Quantitation using GC-TOF-MS: example of bromazepam

Time-of-flight mass spectrometry (TOF-MS) offers new perspectives for forensic toxicology. Qualitative and quantitative analyses of a mixture of three selected benzodiazepines (diazepam, nordazepam and bromazepam) were used to compare gas chromatography (GC-TOF-MS, quadrupole GC-MS, GC-ECD) and liquid chromatography (HPLC-DAD) data. Method validation parameters like LOD, LOQ, S/N-ratios reflect the capabilities of GC-TOF-MS. Five-point calibrations for bromazepam in human peripheral blood (50, 100, 160, 200, 300 ng/ml) using medazepam as internal standard (1000 ng/ml) were performed. The calibrations using GC-TOF-MS (using the fragments of m/z 236 and 288), GC-ECD (dual system) and HPLC-DAD (at 235 nm) all showed correlation coefficients close or superior to 0.99. Quadrupole GC-MS data was not used in the comparison of extracted samples due to the low sensitivity in the full scan mode. Two analyses of real cases concerning bromazepam are presented. In the first case, the presence or absence of bromazepam could not be established with both HPLC-DAD and GC-ECD due to background signals. The extracted ion chromatograms and spectrum traces after the analysis with the GC-TOF-MS could clearly excluded the presence of bromazepam. The second case illustrates the quantitation of bromazepam, where both HPLC-DAD and GC-ECD were unable to give satisfactory results, again due to interfering background signals. The analyses performed on the GC-TOF-MS-system demonstrated high sensitivity and also high selectivity due to the high quality of mass spectra obtained. The advantages of GC-TOF-MS make it a promising analytical technique for forensic toxicology.     

Forensic medicine experiences in cremation autopsy in Bonn

2016 coroner's crematorial inquests of the Bonn Institute of Legal Medicine over a 3 years' period were evaluated and the statements in the death certificates were compared to those ones of the Statistical Country Office of Northrhine-Westphalia. No significant differences were found with respect to age, gender and reason of death. However, the number of insufficiently classified deaths with respect to the reason of death were overrepresented especially in cases of older women, while the number of unnatural deaths was generally underrepresented. According to legal medical criteria 2 1/4 per cent of the deaths were falsely classified as natural. Only in 2 cases an autopsy was accomplished. Other author's statements on the validity of death certificates were confirmed.     

An unusual case of self-harm with petroleum distillate

After surgical treatment of an inguinal hernia, a 26-year-old male was discharged from hospital with a well healing surgical wound. Four weeks later he was admitted again with a major localized painful swelling of the soft tissues, whereas the clinical examination revealed no further local or systemic signs of inflammation. The immediate operation disclosed no abscess formation, but a large colliquation necrosis of the subcutaneous tissue with an outstanding petroleum-like odour. On toxicological examination of the necrotic tissue, a respective petroleum distillate was found. The patient was confronted with the suspicion of having injected the petroleum preparation himself to prolong the wound healing and finally admitted the self-mutilating behaviour. The pathophysiology, clinical history and morphological findings are discussed in comparison with case reports of chemically induced self-mutilation.     

Population genetics of the D1S1656, D12S391, and D18S535 loci in Asturias (North Spain)

Allele and genotype frequencies for three recently described short tandem repeat loci D1S1656, D12S391, and D18S535 were determined in a population sample from Asturias (North Spain). The loci were amplified using a fluorescence based PCR method and were typed automatically. No deviation from Hardy-Weinberg expectations were observed. The three loci proved to be highly discriminating and the allele frequencies observed are similar to those of the other European populations that have been typed for these loci to date.     

Threat mail and forensic science: DNA profiling from items of evidence after treatment with DFO

Two cases of threatening letters with their accompanying envelopes were received to the Division of Forensic Identification unit of the Israel Police. The envelopes, including the stamps, and the letters were initially examined for latent fingerprints by the DFO reagent, known to cause degradation of DNA. Although no latent fingerprints could be visualized on any of the items, the biology laboratory using organic DNA extraction, was successful in defining genetic profiles from all the items employing six STR loci, even after treatment with DFO. In a controlled experiment, a known donor attached a stamp, by licking, to an envelope. This item was treated with DFO and then profiled using STR loci. The results showed that previous DFO treatment on the control stamp before DNA analysis had no negative effects on obtaining the DNA profile of the known donor using STR loci.     

The first mail bomb?

A news item in the newspaper, the Evening Star, for the Country, October 18, 1833, probably is the first report of a mail bomb in the United States. It is also likely to be the first defusing of a mail bomb. The device appeared to be extraordinarily sophisticated and would have likely worked had it not been for suspicions of the recipient of the bomb.     

The Slovenian population data on the PCR based loci HLA-DQA1, LDLR, GYPA, HBGG, D7S8, GC, and D1S80

Allele frequencies for the loci HLA-DQA1, LDLR, GYPA, HBGG, D7S8, GC, and D1S80 were determined for a sample population of unrelated individuals from Slovenia. All loci meet Hardy-Weinberg expectations, except the loci GYPA (p = 0.041) and D1S80 (p = 0.009). There is little evidence for association of alleles among the seven loci. Only one out of 21 pairwise comparisons demonstrated departures from independence (HLA-DQA1/HBGG, p = 0.008). The allelic frequency data generally are similar to that of U.S. Caucasians.