Wakako Hikiji

Pattern of poisoning in Japan: selection of drugs and poisons for systematic toxicological analysis

Patterns of poisoning are known to be different in different countries, because of the local environmental, cultural, and religious situations. Therefore, in Japan, it is important to know the pattern of poisoning in our own country and to prepare for every poisoning case by establishing an efficient systematic toxicological analysis system in forensic practice. We conducted a retrospective study of the kinds of compounds causing poisonings and the frequency of their use based on two series of reports dealing with poisoning cases in Japan prepared by the National Research Institute of Police Science and the Japanese Society of Legal Medicine for 2003 to 2006. From these reports, 459 and 177 compounds, respectively, were extracted as poisonous compounds over the study period. After data analysis, we selected 314 drugs and poisons as important target compounds for systematic drug analysis in Japan; they included 36 volatile compounds, 14 abused drugs, 170 medical drugs, 60 pesticides, 13 natural toxins, and 21 others. This is the first study to show the toxic drugs and poisons to be analyzed in Japan based on frequency of use, and as such the list will be useful in establishing the most efficient screening system in forensic practice.

Construction of calibration-locking databases for rapid and reliable drug screening by gas chromatography-mass spectrometry

Unique calibration-locking databases were constructed for rapid and semiquantitative drug screening by gas chromatography-mass spectrometry (GCMS). In addition to the free-drug database of 127 drugs, a drug database with acetylating reagents was constructed to increase the number of detectable compounds in the analysis by GC-MS; 156 drugs, including 30 drugs of abuse, 42 hypnotics and their metabolites, 18 antipsychotic drugs, 15 antidepressants, and 12 antipyretic analgesic agents, were registered with parameters, such as the mass spectrum, retention time, qualifier ion/target ion percentage, and calibration curve using the novel GC-MS software NAGINATA. Diazepam-d5 was used as internal standard for construction of each calibration curve in the range of 0.01–5.0 μg/ml for most drugs. We examined the applicability of the constructed database to analyzing whole blood samples spiked with 40 drugs most commonly encountered in toxicological cases in Japan. The drugs in blood were extracted using enhanced polymer columns (Focus), subjected to GC-MS after incubation with acetylating reagents, and screened by the drug database. Among the 40 drugs examined, 38 and 30 drugs were successfully identifi ed at the level of 1 and 0.1 μg/ml, respectively, without using standard compounds. The time required for data analysis was less than 1 min, and semiquantitative data were also obtained simultaneously. Because new drugs and metabolites can easily be added to the databases, we can recommend them as useful tools in clinical and forensic toxicological screening.

Electrolyte analysis of pleural effusion as an indicator of drowning in seawater and freshwater

It is important for forensic pathologists to determine the diagnosis of drowning as well as the site of drowning. In a previous study, we propose that analysis of electrolytes in pleural effusion from rats may be useful for determining whether drowning has occurred in seawater or freshwater. To test this proposal, we measured the concentration of sodium, potassium and chloride ions and total protein in pleural effusion from 40 autopsy cases: 24 involving seawater drowning, 9 freshwater drowning and 7 no drowning. The concentrations of sodium and chloride ions in pleural effusion showed a significant difference between seawater drowning and freshwater drowning. The concentration of potassium ions and total protein showed no difference between each group, although they increased in proportion to the postmortem interval in cases of both seawater and freshwater drowning. These results are almost same as our previous study and, thus, the quantitative analysis of electrolytes in pleural effusion may be useful for determining whether drowning has occurred in seawater or freshwater.

A silent allele in the locus D19S433 contained within the AmpFℓSTR® Identifiler™ PCR Amplification Kit

We present two cases where a single locus mismatch was found in the locus D19S433 using the AmpFlSTR Identifiler PCR Amplification Kit (Applied Biosystems) (Identifiler Kit) during paternity and maternity tests. This mismatch differed from the mismatch pattern where there is usually a one repeat difference. We designed forward and reverse primers so that they were positioned further away from the primer set contained in the Identifiler Kit. The results showed the existence of a silent allele 13 in both families, due to a point mutation that changed guanine to adenine at 32 nucleotides downstream from the 3 end of the AAGG repeat sequences in all four members. A single locus mismatch due to a silent allele may occur in any locus using any kit. Accordingly, we should pay attention to this silent allele when carrying out human identification and parentage analysis.

