RESUMO
RATIONALE: Benzodiazepines (BZDs) construct a large group of psychoactive drugs acting as depressants of the central nervous system (CNS) and used in medicine as sedatives and anxiolytic and antiepileptic agents. The illicit use of these materials is a worldwide problem, and for many years, part of the benzodiazepines have been abused as rape drugs. For example, flunitrazepam (Rohypnol) is most commonly linked by media reports to drug-facilitated sexual assaults, more commonly referred to as "date rape." Furthermore, there are growing concerns for other misuses of these drugs. Over the last few years, there was an increase in the number, type, and availability of new psychoactive substances (NPS) belonging to the benzodiazepine group, challenging standard forensic labs to fully identify the chemical structure of new, unknown benzodiazepines. METHODS: This work demonstrates a new application of the automated tool for the detection and identification of benzodiazepine analogues using high-resolution-accurate-mass LC-MS analysis, followed by "Compound Discoverer" (CD) software data processing, to automatically detect various benzodiazepine analogues by picking peaks and compare them to in silico calculated modifications made on a predefined basic backbone. Subsequently, a complete structural elucidation for the proposed molecular formula is obtained by MS/MS data analysis of the suspected component. RESULTS: This method was found to be useful for the automated detection and putative identification of a series of nine "unknown" benzodiazepine analogues, at concentrations in the low ng/mL range. CONCLUSIONS: We hereby present a general demonstration of this powerful tool for the forensic community in the detection and identification of hazardous unknown compounds.
Assuntos
Benzodiazepinas , Software , Benzodiazepinas/análise , Benzodiazepinas/química , Cromatografia Líquida/métodos , Espectrometria de Massas/métodos , Espectrometria de Massas em Tandem/métodos , Espectrometria de Massa com Cromatografia LíquidaRESUMO
Receptor occupancy is an indicator of antipsychotic efficacy and safety. It is desirable to simultaneously determine the occupancy of multiple brain receptors as an indicator of the efficacy and central side effects of antipsychotics because many of these drugs have binding affinities for various receptors, such as dopamine 2 (D2), histamine 1 (H1), and muscarinic acetylcholine (mACh) receptors. The purpose of this study was to develop a method for the simultaneous measurement of multiple receptor occupancies in the brain by the simultaneous quantification of unlabeled tracer levels using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Rats were pre-administered with a vehicle, displacer, or olanzapine, and mixed solutions of raclopride, doxepin, and 3-quinuclidinyl benzilate (3-QNB) were administered (3, 10, and 30 µg/kg). The brain tissue and plasma tracer concentrations were quantified 45 min later using LC-MS/MS, and the binding potential was calculated. The highest binding potential was observed at 3 µg/kg raclopride, 10 µg/kg doxepin, and 30 µg/kg 3-QNB. Tracer-specific binding at these optimal tracer doses in the cerebral cortex was markedly reduced by pre-administration of displacers. D2, H1, and mACh receptor occupancy by olanzapine increased in a dose-dependent manner, reaching 70-95%, 19-43%, and 12-45%, respectively, at an olanzapine dose range of 3-10 mg/kg. These results suggest that simultaneous determination of in vivo D2, H1, and mACh receptor occupancy is possible using LC-MS/MS.
Assuntos
Antipsicóticos , Olanzapina , Ratos Sprague-Dawley , Receptores de Dopamina D2 , Receptores Histamínicos H1 , Receptores Muscarínicos , Espectrometria de Massas em Tandem , Animais , Espectrometria de Massas em Tandem/métodos , Ratos , Masculino , Antipsicóticos/administração & dosagem , Cromatografia Líquida/métodos , Receptores de Dopamina D2/metabolismo , Receptores Muscarínicos/metabolismo , Receptores Muscarínicos/efeitos dos fármacos , Receptores Histamínicos H1/metabolismo , Olanzapina/farmacocinética , Olanzapina/administração & dosagem , Encéfalo/metabolismo , Encéfalo/efeitos dos fármacos , Benzodiazepinas/análise , Benzodiazepinas/metabolismo , Benzodiazepinas/farmacocinética , Racloprida/metabolismo , Doxepina/farmacocinética , Quinuclidinil Benzilato/metabolismo , Relação Dose-Resposta a DrogaRESUMO
Wastewater-based epidemiology (WBE) can provide objective and real time information about the use of addictive substances. A national study was conducted by measuring the most consumed illicit drugs, other drugs whose consumption is not so widespread but has increased significantly in recent years, and benzodiazepines in untreated wastewater from seven wastewater treatment plants (WWTPs) in six Spanish cities. Raw composite wastewater samples were collected from December 2020 to December 2021, a period in which the Spanish and regional governments adopted different restriction measures to contain the spread of the COVID-19 pandemic. Samples were analyzed using a validated analytical methodology for the simultaneous determination of 18 substances, based on solid-phase extraction and liquid-chromatography tandem mass spectrometry. Except for heroin, fentanyl, 6-acetylmorphine and alprazolam, all the compounds were found in at least one city and 9 out of 18 compounds were found in all the samples. In general, the consumption of illicit drugs was particularly high in one of the cities monitored in December 2020, when the restrictions were more severe, especially for cannabis and cocaine with values up to 46 and 6.9 g/day/1000 inhabitants (g/day/1000 inh), respectively. The consumption of MDMA, methamphetamine and mephedrone was notably higher in June 2021, after the end of the state of alarm, in the biggest population investigated in this study. Regarding the use of benzodiazepines, the highest mass loads corresponded to lorazepam. This study demonstrates that WBE is suitable for complementing epidemiological studies about the prevalence of illicit drugs and benzodiazepines during the COVID-19 pandemic restrictions.
