GHB. Club drug or confusing artifact?

GHB can be produced either as a pre- or postmortem artifact. The authors describe two cases in which GHB was detected and discuss the problem of determining the role of GHB in each case. In both cases, NaF-preserved blood and urine were analyzed using gas chromatography. The first decedent, a known methamphetamine abuser, had GHB concentrations similar to those observed with subanesthetic doses (femoral blood, 159 microg/ml; urine, 1100 microg/ml). Myocardial fibrosis, in the pattern associated with stimulant abuse, was also evident. The second decedent had a normal heart but higher concentrations of GHB (femoral blood, 1.4 mg/ml; right heart, 1.1 mg/ml; urine, 6.0 mg/ml). Blood cocaine and MDMA levels were 420 and 730 ng/ml, respectively. Both decedents had been drinking and were in a postabsorptive state, with blood to vitreous ratios of less than 0.90. If NaF is not used as a preservative, GHB is produced as an artifact. Therefore, the mere demonstration of GHB does not prove causality or even necessarily that GHB was ingested. Blood and urine GHB concentrations in case 1 can be produced by a therapeutic dose of 100 mg, and myocardial fibrosis may have had more to do with the cause of death than GHB. The history in case 2 is consistent with the substantial GHB ingestion, but other drugs, including ethanol, were also detected. Ethanol interferes with GHB metabolism, preventing GHB breakdown, raising blood concentrations, and making respiratory arrest more likely. Combined investigational, autopsy, and toxicology data suggest that GHB was the cause of death in case 2 but not case 1. Given the recent discovery that postmortem GHB production occurs even in stored antemortem blood samples (provided they were preserved with citrate) and the earlier observations that de novo GHB production in urine does not occur, it is unwise to draw any inferences about causality unless (1) blood and urine are both analyzed and found to be elevated; (2) blood is collected in NaF-containing tubes; and (3) a detailed case history is obtained.     

A novel hunting accident. Discharge of a firearm by a hunting dog

The authors report the case of a 21-year-old man who was killed while duck hunting when a shotgun accidentally discharged, shooting him in the head. The loaded weapon, which had been lying on the ground with the safety off and the muzzle pointed toward a river a few feet away, discharged when a hunting dog stepped on the trigger. Scene investigation confirmed that the victim had been standing in the river, planting decoys, with his head approximately level with the adjacent bank. Autopsy examination and ballistic testing confirmed a range of fire consistent with the witness' statements. Examination of the weapon in question documented a light trigger pull but no mechanical defects. The authors review the epidemiology and causality of hunting accidents and discuss the various safety rules that were violated in this highly unusual case. The importance of a complete death investigation, including autopsy, when dealing with a firearm death is emphasized.     

Sex determination and estimation of stature from the long bones of the arm

The determination of sex and the estimation of stature from bones play an important role in identifying unknown bodies, parts of bodies or skeletal remains. In medico-legal practice statements on the probable sex of a decomposed body or part of a body are often expected even during autopsy. The present study was, therefore, restricted to few easily accessible dimensions from bones which were prepared only by mechanically removing soft tissues, tendons and ligaments. The specimens came from the Anatomical Institutes in Munich and Cologne from the years 1994-1998 including a total of 143 individuals (64 males and 79 females). The mean age was 79 years (46-108), the mean body height 161cm (134-189). The following measurements were taken: maximum humeral length (mean: 33.4cm in males; 30.7cm in females), vertical humeral head diameter (mean: 5.0cm in males, 4.4cm in females), humeral epicondylar width (mean: 6.6cm in males; 5.8cm in females), maximum ulnar length (mean: 26.5cm in males, 23.8cm in females), proximal ulnar width (mean: 3.4cm in males, 2.9cm in females), distal ulnar width (mean: 2.2cm in males; 1.8cm in females), maximum radial length (mean: 24.6cm in males; 22.0cm in females), radial head diameter (mean: 2.6cm in males, 2.2cm in females) and distal radial width (mean: 3.6cm in males; 3.2cm in females). The differences between the means in males and females were significant (P<0.0005). A discriminant analysis was carried out with good results. A percentage of 94.93% of cases were correctly classified when all measures of the radius were applied jointly, followed by humerus (93.15%) and ulna (90.58%). Applied singly, the humeral head diameter allowed the best distinction (90.41% correctly grouped cases), followed by the radial length (89.13%), the radial head diameter (88.57%) and the humeral epicondylar width (88.49%). The linear regression analysis for quantifying the correlation between the bone lengths and the stature led to unsatifactory results with large 95%-confidence intervals for the coefficients and high standard errors of estimate.     

