Validation of blood and liver oxymorphone analysis using LC-MS-MS: concentrations in 30 fatal overdoses.

A sensitive liquid chromatography-tandem mass spectrometry method was developed and validated for the quantitation of oxymorphone (OM) in human whole blood and liver. Sample preparation was done by solid-phase extraction, using deuterated OM as the internal standard. Separation was achieved using a Waters Aquity UPLC HSS T3 column. Analysis utilized positive electrospray ionization and multiple reaction monitoring. As part of the validation, studies were conducted to determine potential interference, selectivity, ion suppression/enhancement and carryover. Calibration model, limit of detection (LOD), lower limit of quantitation (LLOQ), precision and accuracy were also established. The linear range of the method was 2-500 ng/mL in blood and 5-500 ng/g in the liver. The LOD and LLOQ were 2 ng/mL for blood and 5 ng/g for the liver. Blood and/or liver specimens from 30 cases were analyzed. OM concentrations ranged from 23 to 554 ng/mL ( , n = 26) in blood and 48 to 1740 ng/g ( , n = 30) in the liver.

Characteristics of alprazolam-related deaths compiled by a centralized state medical examiner.

BACKGROUND AND OBJECTIVE: Unintentional drug poisoning deaths represent a major health concern, particularly in rural areas. Although alprazolam is frequently detected in drug-related deaths, characterization of its involvement is limited. Our objective was to compare the characteristics of alprazolam-related deaths with nonalprazolam deaths in a predominantly rural state. METHODS: A comprehensive forensic drug database (FDD) was developed in 2005 to compile demographic, toxicology, and co-morbidity information from all West Virginia (WV) drug-related deaths. All FDD data from 2005 to mid-November 2007 were analyzed. RESULTS: Alprazolam contributed to 204 (17.0%) of the 1,199 drug-related deaths and was identified in 7.2% of the 363 deaths occurring during 2005 and in 27.5% of the 422 deaths entered in the database during 2007. At least one other drug, predominantly an opioid, was identified in 97.5% of the alprazolam cases, with concurrent benzodiazepines also found. Compared to nonalprazolam deaths, alprazolam decedents were significantly more likely to be obese and to have preexisting cardiovascular disease, but were less likely to have documented substance abuse. An alprazolam prescription existed in 52.5% of the alprazolam deaths, with 77.6% having a prescription for all drugs identified. CONCLUSIONS: Alprazolam was a contributing cause of death in a substantial and increasing number of drug-related deaths. Prescriptions for alprazolam and the other drugs detected were often present in these cases. SCIENTIFIC SIGNIFICANCE: Controlled substance monitoring programs should be routinely used as one mechanism to help prevent potential drug misuse/abuse. Our findings provide a baseline for ongoing alprazolam-related death surveillance.

Comparison of oxycodone in vitreous humor and blood using EMIT screening and gas chromatographic-mass spectrometric quantitation.

Vitreous humor may serve as a useful alternative specimen for oxycodone analysis in death investigations where blood samples are not available or are of poor quality or limited quantity. The purpose of this study was to investigate the relationship between immunoassay results and gas chromatography-mass spectrometry (GC-MS) quantitation of oxycodone in postmortem vitreous humor and blood. When used with vitreous humor calibrators, the Microgenics DRI Oxycodone (EMIT) Assay was found to be linear from 25 to 500 ng/mL with an limit of detection of 25 ng/mL. Vitreous humor and postmortem blood precipitate immunoassay responses in 57 oxycodone-positive cases were found to be correlated (r(2) = 0.69, p < 0.01). Confirmation and quantitation of oxycodone in vitreous humor by GC-MS was linear from 50 to 1000 ng/mL with a limit of detection of 10 ng/mL and a limit of quantitation of 50 ng/mL. In 30 cases, oxycodone vitreous humor concentrations ranged from less than 50 to 945 ng/mL, and blood concentrations ranged from 103 to 768 ng/mL. The average vitreous humor/blood ratio was 1.16 and ranged from 0.12 to 3.26. Disparities between vitreous fluid and blood oxycodone concentrations were seen in a few cases.

Cocaine detection in postmortem samples following therapeutic administration.