Assuntos
Benzodiazepinas , COVID-19 , Cidades , Drogas Ilícitas , Drogas Ilícitas/análise , COVID-19/epidemiologia , Espanha/epidemiologia , Benzodiazepinas/análise , Humanos , Águas Residuárias/química , Poluentes Químicos da Água/análise , Pandemias , Detecção do Abuso de Substâncias/métodos , SARS-CoV-2RESUMO
The NC Office of the Chief Medical Examiner regularly assumes jurisdiction over deaths that are suspicious, unusual or unattended by a medical professional. In recent years, the presence of counterfeit pills is occasionally suggested by investigatory notes and/or scene findings that document reported consumption of prescription drugs, or prescription drugs on scene, which are not reflected in the final autopsy findings after toxicological analysis of the decedent's blood samples. Counterfeit pill consumption is a major public health hazard worthy of attention from the forensic toxicology community. Seventy-five cases from January 2020 to December 2022 serve as a convenience sample of cases where prescription pills including formulations of alprazolam, oxycodone and hydrocodone were specifically referenced during the death scene investigation as recently consumed, yet an unexpected substance was found during toxicological analysis rather than the expected pharmaceutical drug. Of note, novel benzodiazepines detected included flualprazolam, etizolam, clonazolam metabolite (8-aminoclonazolam), bromazolam, flubromazolam and desalkylflurazepam. Decedents' ages ranged from 16 to 69, across 33 different NC counties. Case notes indicated that eight of the decedents obtained pills through direct personal relationships, six decedents obtained them from "the street" and one decedent likely purchased pills online. Pills were largely consumed orally or through insufflation. Seven case reports contained indication that decedents knew or suspected the counterfeit nature of their pills. This study describes the context and characteristics of 2020-2022 suspected counterfeit pill-involved deaths in NC to further the understanding of the forensic science community, law enforcement partners, public health stakeholders and those potentially at risk through the consumption of counterfeit pills.
Assuntos
Medicamentos Falsificados , Toxicologia Forense , Humanos , Adulto , Masculino , Pessoa de Meia-Idade , Feminino , Adulto Jovem , Idoso , Benzodiazepinas/análise , Adolescente , Oxicodona/análise , Medicamentos sob Prescrição , Detecção do Abuso de Substâncias/métodos , Alprazolam/análise , HidrocodonaRESUMO
BACKGROUND: Legally prescribed benzodiazepines (BZDs) and designer BZDs are widely misused and must be determined in multiple contexts (eg, overdose, drug-facilitated sexual assaults, or driving under the influence of drugs). This study aimed to develop a method for measuring serum BZD levels using probe electrospray ionization (PESI) mass spectrometry and an isotope dilution approach. METHODS: A tandem mass spectrometer equipped with a probe electrospray ionization source in multiple reaction monitoring mode was used. Isotope dilution was applied for quantification using a deuterated internal standard at a fixed concentration for alprazolam, bromazepam, diazepam, nordiazepam, oxazepam, temazepam, zolpidem, and zopiclone. This method included designer BZDs: clonazolam, deschloroetizolam, diclazepam, etizolam, flualprazolam, flubromazepam, flubromazolam, meclonazepam, nifoxipam, and pyrazolam. Sample preparation was done by mixing 10 µL of serum with 500 µL of an ethanol/ammonium formate 0.01 mol/L buffer. Complete validation was performed, and the method was compared with liquid chromatography coupled with mass spectrometry (LC-MS/MS) and immunoassays (IC) by analyzing 40 real samples. RESULTS: The analysis time for identification and quantification of the 18 molecules was 2.5 minutes. This method was fully validated, and the limits of quantification varied from 5 to 50 mcg/L depending on the molecule. In the 40 real samples, 100% of molecules (n = 89) were detected by both LC-MS/MS and PESI-MS/MS, and regression analysis showed excellent agreement between the 2 methods (r 2 = 0.98). On IC, bromazepam and zolpidem were not detected in 2 and 1 cases, respectively. CONCLUSIONS: PESI-MS/MS allows serum BZD detection and measurement. Given the isotope dilution approach, a calibration curve was not required, and its performance was similar to that of LC-MS/MS, and its specificity was higher than that of IC.
