Methandriol [BSA]

Due to the potential for misuse of a wide range of anabolic steroids in horse racing, a screening test to detect multiple compounds, via a common class of metabolites, would be a valuable forensic tool. An enzyme-linked immunosorbent assay (ELISA) has been developed to detect 17 alpha-alkyl anabolic steroid metabolites in equine urine. 16 beta-Hydroxymestanolone (16 beta,17 beta-dihydroxy- 17 alpha-methyl-5 alpha-androstan-3-one) was synthesised in six steps from commercially available epiandrosterone (3 beta-hydroxy-5 alpha-androstan-17-one). Polyclonal antibodies were raised in sheep, employing mestanolone (17 beta-hydroxy-17 alpha-methyl-5 alpha-androstan-3-one) or 16 beta-hydroxymestanolone conjugated to human serum albumin, via a 3-carboxymethyloxime linker, as antigens. Antibody cross-reactivities were determined by assessing the ability of a library of 54 representative steroids to competitively bind the antibodies. Antibodies raised against 16 beta-hydroxymestanolone showed excellent cross-reactivities for all of the 16 beta, 17 beta-dihydroxy-17 alpha-methyl steroids analysed and an ELISA has been developed to detect these steroid metabolites. Using this 16 beta-hydroxymestanolone assay, urine samples from horses administered with stanozolol (17 alpha-methyl-pyrazolo[4',3':2,3]-5 alpha-androstan-17 beta-ol), were analysed raw, following beta-glucuronidase hydrolysis, and following solid-phase extraction (SPE) procedures. The suppressed absorbances observed were consistent with detection of the metabolite 16 beta-hydroxystanozolol. Positive screening results were confirmed by comparison with standard LCMS analyses. Antibodies raised against mestanolone were also used to develop an ELISA and this was used to detect metabolites retaining the parent D-ring structure following methandriol (17 alpha-methylandrost-5-ene-3 beta,17 beta-diol) administration. The ELISA methods developed have application as primary screening tools for detection of new and known anabolic steroid metabolites. (c) 2005 Elsevier Ltd. All rights reserved.

In this paper, a convenient and self-assembled hollow fiber solvent-stir bar microextraction (HF-SSBME) device was developed, which could stir by itself. In the extraction process, the proposed device made the solvent "bar" not floating at the sample solution and exposing to air while organic solvents outside hollow fiber always wrapped with donor phase solvent, which reduced the vaporization of organic solvents. This design could improve the precisions and recoveries of experiments. For evaluating the device, seven anabolic steroids (prasterone, 5 alpha-androstane-3 alpha, 17 beta-diol, methandriol, 19-norandrostenediol, androstenediol, methyltestosterone and methandienone) were used as model analytes and extraction conditions such as type and volume of organic solvents, agitation speed, extraction time, extraction temperature and salt addition were studied in detail. Under the optimum conditions (15 mu L toluene, 40 degrees C, stirring at 750 rpm for 30 min with 1.5g sodium chloride addition in 20.0 mL donor phase), the linear ranges of anabolic steroids were 0.25-200 ng mL(-1) with gas chromatography-mass spectrometry. The limits of detection were lower than 0.10 ng mL(-1). The recoveries and precisions in spiked urine and hair samples were between 73.97-93.56% and 2.18-4.47% (n = 5). HF-SSBME method combined the intrinsical merits of hollow fiber with the superiority of the proposed self-stirring device which can be developed to two-phase, three-phase and in situ derivatization modes with wide prospect of application. Besides, the pedestal of this proposed device can be converted to fix stir bar in stir bar sorptive extraction (SBSE) method. (C) 2012 Elsevier B.V. All rights reserved.