11-Nor-9-Carboxy Tetrahydrocannabinol Distribution in Fluid from the Chest Cavity in Cannabis-Related Post-Mortem Cases.

In this study, the presence of 11-nor-Δ9-carboxy tetrahydrocannabinol (THC-COOH) in postmortem fluid obtained from the chest cavity (FCC) of postmortem cases collected from drug-related fatalities or criminal-related deaths in Jeddah, Saudi Arabia, was investigated to evaluate its suitability for use as a complementary specimen to blood and biological specimens in cases where no bodily fluids are available or suitable for analysis. The relationships between THC-COOH concentrations in the FCC samples and age, body mass index (BMI), polydrug intoxication, manner, and cause of death were investigated. METHODS: Fifteen postmortem cases of FCC were analyzed using fully validated liquid chromatography-positive-electrospray ionization tandem mass spectrometry (LC-MS/MS). RESULTS: FCC samples were collected from 15 postmortem cases; only THC-COOH tested positive, with a median concentration of 480 ng/mL (range = 80-3010 ng/mL). THC-COOH in FCC were higher than THC-COOH in all tested specimens with exception to bile, the median ratio FCC/blood with sodium fluoride, FCC/urine, FCC/gastric content, FCC/bile, FCC/liver, FCC/kidney, FCC/brain, FCC/stomach wall, FCC/lung, and FCC/intestine tissue were 48, 2, 0.2, 6, 4, 6, 102, 11, 5 and 10-fold, respectively. CONCLUSION: This is the first postmortem report of THC-COOH in the FCC using cannabinoid-related analysis. The FCC samples were liquid, easy to manipulate, and extracted using the same procedure as the blood samples. The source of THC-COOH detected in FCC could be derived from the surrounding organs due to postmortem redistribution or contamination due to postmortem changes after death. THC-COOH, which is stored in adipose tissues, could be a major source of THC-COOH found in the FCC.

Metabolomics Analysis as a Tool to Measure Cobalt Neurotoxicity: An In Vitro Validation.

In this study, cobalt neurotoxicity was investigated in human astrocytoma and neuroblastoma (SH-SY5Y) cells using proliferation assays coupled with LC-MS-based metabolomics and transcriptomics techniques. Cells were treated with a range of cobalt concentrations between 0 and 200 µM. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay revealed cobalt cytotoxicity and decreased cell metabolism in a dose and time-dependent manner was observed by metabolomics analysis, in both cell lines. Metabolomic analysis also revealed several altered metabolites particularly those related to DNA deamination and methylation pathways. One of the increased metabolites was uracil which can be generated from DNA deamination or fragmentation of RNA. To investigate the origin of uracil, genomic DNA was isolated and analyzed by LC-MS. Interestingly, the source of uracil, which is uridine, increased significantly in the DNA of both cell lines. Additionally, the results of the qRT-PCR showed an increase in the expression of five genes Mlh1, Sirt2, MeCP2, UNG, and TDG in both cell lines. These genes are related to DNA strand breakage, hypoxia, methylation, and base excision repair. Overall, metabolomic analysis helped reveal the changes induced by cobalt in human neuronal-derived cell lines. These findings could unravel the effect of cobalt on the human brain.

Heroin-Related Fatalities in Jeddah, Saudi Arabia, between 2008 and 2018.

To date, epidemiological studies have not evaluated heroin-related deaths in the Middle East and North African regions, especially Saudi Arabia. All heroin-related postmortem cases reported at the Jeddah Poison Control Center (JPCC) over a 10-year period (21 January 2008 to 31 July 2018) were reviewed. In addition, liquid chromatography electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) was utilized to determine the 6-monoacetylmorphine (6-MAM), 6-acetylcodeine (6-AC), morphine (MOR), and codeine contents in unhydrolyzed postmortem specimens. Ninety-seven heroin-related deaths were assessed in this study, and they represented 2% of the total postmortem cases at the JPCC (median age, 38; 98% male). In the blood, urine, vitreous humor, and bile samples, the median morphine concentrations were 280 ng/mL, 1400 ng/mL, 90 ng/mL, and 2200 ng/mL, respectively; 6-MAM was detected in 60%, 100%, 99%, and 59% of the samples, respectively; and 6-AC was detected in 24%, 68%, 50%, and 30% of the samples, respectively. The highest number of deaths (33% of total cases) was observed in the 21-30 age group. In addition, 61% of cases were classified as "rapid deaths," while 24% were classified as "delayed deaths." The majority (76%) of deaths were accidental; 7% were from suicide; 5% were from homicide; and 11% were undetermined. This is the first epidemiological study to investigate heroin-related fatalities in Saudi Arabia and the Middle East and North African region. The rate of heroin-related deaths in Jeddah remained stable but increased slightly at the end of the study period. Most patients were heroin-dependent abusers and from the middle-aged group. The availability of urine, vitreous humor, and bile specimens provided valuable information regarding the opioids that were administered and the survival time following heroin injection.

