Bioanalytical studies of designer benzodiazepines

The fast appearance of benzodiazepine analogues, referred to as new psychoactive substance (NPS) or designer benzodiazepines, requires the continuous update of detection methods in order to keep up with the latest drugs on the recreational drug market. Moreover, as usually only limited information on toxicity and excretion patterns of these new drugs exists, this needs to be evaluated to report on adverse effects and to determine suitable targets for drug testing. Urine drug testing usually involves screening using immunoassay followed by confirmation of positive screening results using mass spectrometric (MS) methods. We studied the detectability of designer benzodiazepines in urine using commercial immunoassays and demonstrated that most designer benzodiazepines can be detected by immunoassay. It is thus important to update confirmation methods to include designer benzodiazepines. We also developed a liquid chromatographic–tandem MS (LC–MS/MS) confirmation method for designer benzodiazepines in urine using direct dilution of samples and hydrolysis of conjugates. Subsequently, a further improved screening and confirmation method using LC–high-resolution MS (LC–HRMS(/MS)) was developed. HRMS screening is performed in full scan and is a generic method that can easily include new analytes. LC–HRMS/MS confirmation only requires re-injection of the sample. Many samples from drug dependent patients with a positive immunoassay screening result for benzodiazepines but not containing prescription medicines detected a designer benzodiazepine instead. Comparable results were obtained for acute intoxication cases from emergency wards in the STRIDA project. In total, 28 designer benzodiazepines were covered by the analytical method and 17 of these were detected in the samples. Classification of a designer benzodiazepine as a narcotic substance generally meant that it was removed from the NPS market and replaced with another novel benzodiazepine. It was further demonstrated that intoxications by designer benzodiazepines might cause central nervous system depression. Studies on metabolic patterns of five designer benzodiazepines using urine samples from confirmed intoxication cases identified suitable analytical targets for urine drug testing, instead of or together with the parent compounds and both with and without hydrolysis of conjugated forms. In summary, the results demonstrated frequent use of designer benzodiazepines in Sweden and in cases of acute intoxication that they might cause serious adverse effects. This underlines the importance of including designer benzodiazepines and/or metabolites thereof in drug testing. Screening for designer benzodiazepines can be performed by immunoassay or LC–HRMS, and confirmation methods can make use of direct dilution of urine samples followed by hydrolysis and direct injection into LC–MS or LC–HRMS systems

In vitro metabolic fate of nine LSD-based new psychoactive substances and their analytical detectability in different urinary screening procedures

The market of new psychoactive substances (NPS) is characterized by a high turnover and thus provides several challenges for analytical toxicology. The analysis of urine samples often requires detailed knowledge about metabolism given that parent compounds may either be present only in small amounts or may not even be excreted. Hence, knowledge of the metabolism of NPS is a prerequisite for the development of reliable analytical methods. The main aim of this work was to elucidate for the first time the pooled human liver S9 fraction metabolism of the nine d-lysergic acid diethylamide (LSD) derivatives 1-acetyl-LSD (ALD-52), 1-propionyl-LSD (1P-LSD), 1-butyryl-LSD (1B-LSD), N6-ethyl-nor-LSD (ETH-LAD), 1-propionyl-N6-ethyl-nor-LSD (1P-ETH-LAD), N6-allyl-nor-LSD (AL-LAD), N-ethyl-N-cyclopropyl lysergamide (ECPLA), (2’S,4’S)-lysergic acid 2,4-dimethylazetidide (LSZ), and lysergic acid morpholide (LSM-775) by means of liquid chromatography coupled to high resolution tandem mass spectrometry. Identification of the monooxygenase enzymes involved in the initial metabolic steps was performed using recombinant human enzymes and their contribution confirmed by inhibition experiments. Overall, N-dealkylation, hydroxylation, as well as combinations of these steps predominantly catalyzed by CYP1A2 and CYP3A4 were found. For ALD-52, 1P-LSD, and 1B-LSD deacylation to LSD was observed. The obtained mass spectral data of all metabolites is essential for reliable analytical detection particularly in urinalysis and for differentiation of the LSD-like compounds as biotransformations also led to structurally identical metabolites. However, in urine of rats after the administration of expected recreational doses and using standard urine screening approaches, parent drugs or metabolites could not be detected

In vitro metabolic fate of nine LSD-based new psychoactive substances and their analytical detectability in different urinary screening procedures

