Automated headspace solid-phase dynamic extraction for the determination of cannabinoids in hair samples

This article describes a fully automated procedure for detecting cannabinoids in human hair samples. The procedure uses alkaline hydrolysis and headspace solid-phase dynamic extraction (HS-SPDE), followed by on-coating derivatization and gas chromatography-mass spectrometry (GC-MS). SPDE is a further development of solid-phase microextraction (SPME), based on an inside needle capillary absorption trap. It uses a hollow needle with an internal coating of polydimethylsiloxane as extraction and pre-concentration medium.Ten mg of hair were washed with deionised water, petroleum ether and dichloromethane. After adding deuterated internal standards, the sample was hydrolyzed with sodium hydroxide and directly submitted to HS-SPDE. After absorption of analytes for an on-coating derivatization procedure, the SPDE-needle was directly placed into the headspace of a second vial containing N-methyl-N-trimethylsilyl-trifluoroacetamide before GC-MS analysis. The limit of detection was 0.14 ng/mg for Delta(9)-tetrahydrocannabinol, 0.09 ng/mg for cannabidiol, and 0.12ng/mg for cannabinol. Absolute recoveries were in the range of 0.6 to 8.4%. Linearity was verified over a range from 0.2 to 20 ng/mg, with coefficients of correlation between 0.998 and 0.999. Intra- and inter-day precision were determined at two different concentrations and resulted in ranges between 2.3 and 6.0% (intra-day) and 3.3 and 7.6% (inter-day). Compared with conventional methods of hair analysis, this automated HS-SPDE-GC-MS procedure is substantially faster. It is easy to perform without using solvents and with minimal sample quantities, and it yields the same sensitivity and reproducibility. Compared to SPME, we found a higher extraction rate, coupled with a faster automated operation and greater stability of the device.     

Suicidal yew poisoning--from Caesar to today--or suicide instructions on the internet

Already the Celts and ancient Germanic peoples knew about the poisonousness of the yew, which played an important part in the mythology of these civilizations. For hunting, the arrows were made poisonous with yew juice, and yew leaves were used for homicide and suicide. In modern times taxine is rarely used with suicidal intent, although this method is actually recommended on the respective Websites. After a 14-year-old boy had intensively studied poisonous plants and methods of suicide on various Websites, he cut leaves from a yew tree (taxus baccata) in his parents' garden, crushed and ingested them and died soon afterwards. At the forensic autopsy pieces of the partially crushed, partially completely preserved yew leaves were found in the stomach. The histological findings were unspecific, e.g. marked general blood congestion of the internal organs and pronounced cerebral and pulmonary edema. When the tree leaves found in the stomach were viewed under the light microscope, a stoma typical of taxus was observed; chemical-toxicological investigations revealed 3,5-dimethoxyphenol in the gastric content, which is considered a marker for the ingestion of taxus.     

An unusual area of injury in postmortem animal feeding beneath clothing

The authors report on a woman who died on the age of 75 years and who was found dead in her dwelling with an extensive injury of the left thigh two days after the last sign of life. Together with the corpse a poodle was present in the dwelling. Due to the unusual localization totally beneath the clothes (trousers) and the morphology of the injury an unnatural cause of death was suspected. According to the autopsy findings and the histological results, postmortal animal scavenging caused by the poodle remained the only explanation for the injury. Supplementary DNA testing yielded final certainty.     

Immunochemical screening: manipulation, handling and interpretation errors, exemplified by benzodiazepines

The article describes problems related to immunochemical screening-tests with special regard to benzodiazepines, which are commonly used for therapeutic purposes. Focal points are adulteration, manipulation and handling of samples, the selection of cut-off-values, specificity and cross-reactivity, influence of biotransformation and frequently observed pitfalls in the interpretation of the analytical results.     

Chronology of a homicide and an attempted suicide by DNA analysis of selected blood stains on the weapon (knife)]

A five-year-old boy was fatally injured by a stab and a cut to the throat. The suspected offender, an acquaintance of the victim's mother, stated that he had been disturbed by the boy while attempting suicide and had therefore killed the boy. The suspect showed superficial cuts on both wrists and three shallow stabs on the right-hand side of the abdomen. The purpose of the forensic investigations was to answer the question if the suspect's statement could be confirmed or if the attempted suicide took place after killing the boy. The blood-smeared blade of the knife was divided into different sectors with the cutting edge and the tip of the blade being examined separately from the sides. The blood traces from the different blade sectors were then typed in 12 different DNA regions by PCR (polymerase-chain-reaction). It could be demonstrated that the strong blood traces on the sides of the blade originated from the victim alone, whereas in the area of the edge and the tip of the blade the amount of the victim's blood was negligible compared with that of the suspected offender. The explanation for this finding is that in the areas of the edge and the tip of the knife the blood of the victim must have been replaced by the blood of the suspected perpetrator, which means that the suicidal injuries were the last to be caused by the knife.     

Morbidity and cause of death in homeless persons

The Hamburg Institute of Forensic Medicine analysed 388 deaths of homeless people between 1990 and 1998 considering demographic data, post-mortem results and autopsy findings. The manner of death was either unnatural or unascertained; the autopsy rate amounted to 43.8%. The results confirm the observation discussed in the last few years that homeless people are usually in a poor state of health. In relation to the average age of death, which is 44.5 years, the number of previous diseases is high. Unnatural causes (excluding homicides) account for a high percentage, viz. 62.6%, primarily intoxications. The most frequent natural cause of death is infection (16.8%). Women account for 8.5% of the deaths with their organ findings and causes of deaths being different from those of homeless men in terms of distribution and incidence. The data presented in this report may be helpful for the improvement of the present system of medical help in Hamburg.     

Immunostaining by complement C9: a tool for early diagnosis of myocardial infarction and application in forensic medicine

Before the first 12 hours, diagnosis of early myocardial infarctions is always difficult for forensic pathologists. We tested complement C9 expression in 121 formalin-fixed and paraffin-embedded heart samples by an immunohistochemical procedure. The heart specimens were separated into four groups: 33 cases in group 1 with typical ischemic damages histologically located, 20 cases in group 2 with death related to myocardial infarction on the basis of ischemic presentation on electrocardiogram but no obvious histological ischemic damage, 35 cases in group 3 with severe coronary disease without cause of death found at the autopsy, and 33 cases in group 4 without sign of myocardial infarction and without coronary disease. In the first group, all 33 heart samples showed a well-defined C9 expression in the necrotic areas. The second group in 17 of 20 cases showed positive areas for C9 expression. In the other three heart specimens, only few stained cells were observed whereas the painful symptoms had begun less than 1 h before death. The third group showed C9 immunopositive areas in six of 35 cases, few stained cells in 8 cases, and no C9 deposition in the 21 other cases. The last group showed no staining area. To avoid nonspecific C9 staining due to tissue autolysis, we studied C9 expression during a controlled putrefactive process in four cases included in group 1; staining was found only in infarcted myocardial areas, and was observed up to ten days. Specificity of C9 expression was evaluated to be 100% [89.4 to 100%] and sensitivity to be 85% [62.11 to 96.79%]. In conclusion, evaluation of immunohistochemical expression of C9 appears to be a highly sensitive and specific marker of early myocardial infarction, useful in forensic medicine if survival is more than 1 h after the beginning of myocyte damage.