Bueno Lívia Maronesi, Manoel Joanna Wittckind, Giordani Camila Ferrazza Alves, Mendez Andreas Sebastian Loureiro, Volpato Nadia Maria, Schapoval Elfrides Eva Scherman, Steppe Martin, Garcia Cássia Virginia
Programa de Pós-Graduação em Ciências Farmacêuticas, Universidade Federal do Rio Grande do Sul (UFRGS), Av. Ipiranga 2752, Porto Alegre, RS, Brazil.
Programa de Pós-Graduação em Ciências Farmacêuticas, Universidade Federal do Rio Grande do Sul (UFRGS), Av. Ipiranga 2752, Porto Alegre, RS, Brazil.
Eur J Pharm Sci. 2017 Jan 15;97:22-29. doi: 10.1016/j.ejps.2016.11.004. Epub 2016 Nov 3.
A simple, fast and sensitive analytical method by high-performance liquid chromatography (HPLC) was developed and validated for the simultaneous determination of ticagrelor and two synthesis impurities. The HPLC method was established using an Agilent 1200 Series equipment coupled to photodiode array detector (PDA) at 270nm with a Zorbax Plus C column (150×4.6mm, 5.0μm), injection volume of 20μL, and a constant temperature of 25°C. The mobile phase consisted of acetonitrile: ammonium acetate 50mM (57:43, v/v) and pH adjusted to 8.2 with ammonium hydroxide 6M, at a flow rate of 0.7mL/min. No interference peaks from excipients and diluent system indicated the specificity of the method. The calibration curves showed determination coefficients (r)>0.99, calculated by linear regression. The limit of quantitation (LOQ) for impurities 1 and 2 were 2.0 and 0.2μg/mL, respectively. Intra and interday relative standard deviations (RSDs) were <2% for ticagrelor and <6% for the impurities, proving the precision of the method. Besides, two mayor degradation products formed when sample solutions of ticagrelor were exposed to UVC radiation were elucidated and the mechanisms involved in the photolytic degradation of ticagrelor were proposed.
建立了一种简单、快速且灵敏的高效液相色谱（HPLC）分析方法，并对其进行了验证，用于同时测定替格瑞洛及其两种合成杂质。该HPLC方法采用安捷伦1200系列设备，配备光电二极管阵列检测器（PDA），检测波长为270nm，使用Zorbax Plus C柱（150×4.6mm，5.0μm），进样体积为20μL，柱温为25°C。流动相由乙腈：50mM醋酸铵（57:43，v/v）组成，并用6M氢氧化铵将pH值调至8.2，流速为0.7mL/min。辅料和稀释剂系统无干扰峰，表明该方法具有特异性。校准曲线的测定系数（r）>0.99，通过线性回归计算得出。杂质1和杂质2的定量限（LOQ）分别为2.0和0.2μg/mL。替格瑞洛的日内和日间相对标准偏差（RSD）<2%，杂质的RSD<6%，证明了该方法的精密度。此外，还阐明了替格瑞洛样品溶液暴露于UVC辐射时形成的两种主要降解产物，并提出了替格瑞洛光解降解的相关机制。
