RESUMEN
The detection of testosterone intake is facilitated by monitoring the urinary steroid profile in the athlete biological passport. This technique can be used with confidence to identify target samples for isotope ratio mass spectrometry. Regrettably, most research has been performed on male subjects resulting in a method that does not account for females' steroid concentration and/or variation. This study evaluates the usefulness of the carbon isotope ratio (CIR) in serum of female subjects. Two steroid sulphates are targeted in serum, androsterone and epiandrosterone. Both exhibit statistically significant depletion of their CIR after 10 weeks of daily (10 mg) transdermal testosterone administration. Of the 21 female subjects, samples from six individuals were identified as adverse analytical findings; additionally, four were found atypical considering the serum CIR. The urinary athlete biological passport was not sufficiently sensitive to identify target serum samples for isotope ratio mass spectroscopy. Of the six with a suspicious passport, only two could be confirmed using the serum CIR of androsterone and epiandrosterone. This study shows that CIR analysis in serum cannot be considered the sole confirmatory solution to detect testosterone doping in women due to low sensitivity. However, this analysis has the potential to be used as a complementary method in certain situations to confirm exogenous testosterone in women.
Asunto(s)
Doping en los Deportes , Testosterona , Humanos , Masculino , Femenino , Testosterona/análisis , Andrógenos/análisis , Androsterona , Cromatografía de Gases y Espectrometría de Masas/métodos , Espectrometría de Masas , Esteroides , Isótopos de Carbono/análisis , Suplementos Dietéticos/análisis , Detección de Abuso de Sustancias/métodosRESUMEN
Selective androgen receptor modulators (SARMs) are compounds with specific androgenic properties investigated for the treatment of conditions such as muscle wasting diseases. The reported androgenic properties have resulted in their use by athletes, and consequently they have been on the World Anti-Doping Agency prohibited list for more than a decade. SARMs have been investigated by pharmaceutical companies as potential drug candidates, but to date no SARM has demonstrated sufficient safety and efficacy to gain clinical approval by either the European Medicines Agency or the U.S. Food and Drug Administration. Despite their lack of safety approval, SARMs are often illegally marketed as dietary supplements, available for consumers to buy online. In this study, a range of supplement products marketed as SARMs were purchased and analyzed using high resolution accurate mass - mass spectrometry to evaluate the accuracy of product claims and content labeling. This study found discrepancies ranging from a supplement in which no active ingredients were found, to supplements containing undeclared prohibited analytes. Where SARMs were detected, discrepancies were observed between the concentrations measured and those detailed on the product packaging. The outcome of this experiment highlights the high risk of such supplement products to consumers. The inaccurate product claims give rise to uncertainty over both the dose taken and the identity of any of these unapproved drugs. Even for supplements for which the product labeling is correct, the lack of complete toxicity data, especially for combinations of SARMs taken as stacks, means that the safety of these supplements is unknown.
Asunto(s)
Andrógenos/análisis , Suplementos Dietéticos/análisis , Drogas Ilícitas/análisis , Doping en los Deportes , Humanos , Detección de Abuso de Sustancias , Reino UnidoRESUMEN
Anabolic-androgenic steroids (AASs) are very potent muscle builders, and professional sportsmen often take protein supplements to improve their performance. Several studies have emphasised that protein supplements may contain undeclared AASs banned by the International Olympic Committee/World Anti-Doping Agency. The widespread occurrence and abuse of contaminated protein supplements is extremely dangerous because of their side effects. To minimise the chances of an unattended positive doping test or to avoid serious health problems, adequate screening methods for the detection of a wide range of steroids is essential. To address this requirement, a rapid and effective modified QuEChERS (quick, easy, cheap, effective, rugged and safe) method was developed and validated to screen and quantify the simultaneous analysis of twenty-eight AASs in protein supplements using LC-MS/MS. The validated method was applied to 198 protein supplements collected from on-line and, off-line markets, and direct purchase from overseas between 2019 and 2020. Of the 198 samples, two samples contained testosterone and stanozolol at concentrations of 0.27 µg/g and 0.023 µg/g, respectively. In addition, 5α-hydroxylaxogenin was detected for the first time in three products purchased in Korea from overseas. The modified QuEChERS method was established and successfully applied to screen and determine AASs as a measure of continuous control and supervision in protein supplements.
