BACKGROUND:In 1975, the mummified body of a female has been found in the Franciscan church in Basel, Switzerland. Molecular and genealogic analyses unveiled her identity as Anna Catharina Bischoff (ACB), a member of the upper class of post-reformed Basel, who died at the age of 68 years, in 1787. The reason behind her death is still a mystery, especially that toxicological analyses revealed high levels of mercury, a common treatment against infections at that time, in different body organs. The computed tomography (CT) and histological analysis showed bone lesions in the femurs, the rib cage, and the skull, which refers to a potential syphilis case.RESULTS:Although we could not detect any molecular signs of the syphilis-causing pathogen Treponema pallidum subsp. pallidum, we realized high prevalence of a nontuberculous mycobacterium (NTM) species in brain tissue sample. The genome analysis of this NTM displayed richness of virulence genes and toxins, and similarity to other infectious NTM, known to infect immunocompromised patients. In addition, it displayed potential resistance to mercury compounds, which might indicate a selective advantage against the applied treatment. This suggests that ACB might have suffered from an atypical mycobacteriosis during her life, which could explain the mummy's bone lesion and high mercury concentrations.CONCLUSIONS:The study of this mummy exemplifies the importance of employing differential diagnostic approaches in paleopathological analysis, by combining classical anthropological, radiological, histological, and toxicological observations with molecular analysis. It represents a proof-of-concept for the discovery of not-yet-described ancient pathogens in well-preserved specimens, using de novo metagenomic assembly.

While hair analysis is important and accepted in forensic applications, fundamental knowledge gaps still exist, exacerbated by a lack of knowledge of the incorporation mechanisms of substances into hair. The influence of the hair sampling location on the head on ethyl glucuronide (EtG) and cocaine concentrations was investigated by measuring the complete scalp hair of 14 (2 EtG, 4 cocaine, 8 both EtG and cocaine) study participants in a grid pattern for EtG, drugs of abuse, and benzodiazepines. Head skin perfusion and sweating rates were investigated to rationalize the concentration differences. For EtG, ratios between maximum and minimum concentrations on the scalp ranged from 2.5 to 7.1 (mean 4.4). For cocaine, the ratios ranged from 2.8 to 105 (mean 17.6). EtG concentrations were often highest at the vertex, but the distribution was strongly participant dependent. Cocaine and its metabolites showed the lowest concentrations at the vertex and the highest on the periphery, especially at the forehead. These differences led to hair from some head parts being clearly above conventional cut-offs and others clearly below. In addition to EtG and cocaine, the distributions of 24 other drugs of abuse and benzodiazepines/z-substances and metabolites are described. No clear pattern was observed for the head skin perfusion. Sweating rate measurements revealed higher sweating rates on the periphery of the haircut. Therefore, sweat could be a main incorporation route for cocaine. Concentration differences can lead to different interpretations depending on the sampling site. Therefore, the results are highly relevant for routine forensic hair analysis.

Often in hair analysis, a small hair sample is available while the analysis of a multitude of structurally diverse substances with different concentration ranges is demanded. The analysis of the different substances often requires different sample preparation methods, increasing the amount of required hair sample. When segmental hair analysis is necessary, the amount of hair sample needed is further increased. Therefore, the required sample amount for a full analysis can quickly exceed what is available. To combat this problem, a method for the combined hair sample preparation using a single extraction procedure for analysis of ethyl glucuronide with liquid chromatography-multistage fragmentation mass spectrometry/multiple reaction monitoring (LC-MS3 /MRM) and common drugs of abuse with LC-MRM was developed. The combined sample preparation is achieved by separating ethyl glucuronide from the drugs of abuse into separate extracts by fractionation in the solid-phase extraction step during sample clean-up. A full validation for all substances for the parameters selectivity, linearity, limit of detection, limit of quantification, accuracy, precision, matrix effects, and recovery was successfully completed. The following drugs of abuse were included in the method: Amphetamine; methamphetamine; 3,4-methylenedioxy-N-methylamphetamine (MDMA); 3,4-methylenedioxyamphetamine (MDA); 3,4-methylenedioxy-N-ethylamphetamine (MDE); morphine; 6-monoacetylmorphine; codeine; acetylcodeine; cocaine; benzoylecgonine; norcocaine; cocaethylene; methadone; 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) and methylphenidate. In conclusion, as only 1 sample preparation is needed with 1 aliquot of hair, the presented sample preparation allows an optimal analysis of both ethyl glucuronide and of the drugs of abuse, even when the sample amount is a limiting factor.

