that can be utilized for interpretation of DUID and death investigation cases. In addition to the validation of novel methods for the analysis of emerging drugs, it is important to evaluate and improve upon current methods. There are limited methods available for the simultaneous analysis of cocaine and opioids using LC-MS-MS for whole blood and other biological matrices. Given the nature of limited sample volume in forensic toxicology applications, establishing multi-drug class methods, with decreased sample volume, will have a significant impact on laboratory efficiency. Increased efficiency will directly impact the criminal justice system and death investigations by reducing turnaround times for case completion by enabling the result to be conveyed more rapidly. The proposed validations will include instrumental method development using an LC-MS-MS as well as three sample preparation techniques. The sample preparation techniques evaluated will be a solid phase extraction, liquid-liquid extraction, and protein precipitation. This will enable a comparison of each validation parameter for all extraction procedures as well as an evaluation of the most cost effective and efficient procedure. The anticipated timeline for this project is a total of twenty-four months with the anticipated outcome of at least two quantitative LC-MS-MS methods that will exceed the generally accepted toxicology validation guidelines. This project will have multiple interim reports including an initial report after the completion of method development for each method and a final report after the validations are completed. In addition, it will provide new methods that modernize the analysis of biological matrices using LC-MS-MS technology and will be of significant value to the toxicology community and criminal justice system. ca/ncf