ABSTRACT Doping control analytics is already considered a highly integrative branch of science. In its development, analytical chemistry, biochemistry and pharmacology have merged in a sublime way, all strictly amalgamated and controlled by a quality management system. The objective of the work was to describe the development of this science in Cuba and at the same time it is a tribute to its protagonists. A broad search was carried out on doping control activities in Cuba, interviews were carried out with persons involved in the activity and the archived documents of the Quality Management System of the Havana Anti-Doping Laboratory were reviewed. The present work describes the main antecedents and the evolution of the analysis of doping control in Cuba. It describes the evolution of the instruments and the personnel involved from 1973 to 2020, as well as the inter-center relations, research, and the accreditation process. It was divided into three main parts. The first describes the situation of doping analytics in the 1970s and 1980s; the second part describes the activities carried out in Cuba between 1973 and 1991 and the third part describes the creation and evolution of the Havana Antidoping Laboratory from 1999 to the present.

RESUMEN La analítica del control del dopaje es considerada ya una rama de las ciencias altamente integradora. En su desarrollo se ha fusionado la química analítica, la bioquímica y la farmacología, todo ello amalgamado de manera estricta y controlada por un sistema de gestión de la calidad. El objetivo del trabajo fue describir el desarrollo de esta ciencia en Cuba y al mismo tiempo constituye un homenaje a sus protagonistas. Se realizó una búsqueda amplia sobre las actividades de control del dopaje en Cuba, se realizaron entrevistas a personal involucrado en la actividad y se utilizaron los documentos archivados del Sistema de Gestión de la Calidad el Laboratorio Antidoping de la Habana. El trabajo describe los principales antecedentes y la evolución de la analítica del control del dopaje en Cuba. Describe la evolución de los instrumentos y el personal involucrado desde 1973 hasta el 2020, así como las relaciones inter-centros, investigaciones, y proceso de acreditación. Se dividió en tres partes fundamentales: situación de la analítica del dopaje en las décadas de 1970 y 1980; actividades ejecutadas en Cuba entre los años 1973 y 1991 y creación y evolución del Laboratorio Antidoping de la Habana desde el año 1999 hasta la actualidad.

ABSTRACT The World Anti-Doping Agency has, among its functions, the accreditation of laboratories for doping control and to maintain this category it is necessary to comply with international standards. One of the requirements is the constant increasing of the assays sensitivity and with it the increasing of the analytical performance of the laboratory. The present work describes the introduction of a new instrumental method using tandem mass spectrometry to detect forbidden substances, mainly steroids. The assay was validated for 23 analytes, out of them 21 were anabolic steroids, one β2 agonist and one antiestrogenic substance. The validation results showed that the proposed method is highly selective and precise with intra-day variations between 1-17 % for the lowest concentrations and between 2,1-13,9 % for the highest one. The limits of detection were settled between 0,1-2,5 ng/mL except for clenbuterol (0,02 ng/mL). The detector showed no carryover after the analysis of concentrated samples, and the recovery and robustness tests were adequate according to the purpose, screening or confirmation process. Finally, the application of the validated assay to the evaluation of samples from a study of urinary excretion of methyltestosterone allowed to expand the detection window of this steroid, in time.

RESUMEN La Agencia Mundial Antidopaje posee, entre sus funciones, la acreditación de los laboratorios dedicados al control del dopaje y para mantener esta categoría es necesario el cumplimiento de estándares internacionales. Una de las exigencias es el aumento constante de la sensibilidad de los ensayos y con ello el aumento de la capacidad analítica del laboratorio. El presente trabajo describe la introducción de un método instrumental para la detección de sustancias prohibidas, fundamentalmente esteroides Se validó el ensayo para 23 analitos, de ellos 21 esteroides y otros dos compuestos no esteroideos anabólicos, un β2 agonista y un antiestrogénico. Los resultados del proceso de validación mostraron que el método propuesto es selectivo y preciso con variaciones intra-día entre 1-17 % para las concentraciones más bajas y entre 2,1-13,9 % para las más altas. Los límites de detección se mantuvieron entre 0,1-2,5 ng/mL excepto para el clenbuterol (0,02 ng/mL). El detector no mostró arrastre después del análisis de muestras concentradas, y los ensayos de recobrado y robustez fueron adecuados según el propósito que se persiga de pesquizaje o confirmación. Por último, la aplicación del ensayo validado a la evaluación de muestras provenientes de un estudio de excreción urinaria de metiltestosterona permitió ampliar la ventana de detección de este esteroide.

Os moduladores seletivos do receptor de estrógeno são classificados como substâncias proibidas pelo Comitê Olímpico Internacional e pela Agência Mundial Anti-doping e, estão banidas de acordo com a listagem de substâncias proibidas nos esportes. Clomifeno, (2-(4-(2-cloro-1,2-difeniletenil)fenóxi)-N,N-dietil-etanamina) é usado principalmente no tratamento contra a infertilidade porque age na indução da ovulação por estímulos diretos da atividade gonadotrópica. Citrato de clomifeno foi administrado por via oral (50 mg por dia, por um período de 5 dias) a uma mulher saudável, como tratamento contra a infertilidade. As amostras de urina foram coletadas na frequência de uma dose por semana. Um metabólito novo foi identificado por cromatografia gasosa acoplada à espectrometria de massas, além do hidróxi-clomifeno relatado previamente. Propôs-se a estrutura do novo metabólito como sendo o hidroximetoxiclomifeno. A monitoração deste metabólito foi investigada e o hidroxiclomifeno foi excretado da urina relativamente mais rápido do que o hidroximetoxiclomifeno. Mudanças significativas no perfil dos esteróides após doses múltiplas foram observadas e a principal variação observada foi na concentração absoluta e na razão dos estrogênios.

Selective estrogen receptor modulators are classified as prohibited substances according to the list of forbidden substances in sports by the International Olympic Committee and the World Anti-doping Agency. Clomiphene (2-(4-(2-chloro-1,2-diphenylethenyl)phenoxy)-N,N-diethyl-ethanamine) is mainly used for the treatment of infertility because it acts inducing the ovulation by direct stimulations of gonadotropic activity. Clomiphene citrate (50 mg per day for 5 days) was orally administered to one healthy female with infertility treatment. Urine samples were collected up to 1 week post-dose. A new metabolite using gas chromatography-mass spectrometry has been identified in addition to the hydroxyclomiphene previously reported. It has been proposed as hydroxymethoxyclomiphene. Monitorization of this metabolite have been proposed for doping control keeping in mind that the main metabolite hydroxyclomiphene is excreted relatively faster than hydroxymethoxyclomiphene from urine. On the other hand, significant changes on steroids profile after multiple doses were observed. The main variation was observed on absolute concentration of oestrogens and their ratios.