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Marilyn A. Huestis - One of the best experts on this subject based on the ideXlab platform.
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A review of synthetic Cathinone-related fatalities from 2017 to 2020.
Therapeutic drug monitoring, 2020Co-Authors: Nunzia La Maida, Annagiulia Di Trana, Raffaele Giorgetti, Adriano Tagliabracci, Francesco Paolo Busardò, Marilyn A. HuestisAbstract:Purpose Synthetic Cathinones are designer analogs of the natural active principle of khat. Since their appearance on the black market in 2003, their popularity has increased annually, and they have become the most seized class of new psychoactive substances (NPS) reported to the UNODC Early Warning Advisory system. The constant introduction of newly synthesized molecules makes this issue difficult to monitor. The authors reviewed the most recent synthetic Cathinone-related fatalities worldwide to highlight new trends of consumption, reporting acute pharmacological and toxicological symptoms, scene investigations, analytical methods, and reported synthetic Cathinones concentrations in diverse biological matrices. Methods A literature search was performed using scientific databases such as PubMed, Scopus, Science Direct, Web of Science, and Research Gate to identify relevant scientific publications from 2017 to 2020. In addition, a search was conducted through the EU Early Warning System. Results From 2017 to 2020, 31 different synthetic Cathinones were identified in 75 reported fatal intoxications in the literature, alone or in combination with other substances. The most abused synthetic Cathinones were N-ethylpentylone, N-ethylhexedrone, and 4-chloromethCathinone. The EU Early Warning System included less detail on 72 additional synthetic Cathinone-related fatalities from 2017 to 2020. Conclusion New synthetic Cathinones continuously replace older natural and synthetic stimulant drugs, making determining the cause of death difficult. Analytical methods and high-performance mass spectrometry instruments are essential to detect the low concentrations of these potent new synthetic Cathinones. Little data is available on the pharmacology of these new drugs; the evaluation of toxicological antemortem and postmortem findings provides critical data on the drug's pharmacology and toxicology and for the interpretation of new synthetic Cathinone cases.
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synthetic Cathinone pharmacokinetics analytical methods and toxicological findings from human performance and postmortem cases
Drug Metabolism Reviews, 2016Co-Authors: Kayla N Ellefsen, Marta Concheiro, Marilyn A. HuestisAbstract:Synthetic Cathinones are commonly abused novel psychoactive substances (NPS). We present a comprehensive systematic review addressing in vitro and in vivo synthetic Cathinone pharmacokinetics, analytical methods for detection and quantification in biological matrices, and toxicological findings from human performance and postmortem toxicology cases. Few preclinical administration studies examined synthetic Cathinone pharmacokinetic profiles (absorption, distribution, metabolism, and excretion), and only one investigated metabolite pharmacokinetics. Synthetic Cathinone metabolic profiling studies, primarily with human liver microsomes, elucidated metabolite structures and identified suitable biomarkers to extend detection windows beyond those provided by parent compounds. Generally, Cathinone derivatives underwent ketone reduction, carbonylation of the pyrrolidine ring, and oxidative reactions, with phase II metabolites also detected. Reliable analytical methods are necessary for Cathinone identification in biological matrices to document intake and link adverse events to specific compounds and concentrations. NPS analytical methods are constrained in their ability to detect new emerging synthetic Cathinones due to limited commercially available reference standards and continuous development of new analogs. Immunoassay screening methods are especially affected, but also gas-chromatography and liquid-chromatography mass spectrometry confirmation methods. Non-targeted high-resolution-mass spectrometry screening methods are advantageous, as they allow for retrospective data analysis and easier addition of new synthetic Cathinones to existing methods. Lack of controlled administration studies in humans complicate interpretation of synthetic Cathinones in biological matrices, as dosing information is typically unknown. Furthermore, antemortem and postmortem concentrations often overlap and the presence of other psychoactive substances are typically found in combination with Cathinones derivatives, further confounding result interpretation.
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validation of the only commercially available immunoassay for synthetic Cathinones in urine randox drugs of abuse v biochip array technology
Drug Testing and Analysis, 2014Co-Authors: Sebastien Anizan, Kayla N Ellefsen, Marisol S Castaneto, Nathalie A Desrosiers, Thomas M Martin, Kevin L Klette, Marilyn A. HuestisAbstract:Deterrence of synthetic Cathinone abuse is hampered by the lack of a high-throughput immunoassay screen. The Randox Drugs of Abuse V (DOA-V) Biochip Array Technology contains two synthetic Cathinone antibodies: Bath Salt I (BSI) targets mephedrone/methCathinone and Bath Salt II (BSII) targets 3',4'-methylenedioxypyrovalerone (MDPV)/3',4'-methylenedioxy-α-pyrrolidinobutiophenone (MDPBP). We evaluated DOA-V synthetic Cathinones performance and conducted a full validation on the original assay with calibrators reconstituted in water, and the new assay with calibrators prepared in lyophilized urine; both utilized the same antibodies and were run on the fully automated Evidence® Analyzer. We screened 20 017 authentic military urine specimens and confirmed positives by liquid chromatography-tandem mass spectrometry (LC-MS/MS) for 28 synthetic Cathinones. Limits of detection (LOD) for the original and new assays were 0.35 and 0.18 (BSI), and 8.5 and 9.2 µg/L (BSII), respectively. Linearity was acceptable (R(2) >0.98); however, a large negative bias was observed with in-house prepared calibrators. Intra-assay imprecision was <20% BSI-II, while inter-assay imprecision was 18-42% BSI and <22% BSII. Precision was acceptable for Randox controls. Cross-reactivities of many additional synthetic Cathinones were determined. Authentic drug-free negative urine pH <4 produced false positive results for BSI (6.3 µg/L) and BSII (473 µg/L). Oxidizing agents reduced BSI and increased BSII results. Sensitivity, specificity, and efficiency of 100%, 52.1%, and 53.0% were obtained at manufacturer's proposed cut-offs (BSI 5 µg/L, BSII 30 µg/L). Performance improved if cut-off concentrations increased (BSI 7.5 µg/L, BSII 40 µg/L); however, there were limited confirmed positive specimens. Currently, this is the first and only fully validated immunoassay for preliminary detection of synthetic Cathinones in urine. Published 2014. This article is a U.S. Government work and is in the public domain in the USA.
