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
[email protected]
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
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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
[email protected]
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, [email protected]
Summer 2019 LAB MATTERS
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