Ethyl glucuronide and ethyl sulfate: a review of their roles in forensic toxicology analysis of alcohol postmortem.

PURPOSE: This review presents the current methods used for determining ethyl glucuronide (EtG) and ethyl sulfate (EtS) concentrations in postmortem specimens, including sample preparation, analysis and the role of EtG and EtS in the postmortem assessment of the extent of alcohol abuse. METHODS: Papers pertaining to postmortem investigation were collected from scientific databases and reviewed. The papers were published between January 2006 and October 2020. RESULTS: Most of the analyses involved postmortem blood and urine samples, with a few reports using other bodily specimens and tissues. The method validation was not conducted for all applications. These reports were mostly intended to present data rather than interpret them, and the lack of effort in relating these ethanol biomarkers with the cause of death and/or determination of the time of deaths due to ethanol intoxication might decrease the applicability of these makers after a promising start between 2006 and 2010. Nevertheless, by the beginning of 2020, papers investigating ethanol biomarkers were still increasing. A considerable number of methods used liquid chromatography coupled with mass spectrometry (LC-MS) techniques that require less sample preparation (e.g., protein precipitation extraction, dilution, filtration, and centrifugation). Although solid-phase extraction can be applied, only three applications were reported. CONCLUSIONS: Matrix effects can be a substantial challenge in analytical methods based on LC-MS because they directly affect the ionization of analytes. However, these problems can be avoided due to the high cutoff values used to identify positive results for these ethanol biomarkers, which are often above 0.1-1 mg/L, and using internal standards. Research on using tissue specimens is recommended as most of the reported results on this type of specimen were promising.

Post-Mortem Analysis of Heroin Biomarkers, Morphine and Codeine in Stomach Wall Tissue in Heroin-Related Deaths.

Toxicological analysis of some cases can be complicated by poor sample quality caused by decomposition. Although heroin-related deaths have been researched extensively, the interpretation of toxicology findings in these cases is challenging, especially in instances where blood samples are unavailable. Thus, it is important to develop analytical methods for different sample types. In this study. a method for the quantification of 6-monoacetylmorphine, 6-acetylcodeine, morphine, and codeine in postmortem stomach wall tissue using liquid chromatography coupled with tandem mass spectrometry was developed and validated. All calibration curves prepared with the stomach wall tissue were linear and ranged from 0.5−1000 ng/g with determination coefficients of >0.99 and a lower limit of quantification of 1.0 ng/g. The coefficients of variation for within-run precision and between-run precision were <9%. Matrix effects of stomach wall tissues and their extraction recoveries were investigated and ranged from −19% to +17% and 76% to 80%, respectively. Among the 16 analyzed heroin-related death cases, 6-monoacetylmorphine, 6-acetylcodeine, morphine, and codeine were detected in 75%, 31%, 100%, and 94% of all stomach wall tissues with median concentrations of 90 ng/g, 20 ng/g, 140 ng/g, and 30 ng/g, respectively. This study provides new data on the distribution of 6-monoacetylmorphine, 6-Acetylcodeine, morphine, and codeine in postmortem stomach wall tissue and suggests the usefulness of alternative matrices for investigating heroin-related fatalities when blood samples are unavailable. In addition, the prevalence of 6-monoacetylmorphine in the stomach wall tissue was higher than that in the liver and kidney tissues.

Effects of postmortem interval, putrefaction, diabetes, and location of death on the analysis of ethyl glucuronide and ethyl sulfate as ethanol biomarkers of antemortem alcohol consumption.

