反相高效液相色谱测定依替巴肽注射液中氨基酸的含量

Quantitative Analysis of Amino Acids in Eptifibatide Injection by RP-HPLC

采用异硫氰酸苯酯为柱前衍生化试剂,结合反相高效液相色谱技术,建立了定量分析依替巴肽注射液中氨基酸组分的检测方法。优化色谱条件,选用Agilent XDB-C18色谱柱(5μm,150 mm×4.6 mm),以0.1 mol/L醋酸钠(冰醋酸调至pH 6.0)-乙腈(90∶10)为流动相A,乙腈为流动相B,梯度洗脱方式,检测波长为254 nm,流速为1.0 mL/min,进样体积为5μL,柱温为30℃。结果表明,在优化条件下,各氨基酸能实现有效分离,且在1.23～3.68 mmol/L浓度范围内,各氨基酸的线性关系良好,相关系数(r)为0.999 1～0.999 6。该方法显示了良好的准确性、重复性、稳定性及选择性,能对依替巴肽注射液中的氨基酸实现准确定量分析。

A reversed-phase high performance liquid chromatographic method was developed for the determination of amino acids in eptifibatide injection using phenyl isothiocyanate as pro-column deri vatization reagent. The chromatographic parameter was optimized, and the analysis of amino acids was performed on a Agilent XDB - Cls column（5 μm, 150 mm x4. 6 mm） , using 0. 1 mol/L sodi um acetate（adjusting the pH to 6.0 with acetic acid） and acetonitrile （90 ： 10, A ） - acetonitrile （B） as mobile phase by gradient elution at a flow rate of 1.0 mL/min and an injection volume of 5 μL. The detection wavelength was set at 254 nm, and the column temperature was set at 30 ℃, Under the optimal conditions, the amino acids were effectively separated, good linearities were ob- tained in the range of 1.23 - 3.68 mmol/L for all amino acids, with correlation coefficients （r） of 0. 999 1 - 0. 999 6. The method showed high selectivity, good repeatability, accuracy and stability, and could be applied in the accurate quantification of amino acids in eptifibatide injection.