Jessica Boyd (Presenter)
Calgary Laboratory Services/University of Calgary
Bio: Jessica Boyd is a clinical biochemist and co-director of Analytical Toxicology at Calgary Laboratory Services (CLS). She received her Ph.D. in Analytical and Environmental Toxicology at the University of Alberta in 2012. She was accepted into the Clinical Biochemistry Fellowship program at the University of Calgary and Calgary Laboratory Services (CLS) and completed her training in 2015.
Authorship: Jessica Boyd (1), Valerie Simon (1), Joseph Bacani (1), S.M. Hossein Sadrzadeh (1)
(1) Department of Pathology and Laboratory Medicine, University of Calgary and Calgary Laboratory Services
| Short Abstract Dried urine spots are not commonly used in the clinical toxicology laboratory despite being a convenient and economical alternative to traditional liquid specimens. Here we describe the development of an LC-MS/MS assay for detection of 25 opioids (including pain management drugs, drugs of abuse and designer drugs) from DUS. |
Long Abstract
Introduction: Dried specimens offer a convenient and economical approach for collection, storage and transport of clinical specimens. Dried blood spots have been used as the specimen of choice in clinical laboratories for years for applications such as new born screening. However, other dried specimen types, such as dried urine spots (DUS) have not been established for routine use. Analysis of DUS for drugs of abuse testing has several advantages over use of a liquid specimen in that 1). DUS can be collected and spotted anywhere, removing the need for patients to travel to a laboratory collection centre; and 2). DUS cards can be sent to the lab through the regular mail, making it an attractive option for patients who live far from the lab. Here we describe the development of a LC-MS/MS method for a 25 opioid panel using DUS as the specimen of choice.
Methods: Twenty-five opioids and metabolites including those used for pain management, drugs of abuse and designer drugs were included in the panel (morphine, morphine-3β-glucuronide, hydromorphone, oxymorphone, codeine, norcodeine, hydrocodone, oxycodone, heroin, 6- monoacetylmorphine (6-MAM), fentanyl, norfentanyl, naloxone, tramadol, desmethyltramadol, meperidine, normeperidine, buprenorphine, norbuprenorphine, methadone, EDDP, acetylfentanyl, acetyl norfentanyl, desomorphine, AH-7921). Briefly, DUS extractions were performed by spotting 15 µL of de-identified urine samples containing the above drugs onto Whatman 903 Protein Saver Cards. Drugs were extracted using a mixture of methanol, acetonitrile and water (4:4:1). Extracts were evaporated and reconstituted in 5% acetonitrile before injection onto the LC-MS/MS. Separation was performed using an Agilent 1290 HPLC with a Restek Ultra Biphenyl column (100mm x 2.1 mm x 5µm). Flow rate was 0.5 mL/min and the injection volume was 10 µL.
Sample analysis was performed with an Agilent 6460 triple quadrupole mass spectrometer in positive electrospray ionization mode using multiple reaction monitoring (MRM) mode. Two MRM transitions were selected per analyte. Data analysis was performed using Agilent MassHunter Quantitative analysis software
Results: DUS extraction recoveries for the 25 opioids ranged between 78-115%. Within run precision ranged from 3-8% and between day precision ranged from 5-15%. The developed method compared well with both our in house GC-MS comprehensive screen (qualitative) and LC-MS/MS opioid confirmation assay (quantitative). In some cases, opioids were reported positive using the GC-MS but were below the cutoff chosen for the DUS-LC-MS/MS method, suggesting that lower cutoffs should be evaluated in future. Stability studies suggest that the DUS are stable at room temperature for 1 month.
Conclusions: We developed an LC-MS/MS method to simultaneously detect 25 opioids from DUS which offers a convenient alternative for urine toxicology testing.
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