Anabolic steroids, in urine

C. K. Hatton and D. H. Catlin, Detection of androgenic anabolic steroids in urine, Therapeut. Drug Monit.-II Clin. Lab. Med., 7 655(1987). 

Anabolic steroids are generally used as therapeutic agents in clinical practice, but are also widely abused as performance enhancing drugs. The undesirable side effects ascribed to this practice have led to strict regulation or total prohibition in most countries. Analytical methodologies suitable for the determination of anabolic steroids in urine should meet two criteria ... 

Huang, G., Chen, H., Zhang, X., Cooks, R.G., Ouyang, Z. (2007) Rapid Screening of Anabolic Steroids in Urine by Reactive Desorption Electrospray Ionization. Anal. Chem. 79 8327-8332. 

Pozo, O.J., Van Eenoo, P., Deventer, K., and Delbeke, F.T. (2007) Development and validation of a qualitative screening method for the detection of exogenous anabolic steroids in urine by liquid chromatography-tandem mass spectrometry. Analytical and Bioanalytical Chemistry, 389,1209-1224. 

D. BaiTon, J. Barbosa, J. A. Pascual and J. Segura, Dkect deteimination of anabolic steroids in human urine by online solid-phase extraction/liquid cliromatography/mass specti ometi y , 7. Mass Spectrom. 31 309-319 (1996). 

Boldenone (17 -boldenone) is an androgenic steroid with known anabolic properties. As the oxidation of 17-ol to 17-one steroids is a recurring pathway both in vivo and in vitro, boldenone studies in cattle liver microsomes performed in vitro showed that the most prominent metabolite formed was androst-l,4-diene-3,17-dione 111). Not long ago, it was assumed drat the presence of boldenone or its main metabolite in the urine implied illegal administration of this steroid to the animal. Evidence has been recently presented that the presence of only the boldenone metabolite in urine cannot be taken as a proof of the illegal use of tills compound because boldenone is a naturally occurring steroid in urine of cattle (12). Nevertheless, the presence of 17 -boldenone in urine at levels above... 

Identification of Anabolic Steroids in Horse Urine Solvate a Sep-Pak C-18 cartridge (see Reagent Appendix) with 5 ml of methanol, followed by a wash with 5 ml of water. Pass 10 ml of the urine sample through the cartridge, wash with 10 ml of water, and elute with 10 ml of methanol. Evaporate the eluate at 50° in a rotary film evaporator and dissolve the residue in 1 to 2 ml of a mixture of chloroform and methanol (17 3). 

Homoeopathic preparations, 51, 53 Homogeneous fluoro immunoassays, 154 Homogeneous immunoassays, 149 Homosulfaminum, 717 Homosulphanilamide, 716 Homovanillic acid, 322, 571, 702 Honvan, 975 Honvol, 975 Hordenine, 99, 661 Hordenine sulphate, 662 Hormoestrol, 657 Hormofemin, 537 Hormomed, 831 Hormonin, 830, 831 Horse plasma, phenylbutazone in, 96 Horse urine, anabolic steroids in, 95 examination of, 92 phenylbutazone in, 96 salicylic acid in, 96... 

C. Van Poucke, C. Van Peteghem, Development and validation of a multi-analyte method for the detection of anabolic steroids in bovine urine with LC-MS-MS, J. Chromatogr. B, 772 (2002)211. 

Yu et al. (2005) developed an LC-MS-MS screen for deconjugated anabolic steroids in horse urine and characterized the method using horse urine samples spiked with 15 prohibited anabolic steroids. In an excretion study in two horses, methenolone acetate was administered by mouth, methenolone (Figure 2.1) was detected in urine and the 17-epimethenolone metabolite was detected for a longer time. 

Leinonen et al. (2002) compared LC-MS-MS with electrospray ionization (ESI), atmospheric pressure chemical ionization (APCI) and atmospheric pressure photoionization (APPI) for unconjugated (free) anabolic steroids in human urinary extracts. The selected analytes were synthetic steroids or their metabolites, known to be misused in sports and known to be excreted in urine unconjugated, namely oxandrolone (Figure 2.1), the 3 -hydroxy metabolite of stanozolol and the 6p-hydroxy metabolite of 4-chlorodehydro-methyltestosterone. 

Buiarelli et al. (2004) extended the above analytical approach to many more related steroids when they published a method for the direct analysis of 15 urinary anabolic steroids in a single run, namely T, epitestosterone, dehydroepiandrosterone (DHEA), androsterone, etiocholanolone, their sulfates and their glucuronides (Figure 2,2), They extracted 2 mL of human urine by solid-phase extraction with methanol elution and reconstituted the residue in aqueous methanol in the presence of deuterated internal standards (da-epitestosterone glucuronide, [16,16,17-"H3 testosterone sulfate and [16,16,17-2H3]testosterone), then monitored, for example, mJz. 289-97 and 109 for T and epitestosterone, miz 367-97 for their sulfates, and m/z 463-113 and 287 for their glucuronides. The method does not achieve quantitation, but it allows the estimation of ratios, which makes it possible to monitor the urinary steroid profile, which is useful for monitoring the abuse of anabolic steroids. 