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validation of the only commercially available immunoassay for synthetic Cathinones in urine randox drugs of abuse v biochip array technology
Drug Testing and Analysis, 2014Co-Authors: Sebastien Anizan, Kayla N Ellefsen, Marisol S Castaneto, Nathalie A Desrosiers, Thomas M Martin, Kevin L Klette, Marilyn A. HuestisAbstract:Deterrence of synthetic Cathinone abuse is hampered by the lack of a high-throughput immunoassay screen. The Randox Drugs of Abuse V (DOA-V) Biochip Array Technology contains two synthetic Cathinone antibodies: Bath Salt I (BSI) targets mephedrone/methCathinone and Bath Salt II (BSII) targets 3’,4’-methylenedioxypyrovalerone (MDPV)/3’,4’-methylenedioxy-αpyrrolidinobutiophenone (MDPBP). We evaluated DOA-V synthetic Cathinones performance and conducted a full validation on the original assay with calibrators reconstituted in water, and the new assay with calibrators prepared in lyophilized urine; both utilized the same antibodies and were run on the fully automated Evidence® Analyzer. We screened 20 017 authentic military urine specimens and confirmed positives by liquid chromatography-tandem mass spectrometry (LC-MS/MS) for 28 synthetic Cathinones. Limits of detection (LOD) for the original and new assays were 0.35 and 0.18 (BSI), and 8.5 and 9.2 μg/L (BSII), respectively. Linearity was acceptable (R 2 >0.98); however, a large negative bias was observed with in-house prepared calibrators. Intra-assay imprecisionwas<20% BSI-II, while inter-assay imprecision was 18-42% BSIand <22% BSII. Precision was acceptable for Randox controls. Cross-reactivities of many additional synthetic Cathinones were determined. Authentic drug-free negative urine pH < 4p roduced false positive results for BSI (6.3 μg/L) and BSII (473 μg/L). Oxidizing agents reduced BSI and increased BSII results. Sensitivity, specificity, and efficiency of 100%, 52.1%, and 53.0% were obtained at manufacturer’s proposed cut-offs (BSI 5 μg/L, BSII 30 μg/L). Performance improved if cut-off concentrations increased (BSI 7.5 μg/L, BSII 40 μg/L); however, there were limited confirmed positive specimens. Currently, this is the first and only fully validated immunoassay for preliminary detection of synthetic Cathinones in urine. Published 2014. This article is a U.S. Government work and is in the public domain in the USA. Additional supporting information may be found in the online version of this article at the publisher’s web site.
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Simultaneous quantification of 28 synthetic Cathinones and metabolites in urine by liquid chromatography-high resolution mass spectrometry
Analytical and Bioanalytical Chemistry, 2013Co-Authors: Marta Concheiro, Sebastien Anizan, Kayla N Ellefsen, Marilyn A. HuestisAbstract:Synthetic Cathinones are novel stimulants derived from Cathinone, with amphetamines or cocaine-like effects, often labeled “not for human consumption” and considered “legal highs”. Emergence of these new designer drugs complicate interpretation of forensic and clinical cases, with introduction of many new analogs designed to circumvent legislation and vary effects and potencies. We developed a method for the simultaneous quantification of 28 synthetic Cathinones, including four metabolites, in urine by liquid chromatography coupled to high resolution mass spectrometry (LC-HRMS). These Cathinones include Cathinone, methCathinone, and synthetic Cathinones position-3’-substituted, N-alkyl-substituted, ring-substituted, methylenedioxy-substituted, and pyrrolidinyl-substituted. One mL phosphate buffer pH 6 and 25 μL IStd solution were combined with 0.25 mL urine, and subjected to solid phase cation exchange extraction (SOLA SCX). The chromatographic reverse-phase separation was achieved with a gradient mobile phase of 0.1 % formic acid in water and in acetonitrile in 20 min. We employed a Q Exactive high resolution mass spectrometer, with compounds identified and quantified by target-MSMS experiments. The assay was linear from 0.5–1 to 100 μg/L, with limits of detection of 0.25–1 μg/L. Imprecision ( n = 20) was
Márcia Carvalho - One of the best experts on this subject based on the ideXlab platform.
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The interplay between autophagy and apoptosis mediates toxicity triggered by synthetic Cathinones in human kidney cells.
