Analysis of Forty-Two Psychoactive Substances in a Single Hair by Micro-Segmental Technique / 法医学杂志

OBJECTIVES@#To establish an LC-MS/MS method based on single hair micro-segmental technique, and verify the detection of 42 psychoactive substances in 0.4 mm hair segments.@*METHODS@#Each piece of single hair was cut into 0.4 mm segments and extracted by sonication and the segments were immersed in dithiothreitol-containing extraction medium. Mobile phase A was the aqueous solution containing 20 mmol/L ammonium acetate, 0.1% formic acid, and 5% acetonitrile. Mobile phase B was acetonitrile. An electrospray ionization source in positive ion mode was used for data acquisition in multiple reaction monitoring (MRM) mode.@*RESULTS@#The 42 psychoactive substances in hair had a good linear relationship within their respective linear ranges (r>0.99), the limits of detection were 0.2-10 pg/mm, the limits of quantification were 0.5-20 pg/mm, the intra-day and inter-day precisions were 1.5%-12.7%, the intra-day and inter-day accuracies were 86.5%-109.2%, the recovery rates were 68.1%-98.2%, and the matrix effects were 71.3%-111.7%. The method was applied to hair samples collected from one volunteer at 28 d after a single dose of zolpidem, with zolpidem detected in 5 hairs was 1.08-1.60 cm near the root tip, and the concentration range was 0.62-20.5 pg/mm.@*CONCLUSIONS@#The micro-segmental technique of single hair analysis can be applied to the investigation of drug-facilitated sexual assault cases.

Abuse of Xylazine by Human and its Emerging Problems: A Review from Forensic Perspective

@#Xylazine is a sedative, analgesic and muscle relaxant widely applied in the veterinary field. However, owing to its depressant effect, xylazine has become a substance of abuse by humans. Misuse of xylazine not only triggers unwanted consequences (death), but also linked with various crimes. Google Scholar, PubMed and SciFinder were used to retrieve articles and case reports in relation to the misuses of xylazine and established analytical methods for forensic investigation until November 2021. Literatures reported the accidental and intended poisoning of xylazine, recreational use of xylazine and as an adulterant in recreational drugs. In addition to being a facilitator of crime and sexual assault, it is administered illegally to food producing animals as a sedative and to sports animals as a doping agent. Problems associated with the abuse of xylazine were highlighted in this review, covering the unknown prevalence of xylazine abuse and the need to revise the regulatory status of xylazine. In addition, limited screening and confirmatory methods that can be readily utilised to detect xylazine either alone or simultaneously with other substances of abuse, particularly useful for forensic toxicology and narcotic section were available in the literature. As a conventionally used veterinary drug, xylazine is undoubtedly a potentially hazardous drug, and the investigations on its potential abuse would enhance routine forensic examination to keep pace with the status of illicit drugs.

Identification of Tiletamine, Zolazepam and Their Metabolites in Drug Facilitated Sexual Assault by GC-QTOF-MS / 法医学杂志

Objective To identify tiletamine, zolazepam and their metabolites in samples from drug facilitated sexual assault by gas chromatography-quadrupole time of flight mass spectrometry （GC-QTOF-MS）. Methods Urine samples of victims were collected, and detected by GC-QTOF-MS after liquid-liquid extraction and concentration. The molecular formula of fragments ions was identified by determination of accurate mass numbers, to detect related substances. Results Tiletamine, zolazepam, three metabolites of tiletamine and two metabolites of zolazepam were identified in urine samples from actual cases. Conclusion GC-QTOF-MS provides abundant and accurate information of fragment ions mass numbers, which can be used for qualitative identification of tiletamine, zolazepam and their metabolites in drug facilitated sexual assault.

Identification of New Designer Benzodiazepine Diclazepam in Drug Facilitated Sexual Assault / 法医学杂志

OBJECTIVES@#To identify the new designer drugs which are totally unknown and not in the routine testing list by the technologies such as high-resolution mass spectrometry in drug facilitated sexual assault, in order to solve the problem in actual cases.@*METHODS@#The milky fluid from an actual case was extracted and analyzed using LC-QE, ¹H-NMR and GC-MS, respectively. The accurate masses and cluster ions isotope patterns of unknown compound were obtained by LC-QE. The molecular formula was confirmed as C₁₆H₁₂C₂N₂O based on the protons number of ¹H-NMR. The isomers diclazepam and 4-chlorodiazepam were separated and detected with GC-MS.@*RESULTS@#The new designer benzodiazepine as diclazepam in the milky fluid was identified. The results provided direct evidence for the investigation and qualitative analysis of such cases.@*CONCLUSIONS@#The combined application of various methods, including LC-QE, ¹H-NMR and GC-MS, can be used to detect unknown new psychoactive substances.