This study evaluated the forensic value of ethanol biomarkers ethyl glucuronide (EtG) and ethyl sulfate (EtS) under different conditions, including diabetes mellitus, drug abuse, and advanced decomposition. In addition, we explored whether ethanol, EtG, or EtS formation occurred in patients who died as a result of diabetes mellitus. Fifty-two routine postmortem cases were divided into three groups. Group 1 included only the post-mortem cases in which at least blood samples were available (n=47). Group 2 included all cases with positive BAC (n=28). Group 3 included the cases with negative BAC while information surrounding the cases suspected antemortem alcohol consumption and cases that tested negative for ethanol but positive for EtG and EtS. We analyzed multiple bodily fluid specimens, including the vitreous humor, for ethanol biomarker analysis and accurately identified antemortem ethanol consumption or postmortem ethanol synthesis. We also determined the utility of urine samples for analyzing ethanol and its metabolites in putrefaction cases. If no urine sample was available at autopsy due to urination before death or diabetes-associated glucosuria, vitreous humor samples were an appropriate alternative for ethanol biomarker testing. We observed postmortem ethanol synthesis in diabetic individuals even with a short postmortem interval (PMI), however, glucose did not increase postmortem ethanol production in individuals with diabetes under appropriate preservation. The shorter the PMI, the better the ethanol source can be determined. Postmortem ethanol production occurred in all body fluid specimens analyzed herein, including the vitreous humor. EtG and EtS levels were stable and provided accurate insight into ethanol sources, even in cases of postmortem ethanol production. While the present study focused on the use of vitreous humor for the analysis, it is expected such samples may not be available in cases of advanced decay. In cases where no other bodily fluid specimens are available, solid tissue specimens are highly preferred.

Viridiflorol induces anti-neoplastic effects on breast, lung, and brain cancer cells through apoptosis.

All active natural molecules are not fully exploited as therapeutic agents, causing delays in the advancement of anticancer drug discovery. Viridiflorol is a natural volatile element that may work as anti-cancer compound. We tested the anticancer properties of viridiflorol at different concentrations ranging from 0.03 to 300 µM in vitro on three cancer cells including breast (MCF-7), lung (A549) and brain (Daoy). The cancer cells responses were documented after treatment using MTT and Annexin V assays. Viridiflorol showed cytotoxic effects against all tested cell lines, reducing cell viability in a concentration-dependent manner with variable IC50 values. Daoy and A549 cell lines were more sensitive to viridiflorol when compared with temozolomide and doxorubicin, respectively. Viridiflorol demonstrated the highest anticancer activity against the Daoy cells with an estimated IC50 of 0.1 µM followed by MCF-7 at 10 µM, and A549 at 30 µM. In addition, upon exposure to concentrations ranging from 30 µM to 300 µM of viridiflorol, early and late apoptotic cell death was induced in a concentration dependent manner in Daoy (55.8%-72.1%), MCF-7 (36.2%-72.7%) and A459 (35%-98.9%) cell lines, respectively. In conclusion, viridiflorol demonstrates cytotoxic and apoptotic ability in three different cancer cell lines (brain, breast and lung).

Statistical analysis, source apportionment, and toxicity of particulate- and gaseous-phase PAHs in the urban atmosphere.

Introduction: The concentrations of particulate and gaseous Polycyclic Hydrocarbons Carbon (PAHs) were determined in the urban atmosphere of Delhi in different seasons (winter, summer, and monsoon). Methodology: The samples were collected using instrument air metric (particulate phase) and charcoal tube (gaseous phase) and analyzed through Gas chromatography. The principal component and correlation were used to identify the sources of particulate and gaseous PAHs during different seasons. Results and discussion: The mean concentration of the sum of total PAHs (TPAHs) for particulate and gaseous phases at all the sites were found to be higher in the winter season (165.14 ± 50.44 ng/m3 and 65.73 ± 16.84 ng/m3) than in the summer season (134.08 ± 35.0 ng/m3 and 43.43 ± 9.59 ng/m3), whereas in the monsoon season the concentration was least (68.15 ± 18.25 ng/m3 and 37.63 1 13.62 ng/m3). The principal component analysis (PCA) results revealed that seasonal variations of PAHs accounted for over 86.9%, 84.5%, and 94.5% for the summer, monsoon, and winter seasons, respectively. The strong and positive correlation coefficients were observed between B(ghi)P and DahA (0.922), B(a)P and IcdP (0.857), and B(a)P and DahA (0.821), which indicated the common source emissions of PAHs. In addition to this, the correlation between Nap and Flu, Flu and Flt, B(a)P, and IcdP showed moderate to high correlation ranging from 0.68 to 0.75 for the particulate phase PAHs. The carcinogenic health risk values for gaseous and particulate phase PAHs at all sites were calculated to be 4.53 × 10-6, 2.36 × 10-5 for children, and 1.22 × 10-5, 6.35 × 10-5 for adults, respectively. The carcinogenic health risk for current results was found to be relatively higher than the prescribed standard of the Central Pollution Control Board, India (1.0 × 10-6).

