Targeted screening and quantification of synthetic cathinones and metabolites in hair by UHPLC-HRMS.

OBJECTIVE: Synthetic cathinones (SCs) are new psychoactive substances with sympathomimetic effects, which emerged into the illegal drug market to replace controlled stimulants. Since every year more powerful and toxic substances enter the illicit market, there is the need for analytical methodologies able to detect these new compounds in conventional and non-conventional biological matrices. We sought to develop and validate a targeted screening and quantification method for thirty-two parent SCs and two metabolites in hair samples by ultra-high-performance liquid chromatography coupled to high resolution mass spectrometry (UHPLC-HRMS). MATERIALS AND METHODS: 20 mg hair samples were soaked in 250 µL of 2 mM ammonium formate, methanol and acetonitrile mixture (50/25/25, v/v/v) and incubated overnight at 40°C. After incubation, the samples were evaporated to dryness under nitrogen stream and reconstituted with 100 µL of mobile phase mix (A:B, 80:20) and 10 µL were injected into UHPLC-HRMS. A Q ExactiveTM Focus Orbitrap Mass spectrometer with full scan and targeted data-dependent MS/MS scan acquisition was used for the screening and quantitation analysis. RESULTS: The assay was linear from 5 to 500 pg/mg hair for all the analytes under investigation. Intra-day and inter-day precision were always < 15% and matrix effect and analytical recovery were always within acceptable criteria (±25% and >50%, respectively). The developed method was applied to authentic hair samples from SCs consumers. The most prevalent found SCs were 3,4-Methylenedioxy-α-Pyrrolidinohexanophenone with a concentration range of 6.0-1,000.0 pg/mg along with α-Pyrrolidinohexiophenone (54.0 and 554.0 pg/mg, respectively), 3-Methylmetcathinone (556.0 and 5,000.0 pg/mg) and 4-Methylethcathinone (11.5 and 448.0 pg/mg) CONCLUSIONS: The developed method showed good selectivity, specificity, an easy and low-cost sample preparation and an analysis time compatible with a high throughput laboratory.

Commentary - Heroin purity and adulteration: an updated snapshot from the Italian Early Warning System.

Not Available.

Synthetic cathinones related fatalities: an update.

OBJECTIVE: Synthetic cathinones, more commonly known as "bath salts", are synthetic drugs chemically related to cathinone, a psychostimulant found in the khat plant. They are the first most consumed products among new psychoactive substances, which cause psychostimulant and hallucinogenic effects determining a number of fatalities worldwide.  In this paper, we have systematically reviewed cases of synthetic cathinones-related fatalities analytically confirmed, which have occurred in the last few years. MATERIALS AND METHODS: Relevant scientific articles were identified in Medline, Cochrane Central, Scopus, Web of Science and Institutional/government websites up to November 2017 using the following keywords: synthetic cathinones, mephedrone, methylenedioxypyrovalerone, MDPV, methylone, ethylone, buthylone, fatal intoxication, fatalities and death. RESULTS: In total, 20 citations met the criteria for inclusion, representing several fatal cases with analytically confirmed synthetic cathinones in biological sample/s of the deceased. The death was attributed to hyperthermia, hypertension, cardiac arrest and more in general to the classic serotonin syndrome. Only rarely did the concentration of the parent drug causing fatality overcome the value of 1 mg/L in post-mortem biological fluids. CONCLUSIONS: Abuse of synthetic cathinones still represents a serious public health issue. Systematic clinical studies on both the animal and human model are lacking; therefore, the only available data are from the users who experience the possible hazardous consequences. Analytical methodologies for the identification of parent compounds and eventual metabolites both in ante-mortem and post-mortem cases need to be developed and validated. Analytical data should be shared through different communication platforms with the aim of stopping this serious health threat for drug users.

Hepatotoxicity associated with illicit use of anabolic androgenic steroids in doping.

