PITC柱前衍生HPLC测定柴胡炮制前后17种氨基酸含量

Determination of Amino Acids in Bupleuri Radix Before and After Processing by HPLC with Pre-column Derivatization

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 amino acids in raw and processed Bupleuri Radix. Method： HPLC analysis was performed after derivatization by using phenyl isothiocyanate （PITC） as a derivative reagent. Samples were analyzed on Prevail C18 column with gradient elution of acetonitrile-water （4：1 ） and sodium acetate buffer solution （adjusted to pH 6.5 ） -acetonitrile （97：3 ）, detection wavelength was 254 nm. Result： Seventeen amino acids in Bupleuri Radix could be separated well in 32 min. The average recovery was between 93.4% -107. 1% with RSD of 1.8% -3.9%. Conclusion： Compared with other pre-column derivatization method for determination of the content of amino acid, this method is reliable with good accuracy and repeatability, which is useful for determination of amino acids in raw and processed Bupleuri Radix.