Methamphetamine-related postmortem cases in Jeddah, Saudi Arabia.

A more than 500% increase in the number of deaths involving methamphetamine occurred between 2016 and 2018 in Jeddah, Saudi Arabia. As such, this report employed a validated liquid chromatography tandem mass spectrometry method to quantify methamphetamine and its metabolites in bodily fluids from 47 postmortem cases in which methamphetamine was involved. The mean age of the deceased was 33 years old (median: 30, range: 16-63), and 94% were male. Methamphetamine was co-ingested with another drug in 32 of the cases (68%); however, the deaths were only due to the combined toxicity of methamphetamine and another drug in 15 of the cases (32%). Of note, 13 of these deaths (28% of all deaths) involved heroin. When methamphetamine was the sole cause of death (32% of the studied cases), the median concentrations of methamphetamine and amphetamine were 527 and 128 ng/mL. When methamphetamine was combined toxicity with another drug, the median concentrations of methamphetamine and amphetamine decreased to 161 and 53 ng/mL. When deaths were unrelated to methamphetamine, the median concentrations of methamphetamine and amphetamine were 130 and 44 ng/mL, respectively. The highest median methamphetamine concentration was found in urine (5281 ng/mL), followed by stomach contents (878 ng/mL), bile (762 ng/mL), vitreous humor (3 ng/mL), and blood (208 ng/mL). Almost 40% of the studied cases involved violence, 61% were accidental, 21% were suicides, 17% were homicides, and 2% were natural deaths. Methamphetamine is highly addictive. Increases in deaths have been seen in various countries. More awareness, education and treatment programs are required to reduce the likelihood of addiction, crimes, suicide, and other fatalities resulting from methamphetamine abuse.

Systematic Phytochemical Screening of Different Organs of Calotropis procera and the Ovicidal Effect of Their Extracts to the Foodstuff Pest Cadra cautella.

In developing countries, crop deterioration is mainly caused by inappropriate storage conditions that promote insect infestation. Synthetic pesticides are associated with serious adverse effects on humans and the environment. Thus, finding alternative "green" insecticides is a very pressing need. Calotropis procera (Aiton) Dryand (Apocynaceae) growing in Saudi Arabia was selected for this purpose. LC-MS/MS analysis was applied to investigate the metabolic composition of different C. procera extracts. Particularly, C. procera latex and leaves showed a high presence of cardenolides including calactin, uscharidin, 15ß-hydroxy-calactin, 16ß-hydroxy-calactin, and 12ß-hydroxy-calactin. The ovicidal activity of the extracts from different plant organs (flowers, leaves, branches, roots), and of the latex, against Cadra cautella (Walker) (Lepidoptera, Pyralidae) was assessed. Extracts of C. procera roots displayed the most potent activity with 50% of C. cautella eggs not hatching at 10.000 ppm (1%).

Quantification of cannabinoids in human hair using a modified derivatization procedure and liquid chromatography-tandem mass spectrometry.

