Development of a Gas-Tight Syringe Headspace GC-FID Method for the Detection of Ethanol, and a Description of the Legal and Practical Framework for Its Analysis, in Samples of English and Welsh Motorists' Blood and Urine.

Ethanol is the most commonly used recreational drug worldwide. This study describes the development and validation of a headspace gas chromatography flame ionisation detection (HS-GC-FID) method using dual columns and detectors for simultaneous separation and quantitation. The use of a dual-column, dual-detector HS-GC-FID to quantitate ethanol is a common analytical technique in forensic toxicology; however, most analytical systems utilise pressure-balance injection rather than a simplified gas-tight syringe, as per this technique. This study is the first to develop and validate a technique that meets the specifications of the United Kingdom's requirements for road traffic toxicology testing using a Shimadzu GC-2014 gas-tight syringe. The calibration ranged from 10 to 400 mg/100 mL, with a target minimum linearity of r2 > 0.999, using tertiary butanol as the internal standard marker. The method has an expanded uncertainty at 99.73% confidence of 3.64% at 80 mg/100 mL, which is the blood alcohol limit for drink driving in England and Wales. In addition, at 200 mg%­the limit at which a custodial sentence may be imposed on the defendant­the expanded uncertainty was 1.95%. For both the 80 mg% and 200 mg% concentrations, no bias was present in the analytical method. This method displays sufficient separation for other alcohols, such as methanol, isopropanol, acetaldehyde, and acetone. The validation of this technique complies with the recommended laboratory guidelines set out by United Kingdom and Ireland Association of Forensic Toxicologists (UKIAFT), the recently issued Laboratory 51 guidelines by the United Kingdom Accreditation Service (UKAS), and the criteria set out by the California Code of Regulations (CCR), 17 CCR § 1220.1.

Metabolites of Cannabis Induce Cardiac Toxicity and Morphological Alterations in Cardiac Myocytes.

Cannabis is one of the most commonly used recreational drugs worldwide. Rrecent epidemiology studies have linked increased cardiac complications to cannabis use. However, this literature is predominantly based on case incidents and post-mortem investigations. This study elucidates the molecular mechanism of Δ9-tetrahydrocannabinol (THC), and its primary metabolites 11-Hydroxy-Δ9-THC (THC-OH) and 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THC-COOH). Treatment of cardiac myocytes with THC-OH and THC-COOH increased cell migration and proliferation (p < 0.05), with no effect on cell adhesion, with higher doses (250-100 ng/mL) resulting in increased cell death and significant deterioration in cellular architecture. Conversely, no changes in cell morphology or viability were observed in response to THC. Expression of key ECM proteins α-SMA and collagen were up-regulated in response to THC-OH and THC-COOH treatments with concomitant modulation of PI3K and MAPK signalling. Investigations in the planarian animal model Polycelis nigra demonstrated that treatments with cannabinoid metabolites resulted in increased protein deposition at transection sites while higher doses resulted in significant lethality and decline in regeneration. These results highlight that the key metabolites of cannabis elicit toxic effects independent of the parent and psychoactive compound, with implications for cardiotoxicity relating to hypertrophy and fibrogenesis.

Determination of ethanol in micro-volumes of blood by headspace gas chromatography: Statistical comparison between capillary and venous sampling sites.

Ethanol is the most commonly encountered drug in forensic toxicology, with widespread use throughout society. For this reason, it is important that there are a variety of reliable and robust methods to detect and quantify the content of alcohol in blood samples of suspected drink drivers. A common method of detection is gas chromatography with flame ionisation detector, with a number of sample preparation techniques employed. Typically, venous blood is sampled and used in the analysis. However, there is currently no legal specification in the UK of the blood sample source. This study investigates the use of capillary blood as an alternative to venous blood alongside two different sample volumes: 100 and 10 µL. Venous and capillary blood were collected from volunteers who had consumed alcohol. All blood sampling was carried out one hour after cessation of drinking. The results show a statistically significant difference between venous and capillary samples, with an average difference of 3.38 ± 1.99 mg/100 mL at 100 µL and approximately 4.13 ± 2.42 mg/100 mL at 10 µL, respectively. Predominantly, venous blood was detected at higher concentrations than the corresponding capillary samples. The deviations in alcohol samples between venous and capillary blood are consistent with previous studies. However, our research indicates that capillary blood is a viable matrix to test for alcohol, albeit one that underestimates blood-alcohol content in relation to venous sampling. There was no statistically significant difference between the 100 and 10 µL sample preparation methods on an individual basis, which infers that micro-volumes of alcohol are suitable for forensic blood-alcohol analysis.