Cranio-facial morphanalysis: a new method for enhancing reliability while identifying skulls by photo superimposition

Skull-photograph superimposition continues to be the most prevalent method employed for identifying a skull recovered in a criminal case as that belonging to a putative victim whose face photograph is available. The reliability of identification achieved has been shown to be 91%, indicating the possibility of a skull mismatching with a face photograph belonging to a person other than the actual deceased. This lack of reliability dampens the confidence of the expert and in turn confounds the mind of the judge. It has been shown that the variations in the shape of the facial organs are influenced by the corresponding variations in the skeletal elements of the facial skull. "Cranio-facial morphanalysis", a new anthroposcopic method proposed here for evaluating the shape correlations between a skull and a face photograph, when applied conjointly with skull-photograph superimposition is shown to increase the reliability in forensic skull identification.     

Numerical density of mu opioid receptor expressing neurons in the frontal cortex of drug related fatalities

In animal and cell culture experiments, chronic morphine treatment has been followed by 'up'- as well as 'down-regulation' of the mu opioid receptor (mu OR) number. The present postmortem morphometric study of morphine-related fatalities of drug addicts (n=12, and 22-35 years old, with blood unconjugated morphine levels from 27.1 to 458 ng/ml, m.v. 198.5 ng/ml) versus a non-addicted control group (n=13 and 10-44 years old) was intended to examine whether chronic opiate exposure affects the numerical density of mu OR expressing neurons in the human neocortex (area 10 according to Brodmann). For the immunohistochemical procedure, thick (100 microm) vibratome sections were incubated with a monoclonal antibody against the mu OR [Arvidsson et al., J. Neurosci. 15 (1995) 3328] and immunoreactive sites were visualized using an immunoperoxidase protocol. The numerical densities of mu OR-expressing and Nissl-stained neurons were assessed morphometrically (camera lucida-drawings). In both collectives, the anti-mu OR immunoreactivity was mainly found in pyramidal neurons of layers (L) II/III and V and in multiform neurons of L VI. In the drug-related fatalities and the control group, the density of neurons expressing mu OR protein was similar, amounting for 2698 +/- 153 and 2688 +/- 172/mm(3), respectively. These findings extend the binding studies of opioid ligands in postmortem brains of heroin addicts [Gabilondo et al., Psychopharmacology 115 (1994) 135] revealing similar receptor densities and affinities by showing no difference in the density of mu OR-positive neurons.     

Susceptibility of PharmChek drugs of abuse patch to environmental contamination

The key component of the PharmChek sweat patch, the membrane, has been tested for the passage of externally applied materials. Drugs in the uncharged state rapidly penetrated the membrane but charged species were greatly slowed. In basic media, detectable concentrations of cocaine, methamphetamine, and heroin were observed at the earliest collection time (ca. 30 s), after drugs were placed on the outside of the membrane. Drug concentrations increased over the 2 h time course, when amounts detected (1710 ng cocaine, 1060 ng methamphetamine, 550 ng heroin per pad at 2 h) represented 5-17% of the drug deposited on the surface of the sweat patch.Drugs externally applied to human skin were shown to bind readily. Drugs deposited on the skin of drug-free volunteers several days prior to application of the sweat patch were not completely removed by normal hygiene or the cleaning procedures recommended before application of the sweat patch. Even 6 days of normal hygiene did not remove all drugs from externally contaminated skin and positive sweat patches resulted. A mechanism for passage of drugs through the sweat patch membrane, a mechanism for retention of drugs on skin, and a redesign of the sweat patch and modification of its use to reduce external contamination are proposed. Appropriate care should be taken in the interpretation of positive results from a sweat patch test until more research is conducted.     