The aim of this work was to develop and validate a liquid chromatography-tandem mass spectrometry method for detecting of the main cannabinoids, cannabinol (CBN) and tetrahydrocannabinol (THC) and the primary metabolite 11-nor-9-carboxy-Δ9 -tetrahydrocannabinol (THC-COOH) in hair samples. Extraction of the cannabinoids was carried out by a polymeric strong anion mixed-mode solid-phase extraction cartridge and then employing methanolic HCl followed by 2-fluoro-1-methylpyridinium-p-toluenesulfonate (FMP-TS) as a derivatization procedure of carboxyl and phenolic groups, respectively, offering enhanced sensitivity for the detection of THC-COOH in hair matrices. Formation of a methyl ester increased its lipophilicity and removed the negative charge on the carboxyl group. Calibration curves were prepared over the range of 0.02-4 pg/mg of hair for THC and CBN and 0.2-12 pg/mg of hair for THC-COOH. The extraction recovery was between 81% and 105% for all compounds. The limit of detection (LOD) and limit of quantification (LOQ) were 2 and 20 pg/mg, respectively, for both CBN and THC and 0.1 and 0.2 pg/mg, respectively, for THC-COOH, which met the society of hair testing recommendation. Intra-assay and interassay precision were always lower than 4% and 11%, respectively for these cannabinoids, whereas intra-assay and interassay bias were between +14% and -18% and +15% and -12%, respectively. Twenty-seven hair specimens from cannabis users were investigated. The concentrations of CBN, THC and THC-COOH gave ranges of (0.022-2.562 ng/mg), (0.049-0.431 ng/mg) and (0.222-4.867 pg/mg), respectively. This new method of derivatization improves the LOD to ensure detection of the metabolite.

The role of ethanol in fatalities in Jeddah, Saudi Arabia.

In Saudi Arabia, alcohol consumption is prohibited by law, but interpreting postmortem ethanol can be complicated by its postmortem production. This study developed and validated a method using headspace gas chromatography with flame ionization detection and liquid chromatography tandem mass spectroscopy to detect ethanol and its polar metabolites (ethyl glucuronide [EtG] and ethyl sulfate [EtS]) in postmortem blood and urine specimens, respectively. All calibration curves were linear with coefficients of determination greater than 0.999. The limits of detection ranged 4.5-5.0mg/dL for ethanol and 0.05-0.06mg/L for EtG and EtS. The limits of quantification were 10.0mg/dL for ethanol and 0.075mg/L for EtG and EtS. Within-run precision was less than 11% for all analytes of interest. Matrix effects for EtG and EtS ranged 3-47%. After excluding matrix effects, analytical recoveries ranged 72-100%. This validated method was then used for routine postmortem forensic toxicology analyses in 592 routine postmortem cases to distinguish between antemortem ethanol consumption and its postmortem microbial formation. Among them, 98 blood samples (17%) were positive for ethanol or its polar metabolites. Thirty-two of these cases (33%) were positive for EtG and EtS and therefore due to antemortem ethanol consumption. The remaining 66 (67%) cases were negative for both EtG and EtS and therefore due to postmortem ethanol synthesis. Because this is the first study to report the problem of alcohol consumption in Saudi Arabia, further studies are essential for validating these findings.

Method for Postmortem Tissue Quantification of Δ9-Tetrahydrocannabinol and Metabolites Using LC-MS-MS.

A method for analyzing Δ9-tetrahydrocannabinol (THC), 11-hydroxy-Δ9-THC (THC-OH) and 11-nor-Δ9-THC-9-carboxylic acid (THC-COOH) in postmortem solid specimens using liquid chromatography-tandem mass spectrometry was developed and validated. A Stomacher instrument was used to prepare these tissues before extraction. Prior to solid phase extraction, liver, kidney, stomach, lung, brain, muscle, bladder and intestine tissues were pretreated with alkaline hydrolysis. All calibration curves were found to be linear with coefficients of determination greater than 0.99. The limit of quantification was 1.0 ng/g. Using three controls, within-run precision ranged between 1.0 and 12.0%, between-run precision ranged between 1.0 and 6.0%, and accuracy ranged between -7.0 and 8.0%. Matrix effects ranged from -21 to 24%. After matrix effects were excluded, analytical recoveries ranged from 79 to 97%. The distributions of THC, THC-OH and THC-COOH were investigated in 32 postmortem cases that tested positive for cannabinoids. This revealed new information regarding the distribution of THC metabolites in stomach, intestine and bladder. Alkaline hydrolysis was sufficient for the deglucuronidation of THC-COOH-glucuronide to its free form, THC-COOH, in all tissues of interest. In conclusion, measuring THC and its metabolites (THC-OH and THC-COOH) in tissues is crucial for any forensic toxicology detection method, especially when bodies are heavily decomposed, as solid tissues may be the only specimens available for testing.

Method for the identification and quantification of sixty drugs and their metabolites in postmortem whole blood using liquid chromatography tandem mass spectrometry.