Asunto(s)
Anabolizantes/análisis , Andrógenos/análisis , Suplementos Dietéticos/análisis , Análisis de los Alimentos , Esteroides/análisis , Cromatografía Liquida , República de Corea , Espectrometría de Masas en TándemRESUMEN
BACKGROUND AND OBJECTIVE: Our patient cohort revealed that obesity is strongly associated with steroid-5α reductase type 2 (SRD5A2) promoter methylation and reduced protein expression. The underlying mechanism of prostatic growth in this population is poorly understood. Here we addressed the question of how obesity, inflammation, and steroid hormones affect the development of benign prostatic hyperplasia (BPH). MATERIAL AND METHODS: We used preadipocytes, macrophages, primary human prostatic stromal cells, prostate tissues from high-fat diet-induced obese mice, and 35 prostate specimens that were collected from patients who underwent transurethral resection of the prostate (TURP). RNA was isolated and quantified with RT-PCR. Genome DNA was extracted and SRD5A2 promoter methylation was determined. Sex hormones were determined by high-performance liquid chromatography-tandem mass spectrometry. Protein was extracted and determined by ELISA test. RESULTS: In prostatic tissues with obesity, the levels of inflammatory mediators were elevated. SRD5A2 promoter methylation was promoted, but SRD5A2 expression was inhibited. Inflammatory mediators and saturated fatty acid synergistically regulated aromatase activity. Obesity promoted an androgenic to estrogenic switch in the prostate. CONCLUSIONS: Our findings suggest that obesity-associated inflammation induces androgenic to estrogenic switch in the prostate gland, which may serve as an effective strategy for alternative therapies for management of lower urinary tract symptoms associated with BPH in select individuals.
Asunto(s)
Andrógenos/metabolismo , Estrógenos/metabolismo , Obesidad/inmunología , Próstata/patología , Hiperplasia Prostática/inmunología , 3-Oxo-5-alfa-Esteroide 4-Deshidrogenasa/genética , 3-Oxo-5-alfa-Esteroide 4-Deshidrogenasa/metabolismo , Células 3T3-L1 , Adipocitos/inmunología , Adipocitos/metabolismo , Anciano , Anciano de 80 o más Años , Andrógenos/análisis , Animales , Aromatasa/metabolismo , Metilación de ADN , Dieta Alta en Grasa/efectos adversos , Modelos Animales de Enfermedad , Estrógenos/análisis , Ácidos Grasos/metabolismo , Humanos , Mediadores de Inflamación/análisis , Mediadores de Inflamación/metabolismo , Metabolismo de los Lípidos/inmunología , Masculino , Proteínas de la Membrana/genética , Proteínas de la Membrana/metabolismo , Ratones , Persona de Mediana Edad , Obesidad/complicaciones , Obesidad/metabolismo , Cultivo Primario de Células , Regiones Promotoras Genéticas/genética , Próstata/citología , Próstata/inmunología , Próstata/cirugía , Hiperplasia Prostática/patología , Hiperplasia Prostática/cirugía , Células del Estroma , Células THP-1 , Resección Transuretral de la PróstataRESUMEN
The present study was conducted to explores the effects of short-term addition of 17ß-E2 on the growth,gonad development and internal quality of overwintering Whitmania pigra. Before overwintering,0. 0,1. 0,10. 0,25. 0,50. 0,100. 0 µg·L~(-1) of 17ß-E2 were added to the aquaculture water for 6 weeks and then hibernated for 60 days. The changes of growth performance,gonad index,morphological structure of spermary( ovary),endogenous steroid hormones level and internal quality were measured. The results showed that the body weight,weight gain rate,specific growth rate,female gonad index,oocyte development and endogenous estrogen level of the leech increased first and then decreased with the increase of the concentration of exogenous 17ß-E2,which were higher than those of the control group. The body weight,weight gain rate and specific growth rate of the leech at the concentration of 25 µg·L~(-1)17ß-E2 were significantly higher than those of the other groups( P<0. 05),oocyte development and endogenous estrogen levels were significantly higher than those of other groups at the concentration of 50 µg·L~(-1)( P<0. 05). When the concentration of exogenous 17ß-E2 was higher than 50 µg·L~(-1),the levels of male gonad index,spermatocyte development,endogenous androgen and progesterone were significantly inhibited( P< 0. 05). There was no significant difference in endogenous corticosteroid levels among the groups. In conclusion,short-term addition of exogenous 17ß-E2 of 10-25 µg·L~(-1) could promote the growth of overwintering leeches,oocyte development and antithrombin activity without inhibiting the development of male gonads.