Assuntos
Benzodiazepinas , Espectrometria de Massas por Ionização por Electrospray , Espectrometria de Massas em Tandem , Humanos , Espectrometria de Massas por Ionização por Electrospray/métodos , Benzodiazepinas/sangue , Benzodiazepinas/análise , Espectrometria de Massas em Tandem/métodos , Cromatografia Líquida/métodos , Detecção do Abuso de Substâncias/métodos , Reprodutibilidade dos Testes , Limite de DetecçãoRESUMO
The proliferation of novel psychoactive substances (NPSs) continues to challenge toxicology laboratories. In particular, the United Nations Office on Drugs and Crime considers designer benzodiazepines to be a current primary threat among all NPSs. Herein, we report detection of a new emerging designer benzodiazepine, clobromazolam, using high-resolution mass spectrometry and untargeted data acquisition in combination with a "suspect screening" method built from the crowd-sourced HighResNPS.com database. Our laboratory first detected clobromazolam in emergency department presenting intoxications included within the Emerging Drugs Network of Australia-Victoria project in the state of Victoria, Australia, from April 2022 to March 2023. Clobromazolam was the most frequent designer benzodiazepine detected in this cohort (100/993 cases, 10%). No patients reported intentional administration of clobromazolam, although over half reported exposure to alprazolam, which was detected in only 7% of cases. Polydrug use was prevalent (98%), with phenazepam (45%), methylamphetamine (71%) and other benzodiazepines (60%) most frequently co-detected. This is the first case series published in the literature concerning clobromazolam in clinical patients. The identification of clobromazolam in patients presenting to emergency departments in Victoria demonstrates how high-resolution mass spectrometry coupled with the HighResNPS.com database can be a valuable tool to assist toxicology laboratories in keeping abreast of emerging psychoactive drug use.
Assuntos
Benzodiazepinas , Serviço Hospitalar de Emergência , Detecção do Abuso de Substâncias , Humanos , Benzodiazepinas/análise , Detecção do Abuso de Substâncias/métodos , Austrália , Espectrometria de Massas , Bases de Dados Factuais , Masculino , Adulto , Drogas Desenhadas/análise , Feminino , Vitória/epidemiologiaRESUMO
Carbamazepine (CBZ) is a pharmaceutical compound detected in various water resources. With a view to removing this contaminant, the applicability of non-thermal plasma (NTP) oxidation process has been widely tested in recent years. This study utilized NTP from a dielectric barrier discharge reactor in the treatment of CBZ. NTP on the surface of a water sample containing 25 mg.L-1 of CBZ resulted in a removal efficiency of over 90% with an energy yield of 0.19 g. (kWh)-1. On the other hand, a rapid reduction in pH and an increase of conductivity and nitrate/nitrite ions concentration were observed during the degradation. The applied voltage amplitude significantly affected the removal efficiency and the energy yield as the degradation efficiency was 55%, 70%, and 72% respectively with an applied voltage of 8, 10, and 12 kV. The water matrices containing inorganic anions such as chloride and carbonate ions reduced the removal efficiency by scavenging the reactive species. Accordingly, a reduction in the removal efficiency was observed in tap water. The high-resolution mass spectrometry (HRMS) results revealed that both reactive oxygen and nitrogen species take part in the reaction process which yields many intermediate products including aromatic nitro-products. This study concluded that NTP can effectively degrade CBZ in both pure and tap water, but special attention must be paid to changes in the water quality parameters (pH, conductivity, and nitrate/nitrite ions) and the fate of nitro products.