The aim of this work was to develop and validate a liquid chromatography tandem mass spectrometry method for detecting sixty drugs and metabolites that are most commonly encountered in postmortem whole blood analysis. Although a large number of drugs were included in the panel, acceptance criteria for method validation were achieved. All calibration curves were found to be linear with coefficients of determination greater than 0.99. The limits of detection ranged from 0.2ng/mL to 1.0ng/mL and the limits of quantification range from 1.0ng/mL to 5.0ng/mL. Using three controls, within-run precision was 0.7%-10.3% and between-run precision was 0.6%-9.0%. Accuracy was ranged from 95.0%-104.1%. Matrix effects ranged from -15% to +22%. After excluding matrix effects, analytical recoveries ranged from 76% to 100%. Coefficients of variation for matrix effects ranged from 0.5%-13% and coefficients of variation for recovery ranged from 0.9%-13.0%. Over 1000 postmortem blood samples were analyzed. Among them, 435 cases (45%) tested positive for at least one analyte of interest. In conclusion, this study presents a technique for multianalyte screening of sixty drugs and metabolites that are commonly encountered in postmortem toxicology. This technique was then applied in routine analysis of autopsy blood samples in order to assess the applicability of this method. Data from postmortem cases is rarely reported from Saudi Arabia, and one of the current study goals is to present new information from postmortem cases to help prevent wide-spread drug use.

Postmortem Liver and Kidney Tissue Concentrations of Heroin Biomarkers and Their Metabolites in Heroin-Related Fatalities*†.

A method was developed and validated for analyzing 6-monoacetylmorphine, morphine, 6-acetylcodeine, and codeine in routine postmortem liver and kidney specimens using liquid chromatography-tandem mass spectrometry. Samples were prepared with a Stomacher instrument followed by solid-phase extraction. All calibration curves [0.5-1000 ng/g] were linear with coefficients of determination greater than 0.99 and limits of quantification of 1.0 ng/g. Within-run precision ranged between 2.0% and 8.0%, between-run precision ranged between 1.0% and 9.0%, and accuracy ranged between -5.0% and +3.0%. Matrix effects ranged from -18% to +9%. After matrix effects were excluded, analytical recoveries ranged from 76% to 94%. The distributions of 6-monoacetylmorphine, morphine, 6-acetylcodeine, and codeine were investigated in 31 postmortem cases in which heroin was the primary cause of death. In the current study, the median free morphine ratios were calculated for liver to blood and kidney to blood, which were 2.2 and 4.0, respectively. The current report highlights the importance of testing multiple specimens, including liver and kidney, in heroin-related deaths, especially if no blood samples are available. Furthermore, this work presents new information regarding the distribution of heroin metabolites in liver and kidney.

Postmortem Fluid Concentrations of Heroin Biomarkers and Their Metabolites.

Only limited data exist concerning the utility of complementary specimens in heroin-related deaths. As such, this report employed a validated LC-MS-MS method to quantify 6-monoacetylmorphine (6-MAM), 6-acetylcodeine (6-AC), and their metabolites morphine and codeine in blood with (BN) and without preservative (B) and the additional unpreserved specimens of vitreous humor, urine, stomach contents, and bile from 20 postmortem cases in which heroin was the primary cause of death. The median concentration of 6-MAM in BN was 0.011 mg/L, B was 0.008 mg/L, urine was 0.186 mg/L, vitreous humor was 0.022 mg/L, stomach contents was 0.147 mg/L, and bile was 0.012 mg/L. Only one case was found to be positive for 6-AC in B (case 6, 0.002 mg/L), and the median concentration of 6-AC was 0.002 mg/L in BN, 0.012 mg/L in urine, 0.003 mg/L in vitreous humor, 0.057 mg/L in stomach contents, and 0.004 mg/L in bile. These findings present new information on the distribution of these analytes in complementary matrices and support their inclusion for accurately determining the role of heroin in opioid-related deaths.

Method for Postmortem Quantification of Δ9-Tetrahydrocannabinol and Metabolites Using LC-MS-MS.

