柱前衍生RP-HPLC测定淫羊藿中氨基酸含量

目的：建立柱前衍生-反相高效液相色谱法检测淫羊藿中游离氨基酸和水解氨基酸的含量。方法：采用高效液相色谱法，以异硫氰酸苯酯进行柱前衍生，梯度洗脱的方法，检测淫羊藿中游离氨基酸和水解氨基酸的含量。结果：21种氨基酸的检测浓度线性范围为0.0019~2.7000 μmol·mL^-1，r均≥0.9995。21种游离氨基酸的平均加样回收率在86.7%~100.1%之间，RSD在0.61%~2.7%之间（n=6）；水解氨基酸的平均加样回收率在90.3%~100.0%之间，RSD在0.41%~2.0%之间（n=6）。结论：该方法经方法学验证，适合淫羊藿中游离氨基酸和水解氨基酸的含量测定。

Objective: To establish a method for determination of the contents of 21 free and hydrolyzed amino acids from Epimedium brevicornu Maxim.by pre-column derivatization RP-HPLC. Methods: The samples were derivated with phenyl isothiocyanate and HPLC method was adopted.Gradient elution chromatography was applied to determine contents of free and hydrolyzed amino acids in E.brevicornu. Results: The 21 amino acids had good linearity in the range of 0.0019-2.7000 μmol·mL^-1 (the coefficients of correlation were all over 0.9995).The average recoveries of free amino acids were 86.7%-100.1% (RSD=0.61%-2.7%,n=6);the average recoveries of hydrolyzed amino acids were 90.3%-100.0% (RSD=0.41%-2.0%,n=6). Conclusion: The methodology validation showed that the method was suitable for determination of free and hydrolyzed amino acids in E.brevicornu.