The Experts below are selected from a list of 5193 Experts worldwide ranked by ideXlab platform
Marilyn A. Huestis - One of the best experts on this subject based on the ideXlab platform.
-
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.
-
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.
-
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.
-
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
-
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.
Márcia Carvalho - One of the best experts on this subject based on the ideXlab platform.
-
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.
-
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...
-
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.
-
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.
-
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.
-
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.
-
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
-
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.
Sarah Kerrigan - One of the best experts on this subject based on the ideXlab platform.
-
Postmortem distribution and redistribution of synthetic Cathinones
Forensic Toxicology, 2018Co-Authors: Lindsay Glicksberg, Ruth Winecker, Caitlin Miller, Sarah KerriganAbstract:Purpose Synthetic Cathinones are powerful psychostimulants that have been associated with fatal intoxications. Because of changes that take place following death, postmortem toxicology results require careful interpretation. The purpose of this study was to evaluate the distribution of synthetic Cathinones in postmortem specimens in a series of 50 cathinone-positive fatalities. Methods Liquid chromatography–quadrupole time-of-flight-mass spectrometry was used to quantitatively identify Cathinones in central blood ( n = 51), peripheral blood ( n = 31), urine ( n = 33), liver ( n = 22), vitreous humor ( n = 1) and stomach contents ( n = 1). The distribution of Cathinones and the potential for postmortem redistribution was assessed. Results Among the 50 cases investigated, a total of nine synthetic Cathinones (α-PVP, ethylone, methylone, butylone, MDPV, methedrone, pentylone, 4-MEC, and MDPBP) were identified in 139 specimens. The number of specimens per case ranged from one to six. In cases that included central blood or liver, together with a peripheral blood source, the central/peripheral (C/P) or liver/peripheral (L/P) ratio was calculated to estimate the potential for postmortem redistribution ( n = 21 C/P; n = 11 L/P). Methylone and ethylone appeared to exhibit the greatest potential for postmortem redistribution, producing C/P ratios of 4.0 (1.5–6.1) and 2.9 (0.5–9.2), respectively. In contrast, the C/P ratio for α-PVP was 1.1 (0.5–1.9). Differences in C/P ratios between methylone and α-PVP were statistically significant (α = 0.05). Conclusions Although synthetic Cathinones may exhibit low to moderate postmortem redistribution, significant variability exists due to site- and time-dependent factors. This, in combination with their overall instability, necessitates careful interpretation of postmortem toxicology results.
-
Cathinone stability in authentic urine specimens.
Forensic science international, 2018Co-Authors: Lindsay Glicksberg, Sumandeep Rana, Sarah KerriganAbstract:Abstract Purpose Synthetic Cathinones are encountered in a variety of antemortem and postmortem forensic toxicology investigations. Earlier experimental studies using fortified urine have evaluated analyte, temperature and pH-dependent variables associated with their stability. The purpose of this study was to compare experimental findings with those obtained using authentic urine from cathinone users. Methods In this report we compare cathinone concentrations in 180 authentic unpreserved urine specimens, following known periods of refrigerated storage. These findings are compared with previously published experimental data using fortified drug-free urine. Liquid chromatography quadrupole time-of-flight mass spectrometry (LC-Q/TOF-MS) was used to target 22 Cathinones. Quantitative results were compared in urine specimens (pH 4.5–10) following 5–17 months of storage. Results The 180 specimens resulted in 164 quantitative findings involving α-PVP, ethylone, methylone, MDPV and pentylone. Initial drug concentrations ranged from 25 ng/mL to over 100,000 ng/mL. Upon reanalysis, the percentage of drug remaining (0–119%) was correlated with storage time and specimen pH. The ability to reconfirm original results was not correlated with storage time. Instead, specimen pH was far more predictive. The relationship between initial and final drug concentration was highly pH-dependent, yielding significant correlations for α-PVP, ethylone and methylone, particularly under acidic conditions. Conclusions These results are in good agreement with experimental findings and highlight the critical importance of specimen pH, rather than conventional time dependent variables, when considering cathinone stability in biological samples. The potential for pre-analytical changes in cathinone concentrations must be carefully considered when interpreting their results.
-
Postmortem distribution and redistribution of synthetic Cathinones
Forensic Toxicology, 2018Co-Authors: Lindsay Glicksberg, Ruth Winecker, Caitlin Miller, Sarah KerriganAbstract:Purpose Synthetic Cathinones are powerful psychostimulants that have been associated with fatal intoxications. Because of changes that take place following death, postmortem toxicology results require careful interpretation. The purpose of this study was to evaluate the distribution of synthetic Cathinones in postmortem specimens in a series of 50 cathinone-positive fatalities.