Assuntos
Poluentes Químicos da Água , Purificação da Água , Nitratos/análise , Nitritos/análise , Carbamazepina/química , Cloro/química , Benzodiazepinas/análise , Oxirredução , Poluentes Químicos da Água/análise , Purificação da Água/métodosRESUMO
As one of the most widely abused designer benzodiazepines in the world, etizolam has been found in many cases in many countries. In this study, UPLC-Q-Exactive-MS was used for the first time to establish a dynamic change model of etizolam and its metabolites in rats. Compared with previous studies, the detection sensitivity and reproducibility of the instrument were higher. In the experiment, we optimized the traditional pharmacokinetic model based on Gauss function. According to the significant difference of etizolam in the plasma elimination phase of rats, a new pharmacokinetic model based on Lorentz function was established to describe the dynamic changes of etizolam more rigorously, which made the error effects lower and the accuracy of the pharmacokinetic parameters was improved. At the same time, the pharmacokinetic parameters of etizolam were compared with four other designer benzodiazepines reported in previous studies in rats, and we found the direct reason for the popularity of etizolam in the NPS market and explored the future development of etizolam for the first time. In addition, 21 metabolites were found through rat experiments to effectively detect etizolam abuse for a long time, of which 4 metabolites had the longest detection window and could be used as long-acting metabolites for experiments, which greatly prolongs the detection window and extends the time range in which etizolam was detected in real cases. This study is the first to conduct a systematic and comprehensive study on the metabolism and pharmacokinetics of etizolam and find out the direct reason for the prevalence of etizolam abuse, and we also discuss the development trend of etizolam in the future market of new psychoactive substances, which is beneficial for forensic experts to assess the trend of drug abuse and can provide reference for relevant drug control and drug treatment.
Assuntos
Diazepam/análogos & derivados , Espectrometria de Massa com Cromatografia Líquida , Ratos , Animais , Cromatografia Líquida de Alta Pressão , Reprodutibilidade dos Testes , Benzodiazepinas/análiseRESUMO
BACKGROUND: Being under the influence of psychoactive substances increases the risk of involvement in and dying from a traumatic event. The study is a prospective population-based observational study that aims to determine the prevalence of use and likely impairment from psychoactive substances among patients with suspected severe traumatic injury. METHOD: This study was conducted at 35 of 38 Norwegian trauma hospitals from 1 March 2019 to 29 February 2020. All trauma admissions for patients aged ≥ 16 years admitted via trauma team activation during the study period were eligible for inclusion. Blood samples collected on admission were analysed for alcohol, benzodiazepines, benzodiazepine-like hypnotics (Z-drugs), opioids, stimulants, and cannabis (tetrahydrocannabinol). RESULTS: Of the 4878 trauma admissions included, psychoactive substances were detected in 1714 (35 %) and in 771 (45 %) of these, a combination of two or more psychoactive substances was detected. Regarding the level of impairment, 1373 (28 %) admissions revealed a concentration of one or more psychoactive substances indicating likely impairment, and 1052 (22 %) highly impairment. Alcohol was found in 1009 (21 %) admissions, benzodiazepines and Z-drugs in 613 (13 %), opioids in 467 (10 %), cannabis in 352 (7 %), and stimulants in 371 (8 %). Men aged 27-43 years and patients with violence-related trauma had the highest prevalence of psychoactive substance use with respectively 424 (50 %) and 275 (80 %) testing positive for one or more compounds. CONCLUSION: The results revealed psychoactive substances in 35 % of trauma admissions, 80 % of which were likely impaired at the time of traumatic injury. A combination of several psychoactive substances was common, and younger males and patients with violence-related injuries were most often impaired. Injury prevention strategies should focus on high-risk groups and involve the prescription of controlled substances. We should consider toxicological screening in trauma admissions and incorporation of toxicological data into trauma registries.
Assuntos
Etanol , Transtornos Relacionados ao Uso de Substâncias , Masculino , Humanos , Prevalência , Estudos Prospectivos , Transtornos Relacionados ao Uso de Substâncias/complicações , Transtornos Relacionados ao Uso de Substâncias/epidemiologia , Benzodiazepinas/efeitos adversos , Benzodiazepinas/análise , Analgésicos Opioides , Psicotrópicos/efeitos adversosRESUMO
There is an increasing number of people affected worldwide by mental health disorders, such as depression and anxiety. One of the main courses of treatment, along with psychotherapy, is the use of psychoactive medications, like antidepressants and benzodiazepines. Also, the unprescribed use of these substances is a concerning public health issue. Hence, the analysis of psychotropic medications is mandatory in postmortem toxicology and various biological samples can be used for this detection, among them the vitreous humor (VH) stands out. Also, there is a demand for more sustainable and more efficient extraction methodologies according to green chemistry. An example is solid phase microextraction techniques (SPME), which use a solid sorbent and small solvent amounts. Biosorbents are substances of natural origin with sorptive properties, and they have been successfully used in SPME in environmental toxicology for water analysis, mainly. This study aimed to develop a sustainable, fast, cheap and simple SPME methodology using cork sheet strips as a biosorbent, to extract antidepressants, benzodiazepines and others from VH samples by liquid chromatography coupled to tandem mass spectrometry. The extraction was conducted in a 96-well plate using 200 µL of VH and optimization of relevant parameters for extraction was performed. For solvent optimization, two simplex-centroid experiments were planned for extraction and desorption and to evaluate time and pH, a Doehlert design experiment was performed. The analytical method for the determination and quantification of 17 substances was validated. The quantification limits were 5 ng/mL for all analytes and the calibration curves were linear between 5 and 30 ng/mL. This study was able to develop an efficient, cheap, simple and fast microextraction method for 17 analytes in VH, using strips of cork sheet for extraction and a 96-well plate as a container. Furthermore, this approach system could be automated for routine toxicology laboratories.
