Topic: Tox / TDM / Endocrine
Podium Presentation in Room 1 on Wednesday at 14:40 (Chair: Hema Ketha / Melissa Hoffman)
Authors: Yu Zhang, and Kara L. Lynch
INTRODUCTION: The overdose of illicitly manufactured synthetic opioids including fentanyl and analogues represents a significant escalation in the US. In clinical practice, the quick evolution of synthetic fentanyl analogues brings challenges as they could be undetectable by either immunoassay or LC-MS/MS methods with a limited library. In some cases, the symptoms are consistent with opioid overdose and respond to naloxone treatment, however no drug culprits are identified with routine testing. To address these issues, the Centers for Disease Control and Prevention has developed Traceable Opioid Material Kits to support laboratory detection of current and emerging opioids. We develop and validate a high-resolution mass spectral library for fentanyl analogue screening (FAS).
OBJECTIVES: The FAS kit with 120 synthetic fentanyl compounds and FAS emergent panel version 1 with 30 synthetic opioid compounds were obtained from Cayman Chemical. We collected a high-resolution mass spectrum for each compound to build a FAS library. Moreover, retention time, limit of detection (LOD), and matrix effects are determined with our standard high-resolution comprehensive drug testing method. This library can be used to retrospectively and prospectively analyze cases of suspected opioid overdoses.
METHODS: Briefly, chromatographic separation was performed using a Kinetex C18-column with a 10-minute gradient from 2%-100% organic; and data was collected on a SCIEX TripleTOF®5600 operating in positive-ion mode using a TOF-MS survey scan with dedicated product ion scan method for a library spectra or IDA-triggered collection of product ion spectra for LOD and matrix effects. Based on the high-resolution method, mass spectrum and retention time of each compound have been acquired. LOD was evaluated by duplicated injections of different concentrations of compound standards (1, 2.5, 5, 10, and 25 ng/ml) into drug-free urine and serum samples. A compound was called positive if (a) the combined score >70% and (b) signal-to-noise ratio >20:1. LOD was defined as the lowest concentration for which a compound could be identified as positive in duplicated injections. Matrix effects are under determination by calculating the mean signal intensity in urine minus that in water divided by the mean signal intensity in water (N=3, tested in triplicate at 3 concentrations).
RESULTS: A mass spectrum for each analyte in FAS kits has been collected and added to our in-house screening library. Their retention times range from 3.5-7.8 minutes, and LOD in urine mainly ranges from 1-10 ng/ml. LOD in serum samples, matrix effects, and case analysis is currently under investigation, and a full list will be updated soon.
DISCUSSION: The FAS library includes an expanded list of fentanyl analogues and their metabolites. It will be used to analyze suspected fentanyl overdose cases retrospectively and retrospectively to identify potential fentanyl intoxication which was undetectable before.
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