A pilot study on post-mortem determination of drug abuse on dental tissues.

BACKGROUND: Post-mortem toxicology constantly deals with the research of reliable alternative matrices to be applied in case of highly damaged corpses (such us carbonized, skeletonized, human remains, etc.). Teeth represent a promising alternative matrix since dental tissues are endowed by different features, resistance and stability after death. SCOPE: Since scant literature reported on the pharmacokinetics and mechanism of incorporation of xenobiotics into dental tissues, this pilot research aims to investigate whether in the pulp can be detected the same substances found in blood in drug related death cases. Secondly, the study is addressed to disclose the possible deposit of drugs in dental hard tissues (dentine and/or enamel), thus contributing to reconstruct the drug abuse history (timing, e.g.). MATERIALS AND METHODS: The study experimented with a novel method to separately analyse dental enamel, dentin, and pulp, applied to 10 teeth collected during autopsies of drug-related deaths along with blood and hair samples for classic toxicological analyses. Each tooth was prepared by "pulverization technique" and then analysed by gas chromatography paired with mass spectrometry (GC-MS) and ultra high performance liquid chromatography coupled to high resolution mass spectrometry (UHPLC/HR-MS) for searching cocaine, opiates, and metabolites. The results were then compared with those obtained from blood and hair samples. RESULTS: Preliminary results demonstrated that teeth differ from any other classic matrix (blood and hairs) since the qualitative correspondence of the detected substances between pulp and blood as well as dental hard tissues and hair suggests that they can be useful in post-mortem evaluation as a unique matrix for both acute and chronic assumptions of drugs. The mechanism of accumulation of substances in mineralized dental tissues emerged the most significant result, being influenced by the type of molecule and the method of assumption. The main limitation of this study is the limited availability of the sample and the absence of anamnestic information of the time, rates and method of drug assumption during life. Further research is necessary to systematically investigate the distribution of different substances within the different tissues of the tooth.

Case Series: Hepatic and Splenic Titanium Dioxide Deposition in Association With Intravenous Drug Use.

ABSTRACT: Titanium dioxide is a versatile compound that is found in a variety of consumer products, medical hardware, and pharmaceuticals. Although oral and topical ingestion of this compound is common, intravenous introduction is much less common. We present three cases where significant titanium dioxide deposits were identified in liver and splenic tissue of three decedents, all of whom died of illicit drug overdose in the same geographic area and had fentanyl and its metabolites in blood on postmortem toxicologic testing. At autopsy, liver sections had a granular texture with fine white stippling grossly, and histologic examination of hepatic and splenic tissues showed scattered patches of black granular material with pink birefringence. Energy-dispersive x-ray spectroscopy performed on these tissues revealed the presences of clusters of titanium dioxide. Immunohistochemical staining of both the liver and spleen with CD68 confirmed the titanium dioxide clusters were within macrophages. Intravenous titanium dioxide nanoparticle elimination studies in rats suggest a time sensitive period for this elimination, with a transient period of pigment deposition between 1-58 days following injection. If a time-dependent link between titanium dioxide pigment deposition within tissues and intravenous drug use can be shown, this could be a valuable tool for Pathologists.

Pediatric poisoning management: How clinical practice can benefit from forensic approach.

Pediatric population represents the most vulnerable and at risk for unintentional poisoning, with children younger than 6 years old accounting for nearly half of poison exposures. Poisoning is a time-dependent emergency. The need to reach a scientific agreement on diagnostic protocol and treatment seems to be crucial to reduce morbidity and mortality. Starting from a buprenorphine pediatric intoxication case, this article highlights the limits and pitfalls of the traditional diagnostic approach. Diagnosis of drug intoxication was achieved after several days when an in-depth diagnostic investigation became necessary and complete forensic toxicological analyses were performed. Results evidenced an alarming lack of an unequivocal diagnostic protocol in case of suspect intoxication in structures not provided with a forensic toxicological service/unit. Collection of biological specimens according to forensic protocols at hospitalization plays a paramount role in the definitive diagnosis of intoxication. A diagnostic algorithm that focuses on medical history and biological specimen collection timing is herein proposed, in order to unify emergency approaches to the suspected poisoned child.

Contextual bias in forensic toxicology decisions: A follow-up empirical study from China.