Assuntos
Microextração em Fase Sólida , Corpo Vítreo , Humanos , Toxicologia Forense , Corpo Vítreo/química , Microextração em Fase Sólida/métodos , Psicotrópicos/análise , Solventes/análise , Benzodiazepinas/análiseRESUMO
Keratinized matrices, including nails, are among the most resistant matrices that can be analyzed in cases where remains are deeply decomposed and relatively non-invasive for living people. In order to exploit these new matrices in the search for exogenous substances, it is necessary to develop analytical technologies capable of achieving high levels of sensitivity. In this technical note, an easy method is presented for the simultaneous extraction and quantification of three narcotic substances (morphine, codeine and methadone), two benzodiazepines (BDZs) (clonazepam and alprazolam) and an antipsychotic (quetiapine) from nail matrix by analysis in ultra-high-performance liquid chromatography at high-resolution mass spectrometry. The method has been validated following the Standard Practices for Method Validation in Forensic Toxicology of the Scientific Working Group for Forensic Toxicology. Nail specimens from eight authentic postmortem (PM) cases and 13 living donor samples were extracted and analyzed. Of the eight PM samples, five resulted positive for at least one of the three substances searched. Ten of the 13 living donor specimens were positive for at least one of the targeted BDZs or quetiapine.
Assuntos
Alprazolam , Codeína , Humanos , Cromatografia Líquida de Alta Pressão/métodos , Codeína/análise , Alprazolam/análise , Clonazepam , Morfina/análise , Fumarato de Quetiapina/análise , Metadona/análise , Unhas/química , Benzodiazepinas/análiseRESUMO
OBJECTIVES: To establish a method for the detection of carbamazepine and its metabolites 10,11-dihydro-10,11-epoxycarbamazepine and 10,11-dihydro-10-hydroxycarbamazepine in blood samples by liquid chromatography-tandem mass spectrometry (LC-MS/MS). METHODS: The blood samples were treated with 1-butyl-3-methylimidazolium hexafluorophosphate as an extraction solvent. The samples were extracted by ultrasound-assisted extraction and separated by ZORBAX Eclipse Plus C18, 95Å column. The mobile phase A aqueous solution containing 0.1% formic acid and 10 mmol/L ammonium acetate, and mobile phase B mixed organic solvent containing acetonitrile/methanol (Vacetonitrileâ¶Vmethanol=2â¶3) were used for gradient elution at the flow rate of 1.00 mL/min. An electrospray ion source in positive mode was used for detection in the multiple reaction monitoring. RESULTS: The linearities of carbamazepine and its metabolites 10,11-dihydro-10,11-epoxycarbamazepine and 10,11-dihydro-10-hydroxycarbamazepine in blood samples were good within the corresponding range, with correlation coefficients (r) greater than 0.995 6. The limits of detection were 3.00, 0.40 and 1.30 ng/mL, respectively. The limit of quantitation were 8.00, 1.00 and 5.00 ng/mL, respectively. The extraction recoveries ranged from 76.00% to 106.44%. The relative standard deviations of the intra-day and inter-day precisions were less than 16%. Carbamazepine and its main metabolite 10,11-dihydro-10,11-epoxycarbamazepine were detected in blood samples of death cases with a mass concentration of 2.71 µg/mL and 252.14 ng/mL, respectively. CONCLUSIONS: This method has high sensitivity and good selectivity, which is suitable for the detection of carbamazepine and its metabolites in blood samples, and can be used for carbamazepine-related forensic identifications.
