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Toxicology Section - 2015 Toxicology Section - 2015 (GHB) in Human Hair Samples Via an Automated Bead Mill as a Precursor to High Resolution-Gas Chromatography/Time-of-Flight (GC/TOF) and 2D Gas Chromatography/Time-of-Flight (GCxGC/TOF) Brittany M. Watt, BA*, 651 Brooke Road, Apt D44, Glenside, PA 19038; David Alonso, PhD, LECO Corporation, 1850 Hilltop Road, St. Joseph, MI 49085; Joe Binkley, PhD, LECO Corporation, 1850 Hilltop Road, St Joseph, MI 49085; Jeff Patrick, PhD, LECO Corporation, 1850 Hilltop Road, St Joseph, MI 49085; Frank Kero, PhD, Biotage, 10430 Harris Oaks Boulevard, Ste C, Charlotte, NC 28269; Victor Vandell, PhD, Biotage, 10430 Harris Oaks Boulevard, Charlotte, NC 28269; Elena Gairloch, BS, Biotage, 10430 Harris Oaks Boulevard, Charlotte, NC 28269; M. Brad Nolt, MS, Biotage, 10430 Harris Oaks Boulevard, Ste C, Charlotte, NC 28269; Tom Enzweiler, BS, Biotage, 10430 Harris Oaks Boulevard, Charlotte, NC 28269; Rhys Jones, PhD, Biotage GB Limited, Dyffryn Business Park, Ystrad Mynach, Cardiff CF82 7TS, UNITED KINGDOM; Lee Williams, PhD, Biotage GB Limited, Dyffryn Business Park, Ystrad Mynach, Cardiff CF82 7TS, UNITED KINGDOM; and Karen S. Scott, PhD, Arcadia University, 450 S Easton Road, Glenside, PA 19038 After attending this presentation, attendees will be able to describe the usefulness of improved automated instrumentation for the of forensic samples. This creates a challenge when testing hair samples due to interference from other endogenous compounds. One approach to cleaning up the samples is the use of mass spectrometric deconvolution software and multidimensional GC. In addition, it is critical to extract as much of a drug from the hair matrix as possible so this study has investigated the use of a Biotage® Bead Ruptor This presentation will impact the forensic science community by increasing awareness of alternative instrumental techniques that Techniques to interrogate hair samples have proven valuable in detecting human host exposure to drugs of abuse over a long using the laboratory’s standard preparation procedure (cutting the hair into 1mm-2mm segments using scissors) versus automation in the bead mill at the Center for Forensic Science Research and Education in Willow Grove, PA, using a Biotage® Bead Ruptor 24. The interferences. GHB, Bead Ruptor, GCxGC/TOF Copyright 2015 by the AAFS. Unless stated otherwise, noncommercial photocopying of editorial published in this periodical is permitted by AAFS. Permission to reprint, publish, or otherwise reproduce such material in any form other than photocopying must be obtained by AAFS. 1058 * Presenting Author Toxicology Section - 2015 K2 A Novel Extraction Methodology for the Analysis of Lorazepam and Oxazepam Glucuronide Hydrolysis in Meconium Kaylee R. McDonald, BS*, 631 Sumter Street, Columbia, SC 29208; William E. Brewer, PhD, University of South Carolina, Dept of Chem & Biochem, 631 Sumter Street, Columbia, SC 29208; and Stephen L. Morgan, PhD, University of South Carolina, Dept of Chemistry & Biochemistry, 631 Sumter Street, Columbia, SC 29208 in situ This presentation will impact the forensic science community by illustrating how the proposed analytical methodology provides practical advantages over existing methods in terms of rapid sample clean-up and the removal of biological matrix effects that could for cases of suspected drug abuse. 1 2 the use of meconium as a biological matrix to monitor drug use by liquid chromatography/mass spectrometry is problematic because of the potential for large matrix effects.1,3 Reducing matrix effects requires extracting target analytes from the endogenous biological matrix effects and sample preparation time by a hydrolysis step in situ was vortexed and centrifuged. The supernatant was removed and placed into a clean sample vial (~950ul solution). The solution was transfer of the analytes into the cleaner acetonitrile supernatant. The top acetonitrile layer (~500-600ul) was then transferred to a vial suitable for solvent evaporation. This step separates the acetonitrile and water layers and facilitates removal of the analytes in the acetonitrile supernatant. After evaporation in a fresh vial, the residue was reconstituted in 100mL of 10:90 methanol:water. All analyses were performed using a triple quadrupole system with an Agilent® 2.7µm). Sample injections of 20µL were made using an injection valve incorporated on an autosampler. The mobile phase used 0.1% formic acid in water (A) and 0.1% formic acid in acetonitrile (B). The initial gradient was 70% A for 0.25min, which ramped to 5% A glucuronides. Post-hydrolysis, a decrease of the glucuronides and concurrent increase in the parent compounds demonstrated that the method was viable. To test the validity of this method further, a blind study was performed with a collaborative laboratory including 35 92%. In conclusion, the combination of fast hydrolysis, coupled with a simple clean-up scheme, offers an effective analytical approach Copyright 2015 by the AAFS. Unless stated otherwise, noncommercial photocopying of editorial published in this periodical is permitted by AAFS. Permission to reprint, publish, or otherwise reproduce such material in any form other than photocopying must be obtained by AAFS. 1059 * Presenting Author Toxicology Section - 2015 References: 1. Toxicology, 2008, 32, 492-498. 