The distribution of analyte concentrations in hair across the scalp has not been thoroughly investigated. Differences in concentrations depending on sampling location are problematic, especially when measuring a second strand to confirm the result of the first measurement. Aiming at a better understanding of the concentration differences, the distribution of ethyl glucuronide (EtG) and caffeine concentrations in hair across the entire head of one test subject was investigated by dividing the scalp completely into regions of ca 2 cm × 2 cm area, yielding a total of 104 samples. For the quantification of EtG, a novel LC-MS3 /MRM method was developed and validated with a limit of detection and limit of quantification of 2 and 4 pg/mg, respectively. Large variations of the concentration across the head were found, with factors of ca 3.0 and 10.6 for EtG and caffeine, respectively. These differences could not be attributed to measurement error alone. The concentrations were projected onto the subject's head, and concentration patterns were identified for EtG and caffeine. When examining multiple strands from within one 2 cm × 2 cm sampling area, the strands showed similar concentrations. Segmental analysis of selected 3 cm strands showed decreasing concentrations of EtG and caffeine from proximal to distal end, possibly due to wash-out of the analytes. Copyright © 2017 John Wiley & Sons, Ltd.

A 24-year-old man known to consume illegal drugs was found dead in his apartment. A reclosable plastic zipper bag containing several hundred milligrams of a brown powder was found close to the dead body and the first assumption of the investigators was death due to heroin intoxication. Therefore, a legal autopsy was ordered. The following toxicological analysis revealed ocfentanil in urine and in the brown powder. Four different approaches for the determination of the ocfentanil concentrations in peripheral whole blood are described. Enrichment of ocfentanil from the powder was realized. With this reference, it was possible to determine the ocfentanil concentration in the seized powder to be 0.91%. Concentrations of ocfentanil were also determined in the sampled body fluids using the standard addition procedure. In peripheral blood 9.1 µg/L, in heart blood 27.9 µg/L and in urine 480 µg/L were measured. In addition, the antidepressant citalopram, the neuroleptic quetiapine and cannabinoids were found in urine and subsequently quantified in peripheral blood.

We present an unusual case of suicide by intraperitoneal injection of pentobarbital, an overdose of zolpidem and the intake of diazepam, ethanol and other psychoactive substances. The autopsy and specimen collection were conducted in a 10 to 18 h postmortem interval. The toxicological analysis revealed a significantly higher pentobarbital concentration in femoral blood compared to cardiac blood (36 vs. 15 mg/L). On the contrary, zolpidem and diazepam concentrations in cardiac blood (2700 and 590 µ g/L) were found to be significantly higher than in femoral blood (1500 and 230 µ g/L). These findings point to a postmortem redistribution with a distinct gradient from areas of high drug concentrations in the gastrointestinal tract (zolpidem and diazepam) and the injection site (pentobarbital) to peripheral tissue. Ethanol concentration was 0.95 ‰ which amplified the CNS depression. The choice of this unusual suicide method was associated with the deceased’s former job as a veterinarian’s assistant. In veterinary medicine, the intraperitoneal injection of a lethal dose of pentobarbital is quite commonly performed to euthanise small animals. Intraperitoneal injection is rare as route of administration in humans.

When investigating someone's hair a single lock is cut, washed, extracted and analysed. The forensic institutes in Switzerland agreed to retain a second lock for a possible reassessment. We were interested in the reproducibility of the concentrations of analytes in hair locks taken from different areas of the head of the same person covering the same time period. Therefore we analysed ethyl glucuronide and caffeine as model substances in 10 hair locks from three individuals categorised as social drinkers. The individual coefficients of variation varied between 14% and 28% for ethyl glucuronide and between 13% and 62% for caffeine corresponding to factors of 1.6 to 4.2 for the highest to the lowest concentrations between the hair locks. This finding has a significant importance both when the second hair lock has to be analysed in a forensic case and if the interpretation of the result is depending on a cut-off value.