The impact of contextual bias has been demonstrated repeatedly across forensic domains; however, research on this topic in forensic toxicology is very limited. In our previous study, experimental data from only one context version were compared with the actual forensic biasing casework. As a follow-up, this controlled experiment with 159 forensic toxicology practitioners was conducted, to test whether knowledge of different contextual information influenced their forensic decision-making. Participants in different context groups were tasked to identify testing strategies for carbon monoxide and opiate drugs. The results of chi-squared tests for their selections and two context groups exhibited statistically significant differences (p < 0.05 or p < 0.01). These findings show contextual information can bias forensic toxicology decisions about testing strategies, despite it is a relatively objective domain in forensic science.

Stability of Cocaine, Opiates, and Metabolites in Dried Saliva Spots.

Drug abuse still represents a global problem, and it is associated with an increased risk of diseases, injuries, and deaths. Cocaine (COC) and opiates are the most abused drugs and account for a significant number of fatalities. Therefore, it is important to develop methods capable of effectively identifying and quantifying these substances. The present study aims to evaluate the long-term stability of COC, ecgonine methylester (EME), benzoylecgonine (BEG), cocaethylene (COET), norcocaine (NCOC), morphine (MOR), codeine (COD) and 6-monoacetylmorphine (6-MAM) in oral fluid samples. The analytes of interest were isolated from the matrix (50 µL) using the dried saliva spots (DSS) sampling approach and were subsequently analyzed by gas chromatography coupled with tandem mass spectrometry (GC-MS/MS). The parameters that could influence the stability of the target compounds were studied, and these were storage temperature, light, use of preservatives (and respective concentrations), and time. The effects of each parameter were evaluated using the design of experiments (DOE) approach. The stability of the target analytes was improved when the DSS were stored at room temperature, in the presence of light and using 1% sodium fluoride. The best conditions were then adopted for the DSS storage and long-term stability was assessed. COD was only stable for 1 day, EME was stable for 3 days, COC, COET, NCOC and 6-MAM were stable for 7 days, MOR for 14 days and BEG remained stable throughout the study (136 days). This is the first study that evaluates the stability of these compounds in oral fluid samples after application in DSS cards, and optimizes the conditions in order to improve their stability.

Simultaneous Quantification of 25 Fentanyl Derivatives and Metabolites in Oral Fluid by Means of Microextraction on Packed Sorbent and LC-HRMS/MS Analysis.

Fentanyl and fentalogs' intake as drugs of abuse is experiencing a great increase in recent years. For this reason, there are more and more cases in which it is important to recognize and quantify these molecules and related metabolites in biological matrices. Oral fluid (OF) is often used to find out if a subject has recently used a psychoactive substance and if, therefore, the person is still under the effect of psychotropics. Given its difficulty in handling, good sample preparation and the development of instrumental methods for analysis are essential. In this work, an analytical method is proposed for the simultaneous determination of 25 analytes, including fentanyl, several derivatives and metabolites. OF was collected by means of passive drool; sample pretreatment was developed in order to be fast, simple and possibly semi-automated by exploiting microextraction on packed sorbent (MEPS). The analysis was performed by means of LC-HRMS/MS obtaining good identification and quantification of all the analytes in less than 10 min. The proposed method was fully validated according to the Scientific Working Group for Forensic Toxicology (SWGTOX) international guidelines. Good results were obtained in terms of recoveries, matrix effect and sensitivity, showing that this method could represent a useful tool in forensic toxicology. The presented method was successfully applied to the analysis of proficiency test samples.

Review of LC techniques for determination of methadone and its metabolite in the biological samples.

Methadone (MTD) is a synthetic analgesic drug used for treating opioid dependence and effectively used clinically for patients with severe pain. The abuse of MTD may lead to poisoning, disorder in the central nervous system and even death. The regular monitoring of MTD in biological matrices including serum, plasma and urine samples is an effective way to control abuse of MTD. In this manner, the selection of analytical monitoring of MTD in biological matrices is of paramount importance. This study was conducted to review high-performance liquid chromatography (HPLC) techniques carried out on MTD and its main metabolite 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) in the biological samples during 2015-June 2021.

Spatial, temporal and socioeconomic patterns of illicit drug use in New Zealand assessed using wastewater-based epidemiology timed to coincide with the census.