The market of new psychoactive substances (NPS) is characterized by a high turnover and thus provides several challenges for analytical toxicology. The analysis of urine samples often requires detailed knowledge about metabolism given that parent compounds either may be present only in small amounts or may not even be excreted. Hence, knowledge of the metabolism of NPS is a prerequisite for the development of reliable analytical methods. The main aim of this work was to elucidate for the first time the pooled human liver S9 fraction metabolism of the nine d-lysergic acid diethylamide (LSD) derivatives 1-acetyl-LSD (ALD-52), 1-propionyl-LSD (1P-LSD), 1-butyryl-LSD (1B-LSD), N6-ethyl-nor-LSD (ETH-LAD), 1-propionyl-N6-ethyl-nor-LSD (1P-ETH-LAD), N6-allyl-nor-LSD (AL-LAD), N-ethyl-N-cyclopropyl lysergamide (ECPLA), (2′S,4′S)-lysergic acid 2,4-dimethylazetidide (LSZ), and lysergic acid morpholide (LSM-775) by means of liquid chromatography coupled to high-resolution tandem mass spectrometry. Identification of the monooxygenase enzymes involved in the initial metabolic steps was performed using recombinant human enzymes and their contribution confirmed by inhibition experiments. Overall, N-dealkylation and hydroxylation, as well as combinations of these steps predominantly catalyzed by CYP1A2 and CYP3A4, were found. For ALD-52, 1P-LSD, and 1B-LSD, deacylation to LSD was observed. The obtained mass spectral data of all metabolites are essential for reliable analytical detection particularly in urinalysis and for differentiation of the LSD-like compounds as biotransformations also led to structurally identical metabolites. However, in urine of rats after the administration of expected recreational doses and using standard urine screening approaches, parent drugs or metabolites could not be detected

The emergence of new psychoactive substance (NPS) benzodiazepines: a review

The market for new psychoactive substances has increased markedly in recent years and there is now a steady stream of compounds appearing every year. Benzodiazepines consist of only a fraction of the total number of these compounds but their use and misuse has rapidly increased. Some of these benzodiazepines have only been patented, some of them have not been previously synthesised and the majority have never undergone clinical trials or tests. Despite their structural and chemical similarity, large differences exist between the benzodiazepines in their pharmacokinetic parameters and metabolic pathways and so they are not easily comparable. As benzodiazepines have been clinically used since the 1960s many analytical methods exist to quantify them in a variety of biological matrices and it is expected that these methods would also be suitable for the detection of benzodiazepines that are new psychoactive substances. Illicitly obtained benzodiazepines have been found to contain a wide range of compounds such as opiates which presents a problem since the use of them in conjunction with each other can lead to respiratory depression and death. The aim of this review is to collate the available information on these benzodiazepines and to provide a starting point for the further investigation of their pharmacokinetics which is clearly required

Current trends in drug metabolism and pharmacokinetics.

Pharmacokinetics (PK) is the study of the absorption, distribution, metabolism, and excretion (ADME) processes of a drug. Understanding PK properties is essential for drug development and precision medication. In this review we provided an overview of recent research on PK with focus on the following aspects: (1) an update on drug-metabolizing enzymes and transporters in the determination of PK, as well as advances in xenobiotic receptors and noncoding RNAs (ncRNAs) in the modulation of PK, providing new understanding of the transcriptional and posttranscriptional regulatory mechanisms that result in inter-individual variations in pharmacotherapy; (2) current status and trends in assessing drug-drug interactions, especially interactions between drugs and herbs, between drugs and therapeutic biologics, and microbiota-mediated interactions; (3) advances in understanding the effects of diseases on PK, particularly changes in metabolizing enzymes and transporters with disease progression; (4) trends in mathematical modeling including physiologically-based PK modeling and novel animal models such as CRISPR/Cas9-based animal models for DMPK studies; (5) emerging non-classical xenobiotic metabolic pathways and the involvement of novel metabolic enzymes, especially non-P450s. Existing challenges and perspectives on future directions are discussed, and may stimulate the development of new research models, technologies, and strategies towards the development of better drugs and improved clinical practice

”Kan man skydda sig mot någon form av cancer så ska man väl det.-” : Unga vaccinerade kvinnors kunskap om Humant Papillomvirus samt kunskap om och inställning till vaccination mot Humant Papillomvirus

