Ramesh Thippani, Rao Pothuraju Nageswara, Rao Ramisetti Nageswara
Department of Chemistry, National Institute of Technology, Warangal, Andhra Pradesh, India.
J Sep Sci. 2014 Feb;37(4):368-75. doi: 10.1002/jssc.201300959. Epub 2014 Jan 6.
A rapid, specific, and reliable isocratic LC-MS/MS method has been developed and validated for the identification and characterization of the stressed degradation products of Entecavir (ETV). ETV, an antiviral drug, was subjected to hydrolysis (acidic, alkaline, and neutral), oxidation, photolysis and thermal stress, as per the international conference on harmonization specified conditions. The drug showed extensive degradation under oxidative and acid hydrolysis stress conditions. However, it was stable to thermal, acidic, neutral, and photolysis stress conditions. A total of five degradation products were observed and the chromatographic separation of the drug and its degradation products were achieved on a Waters Symmetry C18 (250 mm × 4.6 mm, id, 5 μm) column using 20 mM ammonium acetate (pH 3)/acetonitrile (50:50, v/v) as a mobile phase. The degradation products were characterized by LC-MS/MS and its fragmentation pathways were proposed. The LC-MS method was validated with respect to specificity, linearity, accuracy, and precision. No previous reports were found in the literature regarding the degradation behavior of ETV.
已开发并验证了一种快速、特异且可靠的等度液相色谱-串联质谱（LC-MS/MS）方法，用于恩替卡韦（ETV）强制降解产物的鉴定和表征。ETV是一种抗病毒药物，按照国际协调会议规定的条件，使其经受水解（酸性、碱性和中性）、氧化、光解和热应激。该药物在氧化和酸水解应激条件下显示出广泛降解。然而，它对热、酸性、中性和光解应激条件稳定。总共观察到五种降解产物，并且在Waters Symmetry C18（250 mm×4.6 mm，内径，5μm）柱上，使用20 mM乙酸铵（pH 3）/乙腈（50:50，v/v）作为流动相，实现了该药物及其降解产物的色谱分离。通过LC-MS/MS对降解产物进行了表征，并提出了其裂解途径。该LC-MS方法在特异性、线性、准确性和精密度方面进行了验证。文献中未发现有关ETV降解行为的先前报道。
