高效液相色谱法测定氨碘肽滴眼液中5种氨基酸的含量  

HPLC determination of 5 amino acid of Amiotide eye drops

作　　者：孙炜[1,2] 姚志斌[1,2] 李雪冬[1,2] 杨娜[1,2] 曾之媛 刘金璐[1,2] 孔珺[1,2] 

机构地区：[1]中国医科大学附属第四医院 [2]中国医科大学眼科医院,沈阳110005

出　　处：《中南药学》2014年第9期909-912,共4页Central South Pharmacy

基　　金：国家自然科研基金--青年基金项目(No.81100654)

摘　　要：目的采用HPLC法测定氨碘肽滴眼液中丝氨酸（Ser）、甘氨酸（Gly）、组氨酸（His）、异亮氨酸（Ile）、亮氨酸（Leu）的含量。方法采用Ocean-C18色谱柱（250 mm×4.6 mm,5μm）;流动相A：乙腈-水（4∶1）,流动相B：0.1 mol·L-1乙酸钠-乙腈（97：3）;流速为1.0 mL·min-1;柱温：25℃;检测波长：254 nm;用异硫氰酸苯酯（PITC）作为柱前衍生化试剂,采用柱前衍生HPLC法进行分析。结果各种氨基酸能良好地分离,衍生化试剂对氨基酸分析无干扰;平均回收率在98%～102%,RSD均〈3%。结论该方法简便、准确、可靠,可用于氨碘肽滴眼液中氨基酸含量的检测。Objective To establish a HPLC method to determine the 5 amino acid in Amiotide eye drops. Methods A Ocean-C18 （250 mm × 4.6 mm, 5μm） column was used. The mobile phase A was acetonitrile-water （4 ： 1） and B was 0.1 mol · L -1 sodium acetate-acetonitrile （97 ： 3）. The flow rate was 1.0 mL · min -1. The column temperature was 25 ℃ . The detection wavelength was 254 nm. Phenylisothiocyanate was used as the derivatizing agent. Results Complete separation was achieved among 5 amino acid under optimun conditions. No interference from derivative reagent was found. The average recoveries were at 98% - 102%, and RSDs were less than 3%. Conclusion The method is simple, accurate, reliable and suitable for the quality control of Amiotide eye drops.

关 键 词：氨碘肽滴眼液 异硫氰酸苯酯 柱前衍生化 高效液相色谱法 氨基酸 

分 类 号：R917[医药卫生—药物分析学]