The aim of the present study was to see whether people with a known history of illicit drug abuse (IDRUG) shift towards benzodiazepines, zolpidem and zopiclone (BZD-Z) when they have to prove their abstinence from IDRUG. We analysed 133 hair samples originating from persons in illicit drug AMPs (abstinence monitoring programs). Regranting of driver's licence was the most common motivation for proof of abstinence. To test a shift in drug consumption, hair samples of 87 cases (65 %) with request to analyse for IDRUG were additionally analysed for BZD-Z. The BZD-Z method includes 19 benzodiazepines and Z-drugs. In 46 of the cases (35 %) both IDRUG and BZD-Z analysis was requested by the public principals. An extraction suitable for illicit drugs and BZD-Z was used: 50 – 100 mg hair is washed with water, isopropanol and acetone 4 mL each and cut into small pieces. Then 20 mg of washed hair is pulverised in a ball mill and internal standards are added. Extraction is done by addition of 1.5 mL of a 1:1 mixture of formiate buffer (200 mM pH 5) and methanol followed by overhead shaking during at least four hours. After centrifugation the liquid is separated and evaporated using nitrogen. The dried sample is reconstituted with 500 μL water for injection into the LC-MS/MS system consisting of a Dionex ultimate 3000 HPLC equipped with a Phenomenex kinetex XCB 50 x 2,1 mm and an AB Sciex 4000 Qtrap mass spectrometer operated in MRM mode. Analysis of ethylglucuronide (EtG) was performed only on special request. In 91 (68 %) of the 133 cases none of the investigated substances (IDRUG, BZD-Z, EtG) were found. In 42 (32 %) of the cases 1 – 6 substances were detected. No BZD-Z analysis was ordered in 87 cases – thereof low and medium BZD-Z concentrations were found in 8 (9 %) cases. In 4 of these cases a regular consumption has to be considered. An additional BZD-Z analysis was ordered in 46 cases. 8 (19 %) samples tested positive. The hair concentrations in 3 cases implicated a regular ingestion. Most frequently detected BZD-Z substances were diazepam and its derivatives and zolpidem, followed by lorazepam and midazolam. When looking just at previously defined analytes in AMPs, we conclude that potentially addictive BZD-Z consumption will remain undiscovered in a few cases. It is therefore crucial for road safety that road traffic examination is performed by specially trained medical staff. In the field of traffic medicine, physicians and psychologists are trained to recognise addictive behaviour, which helps to reduce the probability of inadequate AMP settings.

AIM: The nonlinear elimination rate of THC and metabolites in blood complicates the inter- pretation of last cannabis use, which is an important information in DUID cases. We evaluated cannabinoids in blood and hydrolysed urine from 50 DUID cases in order to gain additional information about last cannabis use. METHODS: THC, THC-OH and THC-COOH were analysed in blood and hydrolysed urine (β- glucuronidase) by GC-MS-MS. Cannabinoid concentrations in blood and urine were compa- red. Mann-Whitney-U-Tests were performed to investigate differences between cannabinoid concentrations from recent and older cannabis intake. Blood/urine cannabinoid concentrations of admitted chronic users were compared to occasional users. An approach for cluster analysis is presented. RESULTS: Cannabinoid concentration in blood and urine were strongly correlated. Blood and urine cannabinoid concentrations from recent cannabis consumers differed significantly from the rest of the test group. No difference in blood/urine concentration was found between chronic and occasional users. RESULTS of a cluster analysis approach were promising. CONCLUSION: In individual cases, an interpretation of last cannabis use is only possible by approximation. However, multivariate data analysis of cannabinoid concentration might facilitate the detection of recent cannabis intake. Language: en

Drug Testing and AnalysisVolume 5, Issue 9-10 p. 777-780 Correspondence case report Chloroquine and diazepam for her last sleep Sarah Hangartner, Sarah Hangartner Institute for Legal Medicine of the University of Basel, Forensic Chemistry and Toxicology, Pestalozzistrasse 22, 4056 Basel, SwitzerlandSearch for more papers by this authorSebastian Eggert, Sebastian Eggert Institute of Pathology and Forensic Medicine, Cantonal Hospital of Grisons, Loestrasse 170, 7000 Chur, SwitzerlandSearch for more papers by this authorFranz Dussy, Franz Dussy Institute for Legal Medicine of the University of Basel, Forensic Chemistry and Toxicology, Pestalozzistrasse 22, 4056 Basel, SwitzerlandSearch for more papers by this authorDaniel Wyler, Daniel Wyler Institute of Pathology and Forensic Medicine, Cantonal Hospital of Grisons, Loestrasse 170, 7000 Chur, SwitzerlandSearch for more papers by this authorThomas Briellmann, Corresponding Author Thomas Briellmann Institute for Legal Medicine of the University of Basel, Forensic Chemistry and Toxicology, Pestalozzistrasse 22, 4056 Basel, Switzerland Correspondece to: Thomas Briellmann, Institute for Legal Medicine of the University of Basel, Forensic Chemistry and Toxicology, Pestalozzistrasse 22, 4056 Basel, Switzerland. E-mail: [email protected]Search for more papers by this author Sarah Hangartner, Sarah Hangartner Institute for Legal Medicine of the University of Basel, Forensic Chemistry and Toxicology, Pestalozzistrasse 22, 4056 Basel, SwitzerlandSearch for more papers by this authorSebastian Eggert, Sebastian Eggert Institute of Pathology and Forensic Medicine, Cantonal Hospital of Grisons, Loestrasse 170, 7000 Chur, SwitzerlandSearch for more papers by this authorFranz Dussy, Franz Dussy Institute for Legal Medicine of the University of Basel, Forensic Chemistry and Toxicology, Pestalozzistrasse 22, 4056 Basel, SwitzerlandSearch for more papers by this authorDaniel Wyler, Daniel Wyler Institute of Pathology and Forensic Medicine, Cantonal Hospital of Grisons, Loestrasse 170, 7000 Chur, SwitzerlandSearch for more papers by this authorThomas Briellmann, Corresponding Author Thomas Briellmann Institute for Legal Medicine of the University of Basel, Forensic Chemistry and Toxicology, Pestalozzistrasse 22, 4056 Basel, Switzerland Correspondece to: Thomas Briellmann, Institute for Legal Medicine of the University of Basel, Forensic Chemistry and Toxicology, Pestalozzistrasse 22, 4056 Basel, Switzerland. E-mail: [email protected]Search for more papers by this author First published: 12 July 2013 https://doi.org/10.1002/dta.1509Citations: 5Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Citing Literature Volume5, Issue9-10Special Issue: Case reports in forensic toxicologySeptember‐October 2013Pages 777-780 RelatedInformation