Toxicology Letters, 2020Co-Authors: I. Vaz, Maria Joao Valente, T. Carvalho, A. Castro, Ana Margarida Araújo, M.d.l. Bastos, Márcia CarvalhoAbstract:Abstract Synthetic Cathinones abuse remains a serious public health problem. Kidney injury has been reported in intoxications associated with synthetic Cathinones, but the molecular mechanisms involved have not been explored yet. In this study, the potential in vitro nephrotoxic effects of four commonly abused Cathinone derivatives, namely pentedrone, 3,4-dimethylmethcatinone (3,4-DMMC), methylone and 3,4-methylenedioxypyrovalerone (MDPV), were assessed in the human kidney HK-2 cell line. All four derivatives elicited cell death in a concentration- and time-dependent manner, in the following order of potency: 3,4-DMMC >> MDPV > methylone ≈ pentedrone. 3,4-DMMC and methylone were selected to further elucidate the mechanisms behind synthetic Cathinones-induced cell death. Both drugs elicited apoptotic cell death and prompted the formation of acidic vesicular organelles and autophagosomes in HK-2 cells. Moreover, the autophagy inhibitor 3-methyladenine significantly potentiated cell death, indicating that autophagy may serve as a cell survival mechanism that protects renal cells against synthetic Cathinones toxicity. Both drugs triggered a rise in reactive oxygen and nitrogen species formation, which was completely prevented by antioxidant treatment with N‑acetyl‑L‑cysteine or ascorbic acid. Importantly, these antioxidant agents significantly aggravated renal cell death induced by Cathinone derivatives, most likely due to their autophagy-blocking properties. Taken together, our results support an intricate control of cell survival/death modulated by oxidative stress, apoptosis and autophagy in synthetic Cathinones-induced renal injury.
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neurotoxicity of β keto amphetamines deathly mechanisms elicited by methylone and mdpv in human dopaminergic sh sy5y cells
ACS Chemical Neuroscience, 2017Co-Authors: Maria Joao Valente, Félix Carvalho, Maria De Lourdes Bastos, Paula Guedes De Pinho, Eduarda Fernandes, Márcia CarvalhoAbstract:Synthetic Cathinones (β-keto amphetamines) act as potent CNS stimulants similarly to classical amphetamines, which raise concerns about their potential neurotoxic effects. The present in vitro study aimed to explore and compare the mechanisms underlying the neurotoxicity of two commonly abused Cathinone derivatives, 3,4-methylenedioxymethCathinone (methylone) and 3,4-methylenedioxypyrovalerone (MDPV), with those of 3,4-methylenedioxymethamphetamine (MDMA), using undifferentiated and differentiated SH-SY5Y cells. Following a 24 h exposure period, methylone and MDPV induced loss of cell viability in a concentration-dependent manner, in the following order of potency: MDPV ≈ MDMA > methylone. Dopaminergic differentiated cells evidenced higher sensitivity to the neurotoxic effects of both Cathinones and MDMA than the undifferentiated ones, but this effect was not inhibited by the DAT inhibitor GBR 12909. Intracellular oxidative stress mediated by methylone and MDPV was demonstrated by the increase in reactive...
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editor s highlight characterization of hepatotoxicity mechanisms triggered by designer Cathinone drugs β keto amphetamines
Toxicological Sciences, 2016Co-Authors: Maria Joao Valente, Félix Carvalho, Maria De Lourdes Bastos, Paula Guedes De Pinho, Eduarda Fernandes, Ana Margarida Araújo, Márcia CarvalhoAbstract:The use of Cathinone designer drugs in recreational settings has been associated with severe toxic effects, including liver damage. The precise mechanisms by which Cathinones induce hepatotoxicity and whether they act by common pathways remain to be elucidated. Herein, we assessed the toxicity of the Cathinones methylone, pentedrone, 3,4-methylenedioxypyrovalerone (MDPV) and 4-methylethCathinone (4-MEC) in primary rat hepatocytes (PRH) and HepaRG cells, and compared with that of 3,4-methylenedioxymethamphetamine (MDMA). MDPV and pentedrone were significantly more toxic than MDMA, while methylone was the least cytotoxic compound. Importantly, PRH revealed to be the most sensitive experimental model and was thus used to explore the mechanisms underlying the observed toxicity. All drugs elicited the formation of reactive oxygen and nitrogen species (ROS and RNS), but more markedly for methylone, pentedrone and 4-MEC. GSH depletion was also a common effect at the highest concentration tested, whereas only MDPV and pentedrone caused a significant decrease in ATP levels. The antioxidants ascorbic acid or N-acetyl-L-cysteine partially attenuated the observed cell death. All Cathinones triggered significant caspase activation and apoptosis, which was partially reversed by the caspase inhibitor Ac-LETD-CHO. In conclusion, the present data shows that (1) Cathinones induce in vitro hepatotoxic effects that vary in magnitude among the different analogues, (2) oxidative stress and mitochondrial dysfunction play a role in Cathinones-induced hepatic injury, and (3) apoptosis appears to be an important pathway of cell death elicited by these novel drugs.
