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The article presents a case of multiple casualties following a textile factory fire. The incident required a full DVI team similar to large mass-disaster because of the specific operational aspects and identification difficulties. The autopsy results were consistent with death by fire and the toxicological investigations revealed carbon monoxide poisoning in four cases (HbCO% ranging between 88,05 and 95,77), two deaths by cyanide intoxication (with concentrations between 5,17 and 8,85 mcg/ml), and in one case there was a synergistic effect of the two substances (carbon monoxide and cyanide). The identification, carried out in accordance with INTERPOL protocols, encountered serious difficulties in the AM phase primarily due to a language barrier and the lack of any dental or medical information relating to the victims. Secondary identifiers proved to be very useful in corroborating possible identities. As a result of the combined efforts of a team of experts the identitity of each victim was determined and in all cases at least one primary identifier was used in the identification process. The deployment of DVI teams composed of forensic experts from different fields of expertise and well acquainted with DVI procedures, is essential in events involving multiple casualities that may also include foreign victims. The DVI team should intervene not only in PM examinations but also in the collection of AM data for those individuals not accounted for and by helping police in contacting families of missing people

L’acido gamma-idrossibutirrico (GHB) è un neurotrasmettitore con effetti gaba-mimetici; si tratta di un composto endogeno, ma notoriamente usato per le applicazioni terapeutiche come anestetico per via endovenosa, come sedativo e per il trattamento della narcolessia. Dagli anni ‘80 è comune sostanza usata come doping, quindi emerge come droga da “tempo libero” nei rave-party; più recentemente, i supposti effetti afrodisiaci e la reale capacità di induzione di amnesia retrograda ne hanno indotto l’utilizzazione quale sostanza in grado di ridurre la capacità di resistenza della vittima di uno stupro annoverandola tra le cosiddette “date rape drugs” (Drug facilitated sexual assault, DFSA), e cioè sostanze psicoattive usate per lo stupro cosiddetto “dopo appuntamento”1. Nel presente contributo si riporta l’approccio metodologico relativamente a due differenti scenari di interesse forense: l’abuso del GHB con quadro di dipendenza patologica e l’uso del GHB come droga da stupro; gli Autori illustrano le modalità di indagine per determinare il GHB nelle differenti matrici biologiche. Conclusioni: gli scenari d’abuso qui presentati rafforzano il concetto che l’analitica tossicologica rivolta alla determinazione del GHB, per tutte le evenienze riguardanti l’uso/abuso e le possibili implicazioni legali, deve realizzarsi secondo canoni analitici convalidati.

This document presents a comprehensive overview of biochemistry and physiology of transferrin (Tf) and carbohydrate-deficient transferrin (CDT) and on the role of CDT as alcohol abuse marker. It discusses the characteristics of the available analytical methodologies for CDT determination and finally presents a set of recommendations. CDT measurements should be performed only in accredited laboratories, having a quality system in place and regularly performing IQC and participating in EQAS. Unequivocal patient identification and a documented chain of custody are needed. Non hemolysed serum is the preferred sample and, if it has to be stored for more than 8 h, freezing at -20°C is recommended. Immunological methods are acceptable only as first level testing. A confirmation of positive findings with analytical methods based on different physicochemical principles is recommended. The result should be expressed as percentage of disialo-Tf on total Tf. Each laboratory should establish its own decision limit based on the characteristics of the analytical method used and on the scope for which the measurement is performed. The employed analytical method should be indicated in the final report.