AIMS: A discrete experiment in wastewater-based epidemiology (WBE) timed to coincide with the census was used to investigate the spatial, temporal and socioeconomic patterns of illicit drug consumption in Auckland, Bay of Plenty and Canterbury. METHODS: For seven consecutive days over census week (6 March 2018), wastewater was sampled from seven wastewater treatment plants and analysed for methamphetamine, cocaine (as benzoylecgonine) and 3,4-methylenedioxymethamphetamine (MDMA). Detailed sewer catchment maps were developed and, together with the data, were used to analyse drug consumption. RESULTS: Methamphetamine (mean 22.9 ± 9.9 doses/day/1000 people) was the most consumed drug, followed by MDMA (mean 1.7 ± 1.5 doses/day/1000 people) and cocaine (mean 0.5 ± 0.3 doses/day/1000 people). Methamphetamine consumption (and to a lesser extent MDMA) was high compared to that reported for Western nations, while cocaine consumption was extremely low. Cocaine and MDMA consumption were higher in cities compared to towns. In contrast, methamphetamine was typically higher in towns. Cocaine and MDMA were consumed more at weekends. Methamphetamine use was more consistent throughout the week. MDMA and cocaine were correlated with socioeconomic advantage, whereas methamphetamine was correlated with disadvantage. CONCLUSIONS: This paper contextualises illicit drug use in three New Zealand regions containing 18.3% of the national population and confirms the pervasiveness of methamphetamine consumption in New Zealand towns. This work demonstrates how WBE can be used to explore the socioeconomic dimensions of drug use when duly combined with other data sources like censuses.

Drug-facilitated sexual assault and other crimes: A systematic review by countries.

Drug-facilitated sexual assault (DFSA) and drug-facilitated crime (DFC) constitute a mode of violence that is generally unknown to the population and may go unnoticed by health professionals. The aim of this systematic review was to analyze the victims of DFC, compiling their sociodemographic characteristics, the toxic substances used and their biological matrices and modes of action, in order to identify the substances that are commonly put to criminal use. The aim would be to establish political and health strategies that inform and warn people about possible criminal social behaviors consequent danger to health. This systematic review was conducted following the PRISMA guidelines. Alcohol, benzodiazepines and cocaine were among the most commonly detected substances. In most of the hospitals, immunoassays, liquid chromatography (LC-MS), or gas chromatography-mass spectrometry (GC-MS) analyses were used to identify the substances, while the most frequently used biological matrices were blood and urine. From a judicial point of view, the instrumental protocols and techniques followed for the detection of toxics in different biological matrices must guarantee the reliability and validity of the results for use in a court of law. The recommendations of international organizations should be followed and must be called upon to strengthen their respective national laws against this chemical submission (CS) phenomenon.

Nuclear resonance fluorescence drug inspection.

There is an increasing challenge to prevent illicit drug smuggling across borders and seaports. However, the existing techniques in-and-of-themselves are not sufficient to identify the illicit drugs rapidly and accurately. In the present study, combining nuclear resonance fluorescence (NRF) spectroscopy and the element (or isotope) ratio approach, we present a novel inspection method that can simultaneously reveal the elemental (or isotopic) composition of the illicit drugs, such as widely abused methamphetamine, cocaine, heroin, ketamine and morphine. In the NRF spectroscopy, the nuclei are excited by the induced photon beam, and measurement of the characteristic energies of the emitted [Formula: see text] rays from the distinct energy levels in the excited nuclei provides "fingerprints" of the interested elements in the illicit drugs. The element ratio approach is further used to identify drug elemental composition in principle. Monte Carlo simulations show that four NRF peaks from the nuclei [Formula: see text]C, [Formula: see text]N and [Formula: see text]O can be detected with high significance of 7-24[Formula: see text] using an induced photon beam flux of [Formula: see text]. The ratio of [Formula: see text]/[Formula: see text] and/or [Formula: see text]/[Formula: see text] for illicit drugs inspected are then extracted using the element ratio approach. It is found that the present results of simulations are in good agreement with the theoretical calculations. The feasibility to detect the illicit drugs, inside the 15-mm-thick iron shielding, or surrounded by thin benign materials, is also discussed. It is indicated that, using the state-of-the-art [Formula: see text]-ray source of high intensity and energy-tunability, the proposed method has a great potential for identifying drugs and explosives in a realistic measurement time.