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Chiral enantioresolution of Cathinone derivatives present in “legal highs”, and enantioselectivity evaluation on cytotoxicity of 3,4-methylenedioxypyrovalerone (MDPV)
Forensic Toxicology, 2016Co-Authors: Bárbara Silva, Márcia Carvalho, Paula Guedes De Pinho, Maria Joao Valente, Carla Fernandes, Maria Elizabeth Tiritan, Madalena M.m. Pinto, Fernando RemiãoAbstract:Recently, great interest has been focused on synthetic Cathinones since their consumption has increased exponentially. All synthetic Cathinones exist as chiral molecules; the biological and/or toxicological properties of Cathinones generally differ according to the enantiomers in human body. In this study, a chiral liquid chromatography method was developed to separate and determine the enantiomeric ratio of synthetic Cathinones present in “legal highs” acquired in old smart shops or over the Internet. All the synthetic Cathinones were efficiently enantio-separated with α and Rs ranging from 1.24 to 3.62 and from 1.24 to 10.52, respectively, using polysaccharide-based chiral stationary phases. All synthetic Cathinones, with the exception of 4-methylethCathinone (4-MEC), were present in the commercialized “legal highs” in an enantiomeric proportion of 50:50. One of the studied chiral compounds was 3,4-methylenedioxypyrovalerone (MDPV), one of the most consumed Cathinone derivative worldwide. Our research group has recently reported its hepatotoxicity in the racemic form. Thus, the analytical enantioresolution of the MDPV was scaled up to multi-milligram using a semi-preparative amylose tris -3,5-dimethylphenylcarbamate column (20 cm × 7.0 mm ID, 7 µm particle size). Both enantiomers were isolated with high enantiomeric purity (enantiomeric excess > 99 %). The toxicity of S -(−)-MDPV and R -(+)-MDPV was evaluated, for the first time, using primary cultures of rat hepatocytes. It was also possible to verify that MDPV enantiomers showed hepatotoxicity in a concentration-dependent manner, but displayed no enantioselective toxicity in this cell culture model.
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Khat and synthetic Cathinones: a review
Archives of Toxicology, 2014Co-Authors: Maria Joao Valente, Félix Carvalho, Maria De Lourdes Bastos, Paula Guedes De Pinho, Márcia CarvalhoAbstract:For centuries, ‘khat sessions’ have played a key role in the social and cultural traditions among several communities around Saudi Arabia and most East African countries. The identification of Cathinone as the main psychoactive compound of khat leaves, exhibiting amphetamine-like pharmacological properties, resulted in the synthesis of several derivatives structurally similar to this so-called natural amphetamine. Synthetic Cathinones were primarily developed for therapeutic purposes, but promptly started being misused and extensively abused for their euphoric effects. In the mid-2000’s, synthetic Cathinones emerged in the recreational drug markets as legal alternatives (‘legal highs’) to amphetamine, ‘ecstasyʼ, or cocaine. Currently, they are sold as ‘bath salts’ or ‘plant foodʼ, under ambiguous labels lacking information about their true contents. Cathinone derivatives are conveniently available online or at ‘smartshops’ and are much more affordable than the traditional illicit drugs. Despite the scarcity of scientific data on these ‘legal highs’, synthetic Cathinones use became an increasingly popular practice worldwide. Additionally, criminalization of these derivatives is often useless since for each specific substance that gets legally controlled, one or more structurally modified analogs are introduced into the legal market. Chemically, these substances are structurally related to amphetamine. For this reason, Cathinone derivatives share with this drug both central nervous system stimulating and sympathomimetic features. Reports of intoxication and deaths related to the use of ‘bath salts’ have been frequently described over the last years, and several attempts to apply a legislative control on synthetic Cathinones have been made. However, further research on their pharmacological and toxicological properties is fully required in order to access the actual potential harm of synthetic Cathinones to general public health. The present work provides a review on khat and synthetic Cathinones, concerning their historical background, prevalence, patterns of use, legal status, chemistry, pharmacokinetics, pharmacodynamics, and their physiological and toxicological effects on animals and humans.
Kayla N Ellefsen - One of the best experts on this subject based on the ideXlab platform.
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synthetic Cathinone pharmacokinetics analytical methods and toxicological findings from human performance and postmortem cases
Drug Metabolism Reviews, 2016Co-Authors: Kayla N Ellefsen, Marta Concheiro, Marilyn A. HuestisAbstract:Synthetic Cathinones are commonly abused novel psychoactive substances (NPS). We present a comprehensive systematic review addressing in vitro and in vivo synthetic Cathinone pharmacokinetics, analytical methods for detection and quantification in biological matrices, and toxicological findings from human performance and postmortem toxicology cases. Few preclinical administration studies examined synthetic Cathinone pharmacokinetic profiles (absorption, distribution, metabolism, and excretion), and only one investigated metabolite pharmacokinetics. Synthetic Cathinone metabolic profiling studies, primarily with human liver microsomes, elucidated metabolite structures and identified suitable biomarkers to extend detection windows beyond those provided by parent compounds. Generally, Cathinone derivatives underwent ketone reduction, carbonylation of the pyrrolidine ring, and oxidative reactions, with phase II metabolites also detected. Reliable analytical methods are necessary for Cathinone identification in biological matrices to document intake and link adverse events to specific compounds and concentrations. NPS analytical methods are constrained in their ability to detect new emerging synthetic Cathinones due to limited commercially available reference standards and continuous development of new analogs. Immunoassay screening methods are especially affected, but also gas-chromatography and liquid-chromatography mass spectrometry confirmation methods. Non-targeted high-resolution-mass spectrometry screening methods are advantageous, as they allow for retrospective data analysis and easier addition of new synthetic Cathinones to existing methods. Lack of controlled administration studies in humans complicate interpretation of synthetic Cathinones in biological matrices, as dosing information is typically unknown. Furthermore, antemortem and postmortem concentrations often overlap and the presence of other psychoactive substances are typically found in combination with Cathinones derivatives, further confounding result interpretation.
