Validation of an ultra performance liquid chromatography tandem mass spectrometry (UPLC™/MS/MS) method for forensic toxicological analysis : Confirmation and quantitation of lysergic acid diethylamide (LSD) and its congeners in forensic samples
| dc.contributor.advisor | McKay, Gordon | en_US |
| dc.contributor.advisor | Hudson, Jeff J. | en_US |
| dc.contributor.committeeMember | Blakley, Barry R. | en_US |
| dc.contributor.committeeMember | Ross, Andrew R. S. | en_US |
| dc.creator | Chung, Angela | en_US |
| dc.date.accessioned | 2006-04-20T10:28:06Z | en_US |
| dc.date.accessioned | 2013-01-04T04:29:29Z | |
| dc.date.available | 2007-04-20T08:00:00Z | en_US |
| dc.date.available | 2013-01-04T04:29:29Z | |
| dc.date.created | 2006-04 | en_US |
| dc.date.issued | 2006-04-18 | en_US |
| dc.date.submitted | April 2006 | en_US |
| dc.description.abstract | The Royal Canadian Mounted Police (RCMP) Forensic Laboratory Services (FLS) needed a method to confirm positive lysergic acid diethylamide (LSD) immunoassay screening results. As a result, an ultra performance liquid chromatography tandem mass spectrometry (UPLC™/MS/MS) method was validated for the confirmation and quantitation of LSD, iso-LSD, N-demethyl-LSD (nor-LSD), and 2-oxo-3-hydroxy-LSD (O-H-LSD). The method was validated in urine and whole blood, where linearity, accuracy, precision, sensitivity, stability, selectivity, recovery, matrix effects, and reproducibility were evaluated. The method involved a liquid-liquid extraction (LLE) of the analytes and the deuterated internal standard from 1 mL of urine or whole blood with dichloromethane:isopropyl alcohol after being basified. The average recovery for all analytes was ≥ 62%, and the matrix effect was found to be insignificant. MS/MS analysis was conducted with a triple quadrupole mass spectrometer by positive electrospray ionization (ESI+) in multiple reaction monitoring (MRM) mode. The lowest limit of quantitation (LLOQ) was 20 pg/mL for LSD and iso-LSD, and 50 pg/mL for nor-LSD and O-H-LSD. The method was linear, accurate, precise, selective, and reproducible from 20 to 2000 pg/mL for LSD and iso-LSD, and from 50 to 2000 pg/mL for nor-LSD and O-H-LSD with an r² ≥ 0.99. The refrigerated and frozen long term stability was investigated for 90 days. LSD was stable at all temperatures for 90 days. Iso-LSD in blood was also stable at all temperatures for 90 days, but iso-LSD in urine showed an initial decrease followed by a gradual increase back to day 0 concentrations. Nor-LSD was stable at all temperatures up to day 14, with >43% decrease by day 30, with no additional decrease for the next 60 days. O-H-LSD in urine was stable at all temperatures for 90 days, but by day 90 O-H-LSD in whole blood stored refrigerated decreased in concentration by >37%. Additionally, a case sample that was stored at -50°C for ten years was found to still contain measurable amounts of each compound. The method was applied to blind samples and a case that screened positive with immunoassay. Retention time, relative retention time, and ion ratios were used as identification parameters and found to correctly identify the analytes 100% of the time with no false positives. The case sample showed that the concentration of O-H-LSD was 4 times greater than LSD in urine. Furthermore, both the detection of O-H-LSD in a blood case sample, and LSD in a vitreous humor case sample were the first to be documented. | en_US |
| dc.identifier.uri | http://hdl.handle.net/10388/etd-04202006-102806 | en_US |
| dc.language.iso | en_US | en_US |
| dc.subject | nor-LSD | en_US |
| dc.subject | iso-LSD | en_US |
| dc.subject | O-H-LSD | en_US |
| dc.subject | Whole blood | en_US |
| dc.subject | Urine | en_US |
| dc.title | Validation of an ultra performance liquid chromatography tandem mass spectrometry (UPLC™/MS/MS) method for forensic toxicological analysis : Confirmation and quantitation of lysergic acid diethylamide (LSD) and its congeners in forensic samples | en_US |
| dc.type.genre | Thesis | en_US |
| dc.type.material | text | en_US |
| thesis.degree.department | Toxicology | en_US |
| thesis.degree.discipline | Toxicology | en_US |
| thesis.degree.grantor | University of Saskatchewan | en_US |
| thesis.degree.level | Masters | en_US |
| thesis.degree.name | Master of Science (M.Sc.) | en_US |