Is adipose tissue suitable for detection of (synthetic) cannabinoids? A comparative study analyzing antemortem and postmortem specimens following pulmonary administration of JWH-210, RCS-4, as well as ∆9-tetrahydrocannabinol to pigs.

Examining fatal poisonings, chronic exposure may be reflected by the concentration in tissues known for long-term storage of drugs. Δ9-tetrahydrocannabinol (THC) persists in adipose tissue (AT), but sparse data on synthetic cannabinoids (SC) are available. Thus, a controlled pig study evaluating antemortem (AM) disposition and postmortem (PM) concentration changes of the SC 4-ethylnaphthalene-1-yl-(1-pentylindole-3-yl)methanone (JWH-210) and 2-(4-methoxyphenyl)-1-(1-pentyl-indole-3-yl)methanone (RCS-4) as well as THC in AT was performed. The drugs were administered pulmonarily (200 µg/kg body weight) to twelve pigs. Subcutaneous (s.c.) AT specimens were collected after 15 and 30 min and then hourly up to 8 h. At the end, pigs were sacrificed and s.c., perirenal, and dorsal AT specimens were collected. The carcasses were stored at room temperature (RT; n = 6) or 4 °C (n = 6) and specimens were collected after 24, 48, and 72 h. After homogenization in acetonitrile and standard addition, LC-MS/MS was performed. Maximum concentrations were reached 0.5-2 h after administration amounting to 21 ± 13 ng/g (JWH-210), 24 ± 13 ng/g (RCS-4), and 22 ± 20 ng/g (THC) and stayed at a plateau level. Regarding the metabolites, very low concentrations of N-hydroxypentyl-RCS-4 (HO-RCS-4) were detected from 0.5 to 8 h. PM concentrations of parent compounds did not change significantly (p > 0.05) over time under both storage conditions. Concentrations of HO-RCS-4 significantly (p < 0.05) increased in perirenal AT during storage at RT. These results suggest a rapid distribution and persistence in s.c. AT. Furthermore, AT might be resistant to PM redistribution of parent compounds. However, significant PM increases of metabolite concentrations might be considered in perirenal AT.

Time- and temperature-dependent postmortem concentration changes of the (synthetic) cannabinoids JWH-210, RCS-4, as well as ∆9-tetrahydrocannabinol following pulmonary administration to pigs.

In forensic toxicology, interpretation of postmortem (PM) drug concentrations might be complicated due to the lack of data concerning drug stability or PM redistribution (PMR). Regarding synthetic cannabinoids (SC), only sparse data are available, which derived from single case reports without any knowledge of dose and time of consumption. Thus, a controlled pig toxicokinetic study allowing for examination of PMR of SC was performed. Twelve pigs received a pulmonary dose of 200 µg/kg BW each of 4-ethylnaphthalene-1-yl-(1-pentylindole-3-yl)methanone (JWH-210), 2-(4-methoxyphenyl)-1-(1-pentyl-indole-3-yl)methanone (RCS-4), and Δ9-tetrahydrocannabinol via an ultrasonic nebulizer. Eight hours after, the pigs were put to death with T61 and specimens of relevant tissues and body fluids were collected. Subsequently, the animals were stored at room temperature (n = 6) or 4 °C (n = 6) and further samples were collected after 24, 48, and 72 h each. Concentrations were determined following enzymatic cleavage and solid-phase extraction by liquid-chromatography tandem mass spectrometry applying the standard addition approach. High concentrations of the parent compounds were observed in lung, liver, kidney and bile fluid/duodenum content as well as brain. HO-RCS-4 was the most prevalent metabolite detected in PM specimens. In general, changes of PM concentrations were found in every tissue and body fluid depending on the PM interval as well as storage temperature.

Distribution of the (synthetic) cannabinoids JWH-210, RCS-4, as well as ∆9-tetrahydrocannabinol following pulmonary administration to pigs.

