Methasterone is a designer anabolic steroid that is prohibited for athletes and is monitored by anti-doping laboratories. In this work, our objective is to discover new human phase I metabolites, define their excretion kinetics for 30 days and analyze their phase II metabolism (sulfate, cysteine and N-acetylcysteine conjugates). Urine samples from four volunteers were analyzed by chromatographic techniques. Through gas chromatography coupled to mass spectrometry analysis it was possible to detect methasterone and its nine phase I metabolites in the urine samples after glucuronide enzymatic hydrolysis, from which one were previously unreported. These nine compounds were not excreted in free form. The new proposed metabolite is 17β-hydroxy-2α,17α-dimethyl-5β-androstan-3-one, obtained from the epimerization at C5. The 3α-hydroxy metabolite, currently monitored by anti-doping laboratories, was the most abundant and was detected for the longest time. Furthermore, four other long-term metabolites were identified. By ultra-performance liquid chromatography coupled to tandem mass spectrometry, only the drug and a known metabolite were detected after glucuronide hydrolysis, and phase II metabolites were not found. Thus, our results contribute to elucidating methasterone metabolism, including long-term metabolites besides of the 3α-hydroxy in routine doping analysis, which is very important due to variation in human metabolism.

Ephedrines are widely used in therapy. Because of their stimulant properties, these substances are relevant in different forensic fields. At present, the state of the art for ephedrines quantification relay based on a liquid chromatography mass spectrometry, mainly because of the dilute-and-shoot approach. Notwithstanding, several gas chromatography based methods have already been described, all of them include cleanup steps, with the potential disadvantage of incurring errors and increasing the workload. In this paper, a straightforward method for ephedrine quantification based on gas chromatographic mass spectrometry, without cleanup and based on Doehlert matrix optimization is presented. Only 10 µL of a urine sample is necessary and for N-methyl-N-(trimethylsilyl)trifluoroacetamide/N-methyl-bis-trifluoracetamide derivatives, the intermediate precision was 2.77% for ephedrine, 9.20% for cathine, 8.29% for norephedrine and 4.27% for pseudoephedrine. The limit of detection was 20 ng mL-1 for ephedrine, 30 ng mL-1 for cathine and 40 ng mL-1 for norephedrine and pseudoephedrine.

In this report, a rapid and selective analytical method for the determination of monoterpenes in Alpinia zerumbet essential oil (AZEO) by gas chromatography (GC) flame ionization detection (FID) was developed, optimized and validated. The suitability of six different capillary columns was investigated by polarity phase constants calculated based on the methodology proposed by Rohrschneider and McReynolds. The most suitable column was then used in a central composite design applying the following GC factors to investigate responses based on peak resolutions and analysis time: initial oven temperature, heating rate and carrier gas flow rate. The optimized method has initial oven temperature of 60 ºC, heating rate of 5.1 ºC min-1 and flow rate of 1.0 mL min-1 using the DB-35 capillary column. The validation acceptance criteria were met in all cases. The method was successfully applied for the quantification of major monoterpenes found in AZEO in different samples of Alpinia zerumbet.

<p>Luteinizing hormone (LH) can be used by athletes as an alternative way to take illicit advantage of androgenic effects, since it stimulates the secretion of testosterone by the Leydig cells. As consequence, LH use is illegal for male athletes and the hormone is included in the World Anti-Doping Agency (WADA) prohibited list. Since LH is an endogenous substance, the strategy used to allow the detection of LH abuse was the establishment of a reference value. All analyses were developed by IMMULITE^®, which was submitted to a complete validation performance with urine. The hormone showed to be stable at 4 °C and -20 °C for at least 40 days and for 10 days at room temperature and unstable after the second cycle of freezing and thawing. The reference population of male Brazilian athletes showed a cut-off value of 37.4 mIU mL-1. The results indicate the assay is suitable for application in doping control analysis.</p>

Glicocorticosteroides estão incluídos na lista de substâncias proibidas da Agência Mundial Antidoping como analitos que dispensam quantificação. Entretanto, abordagens semiquantitativas são necessárias antes de relatar um resultado analítico adverso para essa classe de substâncias. Cromatografia líquida acoplada à espectrometria de massa (LC-MS) tem sido empregada com excelente seletividade e especificidade na análise de xenobiontes em misturas complexas. Foram analisadas amostras contendo glicocorticosteroides exógenos, a partir de diferentes vias de administração, utilizando o método proposto. Amostras geradas após administração sistêmica (via oral) apresentaram níveis de prednisolona ou prednisona superiores a 30 ng mL-1, limite mínimo de desempenho requerido usado como parâmetro de diagnóstico. Por outro lado, vias não-sistêmicas parecem não apresentar biodisponibilidade para gerar um caso adverso em análises de controle de dopagem. Considerando o número reduzido de estudos similares na literatura, mais estudos de excreção podem ser realizados usando o método relatado com o objetivo de fornecer mais informações sobre o perfil de excreção de glicocorticosteroides administrados por vias sistêmicas e não sistêmicas.