Assuntos
Metanol , Espectrometria de Massas em Tandem , Cromatografia Líquida/métodos , Carbamazepina/análise , Benzodiazepinas/análise , Solventes , Cromatografia Líquida de Alta Pressão , Extração em Fase SólidaRESUMO
Liquid chromatography coupled with mass spectrometry (LC-MS) has been increasingly used for screening purposes in forensic toxicology. High versatility and low time/resource consumption are the main advantages of this technology. Numerous multi-analyte methods have been validated in order to face the analytical challenge of new psychoactive substances (NPSs). However, forensic toxicologists must focus the attention also on "classical" NPSs and medicines, such as benzodiazepines (BDZs) and prescription opioids. In this paper, a new method for the simultaneous detection of 169 substances (120 NPSs and 49 other drugs) in hair by LC-MS-MS is described. After the decontamination of hair samples with dichloromethane, a 20-mg aliquot of the sample was mixed with 1 mL of methanol (MeOH; 0.1% of formic acid) and then sonicated at room temperature for 2 h. The mixture was then dried under nitrogen stream and reconstituted with 100 µL of MeOH. LC separation was achieved with a 100-mm-long C18 column in 35 min, and mass acquisition was performed in dynamic multiple reaction monitoring mode and in positive ionization. The analysis results were very sensitive, with the limit of quantification ranging from 0.07 to 10.0 pg/mg. Accuracy and precision were always within the acceptable criteria. Matrix effect and recovery rate ranges were from -21.3 to + 21.9% and from 75.0 to 99.3%, respectively. The new method was successfully applied in a preliminary study on the prevalence of NPSs, BDZs and other substances in case of driving license issuance. In 14% of cases, BDZs/antidepressants (mainly trazodone, diazepam/nordiazepam and flunitrazepam) were found. Codeine, ketamine, methylone and mephedrone were also detected.
Assuntos
Detecção do Abuso de Substâncias , Espectrometria de Massas em Tandem , Cromatografia Líquida/métodos , Espectrometria de Massas em Tandem/métodos , Detecção do Abuso de Substâncias/métodos , Benzodiazepinas/análise , Fármacos do Sistema Nervoso Central , Cabelo/química , Limite de DetecçãoRESUMO
"Designer" benzodiazepines (DBZDs) are becoming increasingly available in Europe, with the European Monitoring Centre of Drugs and Drug Addiction currently monitoring â¼30 new benzodiazepines. The following driving under the influence of drug (DUID) case describes the oral fluid (OF) and blood concentrations, as well as the observed effects after the combined use of stimulants and flubromazepam. Both OF, collected via the Intercept i2 collector (Immunalysis, Pomona, CA, USA), and blood (collected in containers with various stabilizers) were screened using a liquid chromatographic (LC) time-of-flight (TOF) mass spectrometric (MS-MS) method. In addition, various LC-MS-MS methods in multi-reaction monitoring mode were applied for confirmation and quantification. The OF and blood samples were taken 2 h 25 min and 9 h 19 min after the accident, respectively. OF contained 789 ng/mL amphetamine, 5,173 ng/mL MDMA, 168 ng/mL benzoylecgonine, 492 ng/mL cocaine, 134 ng/mL 4-methylmethcathinone (4-MMC) and traces of flubromazepam (less than limit of quantification (LLOQ); 2 ng/mL). The sodium-fluoride blood samples contained 19 ng/mL amphetamine, 284 ng/mL MDMA, 20 ng/mL MDA, 38 ng/mL benzoylecgonine, 4 ng/mL methylecgonine, 161 ng/mL flubromazepam and traces of 4-MMC (Assuntos
Estimulantes do Sistema Nervoso Central
, N-Metil-3,4-Metilenodioxianfetamina
, Masculino
, Humanos
, Anfetamina/análise
, Benzodiazepinas/análise
, Detecção do Abuso de Substâncias/métodos
RESUMO
In this work, we optimize and validate a simple, time-saving, and environmentally friendly sample preparation method based on supramolecular solvents (SUPRAS), green nanostructured liquids, for the extraction of selected drug-facilitated sexual assault (DFSA) substances from human urine. The methodology was fast and simple (stirring, centrifugation, and dilution). Cubosomic SUPRAS were formed by the addition of 1,2-hexanediol (200 µL) to 1.0 mL of human urine containing 1 M Na2SO4. SUPRAS extracts were analyzed by LC-MS/MS. The method was fully validated for 23 DFSA compounds including 10 benzodiazepines, 1 z-hypnotic drug, 5 amphetamine derivatives, 3 cocaine metabolites, and 4 miscellaneous compounds. Extraction efficiency varied between 79 and 119%, and matrix effects were acceptable (-14.3/+21.5) for 87% of the compounds. Method detection and quantification limits ranged from 0.003 to 0.75 ng/mL and from 0.01 to 2.50 ng/mL, respectively. These values were low enough for the established minimum required performance limits (MRPL) of these substances. This simple and green method has a great potential to be implemented for the monitoring of illegal drugs involved in DFSA cases by forensic laboratories.