Anabolic Androgenic Steroids (AAS) abuse and misuse is nowadays a harmful habit involving both professional or recreational athletes, as well as general population. AAS are also frequently present in over-the-counter dietary supplements without being declared in the list of ingredients, leaving consumers unaware of the risks of adverse effects. Indeed, health risks of AAS consumption in pharmaceutical preparations or dietary complements seem still underestimated and under-reported. The variety of complications due to AAS misuse involves cardiovascular, central nervous, musculoskeletal and genitourinary systems of both males and females; psychiatric and behavioral effects, damages to metabolic system, skin and mainly liver. For instance, relevant concern has been raised by the AAS hepatotoxicity including adenoma, hepatocellular carcinoma, cholestasis, and peliosis hepatis. The present review reports the information available on the hepatotoxic effects of AAS use in professional and amateur athletes.

"Vehicular homicide", a new Italian offence: law provisions and comparison with some other European countries.

According to the most recent World Health Organization report, road accidents represent a very serious public health issue, claiming each year more than 1.2 million lives worldwide and being the leading cause of death among young people aged between 15 and 29 years. Up to now, the policies to reduce this issue are different, unbalanced and often inadequate not only in Italy, but also in the other European Countries. Specifically, the Italian Parliament has recently introduced a new law (n. 41 of March 23rd, 2016), making vehicular homicide together with road traffic injuries a criminal offense, both to be punished as a result of negligence. The measure came into force on March 25th, 2016. In this paper, the provisions of the above-reported law have been assessed, taking into account the modifications introduced in the Penal Code by this law and the impact it is having and will have on drivers on a day-to-day basis. Similarities and differences with legislative framework of some other European countries were also examined. Finally, some open questions to be solved are named as an open eye for future considerations.

[Alcohol and work. Consensus Document of the La.R.A. (Workers representing a risk for others) group]. / Alcol e Lavoro. Documento di consenso del gruppo La.R.A.

AIM: Current Italian legislation obliges employers to prevent workers who are occupationally at risk or who perform jobs that may be hazardous for the safety or health of third parties from consuming alcohol. The LaRA Group undertook to assess whether the law fully safeguards the health and safety of both workers and third parties, without impinging upon the civil rights of workers. METHOD: A written document expressing agreement was produced following discussions between doctors, lawyers, bioethicists and social partners. RESULTS: There are gaps and inconsistencies in current laws; the differences in local and regional provisions prevent authorities from applying a single strategy at national level. There should be a change in existing rules under which the employer's obligation to enforce the ban on consumption alcohol in the workplace is enacted solely by the "competent" physician whose institutional role is to safeguard and promote health. Some occupational categories that are subject to a ban on alcohol consumption do not currently under-go health surveillance. For example, if road transport drivers are not exposed to a specific occupational risk foreseen under another law, they can be placed under health surveillance only in those regions where the local laws contemplate this type of control. In other cases, the practice of assessing the risk to third parties and providing for compulsory health surveillance in the Risk Assessment Document, is considered by some jurists to be a "consuetudo praeter legem" and therefore acceptable in a field not yet covered by a specific law, but to be "contra legem" or unlawful by other jurists. Moreover, the competent physician who uses a breathanalyser or tests for alcohol addiction faces an ethical dilemma, since by communicating the results to an employer or authorities responsible for the issuing of licenses, he may be violating his professional oath of secrecy. Furthermore, the emphasis placed on testing has induced companies and inspectors to overlook educational and rehabilitation aspects. It is essential to involve general practitioners, educators and specialist services in addressing the problems of alcohol abuse so as to inform/train, recover and rehabilitate. The few studies available indicate that the rules are poorly enforced and that non-compliance may go unobserved. CONCLUSIONS: The Group urges all employers to assess the risk for third parties caused by alcohol abuse and to devise a policy on alcohol. Controlling alcohol-related risks in the workplace calls for a better definition of the roles of Vigilance Bod-ies and Company Physicians together with a shift from a reactive to a proactive attitude of all the parties involved.

Metilfenidato en el tratamiento del trastorno de déficit de atención con hiperactividad en pediatría: monitorización en matrices biológicas / Methylphenidate in the treatment of children with attention-deficit hyperactivity disorder: Monitoring in biological matrices

En los últimos años, el trastorno por déficit de atención e hiperactividad (TDAH) se ha convertido en el trastorno psiquiátrico más frecuentemente diagnosticado y tratado en población pediátrica. En los años 80 fue aprobado en España el metilfenidato (MFD), un fármaco psicoestimulante, para el tratamiento sintomático del TDAH. Desde entonces, se ha convertido en uno de los medicamentos más ampliamente estudiado y prescrito tanto en niños como en adultos. En este artículo se revisan los principios farmacológicos del MFD especialmente su farmacocinética en matrices biológicas convencionales (sangre, orina) y no convencionales (cabello, saliva y sudor), preparados farmacéuticos, concentraciones terapéuticas y efectos indeseables(AU)