The Role of Segmental Analysis of Clonazepam in Hair in Drug Facilitated Cases / 法医学杂志

OBJECTIVES@#To infer the frequency of dosage and medication history investigate of the victims in drug facilitated cases by the segmental analysis of clonazepam in hair.@*METHODS@#Freezing milling under liquid nitrogen environment combined with ultrasonic bath was used as sample pretreatment in this study, and liquid chromatography-tandem mass spectrometry was used for the segmental analysis of the hair samples collected from 6 victims in different cases. The concentrations of clonazepam and 7-aminoclonazepam were detected in each hair section.@*RESULTS@#Clonazepam and its metabolite 7-aminoclonazepam were detected in parts of hair sections from the 6 victims. The occurrence time of drug peak concentration was consistent with the intake timing provided by victims.@*CONCLUSIONS@#Segmental analysis of hair can provide the information of frequency of dosage and intake timing, which shows an unique evidential value in drug facilitated crimes.

The Role of Segmental Analysis of Clonazepam in Hair in Drug Facilitated Cases / 法医学杂志

Objective T o infer the frequency of dosage and m edication history investigate of the victim s in drug facilitated cases by the segm ental analysis of clonazepam in hair. Methods Freezing m illing un-der liquid nitrogen environm ent com bined w ith ultrasonic bath w as used as sam ple pretreatm ent in this study, and liquid chrom atography-tandem m ass spectrom etry w as used for the segm ental analysis of the hair sam ples collected from 6 victim s in different cases. T he concentrations of clonazepam and 7-am in-oclonazepam w ere detected in each hair section. Results C lonazepam and its m etabolite 7-am inoclon-azepam w ere detected in parts of hair sections from the 6 victim s. T he occurrence tim e of drug peak concentration w as consistent w ith the intake tim ing provided by victim s. Conclusion Segm ental analysis of hair can provide the inform ation of frequency of dosage and intake tim ing, w hich show s an unique evidential value in drug facilitated crim es.

Validación de un método rápido en orina por lC/MS/MS para la búsqueda de compuestos empleados en sumisión química y su aplicación a muestras reales / Validation of a fast screening method in urine by lC/MS/MS for compounds used in drug-facilitated crime (DFC) and its application to real samples

La Oficina de Naciones Unidas contra la Droga y el Delito (UNODC) en 2011 señala que "El delito facilitado por drogas (DFD) es una expresión general que abarca la violación y otras agresiones sexuales, el robo con violencia o intimidación, la extorsión de dinero y los malos tratos deliberados de ancianos o niños bajo la influencia de sustancias sicotrópicas". En este trabajo se validó un método cualitativo y rápido a partir de muestras de orina por LC/MS/MS para 39 compuestos comprendidos en los listados de sumisión química. El objetivo fue alcanzar un límite de detección un 50 % por debajo de la concentración propuesta como "Límites mínimos de funcionamiento exigidos (MRPL)" por la UNODC, para poder ser aplicado a muestras reales.

The United Nations Office on Drugs and Crime (UNODC) in 2011, states that "The Drug-facilitated crime (DFC) is a general term that includes rape or other sexual assault, robbery, money extortion, as well as the deliberate maltreatment of the elderly or children under the influence of psychotropic substances". In this work we validated a qualitative and fast method from urine samples by LC/MS/MS for 39 compounds included in the Drug-facilitated crime lists. The aim was to reach a detection limit of 50% below the proposed concentration as "minimum required performance limits (MRPL)" by UNODC in order to be applied in real samples.

Binge drinking: conductas sexuales de riesgo y drogas facilitadoras del asalto sexual en jóvenes españoles / Binge drinking: Risky sexual behaviors and drug facilitated sexual assault in spanish youths

El uso de drogas -alcohol incluido- como facilitadoras del asalto sexual y las conductas sexuales de riesgo son dos fenómenos relevantes desde una perspectiva sociosanitaria. El objetivo de la presente investigación es comparar las actitudes hacia estas variables en una muestra de 164 jóvenes en condiciones de intoxicación etílica frente a sobriedad. La muestra está compuesta por 82 varones y 82 mujeres. Su media de edad era 27.50 ± 63.72. Contestaron un cuestionario en el que se recogen datos sociodemográficos, información del consumo de drogas y la Escala de Conductas Sexuales; además, se emplearon varios etilómetros Digital Zaphir Deluxe para determinar la alcoholemia. Se trata de un estudio cuasiexperimental realizado sobre un diseño intraindividual. El muestreo es no probabilístico de tipo "bola de nieve". Los resultados muestran unas actitudes más positivas y una mayor intencionalidad hacia el empleo de drogas como facilitadoras del asalto sexual y hacia las conductas sexuales de riesgo de los sujetos que se encuentran bajo el efecto del alcohol en contextos recreativos, independientemente del sexo de la muestra. Se destaca, por un lado, la importancia de investigar sobre el uso de drogas como facilitadoras del asalto sexual (tema ampliamente ignorado en España) y se profundiza en cómo el consumo de drogas afecta al uso de métodos anticonceptivos, y por otro, la necesidad de incluir información en los programas preventivos sobre cómo el consumo de alcohol (y otras drogas) está relacionado con la actividad sexual.

