HPLC法同时测定N-芴甲氧羰基-O-叔丁基-L-苏氨酸中的6个杂质  

Determination of six related substances in N-fluorenylmethoxycarbonyl-O-tert-butyl-L-threonine by HPLC

作　　者：杨欣茹 李铁健 胡发红 韩忠丽 张贵民 YANG Xin-ru;LI Tie-jian;HU Fa-hong;HAN Zhong-li;ZHANG Gui-min(National Engineering and Technology Research Center of Chirality Pharmaceutical,Lunan Pharmaceutical Group Co.,Ltd.,Linyi 273400,China)

机构地区：[1]鲁南制药集团股份有限公司,国家手性制药工程技术研究中心,临沂273400

出　　处：《药物分析杂志》2024年第3期468-474,共7页Chinese Journal of Pharmaceutical Analysis

基　　金：山东省重大科技创新工程项目(2019JZZY010516)。

摘　　要：目的:建立高效液相色谱同时测定N-芴甲氧羰基-O-叔丁基-L-苏氨酸中6个特定杂质的方法。方法:采用YMC Triart C18(250 mm×4.6 mm,3μm)色谱柱,以0.1%三氟乙酸水溶液为流动相A,以0.1%三氟乙酸乙腈溶液为流动相B,流速1.0 mL·min^(-1),梯度洗脱,检测波长265 nm,柱温30℃,进样体积10μL。结果:N-芴甲氧羰基-O-叔丁基-L-苏氨酸与相邻杂质峰的分离良好;6个杂质分离度均大于1.5;且在相应质量浓度范围内呈现良好的线性关系(r≥0.999);6个杂质检测限和定量限分别约为0.03μg·mL^(-1)和0.06μg·mL^(-1);6个杂质的平均回收率(n=9)在97.6%~98.8%范围内。3批N-芴甲氧羰基-O-叔丁基-L-苏氨酸测定结果显示,杂质1的含量<0.2%,杂质4的含量<0.1%,其他4种杂质未检出,总杂含量<1%。结论:本方法分离度好,灵敏度高,专属性强,适用于N-芴甲氧羰基-O-叔丁基-L-苏氨酸中有关物质的检测。Objective:To establish an HPLC method for the determination of six related substances in N-fluorenylmethoxycarbonyl-O-tert-butyl-L-threonine.Methods:The analysis was conducted on YMC Triart C_(18)(250 mm×4.6 mm,3μm)column,the mobile phase was consisted with 0.1%trifluoroacetic acid in water(A)and 0.1%trifluoroacetic acid in acetonitrile(B)at the flow rate of 1.0 mL·min^(-1).The column temperature was set 30℃,the detection wavelength was 265 nm and the injection volume was 10μL.Results:N-Fluorenylmethoxycarbonyl-O-tert-butyl-L-threonine had good separation from the adjacent impurity peaks;The resolution of impurity1-6 was greater than 1.5,and showed a good linear relationship(r≥0.999)in the corresponding mass concentration range,the detection limit of impurity 1-6 was 0.03μg·mL^(-1),and the quantitative limit was 0.06μg·mL^(-1),the average recovery rate(n=9)of impurity 1-6 was in the range of 97.6%-98.8%.The results of the three batches of N-fluorenylmethoxycarbonyl-O-tert-butyl-L-threonine showed that the contents of impurity 2,impurity 4 were<0.2%and<0.1%,respectively,and the other four impurities were not detected,and content of the total impurity was<1%.Conclusion:This method has good resolution,high sensitivity and strong specificity,and is suitable for the determination of related substances in N-fluorenylmethoxycarbonyl-O-tert-butyl-L-threonine.

关 键 词：N-芴甲氧羰基-O-叔丁基-L-苏氨酸 高效液相色谱法 杂质 梯度洗脱 

分 类 号：R917[医药卫生—药物分析学]