Asunto(s)
Estradiol/farmacología , Gónadas/crecimiento & desarrollo , Sanguijuelas/efectos de los fármacos , Sanguijuelas/crecimiento & desarrollo , Andrógenos/análisis , Animales , Estrógenos/análisis , Femenino , Gónadas/efectos de los fármacos , Hibernación , Masculino , Progesterona/análisisRESUMEN
Selective androgen receptor modulators (SARMs) are an emerging class of therapeutics targeted to cachexia, sarcopenia, and hypogonadism treatment. LGD-4033 is a SARM which has been included on the Prohibited List annually released by the World Anti-Doping Agency (WADA). The aim of the present work was the investigation of the metabolism of LGD-4033 in a human excretion study after administration of an LGD-4033 supplement, the determination of the metabolites' excretion profiles with special interest in the determination of its long-term metabolites, and the comparison of the excretion time of the phase I and phase II metabolites. The results were also compared to those derived from previous LGD-4033 studies concerning both in vitro and in vivo experiments. Supplement containing LGD-4033 was administered to one human male volunteer and urine samples were collected up to almost 21 days. Analysis of the hydrolyzed (with ß-glucuronidase) as well as of the non-hydrolyzed samples was performed using liquid chromatography-high resolution mass spectrometry (LC-HRMS) in negative ionization mode and revealed that, in both cases, the two isomers of the dihydroxylated metabolite (M5) were preferred target metabolites. The gluco-conjugated parent LGD-4033 and its gluco-conjugated metabolites M1 and M2 can be also considered as useful target analytes in non-hydrolyzed samples. The study also presents two trihydroxylated metabolites (M6) identified for the first time in human urine; one of them was recently reported in an LGD-4033 metabolism study in horse urine and plasma.
Asunto(s)
Andrógenos/metabolismo , Andrógenos/orina , Nitrilos/metabolismo , Nitrilos/orina , Pirrolidinas/metabolismo , Pirrolidinas/orina , Andrógenos/administración & dosificación , Andrógenos/análisis , Cromatografía Liquida/métodos , Suplementos Dietéticos/análisis , Cromatografía de Gases y Espectrometría de Masas/métodos , Humanos , Hidrólisis , Masculino , Espectrometría de Masas/métodos , Nitrilos/administración & dosificación , Nitrilos/análisis , Pirrolidinas/administración & dosificación , Pirrolidinas/análisis , Detección de Abuso de Sustancias/métodosRESUMEN
Both athletes and the general population use nutritional supplements. Athletes often turn to supplements hoping that consuming the supplement will help them be more competitive and healthy, while the general population hopes to improve body image or vitality. While many supplements contain ingredients that may have useful properties, there are supplements that are contaminated with compounds that are banned for use in sport or have been deliberately adulterated to fortify a supplement with an ingredient that will produce the advertised effect. In the present study, we have used yeast cell and mammalian cell androgen bioassays to characterize the androgenic bioactivity of 112 sports supplements available from the Australian market, either over the counter or via the Internet. All 112 products did not declare an androgen on the label as an included ingredient. Our findings show that six out of 112 supplements had strong androgenic bioactivity in the yeast cell bioassay, indicating products spiked or contaminated with androgens. The mammalian cell bioassay confirmed the strong androgenic bioactivity of five out of six positive supplements. Supplement 6 was metabolized to weaker androgenic bioactivity in the mammalian cells. Further to this, Supplement 6 was positive in a yeast cell progestin bioassay. Together, these findings highlight that nutritional supplements, taken without medical supervision, could expose or predispose users to the adverse consequences of androgen abuse. The findings reinforce the need to increase awareness of the dangers of nutritional supplements and highlight the challenges that clinicians face in the fast-growing market of nutritional supplements.