Drug use (alcohol included) as a sexual-assault facilitator and risky sexual behavior are two important issues from a social and health perspective. This study aims to compare the attitudes towards these variables in a sample of 164 young people in conditions of intoxication vs sobriety. The sample consisted of 82 men and 82 women. Their mean age was 27.50 (SD = 3.72). Participants answered a sociodemographic questionnaire, a drug information questionnaire, and the Sexual Behaviors Scale. and moreover, Several Zaphir Deluxe Digital breathalyzers were also used to monitor alcohol level. This is a quasiexperimental study conducted using a within-subject design. Non-probabilistic snowball sampling was used. The results showed more positive attitudes and greater intentions towards the use of drugs as facilitators of sexual assault and risky sexual behaviors in those who are under the influence of alcohol in recreational contexts, regardless of the sex of the sample. These results showed, on the one hand, the importance of studies into the use of drug facilitated sexual assault -a largely ignored issue in Spain- and to determine how drug use affects the use of contraceptive methods. On the other hand, the results show the need to include information in preventive programs about the way alcohol (and other drugs) use is related to sexual activity.

Desenvolvimento de métodos analíticos para a identificação de drogas facilitadoras de crime em amostras de urina / Developing analytical methods for identification of drug-facilitated crime in urine samples

As drogas facilitadoras de crime (DFC) são uma série de substâncias químicas que permitem o ato sexual e/ou roubo com pouca ou nenhuma resistência da vítima. Benzodiazepínicos, gama-hidroxibutirato (GHB), cetamina e etanol são clássicas DFC, porém outras substâncias também têm sido utilizadas. Devido às diferentes classes de DFC e a necessidade de métodos sensíveis, a determinação dessas substâncias é um desafio aos toxicologistas forenses. A proposta do estudo foi desenvolver métodos analíticos para determinação principais analitos alvos de DFC para benzodiazepínicos, cetamina e GHB em amostras de urina. Esta matriz biológica é considerada uma amostra não-invasiva e apresenta um período de detecção maior que o sangue. A preparação das amostras foi avaliada através de microextração em fase líquida (LPME) e extração líquido-líquido (LLE). A LPME é uma técnica de extração de drogas que utiliza menor quantidade de solventes orgânicos, maior praticidade e possibilidade de obtenção de altos valores de recuperação. Os analitos foram determinados por cromatografia gasosa acoplada à espectrometria de massas (GC-MS). A LPME validada para benzodiazepínicos e seus produtos de biotransformação exigiu uma combinação de solventes e dupla derivatização para atingir a sensibilidade exigida, enquanto o método para determinação de cetamina, norcetamina e deidronorcetamina utilizou óleo essencial de eucalipto como meio extrator, caracterizando-se um procedimento ecologicamente correto com alta sensibilidade. A extração de GHB foi efetiva por LLE com redução da quantidade de solvente e tempo de análise sem o prejuízo na sensibilidade. Em geral, os métodos desenvolvidos neste trabalho são sensíveis e confiáveis para todos os analitos relatados e conclui-se que a LPME é uma técnica de preparo de amostra eficiente, versátil de baixo custo. Estas condições permitem que sua implementação em qualquer laboratório de análises toxicológicas, podendo ser aplicada em situações de DFC ou de qualquer outra natureza

Drug-facilitated crime (DFC) are a series of chemicals that allow the sexual act and/or theft with little or no resistance from the victim. Benzodiazepines, gamma-hydroxybutyrate (GHB) and ketamine and ethanol are considered classic DFC, however other substances were also used as the DFC. Due to the different classes of DFC and the need for sensitive methods, the determination of these substances is a challenge to forensic toxicologists. The purpose of this study was to develop analytical methods for determination of the main target analytes of DFC for benzodiazepines, ketamine and GHB in urine samples. This biological matrix is considered a non-invasive sample and shows a larger window of detection than blood. Sample preparation was assessed using liquid phase microextraction (LPME) and liquid-liquid extraction (LLE). The LPME is a drug extraction technique that uses less organic solvents, greater practicality and possibility of obtaining high recovery values. The analytes were determined by gas chromatography - mass spectrometry (GC-MS). The validated LPME technique for benzodiazepines and their metabolites required a combination of solvents and double derivatization to achieve the required sensitivity, while the ketamine, norketamine and dehydronorketamine method used essential oil of eucalyptus as solvent, characterizing a green chemistry approach with high sensitivity. The extraction of GHB was effective by LLE with a reduced amount of solvent and the analysis time without loss in sensitivity. In general, the methods developed in this work using GC-MS are sensitive and reliable for all analytes reported and LPME technique showed to be an efficient sample preparation, versatile and low cost. These conditions allow LPME implementation in any laboratory of toxicological analysis and it can be applied in situations of DFC or any other kind of analysis