The effect of metal salt treatment on the photoluminescence of DFO-treated fingerprints

Ninhydrin developed fingerprints can be enhanced by treatment with a zinc or cadmium salt. The resulting fingerprint luminescence has been attributed to the induced coplanarity of the bicyclic indanedione rings of Ruhemann's purple due to complexation with the metal ions. This paper explores whether this effect also occurs in the 1,8-diaza-9-fluorenone (DFO)-amino acid adduct (1), formed from the reaction of DFO with amino acids. Molecular modeling studies of (1) indicate a relatively small out-of-plane angle of 24 degrees. 1H NMR studies indicate (1) is asymmetric about the C2 axis in contrast to what has been previously reported. Little, if any, enhancement of luminescence was observed with Zn, Cd, Ru or Eu treated DFO developed latent fingerprints. This lack of enhancement was also borne out by solution luminescence studies. Given this lack of enhancement of luminescence, solutions of (1) and the four metal ions above were analyzed by electrospray mass spectrometry (ESMS). This indicated the formation of predominantly 1:1 complexes of (1) with both Zn and Cd, and the 2:1 complex with ruthenium. No evidence of a Eu complex was found by ESMS.     

Colchicine poisoning: case report of two homicides

Deaths resulting from the oral ingestion of colchicine are occasionally associated with suicides and therapeutic toxicity. However, homicidal deaths in which this drug is used are extremely rare. Two homicides in which colchicine was used are reported.     

Sensitive and specific multiresidue methods for the determination of pesticides of various classes in clinical and forensic toxicology

Original and sensitive multiresidue methods are presented for the detection and quantitation, in human biological matrices, of 61 pesticides of toxicological significance in human. These methods involved rapid solid-phase extraction using new polymeric support (HLB and MCX) OASIS cartridges. Gas chromatography-mass spectrometry (GC-MS) was used for volatile (organophosphate, organochlorine, phtalimide, uracil) pesticides and liquid chromatography-ionspray-mass spectrometry (LC-MS) for thermolabile and polar pesticides (carbamates, benzimidazoles). Acquisition was performed in the selected ion monitoring (SIM) mode. Extraction recovery varied owing to the nature of pesticides, but was satisfactory for all. Limits of detection (LODs) and limits of quantitation (LOQs) ranged, respectively, from 2.5 to 20 and from 5 to 50ng/ml. An excellent linearity was observed from LOQs up to 1000ng/ml for all the pesticides studied. The proposed procedures yielded reproducible results with good inter-assay accuracy and precision. A few cases of intoxication are presented to demonstrate the diagnostic interest of these methods: in two cases were determined lethal concentrations of endosulfan and carbofuran; in four other cases, the procedures helped diagnose intoxication with, respectively, parathion-ethyl, the association of bromacil and strychnine, bifenthrin and aldicarb.     

Spectra interference between diquat and paraquat by second derivative spectrophotometry

A rapid and accurate method, combining solid-phase extraction and second-order derivative spectrophotomety approaches, is developed for the simultaneous determination of diquat (DQ) and paraquat (PQ) in blood, tissue and urine samples. Supernatant resulting from the precipitation of protein (with trichloroacetic acid) in plasma and tissue or Amberlite IRA-401 resin treated urine are passed through a mini-column packed with Wakogel gel (Silica gel). Analytes are then eluted with a non-organic solvent, 0.2mol/l HCl solution containing 2mol/l NH(4)Cl. UV spectrum of the eluent in 220-350nm range provides effective screen to detect the presence of DQ and/or PQ. In the presence of DQ or PQ alone, the analyte present is quantitated by conventional zero- or second-order derivative spectrophotometry. The calibration curve in the 0.1-5.0mg/l range for either analyte obeys Beer's law. When both DQ and PQ are present, their concentrations are determined by the peak amplitudes of their respective second-derivative spectra after the addition of alkaline dithionite reagent. Interference is negligible when the DQ/PQ concentration ratio is within the 5.0-0.2 range.Using a 2-ml of sample size, the detection limits for DQ and PQ in plasma are 0.02 and 0.005mg/l. The corresponding detection limits for urine samples (10ml sample size) are 0.004 and 0.001mg/l. Recoveries of DQ and PQ in triplicate plasma and urine samples spiked with 0.5mg/l of analytes are 93 and 85%. The precision of the proposed method resulting from triplicate study of spiked urine samples varies from 3.2 to 4.6% at 0.5mg/l of DQ and PQ, respectively.