The Increasing Prevalence of Fentanyl: A Urinalysis-Based Study Among Individuals With Opioid Use Disorder in New York City.

BACKGROUND AND OBJECTIVES: Opioid-related overdose deaths in North America have increased drastically, partially due to the increased prevalence of illicitly manufactured fentanyl. The current study sought to assess the prevalence and intentionality of fentanyl use among individuals with opioid use disorder (OUD). METHODS: For this secondary analysis (study 1) we screened a total of 1118 urine samples from 316 participants with OUD from 2016 to 2019. Fentanyl knowledge and intentionality of use were assessed in a separate OUD sample (study 2; N = 33). RESULTS: In study 1, 34.6% of all urine samples tested positive for fentanyl. Overall, 149 (47.2%) participants provided more than or equal to one urine sample that tested fentanyl-positive, and 93 (29.4%) provided more than or equal to two fentanyl-positive samples. The number of fentanyl-positive samples, relative to the number of samples tested each year, increased by 330% from year 1 to 3. Study 2 found all participants had pre-existing knowledge that drugs may be adulterated with fentanyl, yet 67% were surprised by their own fentanyl-positive test result. DISCUSSION AND CONCLUSIONS: Like previous studies, our data indicate the high prevalence of fentanyl exposure and low perception of fentanyl-related risk among individuals with OUD, respectively, suggesting that opioid overdose harm reduction efforts may need to focus more on drug users' understanding of risks related to fentanyl use and adulteration of drugs. SCIENTIFIC SIGNIFICANCE: The current studies provide longitudinal data on fentanyl exposure prevalence and risk perception that is uniquely granular by assessing OUD treatment status, and by identifying potential associations between fentanyl exposure with the presence of other drug use and nonfatal overdose. (Am J Addict 2021;30:65-71).

Drugs in bone: Detectability of substances of toxicological interest in different states of preservation.

In forensic contexts of advanced decomposition, when conventional matrices are no longer available for toxicological analyses, finding alternative matrices is necessary. The skeleton, which is fundamental for anthropologists and geneticists, could be useful also for toxicological purposes. The present study aims to examine what kind of information toxicological analysis performed on bones (the cranium and the ribs) in different states of preservation could provide to the forensic practitioner. Thirty cadavers with known pharmacological history, subjected to forensic autopsy at the Institute of Legal Medicine of Milan, were selected. Rib and cranium samples were collected from each body and separated into two parts in order to create two different states of preservation: One was cleaned from soft tissues and analyzed as a well-preserved bone sample; the other was submitted to a long maceration process, simulating complete skeletonization. All specimens were then processed with accelerated solvent extraction and the eluates analyzed using Q-Exactive™ Orbitrap™ Mass Spectrometer. The analysis of blood and skeletal matrices showed positive results for the tested substances in 63% of cases, mainly benzodiazepines, antidepressants, and drug abuse. Significant Pearson correlations were observed between non-macerated vs. macerated bone samples: r = 0.79 for rib samples, r = 0.61 for cranium samples, and r = 0.69 for all bone samples. As a consequence, the positive results confirm the potential of the bone tissue as an alternative matrix in forensic toxicology, even in cases of extremely decomposed bodies. This study also highlighted important elements for reconstructing the biological profile in cases of forensic anthropological concern.

Simultaneous separation and determination of 32 fentanyl-related substances, including seven sets of isomeric fentanyl analogues, by ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry.

A method for separation and determination of 32 fentanyl-related substances, including seven sets of isomeric fentanyl analogues, was developed using ultra-high-performance liquid chromatography coupled with quadrupole-orbitrap high-resolution mass spectrometry. The collision energy, chromatographic column, and mobile phase were optimized. All compounds were efficiently flushed out of a universal C18 column with a soft gradient consisting of solvent A (2 mM ammonium formate and 0.1% formic acid in water) and solvent B (2 mM ammonium formate and 0.1% formic acid in methanol) in only 20 min, achieving excellent resolution. Detection and analysis were carried out simultaneously in the positive ion mode using the full scan and data-dependent tandem mass spectrometry modes with a normalized collision energy of 40. The method was validated in terms of limit of detection, limit of quantification, linearity, accuracy, and precision. For all fentanyl-related substances, the limit of detection (0.5 ng/mL) and limit of quantification (1 ng/mL) were adequate for screening and quantification in daily drug control. Calibration curves for all compounds were established in the range of 1-500 ng/mL. The intra- and interday precision (RSD%) were within 0.4-2.3 and 0.7-2.7%, respectively. The accuracy ranged from 99 to 106%. The method was applied to analyze seized drug samples.