Asunto(s)
Andrógenos/análisis , Suplementos Dietéticos , Análisis de los Alimentos/métodos , Bioensayo , Línea Celular , Doping en los Deportes , Humanos , Progestinas , Levaduras/efectos de los fármacosRESUMEN
Local healthcare providers often question the possible steroidal activity of traditional Chinese medicine (TCM) herbs or herbal products and implicate them as a cause for adrenal insufficiency or Cushing's syndrome in patients with a history of TCM intake. We conducted a comprehensive database search for evidence of potential glucocorticoid, mineralocorticoid, androgenic or oestrogenic activity of herbs or herbal products. Overall, there are not many herbs whose steroidal activity is well established; among these, most cases were based on preclinical studies. Liquorice root may cause pseudoaldosteronism through interference with the steroidogenesis pathway. Although ginseng and cordyceps have some in vitro glucocorticoid activities, the corroborating clinical data is lacking. Deer musk and deer antler contain androgenic steroids, while epimedium has oestrogenic activity. On the other hand, adulteration of herbal products with exogenous glucocorticoids is a recurrent problem encountered locally in illegal products masquerading as TCM. Healthcare providers should stay vigilant and report any suspicion to the relevant authorities for further investigations.
Asunto(s)
Medicamentos Herbarios Chinos/análisis , Medicina Tradicional China/efectos adversos , Esteroides/análisis , Andrógenos/análisis , Animales , Cordyceps , Bases de Datos Factuales , Ciervos , Medicamentos Herbarios Chinos/efectos adversos , Epimedium , Estrógenos/análisis , Ácidos Grasos Monoinsaturados , Glucocorticoides/análisis , Glycyrrhiza uralensis , Humanos , Mineralocorticoides/análisis , Panax , Preparaciones de Plantas/análisis , Riesgo , Singapur , Esteroides/efectos adversos , Extractos de TejidosRESUMEN
Sport supplements containing steroids never approved for therapeutic use have the potential for abuse by athletes. Most are marketed online and may contain undisclosed steroids yet are readily available despite lacking toxicological or pharmacological evaluation. In this study, 18 supplements purchased online underwent organic solvent extraction to isolate any steroids they contained. From the 18 supplements, 19 steroids were identified and for each, its intrinsic androgenic potency was determined by a yeast cell (Saccharomyces cerevisiae) androgen bioassay and its potential androgenic potency was determined by a liver (HuH7) cell androgen bioassay. The yeast bioassay showed that of the 19 steroids tested, 6 demonstrated strong intrinsic bioactivity, with 4 metabolically activated to even stronger androgens. Moreover, 4 steroids with moderate and 1 with intrinsically weak androgenic bioactivity were activated to more potent androgens. Finally, 8 steroids were metabolically inactivated or deactivated into weaker androgens. Our results show that Internet-sourced sport supplements may contain intrinsically strong androgens, or precursors that can be metabolized to them. These potentially potent pharmacologically active steroids are being used without regulatory control or consumer awareness of their potential adverse effects. Copyright © 2016 John Wiley & Sons, Ltd.
Asunto(s)
Andrógenos/análisis , Andrógenos/farmacología , Suplementos Dietéticos/análisis , Animales , Línea Celular , Doping en los Deportes , Evaluación Preclínica de Medicamentos/métodos , Humanos , Internet , Hígado/citología , Hígado/efectos de los fármacos , Hígado/metabolismo , Ratones , Mioblastos/citología , Mioblastos/efectos de los fármacos , Mioblastos/metabolismo , Saccharomyces cerevisiae/efectos de los fármacos , Saccharomyces cerevisiae/metabolismo , Esteroides/análisis , Esteroides/farmacologíaRESUMEN
To ensure fair competition and to protect the horse's welfare, horses have to compete on their own merits, without any unfair advantage that might follow the use of drugs. Therefore, regulatory authorities list all substances that are not allowed in competition, including most anabolic-androgenic steroids. As zero-tolerance is retained, the question arose whether the consumption of mouldy feed could lead to the excretion of steroids, due to the biotransformation of plant phytosterols to steroids. A rapid ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) analytical method, previously validated according to AORC (Association of Official Racing Chemists) and EC (European Commission) guidelines, was used to measure steroids in different sample types. Multiple mouldy feed samples were tested for the presence of steroids. The effect of digestion was tested by in vitro simulation of the horse's hindgut in batch incubations. In most feed samples no steroids were detected, even when the products were mouldy. Mouldy corn however showed to contain up to 3.0 ± 0.4 µg/kg AED (4-androstenedione), the main testosterone precursor. This concentration increased when mouldy corn (with added phytosterols) was digested in vitro. An herbal phytosupplement also showed to contain α-testosterone. These results demonstrate that it is important to caution against the consumption of any feed or (herbal) supplement of which the detailed ingredients and quantitative analysis are unknown. The consumption of mouldy corn should especially be avoided, not only from a horse health and welfare point of view, but also to avoid possible inadvertent positive doping results. Copyright © 2016 John Wiley & Sons, Ltd.
