文献类型：期刊文献

中文题名：PITC柱前衍生HPLC测定复方氨基酸注射液中18种氨基酸的含量

英文题名：Content Determination of 18 Amino Acids in Compound Amino Acid Injection by HPLC with Pre-column PITC Derivatization

作者：汪媛[1];王海龙[2];金高娃[3];章飞芳[1];杨丙成[1];梁鑫淼[1,3]

机构：[1]华东理工大学药学院,上海200237;[2]吕梁学院,吕梁033000;[3]中国科学院大连化学物理研究所,大连116023

年份：2014

卷号：16

期号：6

起止页码：1347

中文期刊名：世界科学技术-中医药现代化

外文期刊名：Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

收录：CSTPCD;;北大核心:【北大核心2011】；

基金：国家自然科学基金委面上项目(21177040):基于分子印迹聚合物固相微萃取原亲水液相色谱分离的氨基糖苷类抗生素残留快速筛查技术研究;负责人:章飞芳

语种：中文

中文关键词：柱前衍生;异硫氰酸苯酯;高效液相色谱法;复方氨基酸注射液

外文关键词：Pre-column derivatization;phenyl isothiocyanate;high performance liquid chromatography;compound amino acid injection

摘要：目的：建立柱前衍生高效液相色谱法同时测定复方氨基酸注射液（18AA）中18种氨基酸含量的方法。方法：样品采用异硫氰酸苯酯（PITC）柱前衍生，以Unitary-C18柱（250 mm×4.6 mm，5μm）为色谱柱，检测波长为254 nm，醋酸钠缓冲盐及乙腈、甲醇和水的混合体系为流动相，梯度洗脱。结果：优化了衍生试剂用量、衍生时间、缓冲盐浓度以及pH值和柱温等影响因素，最终在40 min内实现了18种氨基酸的分离，在9~1021μg·mL-1浓度范围内线性关系良好，且相关系数大于0.9977，平均回收率为92.6%~110.7%，相对标准偏差（RSD）为0.01%~5.68%。其定量限（LOQ，S/N=10）为0.02~13.41μg·mL-1。结论：该方法操作简便、灵敏度高、准确度和精密度好，可用于复方氨基酸注射液中氨基酸的含量测定。
An efficient and sensitive analytical method for the simultaneous content determination of 18 amino acids in compound amino acid injection was developed using high performance liquid chromatography （HPLC） with pre-column derivatization. Phenyl isothiocyanate （PITC） was used as derivatization reagent. The target compounds were separated on a Unitary-C18 column （250 mm í 4.6 mm, 5 μm） in gradient elution mode using sodium acetate and the mixture of acetonitrile, methanol and water as mobile phases. The detection wavelength was 254 nm. The derivatization reagent dosage, derivatization time, salt concentration, the pH and the column temperature of mobile phase were investigated. Finally, 18 amino acids were separated within 40 minutes. The method showed that the good linearity （r2 ≥ 0.997 7） was at a range from 9 μg·mL-1 to 1 021 μg·mL-1. The recoveries ranged from 92.6% to 110.7%. And the relative standard deviations （RSD） ranged from 0.01% to 5.68%. The limits of quantification （LOQ, S/N = 10） ranged from 0.02 μg·mL-1 to 13.41 μg·mL-1. This method, which was simple, sensitive and accurate, can be applied for the content determination of amino acids in compound amino acid injections.