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validation of the only commercially available immunoassay for synthetic Cathinones in urine randox drugs of abuse v biochip array technology
Drug Testing and Analysis, 2014Co-Authors: Sebastien Anizan, Kayla N Ellefsen, Marisol S Castaneto, Nathalie A Desrosiers, Thomas M Martin, Kevin L Klette, Marilyn A. HuestisAbstract:Deterrence of synthetic Cathinone abuse is hampered by the lack of a high-throughput immunoassay screen. The Randox Drugs of Abuse V (DOA-V) Biochip Array Technology contains two synthetic Cathinone antibodies: Bath Salt I (BSI) targets mephedrone/methCathinone and Bath Salt II (BSII) targets 3',4'-methylenedioxypyrovalerone (MDPV)/3',4'-methylenedioxy-α-pyrrolidinobutiophenone (MDPBP). We evaluated DOA-V synthetic Cathinones performance and conducted a full validation on the original assay with calibrators reconstituted in water, and the new assay with calibrators prepared in lyophilized urine; both utilized the same antibodies and were run on the fully automated Evidence® Analyzer. We screened 20 017 authentic military urine specimens and confirmed positives by liquid chromatography-tandem mass spectrometry (LC-MS/MS) for 28 synthetic Cathinones. Limits of detection (LOD) for the original and new assays were 0.35 and 0.18 (BSI), and 8.5 and 9.2 µg/L (BSII), respectively. Linearity was acceptable (R(2) >0.98); however, a large negative bias was observed with in-house prepared calibrators. Intra-assay imprecision was <20% BSI-II, while inter-assay imprecision was 18-42% BSI and <22% BSII. Precision was acceptable for Randox controls. Cross-reactivities of many additional synthetic Cathinones were determined. Authentic drug-free negative urine pH <4 produced false positive results for BSI (6.3 µg/L) and BSII (473 µg/L). Oxidizing agents reduced BSI and increased BSII results. Sensitivity, specificity, and efficiency of 100%, 52.1%, and 53.0% were obtained at manufacturer's proposed cut-offs (BSI 5 µg/L, BSII 30 µg/L). Performance improved if cut-off concentrations increased (BSI 7.5 µg/L, BSII 40 µg/L); however, there were limited confirmed positive specimens. Currently, this is the first and only fully validated immunoassay for preliminary detection of synthetic Cathinones in urine. Published 2014. This article is a U.S. Government work and is in the public domain in the USA.
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validation of the only commercially available immunoassay for synthetic Cathinones in urine randox drugs of abuse v biochip array technology
Drug Testing and Analysis, 2014Co-Authors: Sebastien Anizan, Kayla N Ellefsen, Marisol S Castaneto, Nathalie A Desrosiers, Thomas M Martin, Kevin L Klette, Marilyn A. HuestisAbstract:Deterrence of synthetic Cathinone abuse is hampered by the lack of a high-throughput immunoassay screen. The Randox Drugs of Abuse V (DOA-V) Biochip Array Technology contains two synthetic Cathinone antibodies: Bath Salt I (BSI) targets mephedrone/methCathinone and Bath Salt II (BSII) targets 3’,4’-methylenedioxypyrovalerone (MDPV)/3’,4’-methylenedioxy-αpyrrolidinobutiophenone (MDPBP). We evaluated DOA-V synthetic Cathinones performance and conducted a full validation on the original assay with calibrators reconstituted in water, and the new assay with calibrators prepared in lyophilized urine; both utilized the same antibodies and were run on the fully automated Evidence® Analyzer. We screened 20 017 authentic military urine specimens and confirmed positives by liquid chromatography-tandem mass spectrometry (LC-MS/MS) for 28 synthetic Cathinones. Limits of detection (LOD) for the original and new assays were 0.35 and 0.18 (BSI), and 8.5 and 9.2 μg/L (BSII), respectively. Linearity was acceptable (R 2 >0.98); however, a large negative bias was observed with in-house prepared calibrators. Intra-assay imprecisionwas<20% BSI-II, while inter-assay imprecision was 18-42% BSIand <22% BSII. Precision was acceptable for Randox controls. Cross-reactivities of many additional synthetic Cathinones were determined. Authentic drug-free negative urine pH < 4p roduced false positive results for BSI (6.3 μg/L) and BSII (473 μg/L). Oxidizing agents reduced BSI and increased BSII results. Sensitivity, specificity, and efficiency of 100%, 52.1%, and 53.0% were obtained at manufacturer’s proposed cut-offs (BSI 5 μg/L, BSII 30 μg/L). Performance improved if cut-off concentrations increased (BSI 7.5 μg/L, BSII 40 μg/L); however, there were limited confirmed positive specimens. Currently, this is the first and only fully validated immunoassay for preliminary detection of synthetic Cathinones in urine. Published 2014. This article is a U.S. Government work and is in the public domain in the USA. Additional supporting information may be found in the online version of this article at the publisher’s web site.
