基于UPLC-MS技术的拉坦前列素滴眼液中15S-拉坦前列素分析

目的：建立超高效液相色谱-四极杆-静电场轨道阱高分辨质谱法定量检测拉坦前列素滴眼液中15S-拉坦前列素。方法：采用色谱柱Thermo Accucore-C₁₈（2.1mm×150 mm，2.6 μm）；流动相为甲醇-乙腈-水（用冰乙酸调节pH3.0）（48∶12∶40）；流速为0.3 mL·min^-1；柱温为35℃；进样量为10 μL。采用HESI离子源，正离子扫描模式，扫描范围为m/z 300~600。采用FullMS模式提取离子方式检测，拉坦前列素与15S-拉坦前列素的检测离子均为[M+Na]⁺m/z 455.277±0.003；电喷雾离子源雾化温度为300℃；毛细管电压为3.50 kV；离子传输管温度为320℃。鞘气为50 arb；辅助气为10 arb；分辨率为（R）70 000。结果：15S-拉坦前列素质量浓度在0.998 8~19.98 ng·mL^-1范围内线性关系良好，相关系数为r=0.999 8，平均回收率为96.8%（n=9）；定量限为0.029 96 ng·mL^-1；3批样品中15S-拉坦前列素的含量分别为0.11%、0.24%、0.06%。结论：经方法学验证，本方法可检测拉坦前列素滴眼液中15S-拉坦前列素立体异构体杂质。

Objective: To determine 15S-latanoprost in latanoprost eye drops by UPLC-Q-orbitrap mass spectrometer.Methods: The separation was carried out on an Thermo Accucore-C₁₈(2.1 mm×150 mm, 2.6 μm) column with the mobile phase consisting of methanol-acetonitrile-water (pH was adjusted to 3.0 with acetic acid)(48:12:40), the flow rate was 0.3 mL·min^-1 and the column temperature was 35℃, and the injection volume was 10 μL.HESI was employed, the sample was analyzed in the positive mode, with the probe heater temperature at 300, capillary temperature 320, and m/z range was from 300 to 600, and detected at[M+Na]⁺ m/z 455.277±0.003 for both latanoprost and 15 S-latanoprost.Results: The calibration curve of 15S-latanoprost was a good linearity within concentration range of 0.998 8-19.98 ng·mL^-1, and the correlation coefficients was 0.999 8.The limits of 15S-latanoprost was 0.029 96 ng·mL^-1, the average recovery of 15S-latanoprost was 96.8% (n=9).The contents of 15S-latanoprost in 3 batches of samples were 0.11%, 0.24%, 0.06% respectively.Conclusion: The methodology validation proved that the method is useful as a quality control method for trace analysis of 15S-latanoprost in latanoprost eye drops.