Authenticity assessment of anabolic androgenic steroids in counterfeit drugs by 1H NMR

Abstract

Anabolic steroids are synthetic hormone derivatives of testosterone, a masculine sex hormone. Their main effects include greater muscle size, physical strength, and resistance. The need for safe and reliable quality control of products and/or raw materials in the pharmaceutical industry makes analyses of active ingredients and other constituents of drug formulations extremely important for the manufacturers as well as users of these products. In this work, we propose the use of ¹H NMR spectroscopy to analyze formulations containing anabolic steroids. Six types of anabolic steroids were studied (testosterone propionate, methyltestosterone, acetate methenolone, oxandrolone, nandrolone decanoate, and stanozolol), with analysis of 16 samples of anabolic drugs that are used in injectable, tablet, and capsule forms. The ¹H NMR spectroscopic measurements were performed using a 600 MHz spectrometer, with deuterated chloroform (CDCl₃) containing 0.03% TMS as the solvent. Of the 16 samples analyzed, four did not show the presence of the active principle stated on the label. For quantitative analysis, dimethyl sulfone (DMSO₂) was used as an internal reference standard. All the analytical curves were obtained for concentration ranges of 1.0–16.0 mg mL⁻¹. The limits of detection and quantification varied in the ranges 0.18–0.61 and 1.30–2.19 mg mL⁻¹, respectively. The intra-day and inter-day precision RSD values were in the ranges 1.86–4.22% and 2.17–4.75%, respectively. The recovery values ranged from 95.2 to 104.0%. The spectroscopic results statistically agreed with those obtained by reference methods (HPLC).