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<p>Abstract</p> <p>Background</p> <p>Anabolic androgenic steroids (AAS) are widely misused for the enhancement of performance in sports. Several drugs are available that contain different combinations or individual steroids in different dosage form. This paper describes a TLC densitometric method for simultaneous determination of four AAS of testosterone derivatives including testosterone propionate (TP), testosterone phenyl propionate (TPP), testosterone isocaproate (TI) and testosterone deaconate (TD) in their pharmaceutical products.</p> <p>Results</p> <p>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 <it>R</it>_f 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 <it>λ</it>_max 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.</p> <p>Conclusions</p> <p>The developed TLC method can be applied for the simultaneous routine analysis of testosterone derivatives in their individual and combined pharmaceutical formulations.</p>