高效液相色谱质谱联用法测定氢溴酸沃替西汀有关物质

Determination of Related Substances of Vortioxetine Hydrobromide by HPLC

目的建立测定氢溴酸沃替西汀有关物质的高效液相色谱方法,并用高效液相色谱-质谱联用法(HPLC-MS)鉴定氢溴酸沃替西汀的降解产物。方法色谱柱为Agilent Zorbax SB-CN色谱柱(4.6 mm×250 mm,5μm),流动相为乙腈-0.02 mol·m L^(-1)甲酸铵(50∶50,磷酸调节p H至5.0),流速为1 m L·min^(-1),检测波长为228 nm,柱温为25℃,进样量20μL。结果氢溴酸沃替西汀与有关物质均可有效地分离;氢溴酸沃替西汀及各有关物质检测灵敏度满足测定要求;氢溴酸沃替西汀和各有关物质的线性关系良好;方法重复性良好(RSD=6.89%,n=6),加样回收率(n=9)均在95%~105%之间;在氧化破坏条件下产生的降解产物经质谱分析初步鉴别为有关物质D。结论建立的方法简便、灵敏度高、结果可靠,可用于氢溴酸沃替西汀有关物质的检测和鉴定。

OBJECTIVE To establish an HPLC method to determine the related substances of vortioxetine hydrobromide and identify the degradation products of vortioxetine hydrobromide by HPLC-MS. METHODS An HPLC method was developed by using a CN column（Agilent Zorbax SB-CN,4. 6 mm × 250 mm, 5 μm） ,the mobile phase consisted of CHaCN-HCOONH4 （50： 50, pH adjusted to 5.0 with phosphoric acid） ,the flow rate was 1.0 mL · min^-1 , and the detection wavelength was set at 228 nm. The column temperature was maintained at 25℃, and the injection volume was 20 μL. RESULTS Vortioxetine hydrobromide was well separated from the related substances. The detection sensitivity of vortioxetine hydrobromide and its related substances met the determination requirements. The calibration curves of vortioxetine hydrobromide and related substances had good linearity. The repeatability of the method was good（RSD =6. 89% , n =6）. The average recoveries of the sample were all within 95% - 105%. The degradation product produced by oxidation was identified by mass spectrometry as related substance D. CONCLUSION The developed method proves to be simple, accurate, specific and reliable. It can be applied to the determination of vortioxetine hydrobromide and its related substances.