Asunto(s)
Anabolizantes/análisis , Andrógenos/análisis , Alimentación Animal/análisis , Alimentación Animal/microbiología , Heces/química , Caballos/orina , Esteroides/análisis , Aerobiosis , Anabolizantes/metabolismo , Anabolizantes/orina , Andrógenos/metabolismo , Andrógenos/orina , Animales , Biotransformación , Cromatografía Líquida de Alta Presión/métodos , Suplementos Dietéticos/análisis , Suplementos Dietéticos/microbiología , Doping en los Deportes , Caballos/metabolismo , Mucor/metabolismo , Mycobacterium/metabolismo , Fitosteroles/análisis , Fitosteroles/metabolismo , Fitosteroles/orina , Esteroides/metabolismo , Esteroides/orina , Espectrometría de Masas en Tándem/métodos , Testosterona/análisis , Testosterona/metabolismo , Testosterona/orina , Zea mays/química , Zea mays/microbiologíaRESUMEN
In a previous study, we found a dose-dependent synergistic effect in recombinant yeast stably transfected with the human androgen receptor (AR), in response to co-exposure to testosterone and a commercially-available lubricant (engine) oil for cars. As there is relatively little knowledge on synergistic toxic effects and causative compounds, particularly for the androgenic system, the objective of the present study was to investigate this oil in more detail. The oil was fractionated into SARA fractions (so-called 'saturates', 'aromatics', 'resins', and 'asphaltenes') by open column chromatography. Surprisingly, when exposing the recombinant AR yeast to testosterone in combination with the separate SARA fractions, the synergistic effect could not be reproduced fully. After pooling the fractions again however, the full synergism returned. From subsequent exposures to combinations of two or three SARA fractions, it appeared that both the 'saturates' and the 'resins' fraction were required for obtaining the synergistic response with testosterone. This clearly demonstrates a synergistic effect related to the androgenic system caused by the joint action of at least three chemically-distinct compounds, or groups of compounds (i.e. testosterone, 'resins' and 'saturates'). Although detailed chemical analyses could not reveal the identity of the causative compounds and the in vivo relevance of the present results remains unclear, the results do add to the growing body of evidence on the potentially extremely complex character of mixture effects.
Asunto(s)
Andrógenos/toxicidad , Petróleo/toxicidad , Receptores Androgénicos/genética , Testosterona/toxicidad , Andrógenos/análisis , Fraccionamiento Químico , Sinergismo Farmacológico , Regulación de la Expresión Génica/efectos de los fármacos , Proteínas Fluorescentes Verdes/genética , Humanos , Petróleo/análisis , Testosterona/análisis , Levaduras/genéticaRESUMEN
The concept of phytoandrogens, plants that contain androgens or those that stimulate androgenic activity in men, is relatively new. In traditional Chinese medicine a number of phytoandrogens are classified in medicinal plant restoratives for reinforcing yang, and they find their application in the treatment of the kidney yang deficiency diseases. In this review, the phytoandrogens used in traditional Chinese medicine are listed, and their proven applications in the treatment of kidney yang deficiency diseases, such as sexual disorders, cancer, and obesity and associated metabolic syndromes are presented. As a background, the mechanism of action of androgens, their synthesis and metabolism, the interrelations of androgens and estrogens, as well as the state of art methods to detect and analyze these hormonal activities in medicinal plants are discussed.
