HPLC测定拉坦前列素滴眼液复杂系统中的有关物质及含量

Determination of the Related Substances and Content in the Complex System of Latanoprost Eye Drops by HPLC

关键词:  拉坦前列素滴眼液  高效液相色谱法  有关物质  含量测定

Key words:  Latanoprost Eye drops  HPLC  related substances  content determination

摘要:

目的 建立HPLC测定拉坦前列素滴眼液复杂系统中的有关物质及含量的方法。方法 采用糖涂敷型手性色谱柱（4.6 mm×250 mm，5 μm），有关物质测定以0.005 mol·L^-1磷酸二氢钠溶液（用磷酸调节pH值至2.7）-乙腈（56：44）为流动相，含量测定以0.005 mol·L^-1磷酸二氢钠溶液（用磷酸调节pH值至2.7）-乙腈（54：46）为流动相，检测波长为200 nm，流速为0.5 ml·min^-1。结果 有关物质测定中2个已知杂质与主峰之间的分离度良好，线性关系良好，5，6-反式-拉坦前列素和15S-拉坦前列素的平均回收率分别为101.5%和99.5%，拉坦前列素、5，6-反式拉坦前列素和15S-拉坦前列素的定量限（S/N≈10）分别为0.12，0.13和0.12 μg·mL^-1；含量测定线性关系良好，重复性RSD=0.90%，拉坦前列素平均回收率为98.8%。结论 建立的方法准确可靠，可用于拉坦前列素滴眼液的质量控制。

Abstract:

OBJECTIVE To develop an HPLC method for the determination of the related substances and content in the complex system of Latanoprost Eye drops. METHODS the conditions of detection were Sugar Chiral column C₁₈(250 mm×4.6 mm, 5 μm), related substances measured using the isocratic mobile phase consisted of 0.005 mmol·L^-1 sodium dihydrogen phosphate (adjust with phosphate to pH 2.7)-acetonitrile(56:44), and the mobile phase for content determination was 0.005 mol·L^-1 sodium dihydrogen phosphate solution (adjusted with phosphoric acid to pH 2.7)-acetonitrile(54:46) with a flow rate of 0.5 mL·min^-1, the UV detection wavelength was 200 nm. RESULTS The resolution of 5, 6-trans-Latanoprost and 15S-Latanoprost was good. the method for determination of the related substances of Latanoprost Eye drops had a good linearity. The average recovery of 5,6-trans-Latanoprost and 15S-Latanoprost were 101.5% and 99.5%. The limit of quantitation of Latanoprost, 5,6-trans-Latanoprost and 15S-Latanoprost were 0.12, 0.13 and 0.12 μg·ml^-1. The method for determination of Latanoprost Eye drops had a good linearity, the repeatability was good (RSD=0.90%). The average recovery of Latanoprost was 98.8%. CONCLUSION The method is accurate and reliable, which can be used to control the quality of Latanoprost Eye drops.