HPLC柱前衍生法测定舒血宁注射液中四种氨酸的含量  

Determination of four amino acids in Shuxuening injection by HPLC coupled with precolumn derivatization

作　　者：陈莹[1] 李晋[1] 曹君[2] 何俊[1] 关秀伟 张纲 李志刚 常艳旭[1] CHEN Ying LI Jin CAO Jun HE Jun GUAN Xiu- wei ZHANG Gang LI Zhi- gang CHANG Yan- xu(i . Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, 300193, China College of Material Chemistry and Chemical Engineering, Hangzhou Normal University, Hangzhou 310036, China Hebei Shineway Pharmaceutical Co., Ltd., Langfang 065200, China Hebei Province Engineering Research Center for TCM Injection, Langfang 065200, China Chinese medicine injection new drug development technology national local joint engineering laboratory, Langfang 065200, China)

机构地区：[1]天津中医药大学,天津市现代中药重点实验室,天津300193 [2]杭州师范大学材料与化学化工学院,杭州310036 [3]河北神威药业有限公司,廊坊065200 [4]河北省中药注射液工程技术研究中心,廊坊065200 [5]中药注射剂新药开发技术国家地方联合工程实验室,廊坊065200

出　　处：《天津中医药大学学报》2017年第4期295-298,共4页Journal of Tianjin University of Traditional Chinese Medicine

基　　金：国家重大新药创制项目(2014ZX09201022-004);国家自然科学基金面上项目(81374050)

摘　　要：[目的]建立舒血宁注射液中4种氨基酸(天门冬氨酸、苏氨酸、丙氨酸、甲硫氨酸)的高效液相色谱(HPLC)柱前衍生化法,为舒血宁注射液的质量控制提供方法。[方法]以异硫氰酸苯酯(PITC)为柱前衍生化试剂,采用C18色谱柱,乙腈-水(5∶1)-乙酸铵缓冲溶液(p H 6.5)为流动相,梯度洗脱,检测波长为254 nm,测定舒血宁注射液中天门冬氨酸、苏氨酸、丙氨酸、甲硫氨酸的含量。[结果]4种氨基酸的分离及线性关系良好,精密度,重复性,稳定性RSD均都小于5%,平均回收率在95.6%~98.9%之间且RSD小于5%。舒血宁注射液中4种氨基酸总含量为1 738.47μg/mL。其中甲硫氨酸含量最高,丙氨酸的含量最低。[结论]本方法简便、准确、可靠,适用于舒血宁注射液中氨基酸含量的测定。[Objective] To establish a HPLC method for determining 4 amino acids （ aspartate, threonine, alanine and methionine） in Shuxuening injection. [Methods] HPLC analysis was performed after derivatization by using phenyl isothiocyanate （ PITC） as a derivative reagent. Aspartate, threonine, alanine and methionine in Shuxuening injection were analyzed on C18 column with gradient elution of acetonitrile-water （ 5 ：1） and sodium acetate buffer solution （ pH=6.5）. The detection wavelength was at 254 nm. [Results] Separation and the linear relationships of 4 amino acids were good. All RSDs of precision,repeatability and stability were less than 5%. The sample recoveries were between 95% and 98%,with RSD less than 5%. The total content of four amino acids in Shuxuening injection was 1 738.47 ｜ xg/mL. The content of methionine was the highest in Shuxuening injection, while the content of alanine was the lowest. [Conclusion] The accurate and simple method was reliably applied to determine content of four amino acids in Shuxuening injection which could provide a scientific tool for the quality control of Shuxuening injection.

关 键 词：舒血宁注射液 柱前衍生 天门冬氨酸 苏氨酸 丙氨酸 甲硫氨酸 

分 类 号：R284[医药卫生—中药学]