摘要
目的:建立高效液相色谱法(high-performance liquid chromatography,HPLC)测定曲伏噻吗滴眼液中曲伏前列素有关物质。方法:采用Agilent SB-C18色谱柱(50 mm×2.1 mm,2.7μm);以磷酸溶液(取磷酸2.0 m L,加水稀释并定容至1000 m L,用氢氧化钠溶液调节p H至3.0)为流动相A,乙腈为流动相B,洗脱梯度;流速为每分钟3.0 m L;柱温为30℃;检测波长为220 nm;进样体积100μL。结果:在该色谱条件下,曲伏前列素与各杂质均可良好分离;在酸、碱、氧化、高温和强光破坏条件下,曲伏前列素峰纯度合格,物料守恒。曲伏前列素、5,6-反式曲伏前列素和15-酮曲伏前列素分别在0.041~3.245μg/m L(r=1.0000)、0.040~3.229μg/m L(r=1.0000)、0.039~3.088μg/m L(r=0.9999)范围内线性关系良好,其最低检测限分别为0.020、0.020和0.020μg/m L;6份样品中5,6-反式曲伏前列素的含量相对标准偏差(relative standard deviatio,RSD)为0.2%(n=6),15-酮曲伏前列素的含量RSD为0.3%(n=6),重复性良好;对照品溶液和供试品溶液在室温条件下放置100h稳定,5,6-反式曲伏前列素的平均回收率为95.2%,相对标准偏差RSD为0.5%(n=9),15-酮曲伏前列素的平均回收率为92.7%,RSD为1.2%(n=9)该方法重现性好。结论:本方法简便、快速、准确。适用于检测曲伏噻吗滴眼液中的曲伏前列素有关物质。
Objective:To develop a high-performance liquid chromatography(HPLC)method for the determination of related substances in Travoprost and Timolol Maleate Eye Drops,sepcifically focusing on Travoprost and its impurities.Methods:The analytic column used was an Agilent SB-C18(50 mm×2.1 mm,2.7μm).A phosphoric acid solution(prepared by diluting 2.0 mL of phosphoric acid with wateto final volume of 1000 mL and adjust the pH to 3.0 with sodium hydroxide solution)was used as mobile phase A,while acetonitrile served as mobile phase B.The elution was performed using a gradient program at a flow rate of 3.0 mL/min.The detection wavelength was set at 220 nm,and the column temperature was maintained at 30℃.The injection volume is 100μL.Results:Under the described chromatographic condition,Travoprost and its various impurities were well separated.The purity of the Travoprost peak was qualified,and the material remained stable under conditions of acid,alkali,oxidation,high temperature,and strong light exposure.The linear ranges for Travoprost,5,6-trans-Travoprost,and 15-keto-Travoprost were determined to be 0.041-3.245μg/mL(r=1.0000),0.040~3.229μg/mL(r=1.0000),0.039~3.088μg/mL(r=0.9999),respectively.The lowest detection limits for these compounds were all 0.020μg/mL,The relative standard deviation(RSD)for the content of 5,6-trans-travoprost and 15 keto trovopros in six samples were 0.2%(n=6)and 0.3%(n=6),respectively,indicating good reproducibility.Both the reference solution and the test solution remained stable at room temperature for 100 hours,The average recovery rates for 5,6-trans-Travoprost and 15-keto-Travoprost were 95.2%(RSD 0.5%,n=9)and 92.7%(RSD 1.2%,n=9).respectively,further confirming the high reproducibility of the method.Conclusions:The developed HPLC method is simple,rapid,and accurate,making it suitable for the determination of related substances in Travoprost and Timolol Maleate Eye Drops.
作者
李裕军
陈婕婷
LI Yujun;CHEN Jieting(State Key Laboratory of Ophthalmology,Zhongshan Ophthalmic Center,Sun Yat-sen University,Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science,Guangzhou 510060,China)
出处
《眼科学报》
2025年第6期471-482,共12页
Eye Science
关键词
高效液相色谱法
曲伏噻吗
滴眼液
曲伏前列素
有关物质
high-performance liquid chromatography
travoprost and timolol maleate
eye drops
travoprost
related substance
作者简介
通信作者:陈婕婷,E-mail:1023827727@qq.com。