Glucocorticosteroids are included in the World Anti-Doping Agency (WADA) prohibited list as non-threshold analytes. Nevertheless, at least semi-quantitative approaches are necessary before reporting an adverse analytical finding for this class of substances. The liquid chromatography coupled to mass spectrometry (LC-MS) has been used with excellent selectivity and specificity in xenobiotic analysis in complex matrices. Real samples containing exogenous glucocorticosteroids from different administration routes were analyzed using the method proposed. Samples from systemic administration (oral route) presented levels of prednisolone or prednisone higher than 30 ng mL-1, the minimum requirement performance level used as diagnostic parameter. On the other hand, non-systemic routes seem to not have bioavailability to generate an adverse case in doping control analysis. Considering the low number of similar studies available in the literature, more urinary studies could be done using the method reported aiming to provide more information about the excretion profile of glucocorticosteroids administered by systemic and non-systemic routes.

No mundo das drogas de otimização de desempenho, hormônio gonadotrófico humano (hCG) é crescentemente usado sozinho em também em combinação com ciclos de Agentes Anabólicos Esteróides. Em homens o hCG mimetiza o hormônio Luteinizante (LH) e ajuda a aumentar a produção de testosterona nos testículos. Assim sendo, hCG é comumente usado durante e depois de ciclos de esteróides para restaurar o tamanho testicular assim como a produção de testosterona endógena. Controle de dopagem para hCG requer atenção especial, visto que, deve ser feita diferença entre a origem endógena e exógena de ambas substâncias. Na carência de um padrão mais claro do que seria um nível esperado de normalidade, os laboratórios de controle de dopagem tem aplicado valores de corte entre 5 e 25 mUI/mL. Por causa de grande diferença em desenhos de ensaios e especificidade para diferentes formas de hCG, as concentrações medidas são altamente dependentes do ensaio usado e população analisada. O objetivo do estudo é o estabelecimento do valor de corte para a caracterização do abuso exógeno por atletas obtido com a aplicação do Imunoensaio Enzimático por Micropartículas.

In the world of performance enhancing drugs, human Chorionic Gonadotrophin (hCG) is increasingly used alone and also in combination with various Anabolic Androgenic Steroid cycles. In males, hCG mimics luteinizing hormone (LH) and helps increase testosterone production in the testes. As such, hCG is commonly used during and after steroid cycles to maintain and restore testicular size as well as endogenous testosterone production. Doping control for hCG requires special attention since a difference must be made between the endogenous and exogenous origin of both substances. In the absence of a clearer pattern of what would be an expected level of normality, the doping control laboratories have been applying cut off values between 5 and 25 mUI/mL. Because of the large differences in assay design and specificity for different forms of hCG, the concentrations measured are highly dependent on the assay used and population analyzed. The aim of the study is the establishment of a cut off value for the characterization of exogenous misuse by athletes obtained with the usage of Microparticles Enzyme Immunoassays.

Um procedimento é descrito para a determinação simultânea de substâncias androgênicas, incluindo esteróides e beta2-agonistas. O método envolve a análise de compostos anabólicos em urina hidrolisada, usando-se extração líquido-líquido com subseqüente conversão a derivados trimetilsililéteres, para análise por CG-EM. Injeção com divisão de fluxo de 1/10 dos derivados TMS a 280 °C, em uma coluna capilar, inicialmente mantida a 140 °C e então programada para 180 °C a 40 °C min-1, seguida por 3 ºC min-1 até 230 ºC, e então, 40 ºC min-1 até 300 ºC, resulta em uma boa resolução e formato de pico para todos os compostos. Os limites de detecção da maioria dos esteróides foi de 1 ng mL-1, exceto para a formebolona e trembolona (25 ng mL-1). Quando aplicado a amostras de urinas selecionadas com evidência de degradação bacteriana e metabólitos provenientes de medicação/vitaminas, o método permite a rápida triagem para androgênios e outras substâncias monitoradas em rotina. A resolução foi adequada para avaliar o perfil esteroidal endógeno, relevante para controle de dopagem e aplicações médicas.

A procedure is described for the simultaneous determination of androgenic substances including steroids and beta2-agonists. The method involves analysis of hydrolized urinary anabolic compounds using liquid-liquid extraction, with subsequent conversion to trimethylsilylether derivatives for the analysis by GC-MS. Pulse split injection 1/10 of the TMS derivatives at 280 °C into the capillary column, initially maintained at 140 °C then programmed to 180 °C at 40 °C min-1, followed by 3 ºC min-1 to 230 ºC and then 40 ºC min-1 to 300 ºC, resulted in good resolution and peak shape for all compounds. The detection limits of most of the steroids were 1 ng mL-1 except for formebolone and trenbolone (25 ng mL-1). When applied to selected urine samples with evidence of bacterial degradation and metabolites from usual medications/vitamins, the method allowed rapid screening for androgens and other substances monitored in routine. The resolution was adequate to evaluate the endogenous steroid profile relevant to doping control and medical applications.

Glucocorticosteroids (CT) show a powerful anti-inflammatory effect. In view of this, they are extensively used in sports medicine, but their misuse (systemic administration) can lead to severe injuries on athletes and has been restricted by the IOC in recent years. For GC-MS analysis, a derivatization step is necessary in order to convert them to a stable form, thus preventing thermal breakdown with the loss of the ketone side chain. Derivatization of CT is reviewed and a new approach of silylation is presented for screening of CT in urine.