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Simultaneous quantification of 28 synthetic Cathinones and metabolites in urine by liquid chromatography-high resolution mass spectrometry
Analytical and Bioanalytical Chemistry, 2013Co-Authors: Marta Concheiro, Sebastien Anizan, Kayla N Ellefsen, Marilyn A. HuestisAbstract:Synthetic Cathinones are novel stimulants derived from Cathinone, with amphetamines or cocaine-like effects, often labeled “not for human consumption” and considered “legal highs”. Emergence of these new designer drugs complicate interpretation of forensic and clinical cases, with introduction of many new analogs designed to circumvent legislation and vary effects and potencies. We developed a method for the simultaneous quantification of 28 synthetic Cathinones, including four metabolites, in urine by liquid chromatography coupled to high resolution mass spectrometry (LC-HRMS). These Cathinones include Cathinone, methCathinone, and synthetic Cathinones position-3’-substituted, N-alkyl-substituted, ring-substituted, methylenedioxy-substituted, and pyrrolidinyl-substituted. One mL phosphate buffer pH 6 and 25 μL IStd solution were combined with 0.25 mL urine, and subjected to solid phase cation exchange extraction (SOLA SCX). The chromatographic reverse-phase separation was achieved with a gradient mobile phase of 0.1 % formic acid in water and in acetonitrile in 20 min. We employed a Q Exactive high resolution mass spectrometer, with compounds identified and quantified by target-MSMS experiments. The assay was linear from 0.5–1 to 100 μg/L, with limits of detection of 0.25–1 μg/L. Imprecision ( n = 20) was
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simultaneous quantification of 28 synthetic Cathinones and metabolites in urine by liquid chromatography high resolution mass spectrometry
Analytical and Bioanalytical Chemistry, 2013Co-Authors: Marta Concheiro, Sebastien Anizan, Kayla N Ellefsen, Marilyn A. HuestisAbstract:Synthetic Cathinones are novel stimulants derived from Cathinone, with amphetamines or cocaine-like effects, often labeled “not for human consumption” and considered “legal highs”. Emergence of these new designer drugs complicate interpretation of forensic and clinical cases, with introduction of many new analogs designed to circumvent legislation and vary effects and potencies. We developed a method for the simultaneous quantification of 28 synthetic Cathinones, including four metabolites, in urine by liquid chromatography coupled to high resolution mass spectrometry (LC-HRMS). These Cathinones include Cathinone, methCathinone, and synthetic Cathinones position-3’-substituted, N-alkyl-substituted, ring-substituted, methylenedioxy-substituted, and pyrrolidinyl-substituted. One mL phosphate buffer pH 6 and 25 μL IStd solution were combined with 0.25 mL urine, and subjected to solid phase cation exchange extraction (SOLA SCX). The chromatographic reverse-phase separation was achieved with a gradient mobile phase of 0.1 % formic acid in water and in acetonitrile in 20 min. We employed a Q Exactive high resolution mass spectrometer, with compounds identified and quantified by target-MSMS experiments. The assay was linear from 0.5–1 to 100 μg/L, with limits of detection of 0.25–1 μg/L. Imprecision (n = 20) was <15.9 % and accuracy (n = 20) 85.2–118.1 %. Extraction efficiency was 78.9–116.7 % (CV 1.4–16.7 %, n = 5), process efficiency 57.7–104.9 %, and matrix effects from −29.5 % to 1.5 % (CV 1.9–13.1 %, n = 10). Most synthetic Cathinones were stable at 4 °C for 72 h (n = 27) and after 3 freeze-thaw cycles (n = 26), but many (n = 19) were not stable at room temperature for 24 h (losses up to −67.6 %). The method was applied to authentic urine specimens from synthetic Cathinone users. This method provides a comprehensive confirmation method for 28 synthetic Cathinones in urine, with good selectivity and specificity.
María Ibáñez - One of the best experts on this subject based on the ideXlab platform.
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Rapid tentative identification of synthetic Cathinones in seized products taking advantage of the full capabilities of triple quadrupole analyzer
Forensic Toxicology, 2019Co-Authors: David Fabregat-safont, Juan V. Sancho, Félix Hernández, María IbáñezAbstract:Purpose In this work, a new approach for synthetic Cathinone identification in seized products, consisting of a rapid pseudo-target screening using liquid chromatography coupled to low-resolution tandem mass spectrometry (MS/MS), is proposed based on typical common product ions and neutral losses observed for this drug class. Methods The term “pseudo-target” screening indicates that although a pre-defined target compound list is not used, the search is limited to synthetic Cathinones with expected common moieties. A total of 22 neutral losses and 36 common product ions were monitored and used for Cathinone identification. Results In order to test the approach, 14 blind samples were analyzed and the results compared with high-resolution mass spectrometry data. From the data obtained, the different moieties of the Cathinones (and therefore their structures) could be derived, allowing their tentative identification. Conclusions This methodology will be useful for the first and rapid synthetic Cathinone detection in laboratories that have low-resolution MS/MS instrumentation.
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rapid tentative identification of synthetic Cathinones in seized products taking advantage of the full capabilities of triple quadrupole analyzer
Forensic Toxicology, 2019Co-Authors: David Fabregatsafont, Juan V. Sancho, Félix Hernández, María IbáñezAbstract:In this work, a new approach for synthetic Cathinone identification in seized products, consisting of a rapid pseudo-target screening using liquid chromatography coupled to low-resolution tandem mass spectrometry (MS/MS), is proposed based on typical common product ions and neutral losses observed for this drug class. The term “pseudo-target” screening indicates that although a pre-defined target compound list is not used, the search is limited to synthetic Cathinones with expected common moieties. A total of 22 neutral losses and 36 common product ions were monitored and used for Cathinone identification. In order to test the approach, 14 blind samples were analyzed and the results compared with high-resolution mass spectrometry data. From the data obtained, the different moieties of the Cathinones (and therefore their structures) could be derived, allowing their tentative identification. This methodology will be useful for the first and rapid synthetic Cathinone detection in laboratories that have low-resolution MS/MS instrumentation.