AIMS To investigate the pharmacokinetics and pharmacodynamics of nasal formulations containing midazolam (5-30 mg ml(-1)) complexed with cyclodextrin. METHODS An open-label sequential trial was conducted in eight healthy subjects receiving single doses of 1 mg and 3 mg intranasally and 1 mg midazolam intravenously. Pharmacokinetic parameters were obtained by non-compartmental and two-compartmental models. Pharmacodynamic effects of midazolam were assessed using VAS and a reaction time test. RESULTS Mean bioavailability of midazolam after nasal administration ranged from 76 +/- 12% to 92 +/- 15%. With formulations delivering 1 mg midazolam, mean C(max) values between 28.1 +/- 9.1 and 30.1 +/- 6.6 ng ml(-1) were reached after 9.4 +/- 3.2-11.3 +/- 4.4 min. With formulations delivering 3 mg midazolam, mean C(max) values were between 68.9 +/- 19.8 and 80.6 +/- 15.2 ng ml(-1) after 7.2 +/- 0.7-13.0 +/- 4.3 min. Chitosan significantly increased C(max) and reduced t(max) of midazolam in the high-dose formulation. Mean ratios of dose-adjusted AUC after intranasal and intravenous application for 1'-hydroxymidazolam were between 0.97 +/- 0.15 and 1.06 +/- 0.24, excluding relevant gastrointestinal absorption of intranasal midazolam. The pharmacodynamic effects after the low-dose nasal formulations were comparable with those after 1 mg intravenous midazolam. The maximum increase in reaction time by the chitosan-containing formulation delivering 3 mg midazolam was greater compared with 1 mg midazolam i.v. (95 +/- 78 ms and 19 +/- 22 ms, mean difference 75.5 ms, 95% CI 15.5, 135.5, P < 0.01). Intranasal midazolam was well tolerated but caused reversible irritation of the nasal mucosa. CONCLUSIONS Effective midazolam serum concentrations were reached within less than 10 min after nasal application of a highly concentrated midazolam formulation containing an equimolar amount of the solubilizer RMbetaCD combined with the absorption enhancer chitosan.

In Switzerland, a two-tier system based on impairment by any psychoactive substances which affect the capacity to drive safely and zero tolerance for certain illicit drugs came into force on 1 January 2005. According to the new legislation, the offender is sanctioned if Delta(9)-tetrahydrocannabinol THC is >or=1.5ng/ml or amphetamine, methamphetamine, 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxyethylamphetamine (MDEA), cocaine, free morphine are >or=15ng/ml in whole blood (confidence interval+/-30%). For all other psychoactive substances, impairment must be proven in applying the so-called "three pillars expertise". At the same time the legal blood alcohol concentration (BAC) limit for driving was lowered from 0.80 to 0.50g/kg. The purpose of this study was to analyze the prevalence of drugs in the first year after the introduction of the revision of the Swiss Traffic Law in the population of drivers suspected of driving under the influence of drugs (DUID). A database was developed to collect the data from all DUID cases submitted by the police or the Justice to the eight Swiss authorized laboratories between January and December 2005. Data collected were anonymous and included the age, gender, date and time of the event, the type of vehicle, the circumstances, the sampling time and the results of all the performed toxicological analyses. The focus was explicitly on DUID; cases of drivers who were suspected to be under the influence of ethanol only were not considered. The final study population included 4794 DUID offenders (4243 males, 543 females). The mean age of all drivers was 31+/-12 years (range 14-92 years). One or more psychoactive drugs were detected in 89% of all analyzed blood samples. In 11% (N=530) of the samples, neither alcohol nor drugs were present. The most frequently encountered drugs in whole blood were cannabinoids (48% of total number of cases), ethanol (35%), cocaine (25%), opiates (10%), amphetamines (7%), benzodiazepines (6%) and methadone (5%). Other medicinal drugs such as antidepressants and benzodiazepine-like were detected less frequently. Poly-drug use was prevalent but it may be underestimated because the laboratories do not always analyze all drugs in a blood sample. This first Swiss study points out that DUID is a serious problem on the roads in Switzerland. Further investigations will show if this situation has changed in the following years.