Attention-deficit hyperactivity disorder (ADHD) has emerged in the last few years as the most commonly diagnosed and treated psychiatric disorder in the paediatric population. In 1980's, methylphenidate (MFD) a psychomotor stimulant drug, was approved in Spain for the symptomatic therapy of ADHD. Since then, MFD has become one of the most extensively prescribed and studied treatment for ADHD both in children and adults. In this paper, the main pharmacological issues of MFD are reviewed, focusing on its pharmacokinetics in conventional (blood and urine) and non-conventional (hair, oral fluid and sweat) biological matrices, its pharmaceutical preparations, therapeutic levels and side effects(AU)

[Methylphenidate in the treatment of children with attention-deficit hyperactivity disorder: monitoring in biological matrices]. / Metilfenidato en el tratamiento del trastorno de déficit de atención con hiperactividad en pediatría: monitorización en matrices biológicas.

Attention-deficit hyperactivity disorder (ADHD) has emerged in the last few years as the most commonly diagnosed and treated psychiatric disorder in the paediatric population. In 1980's, methylphenidate (MFD) a psychomotor stimulant drug, was approved in Spain for the symptomatic therapy of ADHD. Since then, MFD has become one of the most extensively prescribed and studied treatment for ADHD both in children and adults. In this paper, the main pharmacological issues of MFD are reviewed, focusing on its pharmacokinetics in conventional (blood and urine) and non-conventional (hair, oral fluid and sweat) biological matrices, its pharmaceutical preparations, therapeutic levels and side effects.

Matrices biológicas alternativas para detectar la exposición prenatal a drogas de abuso en el tercer trimestre de la gestación / Alternative biological materials to detect prenatal exposure to drugs of abuse in the third trimester of pregnancy

Introducción: La detección de la exposición prenatal a drogas de abuso es fundamental para asegurar un adecuado seguimiento de los niños afectados. El cuestionario materno no es una herramienta de cribado eficiente. En los últimos años, se ha descrito la utilidad del cabello materno y del meconio como matrices biológicas para valorar esta exposición. El objetivo de este estudio es comparar ambas matrices alternativas en la detección de la exposición prenatal a drogas en el tercer trimestre del embarazo, con la finalidad de valorar su uso como herramienta de cribado. Pacientes y métodos: Entre enero y marzo de 2010 se recogieron muestras de cabello materno y meconio de 107 parejas madre-recién nacido del Hospital Can Misses de Ibiza. Se determinó en ambas matrices la presencia de opiáceos, cocaína, cannabis y anfetaminas, utilizando técnicas cromatográficas estandarizadas. Resultados: El análisis del cabello materno fue positivo para drogas de abuso en 17 casos (15,9%): 11 a cannabis, 7 a cocaína, uno a cannabis y éxtasis, y uno a cannabis y cocaína. Solo una madre había declarado consumo de cannabis y otra de cocaína. De los 7 casos positivos para cocaína en cabello, 6 se confirmaron en el estudio de meconio, mientras que de los 11 casos positivos para cannabis, solo 3 fueron confirmados en meconio. Se definieron 2 perfiles diferenciados de consumidoras: cocaína y cannabis (solo se detectaron 2 casos de policonsumo). Se detectó un caso con los valores de cocaína en meconio más altos publicados (1.582ng/g). Discusión: Este estudio revela una elevada prevalencia del consumo de drogas de abuso durante el embarazo en esta cohorte. La mejora en los métodos de cribado podría optimizar la prevención y el seguimiento de los recién nacidos expuestos. El cabello materno parece ser más sensible que el meconio para detectar la exposición prenatal a cannabis durante el tercer trimestre, por lo que podría convertirse en una buena herramienta de cribado(AU)