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reporting the novel synthetic Cathinone 5 ppdi through its analytical characterization by mass spectrometry and nuclear magnetic resonance
Forensic Toxicology, 2018Co-Authors: David Fabregatsafont, Juan V. Sancho, Félix Hernández, Xoan Carbon, Cristina Gil, M Ventura, María IbáñezAbstract:In this work, the identification and characterization of the novel synthetic Cathinone 5-PPDI found in a suspect drug sample were performed. The suspect sample was analyzed by gas chromatography–mass spectrometry (GC–MS), Fourier-transformed infrared (FTIR) spectroscopy, ultra-high performance liquid chromatography–high-resolution mass spectrometry (HRMS) and nuclear magnetic resonance (NMR) spectroscopy. The fragmentation observed in GC–MS and the identification of functional groups by FTIR was not enough for compound identification. After an exhaustive analysis of the accurate-mass fragmentation observed in HRMS, the compound was tentatively identified as the novel Cathinone 5-PPDI. Finally, five different NMR experiments were used for the unequivocal identification and complete characterization of the compound. In addition, the origin of this Cathinone was investigated in depth. The analytical data provided in this work will be useful for the identification of 5-PPDI by forensic laboratories. In addition, the origin of this Cathinone has been investigated, which could be of interest for the identification of future synthetic Cathinones prepared following the similar synthesis route.
Maria Joao Valente - One of the best experts on this subject based on the ideXlab platform.
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The interplay between autophagy and apoptosis mediates toxicity triggered by synthetic Cathinones in human kidney cells.
Toxicology Letters, 2020Co-Authors: I. Vaz, Maria Joao Valente, T. Carvalho, A. Castro, Ana Margarida Araújo, M.d.l. Bastos, Márcia CarvalhoAbstract:Abstract Synthetic Cathinones abuse remains a serious public health problem. Kidney injury has been reported in intoxications associated with synthetic Cathinones, but the molecular mechanisms involved have not been explored yet. In this study, the potential in vitro nephrotoxic effects of four commonly abused Cathinone derivatives, namely pentedrone, 3,4-dimethylmethcatinone (3,4-DMMC), methylone and 3,4-methylenedioxypyrovalerone (MDPV), were assessed in the human kidney HK-2 cell line. All four derivatives elicited cell death in a concentration- and time-dependent manner, in the following order of potency: 3,4-DMMC >> MDPV > methylone ≈ pentedrone. 3,4-DMMC and methylone were selected to further elucidate the mechanisms behind synthetic Cathinones-induced cell death. Both drugs elicited apoptotic cell death and prompted the formation of acidic vesicular organelles and autophagosomes in HK-2 cells. Moreover, the autophagy inhibitor 3-methyladenine significantly potentiated cell death, indicating that autophagy may serve as a cell survival mechanism that protects renal cells against synthetic Cathinones toxicity. Both drugs triggered a rise in reactive oxygen and nitrogen species formation, which was completely prevented by antioxidant treatment with N‑acetyl‑L‑cysteine or ascorbic acid. Importantly, these antioxidant agents significantly aggravated renal cell death induced by Cathinone derivatives, most likely due to their autophagy-blocking properties. Taken together, our results support an intricate control of cell survival/death modulated by oxidative stress, apoptosis and autophagy in synthetic Cathinones-induced renal injury.
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neurotoxicity of β keto amphetamines deathly mechanisms elicited by methylone and mdpv in human dopaminergic sh sy5y cells
ACS Chemical Neuroscience, 2017Co-Authors: Maria Joao Valente, Félix Carvalho, Maria De Lourdes Bastos, Paula Guedes De Pinho, Eduarda Fernandes, Márcia CarvalhoAbstract:Synthetic Cathinones (β-keto amphetamines) act as potent CNS stimulants similarly to classical amphetamines, which raise concerns about their potential neurotoxic effects. The present in vitro study aimed to explore and compare the mechanisms underlying the neurotoxicity of two commonly abused Cathinone derivatives, 3,4-methylenedioxymethCathinone (methylone) and 3,4-methylenedioxypyrovalerone (MDPV), with those of 3,4-methylenedioxymethamphetamine (MDMA), using undifferentiated and differentiated SH-SY5Y cells. Following a 24 h exposure period, methylone and MDPV induced loss of cell viability in a concentration-dependent manner, in the following order of potency: MDPV ≈ MDMA > methylone. Dopaminergic differentiated cells evidenced higher sensitivity to the neurotoxic effects of both Cathinones and MDMA than the undifferentiated ones, but this effect was not inhibited by the DAT inhibitor GBR 12909. Intracellular oxidative stress mediated by methylone and MDPV was demonstrated by the increase in reactive...
