Forced degradation study of thiocolchicoside: characterization of its degradation products.
Thiocolchicoside (TCC, N-[1,2-dimethoxy-10-methylsulphanyl-9-oxo-3-(3,4,5-trihydroxy-6-hydroxymethyl-tetrahydropyran-2-yloxy)-5,6,7,9-tetrahydro-benzo[a]heptalen-7-yl]-acetamide) was subjected to hydrolytic, oxidative, and photolytic stresses. TCC underwent degradation in acidic, basic, and oxidative conditions, while it was stable toward other stress conditions. The degradation products (DPs) were detected and their separation was achieved on a SGE Wakosil C18RS 5μm (250*4.6 mm; SGE) column employing a gradient LC-MS method for a total time of analysis of 18 min. The mass fragmentation pathways of both thiocolchicoside and its degradation products were established using LC-MS experiments assigning the structures to the DPs. In particular, five DPs were identified as: D1SO (N-[1,2-dimethoxy-10-methylsulphoxide-9-oxo-3-(3,4,5-trihydroxy-6-hydroxymethyl-tetrahydropyran-2-yloxy)-5,6,7,9-tetrahydro-benzo[a]heptalen-7-yl]-acetamide), D1SO(2) (N-[1,2-dimethoxy-10-methylsulphone-9-oxo-3-(3,4,5-trihydroxy-6-hydroxymethyl-tetrahydropyran-2-yloxy)-5,6,7,9-tetrahydro-benzo[a]heptalen-7-yl]-acetamide), D2 ([1,2-dimethoxy-10-methylsulphanyl-9-oxo-3-(3,4,5-trihydroxy-6-hydroxymethyl-tetrahydropyran-2-yloxy)-5,6,7,9-tetrahydro-benzo[a]heptalen-7-yl]-amine), D3 (N-[1,2-dimethoxy-3-hydroxy-10-methylsulphanyl-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptalen-7-yl]-acetamide or 3-O-demethylthiocolchicine), D4 ([1,2-dimethoxy-3-hydroxy-10-methylsulphanyl-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptalen-7-yl]-amine or N-deacetyl-3-O-demethylthiocochicine). Moreover, the structures of DPs were confirmed by synthesis of the reference standards which were fully characterized by MS, NMR, IR analyses. Finally a comprehensive degradation scheme of TCC was proposed allowing to outline D1SO and D3 as the indicators of its stability for oxidative and hydrolytic stress conditions.
Dipartimento di Scienze Chimiche, Alimentari, Farmaceutiche e Farmacologiche and Drug and Food Biotechnology Centre, Università degli Studi del Piemonte Orientale Amedeo Avogadro, Largo Donegani 2, 28100 Novara, Italy. erika.delgrosso@pharm.unipmn.it
Del Grosso
del G
Erika
Aprile
A
Silvio
Grosa
G
Giorgio
D002853:Chromatography, Liquid / Q000379:methods; D003078:Colchicine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D003078:Colchicine; C004280:thiocolchicoside