New psychoactive substances, especially synthetic cannabinoids (SC), are gaining increasing relevance in postmortem forensic toxicology. Particularly, the interpretation of analytical results is challenging, as usually, no toxicokinetic (TK) data concerning distribution in organs and tissues are available. Thus, a controlled pig TK study allowing for examination of organ and tissue distribution of SC was performed. For this purpose, 12 pigs received a single pulmonary dose of 200 µg/kg body weight each of 4-ethylnaphthalene-1-yl-(1-pentylindole-3-yl)methanone (JWH-210), 2-(4-methoxyphenyl)-1-(1-pentylindole-3-yl)methanone (RCS-4), and Δ9-tetrahydrocannabinol (THC) via an ultrasonic nebulizer. Eight hours after administration, the animals were put to death by the administration of T61. Thereupon, relevant organs, important body fluids such as bile and colon content, and tissues such as muscle tissue were collected. After enzymatic hydrolysis and solid-phase extraction, analysis was performed by liquid chromatography-tandem mass spectrometry. For quantification, a standard addition method was applied. The parent compounds could be detected in every analyzed specimen with the exception of colon content. Regarding JWH-210, the kidneys and lungs are viable matrices for postmortem analysis. In terms of RCS-4, the lungs were found to be an appropriate matrix. Concerning THC, the liver, bile fluid as well as duodenum content were suitable matrices for detection. Metabolites were only detected in tissues/body fluids involved in metabolism and/or elimination. Bile fluid and duodenum content were shown, as the most appropriate specimens for quantification of metabolites.

Effect of anticoagulation therapy on drying times in bloodstain pattern analysis.

In forensic case work, blood stain pattern analysis frequently aids in deducing the chain of actions or parts thereof taking place during an event leading to blood loss. Wiped single blood stains and/or groups of blood stains are seen at a majority of complex crime scenes. The appearance of wiped blood stains depends on droplet volume and stain age (as a function of blood viscosity and the degree of stain skeletonization) and characteristics of the stained surface (i.e., texture, temperature). Furthermore, based on the biochemical and biophysical properties of blood, not only the drying processes, but also complex coagulation cascades are relevant to the assessment of wiped blood stains. This study was designed to determine if anticoagulation therapies markedly affect the wipeability of blood stains over times elapsed since deposition and the overall drying process. A total of 813 blood stains, originating from donors being treated with acetylsalicylic acid (ASA), clopidogrel + ASA, low-molecular-weight heparin, or rivaroxaban, were dropped on common household tiles. Wipeability at an ambient temperature of 20 °C was tested for 22 time periods (1, 2, 3, 5, 10, 15…105 min since deposition). Whereas stains consisting of untreated blood were dried within 55 min, wipeability of all droplets originating from donors with prior anticoagulation treatment showed pronounced delays compared with the control, ranging from 20 min (ASA and clopidogrel + ASA) to 45 min (rivaroxaban). This pronounced effect was not seen in earlier studies, which might be explained by the higher volume of droplets used in this study, which resulted in a shift in relevance from drying to clotting processes. Significant differences between the drying times of the various anticoagulation regimes might be attributed to anticoagulant activity against different targets in the coagulation cascades. In conclusion, anticoagulation treatment prior to blood loss significantly affected the wipeability of blood stains. Anticoagulation therapy should therefore be taken into account in the analysis of blood stain patterns.

[True color accuracy in digital forensic photography]. / Über die Farbdetailtreue in der digitalen forensischen Fotografie.

Forensic photographs not only need to be unaltered and authentic and capture context-relevant images, along with certain minimum requirements for image sharpness and information density, but color accuracy also plays an important role, for instance, in the assessment of injuries or taphonomic stages, or in the identification and evaluation of traces from photos. The perception of color not only varies subjectively from person to person, but as a discrete property of an image, color in digital photos is also to a considerable extent influenced by technical factors such as lighting, acquisition settings, camera, and output medium (print, monitor). For these reasons, consistent color accuracy has so far been limited in digital photography. Because images usually contain a wealth of color information, especially for complex or composite colors or shades of color, and the wavelength-dependent sensitivity to factors such as light and shadow may vary between cameras, the usefulness of issuing general recommendations for camera capture settings is limited. Our results indicate that true image colors can best and most realistically be captured with the SpyderCheckr technical calibration tool for digital cameras tested in this study. Apart from aspects such as the simplicity and quickness of the calibration procedure, a further advantage of the tool is that the results are independent of the camera used and can also be used for the color management of output devices such as monitors and printers. The SpyderCheckr color-code patches allow true colors to be captured more realistically than with a manual white balance tool or an automatic flash. We therefore recommend that the use of a color management tool should be considered for the acquisition of all images that demand high true color accuracy (in particular in the setting of injury documentation).