Assuntos
Delitos Sexuais , Espectrometria de Massas em Tandem , Humanos , Cromatografia Líquida/métodos , Espectrometria de Massas em Tandem/métodos , Hipnóticos e Sedativos , Benzodiazepinas/análise , SolventesRESUMO
The use of vitreous humor (VH) in forensic casework has been growing in the last years due to numerous advantages. Several compounds can be evaluated in this matrix, including benzodiazepines whose determination is essential due to their great availability and potential of dependance and misuse. Postmortem toxicological analyses are required to determine the influence of benzodiazepines in deaths. However, most of the analytical methods which determine these drugs in VH are laborious and time consuming. This article describes a simple method based on protein precipitation for the determination of eight benzodiazepines in VH samples. Samples were prepared through a protein precipitation method and analyzed by liquid chromatography tandem mass spectrometry. Solvent choice and sample and solvent volumes for precipitation were optimized using chemometric approaches. The method was validated for selectivity, lower limit of quantification (LLOQ), linearity, carryover, precision, bias, matrix effect and dilution integrity. In order to verify the applicability, 62 vitreous humor samples were analyzed. LLOQs were 1 ng/mL and calibration curves were linear from 1 to 25 ng/mL (r2 > 0,99) for all analytes. Bias, precision and dilution integrity results were satisfactory according to proper guidelines. Ionization suppression was significant with values ranging from 8 to 37%. Two samples from real cases were positive for diazepam with the following concentrations: 6.80 ng/mL and 47.68 ng/mL, approximately 10 times lower than those found in peripheral blood. The procedure described here can be used as a straightforward and low cost method for the quantitation of multiple benzodiazepines in VH.
Assuntos
Benzodiazepinas , Espectrometria de Massas em Tandem , Benzodiazepinas/análise , Cromatografia Líquida/métodos , Diazepam/análise , Humanos , Reprodutibilidade dos Testes , Solventes/análise , Espectrometria de Massas em Tandem/métodos , Corpo Vítreo/químicaRESUMO
In recent years, identification and analysis of designer benzodiazepines have become a challenge in forensic toxicology. These substances are analogs of the classic benzodiazepines, but their pharmacology is not well known, and many of them have been associated with overdoses and deaths. As a result, there has been a surge in efforts to develop analytical methods to determine these compounds in different biological samples. Our aim was to develop and validate a fast, sensitive and specific method for determining 17 designer benzodiazepines (adinazolam, clobazam, clonazolam, delorazepam, deschloroetizolam, diclazepam, etizolam, flualprazolam, flubromazepam, flubromazolam, flunitrazolam, N-desmethylclobazam, nifoxipam, nitrazolam, meclonazepam, pyrazolam and zolazepam) in hair by liquid chromatography-tandem mass spectrometry (LC-MS-MS). Hair samples were decontaminated and pulverized, and a 20 mg aliquot was incubated in methanol in an ultrasound bath (1 h, 25°C). The supernatant was evaporated and reconstituted in 200 µL of mobile phase, and the extracts were filtered (nano-filter vials) before injection into LC-MS-MS. All analytes were eluted from the chromatographic column in 8 min, and two multiple-reaction monitoring (MRM) transitions were used to identify each compound. The limits of quantification were 5 or 25 pg/mg depending on the analyte, and the calibration functions were linear to 200 pg/mg. Imprecision was <19.2% (n = 15), and bias was from -13.7 to 18.3% (n = 15). All the analytes yielded high extraction efficiencies >70% and displayed ion suppression between -62.8% and -23.9% (n = 10). The method was applied to 19 authentic cases. Five samples were positive for flualprazolam (
Assuntos
Espectrometria de Massas em Tandem , Zolazepam , Benzodiazepinas/análise , Cromatografia Líquida/métodos , Clobazam , Cabelo/química , Metanol/análise , Detecção do Abuso de Substâncias/métodos , Espectrometria de Massas em Tandem/métodos , Zolazepam/análiseRESUMO
Drug-impaired driver detection is a critical element of traffic safety. However, shifting drug use patterns over time and geography may limit the long-term reliability of assay-based screening tools. In this work, we compare qualitative results from the Abbott SoToxa® oral fluid (OF) screening device to Quantisal™ OF and whole blood. Our objective was to examine these three qualitative toxicological approaches, scope applicability of OF collection at the roadside, and compare them with a previous analysis of SoToxa® in Wisconsin. OF specimens were screened with the SoToxa® for six drugs or drug classes including amphetamine, benzodiazepines, cocaine, methamphetamine, opioids and tetrahydrocannabinol (THC). OF and blood specimens were collected from 106 participants. Quantisal™ OF and blood specimens were screened for drugs on ultra-performance liquid chromatography coupled to quadrupole time-of-flight high-resolution mass spectrometry (UPLC-QToF-HRMS) using a data-independent acquisition mode. UPLC-QToF-HRMS data were compared to comprehensive spectral libraries, and drugs were qualitatively identified. Drug Recognition Expert evaluations were performed, and face sheets submitted for 21 participants in this work. In general, the SoToxa® results were consistent with the combined qualitative results observed in Quantisal™ OF specimens and whole blood specimens. Limitations were uncovered for benzodiazepines, opioids and THC. The SoToxa® benzodiazepine assay has high cutoff concentrations for diazepam and clonazepam, limiting its sensitivity and positive predictive value when considering these drugs. SoToxa® opioid screening did not detect fentanyl, which is increasingly prevalent among drug users. Finally, ∆9-THC and its major metabolite 11-nor-9-carboxy-∆9-THC are lipophilic, limiting partitioning into OF. Despite these limitations, the SoToxa® instrument may be useful in assisting law enforcement with identifying individuals driving under the influence of drugs and establishing probable cause at roadside for making impaired driving arrests. Furthermore, Quantisal™ OF may be useful as screening specimens due to their ease of collection and results consistent with whole blood.