2. 3. Fetal and Neonatal Edition, 2006, 91, F291-F292. Meconium, Benzodiazepines, Chromatography Copyright 2015 by the AAFS. Unless stated otherwise, noncommercial photocopying of editorial published in this periodical is permitted by AAFS. Permission to reprint, publish, or otherwise reproduce such material in any form other than photocopying must be obtained by AAFS. 1060 * Presenting Author Toxicology Section - 2015 K3 Analysis of Promethazine, Chlorpromazine, and Selected Metabolites in Decomposed Skeletal Tissues by Microwave-Assisted Extraction/Microplate Solid Phase Extraction/Ultra High-Performance Liquid Chromatography (MAE/ MPSPE/UHPLC) Courtney A. Campbell, BS*, Laurentian University, 935 Ramsey Lake Road, Sudbury, ON P3E 2C6, CANADA; James Watterson, PhD, Laurentian University, 935 Ramsey Lake Road, Sudbury, ON P3E 2C6, CANADA; and Caroline C. Betit, MSc, 935 Ramsey Lake Road, Sudbury, ON P3E 2C6, CANADA After attending this presentation, attendees will understand how to develop a microwave-assisted extraction methodology using bone tissue. An example of this methodology’s practical application using vertebral bone will be presented. 2 ™ 6 microwave oven for a total of 60min, with extraction solvent recovered and replaced with fresh solvent at 15min and 30min. All solvent extracts were recovered, evaporated to then underwent protein precipitation by adding 1mL of PBS along with 3mL of acetonitrile-methanol (1:1) followed by storage at -20oC plates. Wells were conditioned by sequential addition of 3mL methanol, water, and PBS. Samples were loaded by gravity. Wells were 4 were evaporated to dryness and reconstituted in 500µL of mobile phase A (0.1 % formic acid in 90:10 water:acetonitrile). Samples were used was a Raptor™ biphenyl column (150mm x 2.1mm, 1.7µm) with a column temperature set to 50oC. The mobile phase gradient began with 95:5 A:B (B: 0.1%formic acid in 90:10 acetonitrile:water) held for 1min, then increased to 70:30 A:B over 4min, held for sulphoxide metabolites and 250nm for the remaining analytes. was linear from 10ng/mL to 5,000ng/mL (R2 for each analyte. The majority of analytes were recovered after 30min extraction interval. Analytes were stable under the microwave extraction for at least 60min. Promethazine, Chlorpromazine, Bone Copyright 2015 by the AAFS. Unless stated otherwise, noncommercial photocopying of editorial published in this periodical is permitted by AAFS. Permission to reprint, publish, or otherwise reproduce such material in any form other than photocopying must be obtained by AAFS. 1061 * Presenting Author Toxicology Section - 2015 K4 Detection of Trace Buprenorphine and Norbuprenorphine in Human Hair Using Enzyme-Linked Immuno-Sorbent Assay (ELISA) Irene Shu*, 1700 S Mt Prospect Road, Des Plaines, IL 60018; Valencia Sagnia, BS, 1700 S Mount Prospect Road, Des Plaines, IL 60018; and Joseph Jones, MS, 1700 S Mount Prospect Road, Des Plaines, IL 60018 After attending this presentation, attendees will be able to develop, validate, and implement an ELISA method in their forensic toxicology laboratories for detecting buprenorphine and its metabolite, norbuprenorphine, in human hair. This presentation will impact the forensic science community by introducing a sensitive, robust, and short turn-around-time method to detect both the parent drug and the metabolite to support surveillance of compliance with opioid dependence treatments. months. implemented to quantitate both BUP and norBUP in human hair with a Lower Limit Of Quantitation (LLOQ) of 8pg/mg for each analyte. Only ~30% of the hair samples in the laboratory reported out with quantitated results had BUP/norBUP ratio greater than 1.0 (BUP 8 — 1,517pg/mg, norBUP none-detectable — 1,295pg/mg), and the rest had norBUP as the predominant analyte, including those in which only norBUP was quantitated (BUP none-detectable — 775pg/mg, norBUP 19 — 2,192pg/mg). metabolite at the desired analytical sensitivity. and then sonicated with heat in 1.5mL methanol for two hours. The methanol mixture was centrifuged and 1.0mL of the resulting
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