Asunto(s)
Andrógenos/uso terapéutico , Medicamentos Herbarios Chinos/uso terapéutico , Plantas Medicinales/química , Deficiencia Yang/tratamiento farmacológico , Andrógenos/análisis , Animales , Medicamentos Herbarios Chinos/análisis , Humanos , MasculinoRESUMEN
Androgens are the class of sex steroids responsible for male sexual characteristics, including increased muscle mass and decreased fat mass. Illicit use of androgen doping can be an attractive option for those looking to enhance sporting performance and/or physical appearance. The use of in vitro bioassays to detect androgens, especially designer or proandrogens, is becoming increasingly important in combating androgen doping associated with nutritional supplements. The nutritional sports supplement market has grown rapidly throughout the past decade. Many of these supplements contain androgens, designer androgens or proandrogens. Many designer or proandrogens cannot be detected by the standard highly-sensitive screening methods such as gas chromatography-mass spectrometry because their chemical structure is unknown. However, in vitro androgen bioassays can detect designer and proandrogens as these assays are not reliant on knowing the chemical structure but instead are based on androgen receptor activation. For these reasons, it may be advantageous to use routine androgen bioassay screening of nutraceutical samples to help curb the increasing problem of androgen doping.
Asunto(s)
Andrógenos/análisis , Bioensayo/métodos , Drogas de Diseño/análisis , Animales , Suplementos Dietéticos , Estudios de Factibilidad , Humanos , Receptores Androgénicos/metabolismoRESUMEN
Previously developed estrogen and androgen mammalian reporter gene assays (RGAs) were assessed for their potential use as a quantitative screening method in the detection of estrogenic and androgenic endocrine disruptors (EDs) in sport supplements. The validation of both RGAs coupled with dispersive solid phase extraction (dSPE) was performed in accordance with European Commission Decision EC/2002/6579 for biological screening methods. Decision limits (CCα) and detection capabilities (CCß) were established for both the estrogen and androgen RGAs. All samples were compliant with CCα and CCß in both bioassays. Recovery rates were 96 % for 17ß-estradiol and 115 % for dihydrotestosterone as obtained in their corresponding RGA. Both estrogens and androgens were stable in samples for more than 3 weeks, when stored at -20 °C. Specificity, good repeatability (coefficients of variation (CV), 12-25 %), reproducibility and robustness of both bioassays were also observed. Four different ED modes of action were determined for estrogens and androgens in 53 sport supplements, using the validated RGAs. This study revealed that 89 % of the investigated sport supplements contained estrogenic EDs and 51 % contained androgenic compounds. In conclusion, both bioassays are suitable for sport supplement screening of estrogenic and androgenic EDs.
Asunto(s)
Andrógenos/análisis , Andrógenos/farmacología , Suplementos Dietéticos/análisis , Disruptores Endocrinos/análisis , Disruptores Endocrinos/farmacología , Genes Reporteros , Deportes , Animales , Línea Celular , Supervivencia Celular/efectos de los fármacos , Extracción en Fase Sólida , Relación Estructura-ActividadRESUMEN
The increasing availability and use of sports supplements is of concern as highlighted by a number of studies reporting endocrine disruptor contamination in such products. The health food supplement market, including sport supplements, is growing across the Developed World. Therefore, the need to ensure the quality and safety of sport supplements for the consumer is essential. The development and validation of two reporter gene assays coupled with solid phase sample preparation enabling the detection of estrogenic and androgenic constituents in sport supplements is reported. Both assays were shown to be of high sensitivity with the estrogen and androgen reporter gene assays having an EC(50) of 0.01 ng mL(-1) and 0.16 ng mL(-1) respectively. The developed assays were applied in a survey of 63 sport supplements samples obtained across the Island of Ireland with an additional seven reference samples previously investigated using LC-MS/MS. Androgen and estrogen bio-activity was found in 71% of the investigated samples. Bio-activity profiling was further broken down into agonists, partial agonists and antagonists. Supplements (13) with the strongest estrogenic bio-activity were chosen for further investigation. LC-MS/MS analysis of these samples determined the presence of phytoestrogens in seven of them. Supplements (38) with androgen bio-activity were also selected for further investigation. Androgen agonist bio-activity was detected in 12 supplements, antagonistic bio-activity was detected in 16 and partial antagonistic bio-activity was detected in 10. A further group of supplements (7) did not present androgenic bio-activity when tested alone but enhanced the androgenic agonist bio-activity of dihydrotestosterone when combined. The developed assays offer advantages in detection of known, unknown and low-level mixtures of endocrine disruptors over existing analytical screening techniques. For the detection and identification of constituent hormonally active compounds the combination of biological and physio-chemical techniques is optimal.