A specific, sensitive, fast and simple method for analysis of Δ9-tetrahydrocannabinol (THC), 11-hydroxy-Δ9-THC (THC-OH) and 11-nor-Δ9-THC-9-carboxylic acid (THC-COOH) in routine postmortem cases using LC-MS-MS was developed and validated. Prior to solid phase extraction, urine, stomach contents and bile were pretreated using alkaline hydrolysis, while blood and vitreous humor were pretreated with protein precipitation. The distribution of THC, THC-OH and THC-COOH were investigated in 31 postmortem cases that tested positive for cannabinoids. This revealed new information regarding the distribution of THC in stomach contents and vitreous humor. Alkaline hydrolysis was sufficient for the deglucuronidation of THC-COOH-glucuronide to its free form, THC-COOH, in urine, bile and stomach contents. However, the THC-OH concentration in bile reported in this study is considerably high compared to that of previous studies. In conclusion, including THC and its metabolites (THC-OH and THC-COOH) is crucial for any forensic toxicology detection method to most accurately determine the role of cannabinoids in deaths.

Direct determination of ethyl glucuronide and ethyl sulfate in postmortem urine specimens using hydrophilic interaction liquid chromatography-electrospray ionization-tandem mass spectrometry.

This work was aimed at developing and validating a hydrophilic interaction chromatography (HILIC)-electrospray ionization (ESI)-ion trap-tandem mass spectrometric method for identification and quantification of ethyl glucuronide (ETG) and ethyl sulfate (ETS) as ethanol biomarkers and at employing this method for analysis of postmortem urine samples. Analytes of interest were separated on a ZIC-HILIC column (150 x 2.1 mm, 3.5 microm) connected to a Thermo Finnigan LCQ Deca Plus liquid chromatographic- tandem mass spectrometric instrument operated in the ESI-selected reaction monitoring mode. Seventy-nine urine case samples were divided into three groups depending on the ethanol concentration found in blood and analyzed by the developed method: group A with postmortem blood ethanol concentrations higher than 200 mg/100 mL; group B with ethanol concentrations in the range 80-200 mg/100 mL; and group C with ethanol concentrations in the range 10-80 mg/100 mL. ETG and ETS had high recoveries of 98-99%, and the HILIC column produced fine, sharp peak shapes and achieved baseline separation in less than 7 min. Both ethanol markers were detected in all groups with overall median concentrations of 100 and 23 mg/L for ETG and ETS, respectively. It can be concluded that the potential for postmortem production of alcohol increased in the low ethanol concentration group as several cases tested negative for both biomarkers in group C. ETG was detected at low concentrations in some cases for which ETS tested negative. Although ETS is stable after being subjected to many stability conditions, the use of ETS as sole evidence of alcohol ingestion may lead to a false-negative result, as we noticed in groups A and C in the present study. The use of ETG is a more reliable ethanol biomarker. Both ethanol biomarkers should be determined in heavily putrefied cases and when the ethanol concentration in postmortem blood is low.

Oxycodone-related fatalities in the west of Scotland.

The intent of this study was to review fatalities involving oxycodone in the west of Scotland using a liquid chromatography-electrospray ionization-tandem mass spectrometry method developed for the determination of oxycodone and N- and O-demethylated metabolites in unhydrolyzed postmortem specimens. Ten oxycodone positive postmortem cases were detected, and nine were drug-related fatalities. Five cases were attributed solely to oxycodone intoxication and four to polydrug intoxication. Although there was overlap between blood oxycodone levels in deaths attributed to oxycodone only and those due to polydrug intoxication, lower oxycodone levels (< 1 mg/L) were associated with polydrug intoxication when compared with cases due to oxycodone alone (> 1 mg/L). The role of the parent drug in oxycodone fatalities has been fully studied, but the role of oxycodone metabolites (noroxycodone and oxymorphone) was investigated in this report for the first time. Oxycodone was more commonly detected in blood, urine, and vitreous humor followed by noroxycodone. The ratio between oxycodone and its N-demethylated metabolite was evaluated and found to be useful in determining whether death occurred shortly after drug administration or if there was a significant delay. High parent/metabolite ratios were correlated with short survival times after ingestion. The median ratio of oxycodone/noroxycodone was 2.4 and ranged from 0.7 to 49. Oxycodone prescriptions have risen sharply in Scotland in recent years, and the identification of 10 oxycodone-related deaths in the past 18 months highlights the importance of including this drug in routine laboratory screening and confirmation procedures.