Estimation of postmortem interval based on the spectrophotometric analysis of postmortem lividity

The color of postmortem lividity and control skin in 21 adult cadavers whose postmortem interval was within 72h, was measured by spectrophotometry in order to estimate the postmortem interval, objectively. The L *a *b * system, which has been widely used for the digital expression of skin color, was used and linear regression analysis was performed to determine the relationship between the postmortem interval and 31 color factors including L * (Value), a * and b * (Chroma and Hue) and C * (Chroma). The difference in Chroma between postmortem lividity and control skin (DeltaC * and DeltaC( *)/C(c)( *)) was only weakly correlated with the postmortem interval. We propose a new equation for calculating the postmortem interval using several color factors obtained by measurement of postmortem lividity, together with some autopsy findings that are known to affect the formation of postmortem lividity. The new equation makes it possible to estimate the postmortem interval within +/-4.76h.

Acute fatal poisoning with pilsicainide and atenolol

A fatal case of intentional poisoning with two antiarrhythmic agents, pilsicainide, a pure sodium channel blocker, and atenolol, a selective β1 blocker, is presented. A woman in her twenties was found dead at home and empty pill packages of pilsicainide, atenolol, and aspirin were found near by. Hesitation marks were found on the wrist, and strong fibrous degeneration was observed in the cardiomyocytes of the sinoatrial node. The blood concentrations of pilsicainide and atenolol were 7.83 and 4.94xa0μg/ml, respectively, both far above the reported therapeutic levels. According to these results, we concluded that death was due to cardiac arrhythmia caused by poisoning with pilsicainide and atenolol. This is the first report of fatal poisoning attributable to an overdose of the combination of these two antiarrhythmic drugs.

False negative result for amphetamines on the Triage® Drug of Abuse panel?

On-site drug screening devices are widely used today for their simple test procedures and instantaneous results. Among other devices, a Triage® Drug of Abuse panel is considered to be highly reliable for its high specificity and sensitivity of abused drugs. Although it is known that a false positive amphetamine (AMP) result may be obtained from the urine samples containing putrefactive amines or ephedrine-related compounds, no clinical false negative methamphetamine results have been reported to date. However, a false negative Triage® result was obtained from the urine of a fatal methamphetamine poisoning victim taking Vegetamine® tablets. Further experimental analyses revealed that the cross-reactivity of methamphetamine and chlorpromazine metabolites, including nor-2-chlorpromazine sulfoxide, was the cause for a false negative Triage® reaction for AMP. Forensic scientists and clinicians must be aware of the limitations of on-site drug testing devices and the need for the confirmatory laboratory tests for the precise identification and quantification of drugs in suspicious intoxication cases, as also recommended by the manufacturers.

An autopsy case of misdiagnosis based on postmortem computed tomography findings.

A middle-aged man was found lying beside his bicycle on an early winter morning. The cause of death was diagnosed by clinicians as traumatic intracerebral hemorrhage and cerebral contusion with frontal bone fracture based on the findings of Computed Tomography (CT) of the head. However, forensic autopsy revealed that there were no evidences of intracerebral hemorrhage and left frontal bone fracture but the defect of golf ball size on the frontal lobe which was considered to be a complication from the old cerebral contusion and old bone fracture. The bleeding and pooling blood from subarachnoid hemorrhage (SAH) to the frontal lobe defect had the appearance of an intracerebral hemorrhage. Disruption of left renal artery was found and the cause of death was diagnosed as massive hemorrhage due to this rupture. Although postmortem CT is a useful tool for obtaining information on the body prior to conducting an autopsy, it should be used with extreme caution.