The use of illicit and non-illicit substances is widespread in suicides. The toxicological data may help in understanding the mechanism of death. This systematic review aimed to analyze autopsies related to suicides by consuming poison, focusing on the correlation between substance use and the country of origin to create an alarm bell to indicate that suicide maybe attempted and prevent it. The systematic review was conducted according to the PRISMA guidelines, with the primary objective of identifying autopsies conducted in cases of suicide by consuming poison in specific geographic areas. Significant differences in substances were observed between low-income and Western countries that confirm previous literature data. In rural areas and Asian countries, most suicides by consuming poison involve the use of pesticides, such as organophosphates and carbamates. In Western countries, illicit drugs and medically prescribed drugs are the leading cause of suicide by self-poisoning. Future research should shed light on the correlation between social, medical, and demographic characteristics and the autopsy findings in suicides by self-poisoning to highlight the risk factors and implement tailored prevention programs worldwide. Performing a complete autopsy on a suspected suicide by self-poisoning could be essential in supporting worldwide public health measures and policy makers. Therefore, complete autopsies in such cases must be vigorously promoted.

Endogenous nature of GHB represents a critical issue for forensic toxicologists, especially in alleged sexual assaults. Therefore, discrimination between physiologically and additional amounts from exogenous sources of such a substance must be effective and reliable in order to avoid severe misinterpretation. This study aimed to quantify the GHB baseline concentrations in chest and pubic hairs collected from 105 healthy volunteers, non-consumers of any drugs of abuse. The final scope was to investigate if these keratin matrices could represent valid alternative to scalp hair when not available. Moreover, we also evaluated the age and gender influences on the GHB baseline levels. 25 mg of hair were incubated overnight with NaOH at 56 °C. After acidification with H2SO4, the solution was liquid-liquid extracted with ethyl acetate and a trimethylsilyl derivatization was then achieved. Analysis was performed in gas chromatography-mass spectrometry in single ion monitoring mode (m/z 233, 234, 147 for GHB; m/z 239, 240 and 147 for GHB-d6). The endogenous amount in “blank” hair was estimated by the standard addition method (0.301 for chest hair and 0.235 ng/mg for pubic hair). GHB concentration ranged from 0.205 to 1.511 ng/mg for chest hair and from 0.310 to 1.913 ng/mg for pubic hair. These values were consistent with previous studies on scalp hair and on pubic hair. Unfortunately, research on chest hair is not available in literature. T-Test and Linear Regression highlighted no statistically significant differences for the two matrices and for all age/gender sub-groups. However, further studies are required to estimate a reliable cut-off value for these keratin matrices. For the first time, we demonstrated the suitability of chest and pubic hair to detect endogenous levels of GHB.

In this work, the physical and chemical properties of a novel zwitterionic LC stationary phase are applied to the development, validation and application of a new fast and reliable method devoted to the analysis of GHB (gamma-hydroxybutyric acid) and its relatively new discovered glucuronide metabolite in hair. The obtained sensitivity, expressed as limit of detection (LOD) and quantification (LOQ), were 0.033 and 0.10 ng/mg for GHB and 0.11 and 0.37 ng/mg, for GHB-glucuronide respectively. Linearity was assessed between LOQ and 50 ng/mg for both compounds. GHB and GHB-glucuronide extraction from hair matrix was maintained simple and consisted in an acidified-solvent incubation. No samples purification was required before LC–MS/MS analysis. The method was finally applied to 65 real hair sample, 60 adults and 5 children below 2 years old. The obtained results highlighted that GHB concentrations were in the range 0.11–0.96 ng/mg (average 0.38 ± 0.25 ng/mg) in 44 cases (68%) while in 21 samples GHB concentrations were in the range between LOD and LOQ (0.033–0.1 ng/mg). GHB-glucuronide was detected in few samples (n. 3) at levels below LOQ. The interest on these molecules relies on the fact that GHB is both a naturally occurring inhibitory neurotransmitter in the central nervous system and an illicit drug often experienced by victims of drug-facilitated sexual assault. GHB-glucuronide was firstly identified in urine by the group of Petersen in 2013 and, as per analogy to ethyl glucuronide, it was proposed as a longer biomarker for GHB intoxication.