Introduction: Detection of prenatal drug abuse exposure is essential to ensure an appropriate monitoring of affected children. A maternal questionnaire is not an efficient screening tool. The usefulness of maternal hair and meconium as biological materials to assess this exposure has been described in last few years. The aim of this study was to compare both these alternative biological materials for prenatal drug exposure detection in the third trimester of pregnancy, in order to assess its use as a screening tool. Patients and methods: Between January and March 2010, samples of maternal hair and meconium from 107 mother-infant dyads were collected in Can Misses Hospital, Ibiza. The presence of opiates, cocaine, cannabis, and amphetamines, was determined in both materials, using standard chromatographic techniques. Results: Maternal hair analysis showed a 15.9% positivity for drugs of abuse (17 cases): 11 cannabis, 7 cocaine, 1 cannabis and ecstasy, and 1 cannabis and cocaine. Only one mother reported cannabis consumption and another one, cocaine. Of the 7 cocaine positive cases in hair, 6 were confirmed in meconium analysis, while of 11 cannabis positive cases, only 3 were confirmed in meconium. Two different consumer profiles were defined: cocaine consumers and cannabis consumers (with only 2 cases of multiple drug use). The highest level of cocaine ever published was detected (1.582ng/g) in one case. Discussion: This study reveals a high prevalence of drug abuse in this cohort during pregnancy. Improved screening methods may optimize prevention and monitoring of exposed infants. Maternal hair seems to be more sensitive than meconium to detect prenatal exposure to cannabis during the third trimester, so it might become a good screening tool(AU)

Validez del cuestionario de consumo materno de alcohol para detectar la exposición prenatal / Validity of a maternal alcohol consumption questionnaire in detecting prenatal exposure

Introducción: El consumo de alcohol en mujeres embarazadas puede producir graves efectos adversos en el feto y el recién nacido principalmente a nivel de desarrollo neurológico y pondoestatural, englobados en el término FASD (acrónimo en inglés de trastorno del espectro alcohol fetal). El método de cribado más utilizado para detectar la exposición prenatal es el cuestionario, pero un estudio poblacional previo ha cuestionado la fiabilidad del método. El objetivo de este estudio es comparar la detección de la exposición prenatal al alcohol mediante el cuestionario de consumo y la presencia de biomarcadores en meconio. Metodología: Se estudiaron 62 muestras de meconio de recién nacidos cuyas madres negaron el consumo de alcohol durante el embarazo en el cuestionario realizado. Se llevó a cabo una determinación objetiva de la exposición del feto a alcohol utilizando el meconio del recién nacido como matriz biológica y los FAEE (fatty acid ethyl esters) como biomarcadores de exposición. Resultados: En el meconio de 10 de los 62 recién nacidos de mujeres que negaron el consumo de alcohol durante el embarazo en el cuestionario (16,12%), se obtuvieron valores totales de los FAEE analizados positivos (iguales o superiores a 2 nmol/g).Discusión: Los cuestionarios realizados como método de cribado para descartar la exposición a etanol durante el embarazo no deben considerarse una herramienta eficiente. Es necesaria la determinación de biomarcadores en matrices biológicas alternativas de la madre o del recién nacido. La detección precoz de la exposición prenatal permitirá a estos pacientes beneficiarse de un seguimiento y tratamiento con el que alcanzarán el mejor desarrollo neurológico posible(AU)

Introduction: Ethanol consumption by pregnant women can produce severe effects in the foetus and the newborn, mainly in neurological and weight-height development, and are included in the term FASD (Fetal Alcohol Spectrum Disorder). Questionnaires are the most used screening method to detect prenatal exposure, but a previous population study questioned its reliability. The objective of this study was to compare alcohol prenatal exposure detection by questionnaire compared with biomarkers in meconium. Methodology: Sixty two meconium samples from mothers who denied alcohol consumption during pregnancy by questionnaire were analysed. The objective analysis was made by determination of FAEEs (fatty acid ethyl esters) as exposure biomarkers in meconium as biological matrix. Results: In the meconium from 10 of 62 newborns from non-alcohol consuming mothers by questionnaire (16.12%) FAEE values were positive (minor=2 nmol/g). Discussion: Questionnaires as a screening method during pregnancy are not a reliable tool. It is necessary to identify prenatal exposure to alcohol as soon as possible by biomarkers analysis in biological matrices from the newborn or the mother. The early detection will allow these patients to benefit from follow up and treatment to reach the best possible neurological development(AU)

[Alternative biological materials to detect prenatal exposure to drugs of abuse in the third trimester of pregnancy]. / Matrices biológicas alternativas para detectar la exposición prenatal a drogas de abuso en el tercer trimestre de la gestación.