Swiss laboratories performing toxicological road traffic analyses have been authorized for many years by the Swiss Federal Roads Office (FEDRO). In 2003 FEDRO signed a contract with the Swiss Society of Legal Medicine (SSLM) to organize the complete quality management concerning road traffic analyses. For this purpose a multidisciplinary working group was established under the name of "road traffic commission (RTC)". RTC has to organize external quality control, to interpret the results of these controls, to perform audits in the laboratories and to report all results to FEDRO. Furthermore the working group can be mandated for special tasks by FEDRO. As an independent organization the Swiss Center for Quality Control (CSCQ) in Geneva manages the external quality controls in the laboratory over the past years. All tested drugs and psychoactive substances are listed in a federal instruction. The so-called 'zero tolerance substances' (THC, morphine, cocaine, amphetamine, methamphetamine, MDMA and MDEA) and their metabolites have to be tested once a year, all other substances (benzodiazepines, zolpidem, phenobarbital, etc.) periodically. Results over the last years show that all laboratories are generally within the confidence interval of +/-30% of the mean value. In cases of non-conformities measures have to be taken immediately and reported to the working group. External audits are performed triennially but accredited laboratories can combine this audit with the approval of the Swiss Accreditation Service (SAS). During the audits a special checklist filled in by the laboratory director is assessed. Non-conformities have to be corrected. During the process of establishing a new legislation, RTC had an opportunity of advising FEDRO. In collaboration with FEDRO, RTC and hence SSLM can work actively on improving of quality assurance in road traffic toxicological analyses, and has an opportunity to bring its professional requests to the federal authorities.

The validation of a qualitative ion mobility spectrometry (IMS) procedure for the detection of trace amounts of heroin and cocaine on incriminated material using a vacuum cleaner for sampling is presented. The limit of detection, the limit of decision, selectivity and robustness were determined. As an approach, robustness was determined using ionizational interferences and matrix effects. By using this simple sampling procedure, a positive result for incriminated clothes needs a contamination of 250ng cocaine and 1000ng heroin, respectively.

A high-performance liquid chromatography method for the determination of benzodiazepines and their metabolites in whole blood and serum using mass spectrometry (MS) and photodiode array (PDA) detection is presented. The combination of both detection types can complement each other and provides extensive case relevant data. The limits of quantification (LOQ) with the MS detection lie between 2 and 3 μg/l for the following benzodiazepines or metabolites: 7-amino-flunitrazepam, alprazolam, desalkyl-flurazepam, desmethyl-flunitrazepam, diazepam, flunitrazepam, flurazepam, α-hydroxy-midazolam, lorazepam, midazolam, nitrazepam, nordazepam and oxazepam, respectively 5 μg/l for lormetazepam and 6 μg/l for bromazepam. The LOQ of clobazam determined with the PDA detector is 10 μg/l. A convenient approach for determining the measurement uncertainty of the presented method—applicable also for other methods in an accreditation process—is presented. At low concentrations (<10 μg/l), measurement uncertainty was estimated to be about 50%, and at concentrations >180 μg/l, it was estimated to be about 15%. One hundred and twenty-eight case data acquired over 1 year are summarised.

In 1936 Randall and Melkin1 reported that papillary tip calcification occurred more frequently in patients with stone disease than in non-stone formers. Since then many authors2−7 have regarded so-called Randall’s plaques as the first manifestation of urinary stones; yet, little is known about intrarenal crystallization8,9. The aim of this study was to attempt to detect primary sites of intrarenal crystallization and to describe any differences that there might be between stone formers and healthy persons10,11.