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editor s highlight characterization of hepatotoxicity mechanisms triggered by designer Cathinone drugs β keto amphetamines
Toxicological Sciences, 2016Co-Authors: Maria Joao Valente, Félix Carvalho, Maria De Lourdes Bastos, Paula Guedes De Pinho, Eduarda Fernandes, Ana Margarida Araújo, Márcia CarvalhoAbstract:The use of Cathinone designer drugs in recreational settings has been associated with severe toxic effects, including liver damage. The precise mechanisms by which Cathinones induce hepatotoxicity and whether they act by common pathways remain to be elucidated. Herein, we assessed the toxicity of the Cathinones methylone, pentedrone, 3,4-methylenedioxypyrovalerone (MDPV) and 4-methylethCathinone (4-MEC) in primary rat hepatocytes (PRH) and HepaRG cells, and compared with that of 3,4-methylenedioxymethamphetamine (MDMA). MDPV and pentedrone were significantly more toxic than MDMA, while methylone was the least cytotoxic compound. Importantly, PRH revealed to be the most sensitive experimental model and was thus used to explore the mechanisms underlying the observed toxicity. All drugs elicited the formation of reactive oxygen and nitrogen species (ROS and RNS), but more markedly for methylone, pentedrone and 4-MEC. GSH depletion was also a common effect at the highest concentration tested, whereas only MDPV and pentedrone caused a significant decrease in ATP levels. The antioxidants ascorbic acid or N-acetyl-L-cysteine partially attenuated the observed cell death. All Cathinones triggered significant caspase activation and apoptosis, which was partially reversed by the caspase inhibitor Ac-LETD-CHO. In conclusion, the present data shows that (1) Cathinones induce in vitro hepatotoxic effects that vary in magnitude among the different analogues, (2) oxidative stress and mitochondrial dysfunction play a role in Cathinones-induced hepatic injury, and (3) apoptosis appears to be an important pathway of cell death elicited by these novel drugs.
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Chiral enantioresolution of Cathinone derivatives present in “legal highs”, and enantioselectivity evaluation on cytotoxicity of 3,4-methylenedioxypyrovalerone (MDPV)
Forensic Toxicology, 2016Co-Authors: Bárbara Silva, Márcia Carvalho, Paula Guedes De Pinho, Maria Joao Valente, Carla Fernandes, Maria Elizabeth Tiritan, Madalena M.m. Pinto, Fernando RemiãoAbstract:Recently, great interest has been focused on synthetic Cathinones since their consumption has increased exponentially. All synthetic Cathinones exist as chiral molecules; the biological and/or toxicological properties of Cathinones generally differ according to the enantiomers in human body. In this study, a chiral liquid chromatography method was developed to separate and determine the enantiomeric ratio of synthetic Cathinones present in “legal highs” acquired in old smart shops or over the Internet. All the synthetic Cathinones were efficiently enantio-separated with α and Rs ranging from 1.24 to 3.62 and from 1.24 to 10.52, respectively, using polysaccharide-based chiral stationary phases. All synthetic Cathinones, with the exception of 4-methylethCathinone (4-MEC), were present in the commercialized “legal highs” in an enantiomeric proportion of 50:50. One of the studied chiral compounds was 3,4-methylenedioxypyrovalerone (MDPV), one of the most consumed Cathinone derivative worldwide. Our research group has recently reported its hepatotoxicity in the racemic form. Thus, the analytical enantioresolution of the MDPV was scaled up to multi-milligram using a semi-preparative amylose tris -3,5-dimethylphenylcarbamate column (20 cm × 7.0 mm ID, 7 µm particle size). Both enantiomers were isolated with high enantiomeric purity (enantiomeric excess > 99 %). The toxicity of S -(−)-MDPV and R -(+)-MDPV was evaluated, for the first time, using primary cultures of rat hepatocytes. It was also possible to verify that MDPV enantiomers showed hepatotoxicity in a concentration-dependent manner, but displayed no enantioselective toxicity in this cell culture model.
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Khat and synthetic Cathinones: a review
Archives of Toxicology, 2014Co-Authors: Maria Joao Valente, Félix Carvalho, Maria De Lourdes Bastos, Paula Guedes De Pinho, Márcia CarvalhoAbstract:For centuries, ‘khat sessions’ have played a key role in the social and cultural traditions among several communities around Saudi Arabia and most East African countries. The identification of Cathinone as the main psychoactive compound of khat leaves, exhibiting amphetamine-like pharmacological properties, resulted in the synthesis of several derivatives structurally similar to this so-called natural amphetamine. Synthetic Cathinones were primarily developed for therapeutic purposes, but promptly started being misused and extensively abused for their euphoric effects. In the mid-2000’s, synthetic Cathinones emerged in the recreational drug markets as legal alternatives (‘legal highs’) to amphetamine, ‘ecstasyʼ, or cocaine. Currently, they are sold as ‘bath salts’ or ‘plant foodʼ, under ambiguous labels lacking information about their true contents. Cathinone derivatives are conveniently available online or at ‘smartshops’ and are much more affordable than the traditional illicit drugs. Despite the scarcity of scientific data on these ‘legal highs’, synthetic Cathinones use became an increasingly popular practice worldwide. Additionally, criminalization of these derivatives is often useless since for each specific substance that gets legally controlled, one or more structurally modified analogs are introduced into the legal market. Chemically, these substances are structurally related to amphetamine. For this reason, Cathinone derivatives share with this drug both central nervous system stimulating and sympathomimetic features. Reports of intoxication and deaths related to the use of ‘bath salts’ have been frequently described over the last years, and several attempts to apply a legislative control on synthetic Cathinones have been made. However, further research on their pharmacological and toxicological properties is fully required in order to access the actual potential harm of synthetic Cathinones to general public health. The present work provides a review on khat and synthetic Cathinones, concerning their historical background, prevalence, patterns of use, legal status, chemistry, pharmacokinetics, pharmacodynamics, and their physiological and toxicological effects on animals and humans.