Asunto(s)
Bioensayo/métodos , Suplementos Dietéticos/análisis , Disruptores Endocrinos/análisis , Genes Reporteros , Antagonistas de Andrógenos/análisis , Antagonistas de Andrógenos/aislamiento & purificación , Andrógenos/agonistas , Andrógenos/análisis , Andrógenos/aislamiento & purificación , Línea Celular , Disruptores Endocrinos/aislamiento & purificación , Antagonistas de Estrógenos/análisis , Antagonistas de Estrógenos/aislamiento & purificación , Moduladores de los Receptores de Estrógeno/análisis , Moduladores de los Receptores de Estrógeno/aislamiento & purificación , Humanos , Receptores Androgénicos/genética , Receptores Androgénicos/metabolismo , Receptores de Estrógenos/genética , Receptores de Estrógenos/metabolismo , Extracción en Fase Sólida/métodosRESUMEN
The detection of corticosteroids and sex steroids in samples with no content indication, which are confiscated for forensic investigation, is a challenge in doping analysis. A screening method based on the identification of androgens, estrogens, gestagens, and their esters by means of a mass spectral library, along with a fast ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) method, was recently developed in our lab for the analysis of dietary supplements. However, for forensic investigations, it is important to extend the scope of the method to corticosteroids in various matrices. Therefore, 36 corticosteroids were added to the mass spectral library, and the sample preparation step was modified so that androgens, gestagens, corticosteroids, and their esters could be analyzed with only one injection with the UPLC-MS method. A complementary tool to the existing library identification was found in the extraction of common fragment ions out of the full scan data obtained for the library search. The fragment ion with m/z 147 was found to be a good marker for the detection of steroids. Extra confirmation was obtained from the fragment ions with m/z 135 (for all steroids) and 237 (specific for corticosteroids) or from the fragment ions with m/z 77, 91, and 105. The effectiveness of this approach was evaluated on some samples previously screened for forensic investigation with thin-layer chromatography and confirmed with a targeted gas chromatography-mass spectrometry method. This study shows that the combination of the library identification and the common fragment ions approach can be a valuable tool in the detection of steroids without defining any target at the start of the analysis.
Asunto(s)
Corticoesteroides/análisis , Suplementos Dietéticos/análisis , Espectrometría de Masas/métodos , Andrógenos/análisis , Cromatografía Líquida de Alta Presión/métodos , Doping en los Deportes , Ciencias Forenses , Humanos , Progestinas/análisisRESUMEN
The aim of this study was to collect further data about levels of endogenous hormones in foodstuffs derived from animal production. Because of expected higher concentrations of especially estrogens in colostrum compared to other foodstuffs, our investigation focused on this matrix. For evaluation of endogenous steroid hormones in separated colostrum (fat and defatted fraction) and colostrum powder, the relevant free and conjugated estrogens (estradiol-17ß, estradiol-17α, estrone, and estriol) androgens (androstendione, 19nor-androstendione, 19nor-testosterone-17ß, 19nor-testosterone-17α, testosterone-17ß, and testosterone-17α), and progesterone were determined by means of liquid chromatography-tandem mass spectrometry (LC-MS/MS). Upmost determined concentrations were found in the fat fraction, with 25.56 and 7.59 µg/L for estrone and androstendione, respectively. In defatted milk and colostrum powder, conjugated estrogens dominated, whereas total (free and conjugated) estrone (5.51 µg/L; 15.0 µg/kg) exceeded estradiol-17α (2.66 µg/L; 7.5 µg/kg) and estradiol-17ß (2.28 µg/L; 3.3 µg/kg). Neither 19nor-steroids nor estriol were detected in colostrum fractions or processed colostrum.