-
Stability of Synthetic Cathinones in Urine.
Journal of analytical toxicology, 2017Co-Authors: Lindsay Glicksberg, Sarah KerriganAbstract:In this report, we evaluate the concentration, pH, temperature and analyte-dependent effects on cathinone stability in preserved human urine. A total of 22 synthetic Cathinones were evaluated at 100 ng/mL and 1,000 ng/mL in pH 4 and pH 8 urine over 6 months. Specimens were stored at -20°C, 4°C, 20°C and 32°C. The stability of synthetic Cathinones was highly dependent on urine pH and storage temperature. Cathinones were considerably more stable in acidic urine (pH 4) at low temperature. In alkaline urine (pH 8) at 32°C, significant losses (>20%) were observed within hours for the majority of drugs. In contrast, all drugs were stable in frozen and refrigerated urine at pH 4 for the duration of the study. These results highlight the importance of sample storage and the potential for pre-analytical changes in concentration during routine shipping and handling of specimens. Significant structural influence was also observed. Cathinones bearing a tertiary amine (pyrrolidine group) were significantly more stable than their secondary amine counterparts. The methylenedioxy group also exerted a significant stabilizing effect on both the tertiary and secondary amines. In the absence of the methylenedioxy group, no significant differences in stability were observed between the unsubstituted and ring substituted secondary amines. Half-lives at ambient temperature in pH 8 urine ranged from 9 h (3-fluoromethcathinone) to 4.3 months (methylenedioxypyrovalerone and 3,4-methylenedioxy-α-pyrrolidinobutiophenone), demonstrating the importance of analyte dependence, and the dual stabilizing effect of both the pyrollidine and methylenedioxy groups. Biological evidence may be subjected to a variety of environmental conditions prior to, and during transport to the forensic laboratory. These findings demonstrate the inherent instability of certain cathinone species in biological evidence under some conditions. Moreover, this study highlights the need for quantitative drug findings in toxicological investigations to be interpreted cautiously, and within the context of specimen storage and integrity.
-
Stability of Synthetic Cathinones in Blood.
Journal of analytical toxicology, 2017Co-Authors: Lindsay Glicksberg, Sarah KerriganAbstract:The synthetic Cathinones are powerful psychostimulants that have been associated with impairment, intoxication and fatal overdose. Forensic laboratories must be able to identify these new drugs as part of antemortem and postmortem toxicology investigations. Preliminary reports have indicated that some of the synthetic Cathinones are unstable in biological matrices. It is important to understand drug stability in biological evidence so that analytical findings can be interpreted appropriately. The objective of this study was to systematically evaluate the concentration, temperature and analyte-dependent stability of synthetic Cathinones in preserved blood using liquid-chromatography/quadrupole-time of flight-mass spectrometry (LC/Q-TOF-MS). Cathinone stability was investigated at frozen, refrigerated, ambient and elevated temperature (-20°C, 4°C, 20°C and 32°C). Although no concentration dependent differences in stability were observed, cathinone stability was highly temperature and analyte-dependent. Substituents on the aromatic ring and nitrogen profoundly influenced stability. Tertiary amines (pyrrolidinyl analogs) were significantly more stable than their N-alkylated (secondary amine) counterparts. Furthermore, the methylenedioxy (MD) group also exerted a significant stabilizing effect, for both secondary and tertiary amines. The unsubstituted and ring-substituted secondary amines were the least stable, most notably 3-fluoromethcathinone (3-FMC). Under some conditions, significant losses were observed within hours of storage. Half-lives ranged from a little as 8 h (3-FMC) to 21 days (3,4-methylenedioxy-α-pyrrolidinobutiophenone, MDPBP) at elevated temperature (32°C). In contrast, half-lives ranged from 0.4 to >10 months when refrigerated and demonstrated even greater stability when frozen. Biological evidence may be subjected to a variety of environmental conditions prior to, and during transport to the laboratory. These findings highlight the need to consider the potential for both temperature and analyte-dependent differences. Due to the inherent instability of certain drugs within the class, quantitative drug findings in toxicological investigations must be interpreted with caution, and within the context of specimen storage and integrity.
Maria Joao Valente - One of the best experts on this subject based on the ideXlab platform.
-
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.
-
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...
-
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.
-
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.