Analysis of seized stanozolol formulations in South Brazil by liquid chromatography coupled to quadrupole time-of-flight-mass spectrometry

Abstract

Anabolic-androgenic steroids (AAS) are synthetic derivatives of testosterone which are used medically for several diseases. However, misuse is commonly observed by athletes to promote enhancement of strength and performance. AAS are frequently obtained through online black markets from clandestine drug manufacturing laboratories, without any quality standards, being potentially dangerous for users. The purpose of this work was the development and application of a fast and simple procedure for the quantitation of stanozolol by liquid chromatography coupled to quadrupole time-of-flight-mass spectrometry (LC-QTOF-MS) in tablets packs seized in Rio Grande do Sul state, Brazil. The samples of stanozolol were separated considering its dosage form. The internal standard (methyltestosterone) was added to the aliquots of the samples, dissolved in methanol and 5μL were injected into the analytical system. The newly developed method has been validated for lower limit of quantitation (LLOQ), linearity, accuracy, precision and selectivity. The LLOQ was 0.1 µg/mL. The developed method was successfully applied to 31 samples seized by the Secretaria da Receita Federal do Brasil (a Brazilian federal revenue service agency). According to the results, 90.3% of the suspected medicines (n=31) were adulterated, and 65% exhibited higher concentrations of stanozolol than those indicated in the label. This work successfully established a new method for quantification of stanozolol using LC-QTOF-MS. This method aims at contributing to the identification and quantification of this anabolic androgenic steroid frequently seized by federal inspection agencies.

Fake anabolic androgenic steroids on the black market – a systematic review and meta-analysis on qualitative and quantitative analytical results found within the literature

“…The analysis for substandard substances comes with some challenges. Firstly, defined ranges of declared labels could vary massively between articles, had a quantitative analysis been performed, with defined ranges between 50–200% [ 26 ], 80–130% [ 42 ], 80–120% [ 43 ] or 90–110% [ 44 ]. In some studies, the contained active ingredients in “under-concentrated” preparations was much lower than 50% of that indicated (e.g.…”

“…In some studies, the contained active ingredients in “under-concentrated” preparations was much lower than 50% of that indicated (e.g. 0.5–1.5% [ 45 ], 9% [ 44 ] or 16% [ 46 ]) if quantitative data was available. For “over-concentrated” preparations however, active ingredients could go as much as 200% above that indicated on the label (e.g.…”

“…For “over-concentrated” preparations however, active ingredients could go as much as 200% above that indicated on the label (e.g. 221% [ 25 ] or 225% [ 44 ]) if quantitative data was available. Furthermore, most authors ( n = 7) performed a quantification only in the accurately labeled substances, whereas Weber and colleagues [ 26 ] included mixed samples (accurately labeled and adulterated) for quantitative analysis.…”

“…GC–MS with [ 51 ] or without [ 40 , 46 ] prior derivatization of the AAS has also been successfully used to screen and quantify AAS, based on their fragmentation patterns and retention times. Other complementary techniques like 1H-nuclear magnetic resonance (NMR) [ 39 , 44 ], infrared (IR) spectroscopy [ 34 , 40 , 46 , 55 ], differential scanning calorimetry (DSC) [ 40 ], or high resolution/high accuracy mass spectrometry (LC-HRMS) [ 26 , 32 , 42 ] have also been used to measure AAS. For AAS, both low-resolution and high-resolution mass spectrometers were employed.…”