Bakgrund Humant papillomvirus (HPV) orsakar vårtor och är en vanligt förekommande könssjukdom världen över. Vaccination mot de vanligaste HPV-typerna som kan orsaka kondylom och leda till cancer ingår sedan 2012 i det allmänna vaccinationsprogrammet för flickor och unga kvinnor. Tidigare forskning visar att unga kvinnor trots låg kunskap om viruset, har en positiv inställning till vaccination. Syfte Att undersöka unga vaccinerade kvinnors kunskap om HPV samt deras kunskap om och inställning till HPV-vaccination. Metod En kvalitativ explorativ studie. The Health Belief Model användes som teoretisk modell. Individuella intervjuer med åtta unga kvinnor som vaccinerats mot HPV. Data analyserades med innehållsanalys. Resultat Totalt genomfördes åtta intervjuer med unga kvinnor födda 1993-1998. Tre kategorier skapades: 1) Bristande kunskap om HPV 2) Tillförlitligt skydd mot cancer samt 3) Vaccinet är tillgängligt. Kunskapen om HPV och HPV-vaccin var låg hos de unga kvinnorna. Den främsta anledningen till att de valde att vaccinera sig var rädsla för cancer, andras inflytande till vaccinering, främst från mödrar, en tilltro till hälso- och sjukvården och till vaccinet samt att vaccinet är tillgängligt. Slutsats Det är tydligt att kunskapen om HPV och vaccinet är låg bland de deltagande unga kvinnorna. Inför framtiden behövs anpassad information till unga kvinnor om viruset och vaccinet för att tillgodose behovet av information. Det är viktigt att unga kvinnor som vaccineras mot HPV har kunskap om vaccinet för att veta hur de skyddar sig mot HPV och att de även ska förstå vikten av att gå på gynekologisk cellprovskontroll som en del av prevention av HPV.Background Human papillomavirus (HPV) cause warts and is a common sexually transmitted infection worldwide. Vaccination against the most common HPV types that can cause genital warts and cancer is implemented in the national vaccination programme for girls and young women since 2012. Previous research shows that young women, despite low knowledge about the virus, are in favour of the vaccine. Objective To explore young vaccinated women’s knowledge about HPV and knowledge and attitudes towards HPV-vaccination. Method An qualitative explorative study. The Health Belief Model was the  theoretical framework. Individual interviews were conducted with young women vaccinated against HPV. Data were analyzed with content analyses. Results In total eight interviews were undertaken with young women born in 1993-1998. Three categories were revealed through the interviews: 1) Lack of knowledge about HPV 2) Reliable protection against cancer and 3) The vaccine is available. The young women had low knowledge about HPV and HPV vaccine. The main reasons for vaccination were; fear of cancer, influence from others, especially the mothers, trust in the healthcare and the vaccine and the vaccine is available. Conclusion The knowledge of HPV and the vaccine was low among the included women. In the future the iformation about the virus and the vaccine needs to be adapted to the young women to provide the need of information. It is important that young women who are vaccinated against HPV have knowledge about the vaccine to be able to protect themselves against HPV and that they are aware of the importance of attending future cervical cancer screening controls as a part of the prevention against HPV.

”Kan man skydda sig mot någon form av cancer så ska man väl det.-” : Unga vaccinerade kvinnors kunskap om Humant Papillomvirus samt kunskap om och inställning till vaccination mot Humant Papillomvirus

Bakgrund Humant papillomvirus (HPV) orsakar vårtor och är en vanligt förekommande könssjukdom världen över. Vaccination mot de vanligaste HPV-typerna som kan orsaka kondylom och leda till cancer ingår sedan 2012 i det allmänna vaccinationsprogrammet för flickor och unga kvinnor. Tidigare forskning visar att unga kvinnor trots låg kunskap om viruset, har en positiv inställning till vaccination. Syfte Att undersöka unga vaccinerade kvinnors kunskap om HPV samt deras kunskap om och inställning till HPV-vaccination. Metod En kvalitativ explorativ studie. The Health Belief Model användes som teoretisk modell. Individuella intervjuer med åtta unga kvinnor som vaccinerats mot HPV. Data analyserades med innehållsanalys. Resultat Totalt genomfördes åtta intervjuer med unga kvinnor födda 1993-1998. Tre kategorier skapades: 1) Bristande kunskap om HPV 2) Tillförlitligt skydd mot cancer samt 3) Vaccinet är tillgängligt. Kunskapen om HPV och HPV-vaccin var låg hos de unga kvinnorna. Den främsta anledningen till att de valde att vaccinera sig var rädsla för cancer, andras inflytande till vaccinering, främst från mödrar, en tilltro till hälso- och sjukvården och till vaccinet samt att vaccinet är tillgängligt. Slutsats Det är tydligt att kunskapen om HPV och vaccinet är låg bland de deltagande unga kvinnorna. Inför framtiden behövs anpassad information till unga kvinnor om viruset och vaccinet för att tillgodose behovet av information. Det är viktigt att unga kvinnor som vaccineras mot HPV har kunskap om vaccinet för att veta hur de skyddar sig mot HPV och att de även ska förstå vikten av att gå på gynekologisk cellprovskontroll som en del av prevention av HPV.Background Human papillomavirus (HPV) cause warts and is a common sexually transmitted infection worldwide. Vaccination against the most common HPV types that can cause genital warts and cancer is implemented in the national vaccination programme for girls and young women since 2012. Previous research shows that young women, despite low knowledge about the virus, are in favour of the vaccine. Objective To explore young vaccinated women’s knowledge about HPV and knowledge and attitudes towards HPV-vaccination. Method An qualitative explorative study. The Health Belief Model was the  theoretical framework. Individual interviews were conducted with young women vaccinated against HPV. Data were analyzed with content analyses. Results In total eight interviews were undertaken with young women born in 1993-1998. Three categories were revealed through the interviews: 1) Lack of knowledge about HPV 2) Reliable protection against cancer and 3) The vaccine is available. The young women had low knowledge about HPV and HPV vaccine. The main reasons for vaccination were; fear of cancer, influence from others, especially the mothers, trust in the healthcare and the vaccine and the vaccine is available. Conclusion The knowledge of HPV and the vaccine was low among the included women. In the future the iformation about the virus and the vaccine needs to be adapted to the young women to provide the need of information. It is important that young women who are vaccinated against HPV have knowledge about the vaccine to be able to protect themselves against HPV and that they are aware of the importance of attending future cervical cancer screening controls as a part of the prevention against HPV.