Aerodynamic Thermal Breakup Droplet Ionization in Mass Spectrometric Drug Analysis.

Aerodynamic thermal breakup droplet ionization (ATBDI) in mass spectrometric drug analysis is considered. Cocaine, heroin, and the main alkaloids of opium (morphine, codeine, papaverine) were chosen as the test compounds. The principles of ATBDI ionization are discussed. The dependences of the intensities of the peaks of the target compounds on temperature during ATBDI ionization are also considered. In some cases, a comparison of ATBDI ionization with electrospray ionization (ESI) was performed. In addition, a comparison of methods is demonstrated by the analysis of confiscated opium that was provided by the local police department. Five major alkaloids are found in opium: morphine, codeine, thebaine, papaverine, and narcotine.

TESTING OF DRUGS IN THE IMPLEMENTATION OF CUSTOMS CONTROL IN UKRAINE: LEGAL ASPECTS.

The purpose of the study is to analyze certain aspects of the legal regulation of the examination of medicines containing narcotic drugs, psychotropic substances, or precursors in the field of customs. To achieve this goal, we analyzed the number of customs examinations carried out, new narcotic compounds identified for the first time. An analysis of the understanding of the definition "examination" and "customs examination" in the scientific literature. Two directions of the implementation of expert examinations of medicines containing narcotic drugs, psychotropic substances, or precursors in the field of customs were identified, its concept was defined. It is proposed to attribute the investigative examination to the methods of customs control. It was found out that this examination is not a forensic examination, and it can be attributed to a special class of examinations, since it has characteristic features inherent only in it: subject, tasks, objects, and research methods. Seven stages of the implementation of expert examinations of medicinal products containing narcotic drugs, psychotropic substances, or precursors during customs control are highlighted and their procedural order is analyzed.

Identification and Confirmation of Fentanyls on Paper using Portable Surface Enhanced Raman Spectroscopy and Paper Spray Ionization Mass Spectrometry.

Fentanyl and its analogues play a major role in the current opioid epidemic. In particular, these highly potent opioids have become a health hazard due to their use as additives in street drugs. Consequently, rapid on-site procedures for the analysis of this class of seized drugs are needed, especially considering the reported backlog of drug samples, which must undergo identification and confirmation tests to validate the presence of an illicit substance. Paper based devices are cheap sampling and analysis vehicles that have been shown capable of allowing rapid identification and confirmation of drugs of abuse. Modifying paper substrates by imprinting nanoparticles enables surface enhanced Raman spectroscopy (SERS) as well as a second analysis from the same substrate, namely paper spray ionization mass spectrometry. While such a procedure has been described for laboratory use, these illicit drug samples are typically collected in the field and this is where testing should be done. We combine paper SERS and paper spray MS on field-portable and commercial off-the-shelf (COTS) devices for the rapid and low-cost identification and confirmation of fentanyl and its analogues, enabling in situ analysis at the point of seizure of suspect samples. The commercial nature of both instruments moves this technology from the academic realm to a setting where the criminal justice system can realistically utilize it. The capabilities of this single-substrate dual-analyzer technique are further examined by sampling a variety of surfaces of forensic interest.

Simultaneous Quantitative Determination of Amphetamines, Opiates, Ketamine, Cocaine and Metabolites in Human Hair: Application to Forensic Cases of Drug Abuse.

