异硫氰酸苯酯法测定清开灵注射液中氨基酸的含量

Determination of amino acids in Qingkailing Injection by using phenylisothiocyanate method

目的建立准确测定清开灵注射液中氨基酸含量的方法。方法采用异硫氰酸苯酯为衍生化试剂,清开灵注射液衍生化反应后,进行HPLC分析。色谱柱为Diamonsil C18色谱柱(4.6 mm×250 mm,5μm);流动相A为0.1 mol/L醋酸钠-乙腈(pH 6.5)(93∶7),流动相B为乙腈-水(4∶1),梯度洗脱,流速为1 mL/min;检测波长为254 nm。结果该方法可以对清开灵注射液中20种氨基酸准确定量,其浓度在0.019～24.855 g/L范围内,线性关系良好(r=0.999 7～0.999 9)。方法重复性的RSD(%)在0.766%～4.49%之间;20种氨基酸的加样回收率在92.2%～106.8%之间。结论该方法准确、可靠、灵敏,可用于清开灵注射液中氨基酸的含量测定。

Objective To establish a accurate method for quantitative determination of amino acids in Qingkailing Injection.Methods After derivatization of amino acids in Qingkailing Injection and taking phenylisothiocyanate as derivatization reagent,the amino acids were analysed by using HPLC.The HPLC procedure was performed on the chromatographic column of Diamonsil C18（4.6 mm×250 mm,5 μm）,and the mobile phase A was sodium acetate（0.1 mol/L）-acetonitrile（pH 6.5,93∶ 7）,and mobile phase B was acetonitrile-water（4∶ 1） in gradient elution.The flow velocity was 1 mL/min and detection wavelength was 254 nm.Results This method determined accurately the quantity of 21 kinds of amino acids in Qingkailing Injection,and they showed a good linear relationship at a range from 0.019 g/L to 24.855 g/L（r=0.999 7-0.999 9）.The average recovery of 21 kinds of amino acids was from 92.2% to 106.8%（RSD from 0.766% to 4.49%）.Conclusion The method was accurate,reliable and sensitive,which can be used for the quantitative determination of amino acids in Qingkailing Injection.