”Kan man skydda sig mot någon form av cancer så ska man väl det.-” : Unga vaccinerade kvinnors kunskap om Humant Papillomvirus samt kunskap om och inställning till vaccination mot Humant Papillomvirus

Bakgrund Humant papillomvirus (HPV) orsakar vårtor och är en vanligt förekommande könssjukdom världen över. Vaccination mot de vanligaste HPV-typerna som kan orsaka kondylom och leda till cancer ingår sedan 2012 i det allmänna vaccinationsprogrammet för flickor och unga kvinnor. Tidigare forskning visar att unga kvinnor trots låg kunskap om viruset, har en positiv inställning till vaccination. Syfte Att undersöka unga vaccinerade kvinnors kunskap om HPV samt deras kunskap om och inställning till HPV-vaccination. Metod En kvalitativ explorativ studie. The Health Belief Model användes som teoretisk modell. Individuella intervjuer med åtta unga kvinnor som vaccinerats mot HPV. Data analyserades med innehållsanalys. Resultat Totalt genomfördes åtta intervjuer med unga kvinnor födda 1993-1998. Tre kategorier skapades: 1) Bristande kunskap om HPV 2) Tillförlitligt skydd mot cancer samt 3) Vaccinet är tillgängligt. Kunskapen om HPV och HPV-vaccin var låg hos de unga kvinnorna. Den främsta anledningen till att de valde att vaccinera sig var rädsla för cancer, andras inflytande till vaccinering, främst från mödrar, en tilltro till hälso- och sjukvården och till vaccinet samt att vaccinet är tillgängligt. Slutsats Det är tydligt att kunskapen om HPV och vaccinet är låg bland de deltagande unga kvinnorna. Inför framtiden behövs anpassad information till unga kvinnor om viruset och vaccinet för att tillgodose behovet av information. Det är viktigt att unga kvinnor som vaccineras mot HPV har kunskap om vaccinet för att veta hur de skyddar sig mot HPV och att de även ska förstå vikten av att gå på gynekologisk cellprovskontroll som en del av prevention av HPV.Background Human papillomavirus (HPV) cause warts and is a common sexually transmitted infection worldwide. Vaccination against the most common HPV types that can cause genital warts and cancer is implemented in the national vaccination programme for girls and young women since 2012. Previous research shows that young women, despite low knowledge about the virus, are in favour of the vaccine. Objective To explore young vaccinated women’s knowledge about HPV and knowledge and attitudes towards HPV-vaccination. Method An qualitative explorative study. The Health Belief Model was the  theoretical framework. Individual interviews were conducted with young women vaccinated against HPV. Data were analyzed with content analyses. Results In total eight interviews were undertaken with young women born in 1993-1998. Three categories were revealed through the interviews: 1) Lack of knowledge about HPV 2) Reliable protection against cancer and 3) The vaccine is available. The young women had low knowledge about HPV and HPV vaccine. The main reasons for vaccination were; fear of cancer, influence from others, especially the mothers, trust in the healthcare and the vaccine and the vaccine is available. Conclusion The knowledge of HPV and the vaccine was low among the included women. In the future the iformation about the virus and the vaccine needs to be adapted to the young women to provide the need of information. It is important that young women who are vaccinated against HPV have knowledge about the vaccine to be able to protect themselves against HPV and that they are aware of the importance of attending future cervical cancer screening controls as a part of the prevention against HPV.