INTRODUCTION: Detection of prenatal drug abuse exposure is essential to ensure an appropriate monitoring of affected children. A maternal questionnaire is not an efficient screening tool. The usefulness of maternal hair and meconium as biological materials to assess this exposure has been described in last few years. The aim of this study was to compare both these alternative biological materials for prenatal drug exposure detection in the third trimester of pregnancy, in order to assess its use as a screening tool. PATIENTS AND METHODS: Between January and March 2010, samples of maternal hair and meconium from 107 mother-infant dyads were collected in Can Misses Hospital, Ibiza. The presence of opiates, cocaine, cannabis, and amphetamines, was determined in both materials, using standard chromatographic techniques. RESULTS: Maternal hair analysis showed a 15.9% positivity for drugs of abuse (17 cases): 11 cannabis, 7 cocaine, 1 cannabis and ecstasy, and 1 cannabis and cocaine. Only one mother reported cannabis consumption and another one, cocaine. Of the 7 cocaine positive cases in hair, 6 were confirmed in meconium analysis, while of 11 cannabis positive cases, only 3 were confirmed in meconium. Two different consumer profiles were defined: cocaine consumers and cannabis consumers (with only 2 cases of multiple drug use). The highest level of cocaine ever published was detected (1.582ng/g) in one case. DISCUSSION: This study reveals a high prevalence of drug abuse in this cohort during pregnancy. Improved screening methods may optimize prevention and monitoring of exposed infants. Maternal hair seems to be more sensitive than meconium to detect prenatal exposure to cannabis during the third trimester, so it might become a good screening tool.

Determination of maternal-fetal biomarkers of prenatal exposure to ethanol: a review.

The deleterious effects exerted by prenatal ethanol exposure include physical, mental, behavioural and/or learning disabilities that are included in the term fetal alcohol spectrum disorder (FASD). Objective assessment of exposure to ethanol at both prenatal and postnatal stages is essential for early prevention and intervention. Since pregnant women tend to underreport alcohol drinking by questionnaires, a number of biological markers have been proposed and evaluated for their capability to highlight gestational drinking behaviour. These biomarkers include classical biomarkers (albeit indirect) of alcohol-induced pathology (mean corpuscular volume (MCV), gamma glutamyltransferase (GGT), aspartate aminotransferase (AST) and alanine aminotransferase (ALT)) acetaldehyde-derived conjugates, and finally derivatives of non-oxidative ethanol metabolism (fatty acid ethyl esters (FAEEs), ethyl glucuronide (EtG), ethyl sulphate (EtS) and phosphaditylethanol (PEth)). Since ethanol itself and acetaldehyde are only measured few hours after ethanol intake in conventional matrices such as blood, urine and sweat, they are only useful to detect recent ethanol exposure. In the past few years, the non-oxidative ethanol metabolites have received increasing attention because of their specificity and in some case wide time-window of detection in non-conventional matrices from the pregnant mother (oral fluid and hair) and fetus-newborn (neonatal hair, meconium, placenta and umbilical cord). This article reviews bioanalytical procedures for the determination of these markers of ethanol consumption during pregnancy and related prenatal exposure. In addition, clinical toxicological applications of these procedures are presented and discussed.

[Validity of a maternal alcohol consumption questionnaire in detecting prenatal exposure]. / Validez del cuestionario de consumo materno de alcohol para detectar la exposición prenatal.

INTRODUCTION: Ethanol consumption by pregnant women can produce severe effects in the foetus and the newborn, mainly in neurological and weight-height development, and are included in the term FASD (Fetal Alcohol Spectrum Disorder). Questionnaires are the most used screening method to detect prenatal exposure, but a previous population study questioned its reliability. The objective of this study was to compare alcohol prenatal exposure detection by questionnaire compared with biomarkers in meconium. METHODOLOGY: Sixty two meconium samples from mothers who denied alcohol consumption during pregnancy by questionnaire were analysed. The objective analysis was made by determination of FAEEs (fatty acid ethyl esters) as exposure biomarkers in meconium as biological matrix. RESULTS: In the meconium from 10 of 62 newborns from non-alcohol consuming mothers by questionnaire (16.12%) FAEE values were positive (≥ 2 nmol/g). DISCUSSION: Questionnaires as a screening method during pregnancy are not a reliable tool. It is necessary to identify prenatal exposure to alcohol as soon as possible by biomarkers analysis in biological matrices from the newborn or the mother. The early detection will allow these patients to benefit from follow up and treatment to reach the best possible neurological development.