Drug tampering practices, with the aim to increase availability of drug delivery and/or enhance drug effects, are accessible on Internet and are practiced by some portion of recreational drug users. Not rarely, recreational misuse may result in toxic and even fatal results. The aim of the present study was to assess the tampering risk of medicaments containing different formulations of an opioid in combination with paracetamol or dexketoprofen, following the procedures reported in dedicated forums on the web. Tablets and suppositories containing codeine, tramadol and oxycodone were extracted following the reported “Cold water extraction”; dextromethorphan was extracted from cough syrup following the procedure reported as “Acid/base extraction” and fentanyl was extracted from transdermal patches according the procedure reported in Internet. The tampered products and opportunely prepared calibrators in water were analysed by liquid chromatography coupled to tandem mass spectrometry (LC–MS/MS). The separation of the analytes was carried on Agilent ZORBAX Eclipse Plus C18 (RRHT 2.1 mm × 50 mm, 1.8 μm) by the gradient elution of 0.01% formic acid in water and 0.01% formic acid in methanol. Acquisition was by MRM mode considering at least two transitions for compound. Declared recoveries for these home-made extractions claimed to exceed 99% for the opioid and to complete remove paracetamol, often associated to liver toxicity and thus to obtain a “safer” preparation. In this study, the authors demonstrated that rarely the recoveries for the opioid reached 90% and that up to 60% of the paracetamol amount remained in solution. Thus, high risks for health remained both for the potential lethality of the opioid content, but also for the sub-lethal chronic use of these mixtures, which contained still uncontrolled, ignored, but often important amounts of paracetamol.

Comprehension of illicit drug market's features at local level is useful to plan and to correctly set-up specific informative and contrast activities. In this paper we report trends, purities and consumption estimations of illicit substances available on the Florentine territory from 2006 to 2016. These data were obtained by the analysis of 10,451 samples seized by the Law Enforcement Agencies in case of personal use offence. Analytical procedures consisted in targeted and untargeted analyses by gas chromatography-flame ionization detector, gas chromatography–mass spectrometry and liquid chromatography–tandem mass spectrometry. The most detected substances were: cannabis (78.0%; resin: 51.7%; herb: 26.3%), cocaine (10.4%), opiates (6.6%; heroin: 6.5%; morphine: 0.1%), ketamine (1.4%), amphetamines (1.3%; 3,4-methylenedioxymethamphetamine – MDMA –: 0.7%; methamphetamine: 0.6%; amphetamine

A gas chromatography-mass spectrometry (GC-MS) and a liquid chromatography tandem mass spectrometry (LC-MS/MS) method were validated for quantifying endogenous and exogenous hair concentrations of gamma-hydroxybutyrate (GHB). The GC-MS method is based on overnight extraction of 25 mg hair in NaOH at 56 °C, liquid/liquid extraction in ethylacetate and trimethylsylil derivatization; analysis is by electron ionization and single ion monitoring of three ions. The LC-MS/MS method entails a rapid digestion of 25 mg hair with NaOH at 75 °C for 40 min, liquid/liquid extraction in ethylacetate and reconstitution of the extract in the LC mobile phase; negative ion electrospray ionization and multiple reaction monitoring (MRM) analysis are employed for the LC-MS/MS detection. In both cases, GHB-d6 is used as an internal standard. The endogenous amount in "blank" hair are estimated by the standard addition method. Limits of detection are 0.4 and 0.5 ng/mg for GC-MS and LC-MS/MS respectively, while the limit of quantification (LOQ) is 0.6 ng/mg for both methods; the GC-MS method proved to be linear in the range 1-50 ng/mg whereas linearity was demonstrated from 0.6 to 50 ng/mg for the LC-MS/MS; imprecision and inaccuracy were always lower than 23% for quality controls samples. The two methods were applied to a real case of a man addicted to GHB; the drug concentration in segments from 17 cm hair strand well correlated with self-reported use of GHB in different periods of his life. Performances of the two methods were similar.