Determination of Three Types of New Psychoactive Tryptamines in Blood by QuEChERS Combined with UPLC-MS/MS / 法医学杂志

Objective To establish a method combining QuEChERS and ultra-high liquid chromatography-tandem mass spectrometry （UPLC-MS/MS） for rapid screening and testing of three types of new psychoactive tryptamines in human blood： 5-MeO-DALT, 5-MeO-MiPT and 5-MeO-DiPT. Methods The effects of the type of extractant, the type and dosage of salting-out agent, and the dosage of adsorbent on the test results of the three tryptamines were investigated. Blood samples were processed by QuEChERS method and then determined by UPLC-MS/MS. Results The linear relationships of 5-MeO-DALT, 5-MeO-MiPT and 5-MeO-DiPT in human blood were good in the range of 0.5-100, 0.5-100 and 0.2-100 ng/mL, respectively, with their coefficients higher than 0.99. The limits of detection （LODs） were 0.1-0.2 ng/mg. The recoveries ranged from 84.86% to 94.57%. Intra-day and inter-day precisions were good. Conclusion The method is simple, rapid, easy to operate and has a high recovery. It is suitable for the qualitative and quantitative study of tryptamines in blood and can provide the reference for public security organs to deal with related cases.

Test purchase of new synthetic tryptamines via the Internet: Identity check by GC-MS and separation by HPLC.

Over the past few years, a continuous alteration of the recreational drug market took place. Among other novel psychoactive drugs, new synthetic tryptamine derivatives appeared on the market. These compounds are mainly traded via the Internet, which has become an important marketplace for the sale of recreational drugs. The goal of our research was to check, if 13 new synthetic tryptamines obtained by test purchase via different online vendors meet the promised identity. Analysis was performed by GC-MS, using a common 30 m HP-5MS capillary column as stationary phase. Subsequently, a simple HPLC method for the separation of these tryptamines was developed. Therefore, the aim was to establish a method to separate a broad spectrum of trypamines simultaneously within short time. Measurements were performed by a LiChrospher® RP-18e column and a mobile phase consisting of 0.1% triethylammonium acetate buffer, methanol and acetonitrile. Both presented methods were found to be suitable for the identification as well as separation of tryptamines as the analysis times were short and the selectivity sufficient. Moreover, enantioseparation of 3 chiral tryptamine derivatives purchased via the Internet by HPLC-UV and an Astec® Cyclobond I™ 2000 as CSP was performed. All of them were sold as racemic mixtures.

Ação de metabólitos da via das triptaminas em linhagens de melanomas / Action of tryptamines pathway metabolites in melanoma strains

O triptofano (Trp) é essencial para muitos processos fisiológicos e seu metabolismo apresenta-se alterado em doenças como no câncer. O Trp é degradado por três vias: via das quinureninas; via serotonérgica e via das triptaminas. A primeira está envolvida com a tolerância e o imune escape de células tumorais, já a segunda leva a produção de serotonina e melatonina, com uma diversidade de funções biológicas e que possuem atividades antitumorais reconhecidas. A terceira rota é a menos estudada e a função dos compostos sintetizados ainda é desconhecida. Trabalhos do grupo mostram que a via das triptaminas é ativa em melanomas e que triptamina (TRY) e dimetiltriptamina (DMT), metabólitos produzidos por esta via, também possuem atividade antitumoral. Nosso objetivo é avançar sobre a compreensão de como esta via afeta o metabolismo e crescimento tumoral. Para tanto, estudamos mais detalhadamente a via em linhagens de melanoma. A adição de TRY e DMT nas culturas modificou a produção de compostos das outras rotas de metabolização do Trp. Além disso, TRY e DMT afetaram a invasividade das células tumorais e TRY aumentou a atividade citotóxica de células mononucleares frente a melanomas. Observamos que apesar dos efeitos biológicos da via das quinureninas estar amplamente relacionado a ligação dos metabólitos no receptor de aril hidrocarbonetos, para a via das triptaminas o receptor parece não estar associado com a atividade antitumoral descrita. Nossos resultados apontam para a importância da via das triptaminas na dinâmica tumoral e a necessidade de estudos mais amplos relacionados ao metabolismo do Trp

Tryptophan (Trp) is essential for many physiological processes and its metabolism is modified in several diseases such as in cancer. TRP is broken down into three pathways: kynurenine path, serotonergic path and tryptamine path. The first is involved in tolerance and immune escape of cancer cells, while the second leads to the production of serotonin and melatonin, which have a variety of biological functions and recognized antitumor activity. The third route is the least studied and the biological function of the synthesized compounds is still unknown. Our group shows that tryptamine path is active in melanomas and tryptamine (TRY) and dimethyltryptamine (DMT), metabolites produced by this route, also have antitumor activity. Our goal is make progress on understanding how tryptamine route affects metabolism and tumor growth. Therefore, we studied in detail this pathway in melanoma cell lines. The addition of TRY and DMT into the cultures leds the production of metabolites of other Trp routes. Moreover, TRY and DMT affect the invasiveness of tumor cells and TRY increased antitumor activity of the immune system against melanomas. We observed that despite biological effects of kynurenine path be largely related to metabolites binding aryl hydrocarbon receptor, for tryptamine pathway the receptor seems not to be associated with the described antitumor activity. Our results point the importance of tryptamine pathway in tumor dynamics and the need for broader studies related to Trp metabolism

Effects of rizatriptan on calcitonin gene-related peptide, proenkephalin and cholecystokinin mRNA expressions in the trigeminal ganglia of a rat migraine model / 中华神经科杂志

Objective This study assesses the influence of rizatriptan on calcitonin gene-related peptide (CGRP),proenkephalin (PENK) and cholecystokinin (CCK) mRNA expressions in the trigeminal ganglia of a rat migraine model and investigates the possible mechanisms by which triptans treat migraine.Methods A total of 24 rats were randomly divided into four groups:normal control group (A),migraine model group(B),rizatriptan control group (C) and rizatriptan treatment group(D).Groups C and D were intragastrically perfused with rizatriptan,1 mg/kg per day.After 7 days,nitroglycerin was subcutaneously injected into the buttocks of the groups B and D to induce migraine.Two hours after nitroglycerin injection,the trigeminal ganglia was isolated.CGRP,PENK and CCK mRNA expressions in the trigeminal ganglia were determined using SYBR Green Ⅰ real-time quantitative PCR.Results The copy number of CGRP mRNA (× 107) in 200 ng total RNA of each group was 0.05 ±0.01,1.30 ±0.52,0.23 ±0.12,0.43 ±0.33 ; The copy number of PENK mRNA (× 103) in 200 ng total RNA of each group was 3.30 ± 1.65,0.34 ±0.14,3.91 ± 2.44,0.71 ± 0.13.The copy number of CGRP mRNA in the trigeminal ganglia of group B was significantly higher than that of group A (q =7.854,P ＜ 0.05) ; CGRP mRNA expressions were significantly lower in the trigeminal ganglia of rats in group D compared with group B (q =5.458,P ＜0.05).Compared with group A,PENK mRNA expressions in the trigeminal ganglia of rats were significantly lower in group B (q =4.478,P ＜ 0.05).PENK mRNA expressions were significantly higher in trigeminal ganglia of rats in group C compared with group D (q =4.838,P ＜ 0.05).CCK mRNA expression in trigeminal ganglia of rats was similar among groups.Conelusions Rizatriptan can decrease the expressions of CGRP in the trigeminal ganglia of the migraine rats and exhibits neurogenic inflammation triggered by CGRP.PENK expressions decrease in the trigeminal ganglia of the migraine rats,weaken the analgesic effects of enkephalin.