A validated stability-indicating TLC-densitometric method for the determination of stanozolol in pharmaceutical formulations

Abstract

BackgroundStanozolol is a synthetic derivative of dihydrotestosterone (DHT), and one of the frequently detected anabolic steroids in doping analysis. The current work describes the development and validation of the stability-indicating TLC-densitometric method for sensitive and specific estimation of stanozolol even its degradation product being there. Precoated silica gel TLC aluminium plates were utilized as the stationary phase and the eluent comprised of petroleum ether: acetone (6:4, v/v). Densitometric analysis of stanozolol was achieved at λmax 750 nm in the absorbance mode after staining with phosphomolybdic acid (PMA). Stress degradation of stanozolol was carried out under various reaction conditions including acid, base and neutral hydrolysis, wet and dry heating treatment, oxidation, and photo-degradation. Resulted stress samples and pharmaceutical products were analyzed with the developed TLC method.ResultsThis system showed a compact spot for stanozolol at Rf value of 0.46 ± 0.02. The data of linear regression analysis indicated a good linear relationship over the range of 200–1200 ng/spot concentrations. The method was validated for robustness, precision and recovery. The LOD and LOQ were 1.6 and 5.1 ng/spot, respectively. Under various stressed conditions, stanozolol showed degradation only under acidic hydrolysis. Peak of a degraded product was well resolved from the stanazolol with reasonably different Rf value and identified as 17, 17-dimethyl-l8-nor-5α-androst-13(14)-eno [3,2c] pyrazole through 1D- and 2D-NMR spectroscopic techniques and ESI-QqTOF-MS/MS analysis.ConclusionResult reflected that the stanozolol is majorly affected by the acidic condition. Statistical analysis indicated the application of the developed stability-indicating TLC-densitometric method for routine analysis of stanozolol in the presence of its degradation product.

Advances in the Determination of Anabolic-Androgenic Steroids: From Standard Practices to Tailor-Designed Multidisciplinary Approaches

“…An overview of TLC analysis of steroids, including AASs, is given in [ 74 ]. Of the current steroids, the development of a method for the densitometric determination of stanozolol is worth mentioning [ 75 ]. In this method, the limit of detection (LOD) is 1.6 ng per spot, and a good linear relationship over the range of 200–1200 ng per spot concentrations was achieved on traditional silica-gel-coated aluminum plates using petroleum ether:acetone (6:4, v/v) as the mobile phase.…”

Neurotoxic Effects of Stanozolol on Male Rats‘ Hippocampi: Does Stanozolol cause apoptosis?

“…One of the most popular members of the AAS family is stanozolol which is used by athletes in high doses for boosting their energy and improving their physical appearance and size [5]. Despite being a banned substance by the International Olympic Committee since 1974, stanozolol is one of the most commonly abused synthetic AASs in sport [6].…”

“…Stanozolol (5α-androstane-17α-methyl-17β-ol[3,2-c] pyrazole) is a heterocyclic AAS first synthesised by Clinton in 1959 ( Figure 1). Its chemical structure is different from both intrinsic and most extrinsic steroids [6]. These steroids are transferred to the nucleus as the hormonereceptor complex through the intracellular receptor and induce protein synthesis leading to the construction of new proteins [1].…”

“…These steroids are transferred to the nucleus as the hormonereceptor complex through the intracellular receptor and induce protein synthesis leading to the construction of new proteins [1]. Therapeutic effects have been reported in osteoporosis, protein deficiency, increasing appetite and weight, inherited angioedema and specific kinds of anaemia [6][7][8].…”

Post-derivative TLC densitometric stability indicating assay for mesterolone and quantum chemical calculations

“…In addition to our previous contributions to the development of various chromatographic methods for the analysis of biologically and pharmaceutically important compounds, [21][22][23][24][25][26][27] this study describes a simple stability-indicating TLC-densitometric method for the analysis of mesterolone in pharmaceutical products as per the ICH guidelines 28 and also establishes mesterolone's reactivity under various stress conditions.…”