Assuntos
Cocaína , Metanfetamina , Anfetamina/análise , Analgésicos Opioides/análise , Benzodiazepinas/análise , Clonazepam/análise , Cocaína/análise , Diazepam/análise , Dronabinol/análise , Fentanila/análise , Humanos , Metanfetamina/análise , Reprodutibilidade dos Testes , Saliva/química , Detecção do Abuso de Substâncias/métodos , WisconsinRESUMO
BACKGROUND: The severe acute respiratory syndrome coronavirus 2 outbreak has been classified as a pandemic. Because many coronaviruses are heat sensitive, heat inactivation of patient samples at 56°C before testing reduces the risk of transmission. The aim of this study is to assess the impact of heat inactivation of patient blood samples on plasma concentrations of 5 second-generation antipsychotics and their metabolites. METHODS: Blood samples were collected during routine clinical therapeutic drug monitoring examination between April 3, 2021, and April 19, 2021. Samples were divided into 2 groups: group A, noninactivated raw sample, and group B, inactivated samples. Inactivation was performed by a 30-minute incubation at 56°C. The levels of the 5 drugs and their metabolites before and after sample heat inactivation were measured using liquid chromatography-tandem mass spectrometry and compared. Furthermore, correlation and Bland-Altman analyses were conducted. RESULTS: No statistically significant difference was observed between the levels of the 5 drugs and their metabolites (ie, risperidone, 9-OH-risperidone, aripiprazole, dehydroaripiprazole, olanzapine, quetiapine, norquetiapine, clozapine, and norclozapine) in the noninactivated group A and the inactivated group B ( P > 0.05). Each drug's concentration values in inactivated and noninactivated treatments correlated (Spearman rs > 0.98; P < 0.001). The results of the noninactivated treatment methods and samples alone showed good consistency via Bland-Altman analysis. CONCLUSIONS: Blood sample heat inactivation had no significant effect on the therapeutic drug monitoring of 5 second-generation antipsychotics and their metabolites. This inactivated treatment method should be recommended to effectively protect laboratory staff from virus contamination.
Assuntos
Antipsicóticos , COVID-19 , Aripiprazol , Benzodiazepinas/análise , Monitoramento de Medicamentos/métodos , Temperatura Alta , HumanosRESUMO
Analysis of new psychoactive substances (NPS) still poses a challenge for many institutions due to the number of available substances and the constantly changing drug market. Both new and well-known substances keep appearing and disappearing on the market, making it hard to adapt analytical methods in a timely manner. In this study we developed a qualitative screening approach for serum samples by means of liquid chromatography--quadrupole time-of-flight mass spectrometry. Samples were measured in data-dependent auto tandem mass spectrometry mode and identified by fragment spectra comparison, retention time and accurate mass. Approximately 500 NPS, including 195 synthetic cannabinoids, 180 stimulants, 86 hallucinogens, 26 benzodiazepines and 7 others were investigated. Serum samples were fortified to 1 ng/mL and 10 ng/mL concentrations to estimate approximate limits of identification (LOIs). Samples were extracted using solid-phase extraction with non-endcapped C18 material and elution in two consecutive steps. Benzodiazepines were eluted in the first step, while substances of other NPS subclasses were distributed among both extracts. To determine LOIs, both extracts were combined. Ninety-six percent (470/492) of investigated NPS were detected in 10 ng/mL samples and 88% (432/492) were detected in 1 ng/mL samples. Stimulants stood out with higher LOIs, possibly due to instability of certain methcathinone derivatives. However, considering relevant blood concentrations, the method provided sufficient sensitivity for stimulants as well as other NPS subclasses. Data-dependent acquisition was proven to provide high sensitivity and reliability when combined with an information-dependent preferred list, without losing its untargeted operation principle. Summarizing, the developed method fulfilled its purpose as a sensitive untargeted screening for serum samples and allows uncomplicated expansion of the spectral library to include thousands of targets.