拉坦前列素3种异构体HPLC分析方法建立

Determination of three isomers of latanoprost by HPLC

目的:建立用HPLC法测定拉坦前列素中3种异构体。方法:采用硅胶色谱柱Inertsil SIL 100A(250 mm×4.6 mm,5μm),以正庚烷-异丙醇-乙腈(93∶4∶3)为流动相,流速1 ml/min,检测波长210 nm,柱温30℃;样品室温度为25℃。结果:拉坦前列素与3种异构体峰分离良好。拉坦前列素在1.01~67.26μg/ml浓度范围内呈良好的线性关系(r=0.999 9),异构体15(S)-顺式拉坦前列素、15(S)-反式拉坦前列素及15(R)-反式拉坦前列素分别在0.423~13.53、0.607~4.55和1.042~69.48μg/ml浓度范围内线性关系良好(r为0.999 5~0.999 9),检出限分别为3.2、4.6和7.8 ng,平均回收率分别为99.9%、100.8%和98.8%,RSD分别为0.3%、0.9%和1.1%。结论:该方法专属性好,适用于拉坦前列素3种异构体的测定。

Objective: To develop an HPLC method for separation and determination of three isomers in latanoprost. Methods: A Inertsil SIL 100 A column( 250 mm × 4. 6 mm,5 μm) was adopted. The mobile phase was composed of n-heptane-isopropanol-acetonitrile( 93∶ 4∶ 3),the flow rate of which was 1. 0 ml/min,and the detection wavelength was set at 210 nm.The temperature of the columne and autosampler was set at 30 ℃ and 25 ℃,respectively. Results: Latanoprost and its three isomers were separated satisfactorily. Calibration curves were found to be linear in the ranges of 1. 01 ~ 67. 26 μg/ml of latanoprost,0. 423 ~ 13. 53 μg/ml of 15( S)-cis latanoprost,0. 607 ~ 4. 55 μg/ml of 15( S)-trans latanoprost,and 1. 042 ~ 69. 48 μg/ml of 15( R)-trans latanoprost( r = 0. 999 5 ~ 0. 999 9),respectively. The LOD of 15( S)-cis latanoprost,15( S)-trans latanoprost and 15( R)-trans latanoprost were 3. 2,4. 6 and 7. 8 ng,respectively,The average recovery of the above substances was99. 9%,100. 8% and 98. 8%,respcetively. The RSD of sampling precision was ≤2%. Conclusion: The developed method is specific and accurate for the determination of latanoprost and its three isomers.