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Effective separation and simultaneous analysis of anabolic androgenic steroids (AAS) in their pharmaceutical formulations by a validated TLC-densitometry method

DOI: 10.1186/1752-153x-6-54

Keywords: Testosterone derivatives, TLC-densitometry, Testosterone propionate, Testosterone phenyl propionate, Testosterone isocaproate, Testosterone deaconate

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Separation was carried out on Al based TLC plates, pre-coated with silica gel 60F-254 using hexane and ethyl acetate (8.5:1.5, v/v). Spots at Rf 0.31?±?0.01, 0.34?±?0.01, 0.40?±?0.01 and 0.45?±?0.02 were recognized as TPP, TP, TI and TD, respectively. Quantitative analysis was done by densitometric measurements at λmax 251 nm for all derivatives. The developed method was validated as per ICH guidelines. Method was found linear over the concentration range of 200–1200 ng/spot with the correlation coefficient of 0.995, 0.993, 0.995 and 0.996 for TP, TPP, TI, TD, respectively. Limit of detection for all derivatives were in the range of 16.7-22.3 ng/spot while limit of quantitation were found to be in the range of 55.7-70.9 ng/spot.The developed TLC method can be applied for the simultaneous routine analysis of testosterone derivatives in their individual and combined pharmaceutical formulations.Anabolic androgenic steroids (AAS) are mainly a group of natural or synthetic compounds that are chemically similar to the actions of androgynous testosterone, which is primarily a natural male hormone responsible for androgenic and anabolic effects observed during male adolescence and adulthood, and are modified to improve the anabolic effect of testosterone rather than its androgenic effect [1]. In addition to their medical uses, AAS have been widely misused by variety of athletes with the hope of improving their performance [2,3]. The use of AAS in sports has been banned since the mid-1970s but they are still the most misused class of drugs in sports. Moreover, steroid abuses have also become more and more prevalent outside sports.Because of the short half-life of only one hour, exogenous intake of pure testosterone do not have any effect, since only 2% of oral intake reaches to the muscles. To slow down the metabolism and receive better effect, the testosterone molecule has been modified at its 17-position which creates stronger anabolic effect and weaker androgen effect [4]


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