Lab Matters Summer 2019 - Page 51

APHL 2019 POSTER ABSTRACTS Brodifacoum-laced Synthetic Cannabinoids L. Blum, J. Homan and M. Danno, NMS Labs In early 2018, individuals seeking medical care for unexplained bleeding began to appear in the mid-western states of the US. Initially a blood sample from one of the cases was sent to NMS Labs for an anticoagulant screen, which consisted of an LC-MS/MS based assay to identify brodifacoum, bromadiolone, chlorophacinone, dicumarol, difenacoum, diphacinone, and warfarin. This blood sample tested positive for brodifacoum. Soon, additional samples from patients with similar clinical signs were sent to the lab from various state departments of health and hospitals and tested positive for mostly brodifacoum with some also detecting small amounts of difenacoum and bromadiolone. The only common factor amongst these patients was the use of synthetic cannabinoid products around the time the bleeding symptoms developed. Subsequent testing by law enforcement laboratories confirmed the presence of brodifacoum in some of the synthetic cannabinoid products. The qualitative LC-MS/MS analysis for anticoagulants employed a standard addition analysis with a 10 ng/mL standard to verify the reporting limit. With no deuterated standards available as internal standards, chloro-warfarin was added to each sample as an analog internal standard. This was followed by an acetonitrile crash and then a back-extraction into methyl tert-butyl ether. After dry down, the extract was reconstituted with a mixture of aqueous ammonium hydroxide/methanolic ammonium hydroxide. The LC- MS/MS analysis employed negative-ion electrospray and utilized a C18 column with aqueous ammonium hydroxide/methanolic ammonium hydroxide mobile phase. From mid-March through the end of 2018, we received approximately 424 requests for anticoagulants or brodifacoum testing which included about 351 individuals from 31 states and Washington, DC. Of these samples, 219 qualitatively tested positive for brodifacoum, 59 for difenacoum, 30 for bromadiolone, and 1 for diphacinone. Following the receipt of a deuterated brodifacoum standard from the CDC, a quantitative assay for brodifacoum was developed and then 26 samples were subsequently tested. Twenty-one (21) quantitative samples were positive with a few patients having multiple samples collected over time for testing. The first positive quantitative sample measured from 13 individual patient specimens ranged from 14 to 357 ng/ mL and averaged 108 ± 102 ng/mL with a median of 78 ng/mL. Adulterants are often prevalent in abused drugs and the substances can be wide ranging. Their impact on public health can vary depending on the substances involved. In this case, the coordinated efforts of governmental agencies and the testing laboratory were crucial in identifying the cause of bleeding and the extent of the problem. Presenter: Lee Blum, NMS Labs, Horsham, PA Quantitative Analysis of Fentanyl and Analogues in Human Whole Blood P. Negri 1 , C. Burrows 1 , A. Krotulski 2 , D. Tran 1 , X. He 1 , O. Cabrices 1 , A. Wang 1 , H. McCall 1 , X. Chen 1 ; 1 SCIEX, 2 Center for Forensic Science Research and Education at the Fredric Rieders Family Foundation Fentanyl analogues and their metabolites are a rising concern as thousands die from opioid overdose across the country. Some of these synthetic drugs have very high potency and thus only require a small amount for an accidental overdose. In addition, these high PublicHealthLabs @APHL potency drugs pose a danger to public health and public safety personnel due to the possibility of skin absorption or inhalation of the drug. In order to properly identify these fentanyl analogues in biological matrices, forensic laboratories require sensitive MS systems to accurately quantitate at low concentrations, and highly specific chromatographic methods in order to separate and properly identify isomers. Here, a confirmatory method for fentanyl and its analogues in human blood is demonstrated. Due to the inclusion of isomers that are challenging to monitor based on similar fragmentation patterns, the separation of these isomers within the panel was key in method development, in order to accurately identify all fentanyl analogues. The development of an optimized chromatographic method enabled baseline resolution of 29 fentanyl analogues in a 17-minute runtime. The column used in conjunction with an optimized mobile phase composition produced the separation that was needed to correctly distinguished all isomers. Using this comprehensive workflow, confident identification of fentanyl, its analogues, and metabolites was achieved in a complex biological matrices such as human whole blood. Quantification of the 29 fentanyl analogues resulted in LODs in the sub ng/mL range while maintaining linearity and precision for all compounds across the calibration range. These results demonstrate that the combination of mass spectrometry and highly specific chromatographic methods allows accurate quantification of these new substances while offering confident drug identification at low concentration levels. Presenter: Pierre Negri, SCIEX, Redwood City, CA Regional Environmental Health Meeting – Monterey: A Template for Coordinating Water Emergency Response A. Babatola 1 , D. Ferguson 2 ; 1 City of Santa Cruz (CA) Environmental Laboratory, 2 County of Monterey (CA) Public Health Laboratory The importance of streamlining reporting and accessing resource for managing water contamination emergencies is often demonstrated after the experience of the crises. The consequences of unclear protocols for resource allocation and coordination are preventable by advanced planning and training. The Central Coast Region of California includes private and public water utilities, special districts, and federal facilities including military installations whose roles and responsibilities may be directed by multiple jurisdictions including local and state environmental and public health departments. Response coordination may be complex, given the type and magnitude of contamination. Further, many agencies have overlapping responsibilities; thus, water resource managers may have unclear protocols for communicating contamination emergencies and coordinating laboratory support. Therefore, this area provides favorable conditions to develop and test a template, i.e., quick reference guidance manual for efficient and effective coordination. Representatives of state and local agencies including five counties have committed to this regional environmental health meeting with the purpose of creating a streamlined and effective template for the region and as a template for others. This poster presents excerpts from the meeting including flow charts, checklists, and brief session summaries from the meeting in Salinas on April 30, 2019. Presenter: Akin Babatola, City of Santa Cruz (CA) Environmental Laboratory, Santa Cruz, CA, Summer 2019 LAB MATTERS 49