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目的:建立一种同时测定鳖血柴胡中17种氨基酸含量的方法,为该药材的质量控制提供参考。方法:以异硫氰酸苯酯(PITC)为柱前衍生试剂,采用Prevail C18色谱柱,流动相[乙腈-水(4∶1)]-[乙酸钠缓冲溶液(p H 6.5)-乙腈(97∶3)]梯度洗脱,检测波长254 nm,测定柴胡炮制前后氨基酸的含量。结果:17种氨基酸在32 min内可良好分离;样品平均加样回收率93.4%~107.1%,RSD 1.8%~3.9%。结论:与其他柱前衍生测定氨基酸含量的方法相比,该方法具有可操作性强、成本低、易推广等优势,可作为柴胡及其炮制品中氨基酸的含量测定方法。
Objective: To establish a method for simultaneous determination of 17 kinds of amino acids in Radix Bupleuri, providing reference for the quality control of Radix Bupleuri. Methods: Phenylisothiocyanate (PITC) was used as a pre-column derivatization reagent. Prevail C18 column was used. The mobile phase consisted of acetonitrile and water (4:1) sodium acetate buffer solution (p H 6.5) 97: 3)], the detection wavelength was 254 nm, and the content of amino acids before and after the preparation of Radix Bupleuri was determined. Results: The 17 amino acids were well separated in 32 min. The average recoveries were 93.4% -107.1% and RSD 1.8% -3.9%. Conclusion: Compared with other methods for determination of amino acid content by pre-column derivatization, this method has many advantages such as high operability, low cost and easy popularization. It can be used as a method for determination of amino acids in Bupleurum and its processed products.