Development and validation of a stability-indicating HPLC-UV method for the determination of Thiocolchicoside and its degradation products

• Published in: Journal of pharmaceutical and biomedical analysis 2017-01, Vol.132, p.66-71
• Main Authors: Aprile, Silvio, Canavesi, Rossana, Bianchi, Michele, Grosa, Giorgio, Del Grosso, Erika
• Format: Article
• Language: English
• Subjects: Catalysis; Chromatography, High Pressure Liquid - methods; Colchicine - analogs & derivatives; Colchicine - analysis; Degradation products; Drug Stability; HPLC-UV; Hydrogen-Ion Concentration; Hydrolysis; Limit of Detection; Linear Models; Oxidation-Reduction; Reproducibility of Results; Stability indicating method; Thiocolchicoside; Ultraviolet Rays; Validation
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2016.09.037

[Display omitted] •A stability indicating method for thiocolchicoside has been developed.•The method was validated as for ICH guidelines.•The method was used for the analyses of thiocolchicoside commercial liquid or solid dosage form samples. A stability indicating high performance liquid chromatography method has been developed for the determination of thiocolchicoside (TCC) and its main degradation products thiocolchicoside S-oxide (D1SO) and 3-O-demethylthiocolchicine (D3) in liquid and solid formulations. The method was developed based on a previous forced degradation study showing that TCC underwent chemical degradation by acid/base catalyzed hydrolysis and oxidation being the main degradation products D3 and D1SO respectively. The analytes separation and quantification were achieved on a Synergi™ 4μm Polar-RP 80Å, column 150×4.6mm (Phenomenex) using the mobile phase constituted (flow rate 1mLmin−1) of eluant A: 20mM sodium acetate buffer (pH 5.0) and eluant B: MeOH:CH3CN (20:80); the elution was performed in gradient mode detecting the analytes at 254nm. The method showed linearity for TCC assay in the 5–15μgmL−1, range and for unknown (TCCfu) and known (D1SO and D3) degradation products assay, in the 0.5–10μgmL−1 range: all the square of the correlation coefficients were greater than 0.999. The precision, determined in terms of intra-day and inter-day were expressed as RSDs and resulted to be 1.19, 1.10, 1.37 and 1.04% and 0.95, 0.83, 1.30 and 0.72 for TCC, TCCfu, D1SO and D3, respectively. The method demonstrated also to be accurate; indeed, the average recoveries were 102.1/102.0% for TCC (ampoules and hard capsules respectively), 101.3/100.3% for TCCfu, 101.7/100.2% for D1SO, and 101.4/101.4% for D3. The robustness was also evaluated by variations of mobile phase composition and pH. Finally, the applicability of the method was evaluated by analysis of commercial liquid and solid dosage forms.