Asunto(s)
Bovinos/metabolismo , Cromatografía Liquida , Calostro/química , Hormonas/análisis , Espectrometría de Masas en Tándem , Andrógenos/análisis , Animales , Estrógenos Conjugados (USP)/análisis , Grasas/análisis , Femenino , Leche/química , Progesterona/análisisRESUMEN
Androgenic compounds induce an interaction between the NH(2)- and COOH-terminal regions (N-C interaction) of androgen receptor (AR). We describe a rapid yeast bioassay for androgenic and anti-androgenic compounds based on androgen-dependent ß-catenin-enhanced N-C interaction. The bioassay was also effective at detecting compounds that inhibit the N-C interaction in ways that do not involve binding to the ligand-binding domain.
Asunto(s)
Antagonistas de Andrógenos/análisis , Andrógenos/análisis , Evaluación Preclínica de Medicamentos/métodos , Receptores Androgénicos/química , Animales , Humanos , Receptores Androgénicos/metabolismo , Levaduras , beta CateninaRESUMEN
PURPOSE: Supplemental administration of androgens has been advocated for men with sexual dysfunction (SD) and hypoandrogenism. The preponderance of evidence indicates that most delivery forms of testosterone (T) are effective but the role of dehydroepiandrosterone (DHEA) is controversial. A placebo-controlled, randomized trial of oral androgen (T versus DHEA) supplementation was carried out to determine their efficacy. MATERIALS AND METHODS: Eighty-six men with SD and decreased levels of serum T and/or DHEA, participated in a study receiving oral T undecanoate (OTU) (n = 29) 80 mg twice daily, DHEA (n = 28) 50 mg twice daily, or placebo (n = 29). Outcomes included evaluation of sexual performance by the International Index of Erectile Function (IIEF), the Androgen Deficiency in the Aging Male (ADAM), Aging Male Symtom Scale (AMS), and Global Assessment Questionnaire (GAQ) questionnaires. Biochemical evaluations included measurement of T and DHEA, prolactin, gonadotropins, and PSA. RESULTS: Seventy-nine men completed the study. There were no significant differences in outcomes as assessed by four different instruments: the ADAM, IIEF, AMS, and GAQ in regard to sexual interest or erectile function. Biochemically, a significant increase in serum DHEA between baseline and final visit was documented in the group receiving DHEA. The levels of T, on the other hand, increased insignificantly between entry and final visit in the T cohort. No biochemical changes were observed in the placebo group. Levels of PSA remained stable in all three groups. CONCLUSIONS: This study did not suggest a clinical benefit of OTU or DHEA supplementation in men with hypoandrogenism and SD. The recommended dose of OTU may have been inadequate or poorly absorbed. Increased doses or an alternative T delivery form may result in a different response.
Asunto(s)
Andrógenos/análisis , Deshidroepiandrosterona/administración & dosificación , Disfunciones Sexuales Fisiológicas/tratamiento farmacológico , Testosterona/administración & dosificación , Anciano , Deshidroepiandrosterona/metabolismo , Método Doble Ciego , Humanos , Masculino , Persona de Mediana Edad , Evaluación de Resultado en la Atención de Salud , Disfunciones Sexuales Fisiológicas/fisiopatología , Testosterona/metabolismoRESUMEN
Recently we constructed a recombinant yeast cell that expresses the human androgen receptor (hAR) and yeast enhanced green fluorescent protein (yEGFP), the latter in response to androgens. When exposed to testosterone, the concentration where half-maximal activation is reached (EC(50)) was 50 nM. Eighteen different dietary supplements, already analysed by a liquid chromatography-tandem mass spectrometry method (LC-MS/MS) for the presence of anabolic steroids, were screened for androgenic activity. Eleven samples containing at least one anabolic steroid, with a concentration that was around or above 0.01 mgunit(-1) according to LC-MS/MS, were also positive in the bioassay. Seven samples did not contain any of the 49 compounds screened for in LC-MS/MS. In contrast two of them were positive in the bioassay. Bioassay-directed identification, using the bioassay as an off-line LC-detector and LC-time of flight-MS with accurate mass measurement was carried out in these two samples and revealed the presence of 4-androstene-3beta,17beta-diol and 5alpha-androstane-3beta,17beta-diol in the first and 1-testosterone in the second supplement, showing the added value of the bioassay in comparison with a LC-MS/MS screening method alone.