A method using liquid chromatography-tandem mass spectrometry (LC-MS/MS) to simultaneously quantify amphetamines, opiates, ketamine, cocaine, and metabolites in human hair is described. Hair samples (50 mg) were extracted with methanol utilizing cryogenic grinding. Calibration curves for all the analytes were established in the concentration range 0.05-10 ng/mg. The recoveries were above 72%, except for AMP at the limit of quantification (LOQ), which was 48%. The accuracies were within ±20% at the LOQ (0.05 ng/mg) and between -11% and 13.3% at 0.3 and 9.5 ng/mg, respectively. The intraday and interday precisions were within 19.6% and 19.8%, respectively. A proficiency test was applied to the validated method with z-scores within ±2, demonstrating the accuracy of the method for the determination of drugs of abuse in the hair of individuals suspected of abusing drugs. The hair concentration ranges, means, and medians are summarized for abused drugs in 158 authentic cases.

A new clean-up approach for stomach content toxicological analysis.

Stomach content is a matrix often applied in post-mortem cases. It is especially important in oral intoxication cases. The main advantages are the usually high concentration of analytes and the lack of biotransformation process. Still, even with extensive sample preparation, the final extract is not always suitable for analysis. The aim of this study was to develop an alternative method using QuEChERS for the extraction of drugs and pesticides from postmortem stomach content. Sample preparation started acetonitrile with 1% of acetic acid and QuEChERS salts. Later, the initial extract was cleaned-up using the EMR-Lipid sorbent. Residual water was withdrawn with MgSO4/NaCl in the third step and a final step with MgSO4. Vigorous shaken and centrifugation was performed after each step. The final supernatant was evaporated, re-suspended, and injected into GC-MS in full scan mode. This approach was successfully applied to stomach content, resulting in clean extracts, with low lipid levels. The method was able to detected target drugs and pesticides (cocaine, tramadol, diazepam, amytriptiline, phenobarbital, prochloraz, diazinon, heptachlor, permethrin, malathion and carbaryl) at the limit of detection of 0.1 mg/g or 0.1 mg/L. Recovery was over 70% for the majority of analytes, precision and accuracy was within acceptable range. The method was also applied to real forensic cases and carbofuran, terbuphos and fluoxetine was detected likewise. This work demonstrates that this method can provide an effective clean-up in high lipids samples such as stomach content, and can be used to analyze of pesticides and drugs in forensic cases.

[Comparison of sample preparation techniques in chemical-toxicological analysis of urine for the presence of narcotic drugs, psychotropic substances and their metabolites]. / Sravnenie protsedur probopodgotovki pri khimiko-toksikologicheskom issledovanii mochi na nalichie narkoticheskikh sredstv, psikhotropnykh veshchestv i ikh metabolitov.

The sample preparation procedures within undirected chemical-toxicological urine analysis for GCMS and HPLC-MS/MS methods are presented. These methods are intended for analytical diagnosis of the presence of narcotic drugs, psychotropic substances and their metabolites in humans.

AEME production in cocaine positive hair after thermal hair treatment.

INTRODUCTION: Currently, hair straightening has become a regular hair treatment for women but likewise for men. Several studies have shown that thermal straightening has an influence on the concentration of ethyl glucuronide and of drugs of abuse content in hair. Heat treatment of hair may decrease concentrations of cocaine (COC) and of cocaethylene (CE) in hair and increase concentrations of benzoylecgonine (BZE). The goal of this study was to evaluate the influence of thermal straightening on anhydroecgonine methyl ester (AEME), a known cocaine smoking marker, in hair. METHOD: 42 positive COC hair samples were treated in vitro with iron plates heated to 200°C. During this treatment one lock of hair was put sequentially 30 times in contact with a hair straightener during 2s, the other lock was not treated. The hair samples were analyzed by a validated GC/MS method for AEME, COC and its metabolites BZE, norcocaine (NC), ecgonine methyl ester (EME) and CE. RESULTS: After treatment, a median increase of concentrations was observed for AEME (110.3%) and BZE (27.6%) whereas a median decrease was found for COC (56.9%), NC (46.7%), EME (33.3%) and CE (41.7%). The median BZE/COC ratio of 0.6 in not treated hair increased to 1.5 in treated hair. CONCLUSION: Regarding our in vitro results, AEME may be produced by thermal hair straightening. Therefore, the presence of AEME in hair should not be used as an irrefutable prove of cocaine smoking. Our study shows that for the interpretation of AEME results in hair, potential heat treatment of hair should be considered. A ratio BZE/COC higher 1 appears to be a good marker to identify thermal treatment of hair before collection. Finally, thermal straightening should be documented during hair collection and should also be considered for the interpretation of COC results in hair.