The standardization of results on hair testing for drugs of abuse: An interlaboratory exercise in Lombardy Region, Italy.

Hair testing for drugs of abuse is performed in Lombardy by eleven analytical laboratories accredited for forensic purposes, the most frequent purposes being driving license regranting and workplace drug testing. Individuals undergoing hair testing for these purposes can choose the laboratory in which the analyses have to be carried out. The aim of our study was to perform an interlaboratory exercise in order to verify the level of standardization of hair testing for drugs of abuse in these accredited laboratories; nine out of the eleven laboratories participated in this exercise. Sixteen hair strands coming from different subjects were longitudinally divided in 3-4 aliquots and distributed to participating laboratories, which were requested to apply their routine methods. All the participants analyzed opiates (morphine and 6-acetylmorphine) and cocainics (cocaine and benzoylecgonine) while only six analyzed methadone and amphetamines (amphetamine, methamphetamine, MDMA, MDA and MDEA) and five Δ(9)-tetrahydrocannabinol (THC). The majority of the participants (seven labs) performed acidic hydrolysis to extract the drugs from the hair and analysis by GC-MS, while two labs used LC-MS/MS. Eight laboratories performed initial screening tests by Enzyme Multiplied Immunoassay Technique (EMIT), Enzyme-linked Immunosorbent Assay (ELISA) or Cloned Enzyme Donor Immunoassay (CEDIA). Results demonstrated a good qualitative performance for all the participants, since no false positive results were reported by any of them. Quantitative data were quite scattered, but less in samples with low concentrations of analytes than in those with higher concentrations. Results from this first regional interlaboratory exercise show that, on the one hand, individuals undergoing hair testing would have obtained the same qualitative results in any of the nine laboratories. On the other hand, the scatter in quantitative results could cause some inequalities if any interpretation of the data is required.

Maternal hair testing for the assessment of fetal exposure to drug of abuse during early pregnancy: Comparison with testing in placental and fetal remains.

UNLABELLED: Drug use by pregnant women in the first trimester of pregnancy and subsequent fetal exposure during early gestation can be assessed only by repetitive/systematic maternal blood/urine analysis or segmental hair analysis. No evidence of any relationship between maternal/fetal exposure during this specific period of gestation has been demonstrated to date in a human model. METHODS: To clarify drugs toxicokinetics and transplacental passage during early pregnancy, the presence of the most widely used recreational drugs of abuse and metabolites was investigated in the proximal 4cm hair segments of women undergoing voluntary termination of pregnancy (n=280) during the 12th week of gestation and the results were compared to those from placenta and fetal tissue samples in order to verify whether maternal hair testing can reflect fetal exposure and, if so, to what extent. Hair, placenta and fetal remains were analyzed by validated gas chromatography mass spectrometry assays. RESULTS: Eighty one positive hair samples were identified: 60 were positive for cannabis (74.1%), 28 for cocaine (34.6%), 7 for opiates (8.6%), 3 for MDMA (3.7%) and 18.5% were positive for more than one drug. The positive hair test results were confirmed in placenta/fetal tissues in 10 cases out of 60 for cannabis (16. 7%); in 7 out of 28 for cocaine (25%); and none for the 6 opiates positive cases and 3 MDMA cases, respectively. CONCLUSION: Drugs/metabolites in hair of pregnant women can be used as biomarkers of past drug use (repetitive or sporadic), although the use is not always reflected in fetal/placental tissues. There are several possible hypotheses to explain the results: (1) the use occurred before the start of pregnancy, (2) past sporadic consumption which could be measured in hair but not in fetal and placental remains because of the narrow window of drug detection in placental/fetal tissues; (3) the sensitivity of the analytical methods was not high enough for the detection of the minute amount of drugs of abuse and metabolites which reached these tissues (4) there is a large variability in the transplacental passage of drugs of abuse and in the placenta's metabolizing capacity.