氢溴酸沃替西汀中有关物质的结构表征和测定

Structural characterization and determination of vortioxetine hydrobromide and its related substance

目的建立氢溴酸沃替西汀及其片剂中有关物质的高效液相色谱(HPLC)测定方法。方法 Waters Symmetry?C18色谱柱(250 mm×4.6 mm,5μm);流动相:0.05%三氟乙酸水溶液–[甲醇–乙腈–水–三氟乙酸(60∶35∶5∶0.035)],梯度洗脱;体积流量:1.0 m L/min;柱温:40℃;进样量:10μL;波长:254 nm。采用红外光谱、质谱、核磁对4个主要杂质进行了结构确证,并阐述了各杂质的产生途径。结果氢溴酸沃替西汀和杂质A^G在测定质量浓度范围内线性关系良好,r均大于0.999 4;氢溴酸沃替西汀和杂质A、B、C、E、F、G的定量限浓度均为0.2μg/m L,最小检出限浓度均为0.02μg/m L;杂质D的定量限浓度为0.8μg/m L,最小检出限浓度为0.08μg/m L。平均回收率为100.4%,RSD值为1.06%。氢溴酸沃替西汀中主杂质的质量分数为0~0.01%。结论该方法快速、简便、准确,为氢溴酸沃替西汀及其片剂的质量标准的制定提供了重要依据。

Objective To establish an HPLC method for the determination of vortioxetine hydrobromide and its related substance. Methods Separation was carried out on Waters Symmetry？ C18 column（250 mm × 4.6 μm, 5 μm）. The mobile phase consisted of 0.05% aqueous solution of trifluoroacetic acid – [methyl-acetonitrile-water-trifluoroacetic acid（60∶35∶5∶0.035）] with gradient elution. The flow rate was 1.0 m L/min, temperature of column was set at 40 ℃, and volume of injection was 10 μL. The detection wavelengths were set at 254 nm. The structures of four process-related impurities formed during the synthesis were characterized and confirmed by IR, MS, and NMR spectroscopy analyses. The probable formation mechanisms of the impurities identified were proposed. Results The linear relations of vortioxetine hydrobromide and impurities A — G in the range of mass concentration was good with perfect precision with r all more than 0.999 4. Quantitative limiting concentration of vortioxetine hydrobromide and impurities A, B,C, E, F, and G were 0.2 μg/m L, and minimum detection limit concentration were 0.02 μg/m L. Quantitative limiting concentration of impurity D was 0.8 μg/m L, and minimum detection limit concentration was 0.08 μg/m L. The average recovery rates were 100.4%, and RSD was 1.06%. The content of main impurity in vortioxetine hydrobromide was 0 — 0.01%. Conclusion The method is rapid, convenient, and with precision and accuracy, and provides reference and basis to develop and improve vortioxetine quality standard.