Cocaine is one of the most widely abused drugs and one that is frequently encountered in forensic toxicology laboratories. Most often, the detection of cocaine would lead toxicologists and forensic pathologists to believe that the drug was used illicitly; however, cocaine is an effective local anesthetic and vasoconstrictor and is used clinically in surgeries of the eye, ear, nose, and throat. Therefore, it is important to note that the presence of cocaine and its metabolites in forensic samples cannot always be attributed to abuse and that a thorough investigation and review of medical records is warranted before an informed conclusion can be made. In this case report, a 54-year-old male died three days after an altercation in which he suffered multiple injuries. In addition to natural disease and injuries documented at autopsy, cocaine and its metabolites were detected in the decedent's urine, and a review of surgical records showed that earlier on the day of death, he was administered cocaine clinically during a procedure to repair nasal bone fractures. If not for this comprehensive investigation and review of surgical records, the assumption of cocaine abuse might have otherwise been made and the cause and manner of death incorrectly established.

Direct injection mass spectrometric confirmation of multiple drugs in overdose cases from postmortem blood using electrospray ionization-tandem mass spectrometry and MS(3).

Five cases of confirmed multiple-drug overdose were previously screened and quantified by the West Virginia Office of the Chief Medical Examiner; 26 different drugs and metabolites were identified and quantified in blood at > or = 10 ng/mL. In this study, whole blood from those five case samples was analyzed by a direct injection multi-stage mass spectrometric (MSn) method to confirm the identity of 26 analytes at or above 10 ng/mL using 16 different deuterium-labeled internal standards. Samples were spiked with internal standards, precipitated with acetonitrile, and centrifuged. Samples were further diluted with either 0.1% formic acid or 0.1% ammonium hydroxide in methanol prior to injection into an electrospray ionization ion trap mass spectrometer (MS). Ions were monitored as MS-MS or MS3 product ions. In all cases, analysis by MS-MS confirmed the presence of the drugs and metabolites when the internal standards were detected. Detection of characteristic MS3 ions was used for further confirmation of the presence of parent drugs in all but three instances. Total analysis time was less than 1 h. Although only useful for qualitative or confirmatory purposes, this direct injection MSn method provides a simple and rapid confirmation of multiple drugs that have been previously identified and quantified by gas chromatographic-MS or liquid chromatographic-MS analytical methods.

Validation of a headspace GC method for the analysis of a pyrolytic product of methamphetamine in urine.

A method has been developed and validated using headspace GC-FID for the identification of 1-phenylpropene in urine. This compound is a pyrolytic product of methamphetamine that has been previously proposed as a marker for smoked methamphetamine. The instrumentation used is the same as employed for blood alcohol determination. The extraction-free procedure is rapid, simple, and quantitative using 2-phenylpropene as the internal standard. The method was validated for linearity over a range of 0.1-20 microg/mL with a limit of detection of 0.05 microg/mL, limit of quantification of 0.1 microg/mL, interday accuracy within 3.2 to -5.3%, intraday accuracy better than 7.5%, interday precision of 7.5 to 10.7%, intraday precision of 2 to 8.6%, and recovery above 80%. For the robustness determination in urine, the accuracy of four different sources of urine at the mid control level (1 microg/mL) ranged from 1.6 to 19% error. The % relative standard deviation of the different urine sources ranged from 3.1 to 11%. Urine samples from nine methamphetamine-positive cases investigated by the Office of the Chief Medical Examiner of West Virginia were included in the study. 1-Phenylpropene was found in two methamphetamine-positive cases (0.25 and 0.44 microg/mL).

Potential marker for smoked methamphetamine hydrochloride based on a gas chromatography-mass spectrometry quantification method for trans-phenylpropene.

Residue from smoked methamphetamine hydrochloride contains pyrolytic products that are detectable by gas chromatography-mass spectrometry (GC-MS). A validated GC-MS method was developed for the determination of trans-phenylpropene, a pyrolytic product of methamphetamine HCl, in residue of smoked drug as well as in human urine. trans-Phenylpropene and an isomeric internal standard, 2-phenylpropene, were extracted from urine using n-hexane. The method was validated for linearity over a range of 0.1-10 microg/mL with a limit of detection of 0.05 microg/mL, limit of quantification of 0.1 microg/mL, interday accuracy within 10.5%, intraday accuracy better than 3.7%, interday precision of 15.4%, intraday precision of 14.4%, and recovery of 89.1%. The method was applied to the detection of trans-phenylpropene found in the residue of methamphetamine HCl heated beyond its melting temperature on aluminum foil under simulated smoking conditions. The method is applicable to the detection of trans-phenylpropene in urine as a potential marker for smoked methamphetamine HCl abuse.