C/min), followed by an increase to 230°C (4°C/min), and finally to 300°C (30°C/min), which was held for 5 min. The MS analysis was performed on electron impact (El) mode. The analysis of residual anabolic steroids in meat was reported by Fuh, Huang, and Lin. The isolation was conducted by SPE. To derivatize the isolated steroids, MSTFA" "" " was used. The enzymatic hydrolysis was not conducted, as previous studies have discovered that there was no significant hormone liberation involved in muscle, fatty tissue, and meat. The derivatization products were then subjected to GC-MS/MS analysis. For this purpose, a DB-5 GC column (30 m X 0.25 mm, with 0.25 pm film thickness) was used, with a flow rate of 1.0 ml/min. The oven temperature was initially set at 180°C (1 min), increased to 240°C (6°C/min, for 2 min), and finally increased to 290°C (6°C/min, held for 10 min). The MS analysis was performed on El mode. Impens et al. " used a non-polar 5% phenyl-polysilphenylene-siloxane BPX-5 GC column to analyze anabolic steroids in bovine urine. The sample was derivatized with MSTFA" before being injected for the GC-MS/MS and GC-MS/MS/MS analyses. The oven temperature was initially set to 100°C (1 min), then increased to 250°C (30°C/min), followed by an increase to 290°C (2.5°C/min), and finally to 300 C (10°C/min), which was held for 1.5 min. Interested readers can consult Refs. 3, 8, and 18-25 for details. 

Impens, S. Van Loco, J. Degroodt, J.M. De Brabander, H. A downscaled multi-residue strategy for detection of anabolic steroids in bovine urine using gas chromatography-tandem mass spectrometry (GC-MS ). Anal. Chim. Acta 2007, 586, 43 8. 

Leinonen A, Ruuranne T, Kotiaho T et al (2004) Screening of free 17-alkyl-substituted anabolic steroids in human urine by liquid chromatography-electrospray ionization tandem mass spectrometry. Steroids 69 101-109... 

Anabolic steroids, such as stanozolol, nandrolone, and tetrahydrogestrinone have the same effect on the body as testosterone, but they are more stable, so they are not metabolized as quickly. Tetrahydrogestrinone (also called THG or The Clear), the performance-enhancing drug used by track star Marion Jones during the 2000 Sydney Olympics, was considered a designer steroid because it was initially undetected in urine tests for doping. After its chemical structure and properties were determined, it was added to the list of banned anabolic steroids in 2004. 

Initial studies with Dianabol resulted in the development by Prof. R.V. Brooks of a radioimmunoassay which was suitable for screening for the 17a-methyl and 17a-ethyl groups of anabolic steroids in the urine of athletes (1). Antiserum has recently been raised which is specific for the 19-nor compounds. 

Mazzarino, M., de la Torre X., and Botre, F. (2008) A screening method for the simultaneous detection of glucocorticoids, diuretics, stimulants, anti-oestrogens, beta-adrenergic drugs and anabolic steroids in human urine by LC-ESI-MS/MS. Analytical and Bioanalytical Chemistry, 392, 681-698. 

Deuterated steroids are also valuable for use in metabolic studies. For example, the isolation of di-deuterotestosterone in urine after oral administration in men of [16,16- H2]DHEA demonstrated that DHEA had potential as an anabolic steroid (Dehennin et al., 1998). Similarly, after treatment of rats with [1- H2]ethanol, mono-deuterotestosterone was isolated from plasma (Alomary et al., 2003). 

If you re an athlete and get tested for steroids, you can still use anabolic steroids and possibly beat the cutoff. The body naturally produces testosterone (a steroid), and small amounts of testosterone show up in urine by default. Some athletes are able to keep their steroid intake low enough to... 

There is a way to use substitution even when you are under the strictest supervision. Athletes trying to pass tests for anabolic steroids have been known to empty their bladders, and have the substituted urine injected directly into their bladders via needle. It was shown in a motion picture like Wildcats I believe. While theoretically possible, it s painful and subject to infection. It would certainly be the most senseless way to get clean urine into the testees bladder. If this must be done, catheterization should be used. 

Ho, E.N.M., Leung, D.K.K., Wan, T.S.M. and Yu, N.H. (2006) Comprehensive Screening of Anabolic Steroids, Corticosteroids, and Acidic Drugs in Horse Urine by Solid-Phase Extraction and Liquid Chromatography-Mass Spectrometry. J Chromatopfr A, 1120, 38. 

In 1992, the International Olympic Committee (IOC) produced a list of approximately 500 banned substances that can be classified into four categories of molecules stimulants, analgesics-narcotics, anabolic steroids and beta-blockers. Several of these active molecules can only be detected by their metabolites in urine. ... 

Many Anabolic/Androgenic Steroids (AAS) are available in an oral form. Unfortunately some are also quite toxic to the liver. Orally administered AAS are very susceptible to first pass liver deactivation unless chemical molecular structures are altered to make them harder to deactivate. When an oral AAS is swallowed it enters the stomach where it is partially broken down and passed to the small intestines. The small intestines contain a group of enzymes called CYP-450 s. These enzymes begin to break down the AAS further in an attempt to deactivate it. The AAS is then absorbed through intestinal mucosa cells and transferred to the liver portal vein for further deactivation into inactive chemicals such as etiocholanone. These chemicals are then conjugated with glucuronic acid and excreted in urine. Up to 100% of the original compound can be deactivated in this process which is known as first pass deactivation. 

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