UPLC法同时测定不同产地吴茱萸中绿原酸、吴茱萸碱、吴茱萸次碱的含量

Simultaneous Determination of Chlorogenic Acid,Evodiamine and Rutecarpine in Different Places of Evodia rutaecarpa by UPLC

目的：建立同时测定不同产地吴茱萸中绿原酸、吴茱萸碱、吴茱萸次碱含量的方法。方法：采用超高效液相色谱法。色谱柱为ACQUITYUPLCBEHC18，流动相为乙腈-0.1％磷酸（梯度洗脱），流速为0.40ml/min，检测波长为326、220nm，柱温为30℃，进样量为2μl。结果：绿原酸、吴茱萸碱、吴茱萸次碱的检测质量浓度线性范围分别为7.67～76.67μg/ml（r＝0.9992）、13.33～133.33μg/ml（r＝0.9997）、13.33～133.33μg/ml（r＝0.9998）；定量限分别为0.11、0.05、0.05ng，检测限分别为0.03、0.01、0.01ng；精密度、稳定性、重复性试验的RSD＜3％；加样回收率分别为96.19％～101.90％（RSD＝2.19％，n＝6）、95.35％～101.16％（RSD＝2.27％，n＝6）、95.92％～98.98％（（RSD＝1.33％，n＝6）。结论：该方法快速、准确，适用于同时测定不同产地吴茱萸中绿原酸、吴茱萸碱、吴茱萸次碱的含量。

OBJECTIVE:To establish a method for the simultaneous determination of chlorogenic acid,evodiamine,rutecarpinein different places of Evodia rutaecarpa. METHODS:UPLC was performed on the column of ACQUITY UPLC BEH C18 withmobile phase of acetonitrile-0.1% Phosphoric acid aqueous solution(gradient elution)at a flow rate of 0.40 ml/min,the detectionwavelength was 326 nm and 220 nm,column temperature was 30 ℃,and the volume injection was 2 μl. RESULTS:The linearrange was 7.67-76.67 μg/ml for chlorogenic acid(r=0.999 2),13.33-133.33 μg/ml for evodiamine(r=0.999 7)and 13.33-133.33μg/ml for rutecarpine(r=0.999 8);the limits of quantitation were 0.11 ng,0.05 ng and 0.05 ng,the limits of detection were 0.03ng,0.01 ng and 0.01 ng,respectively;RSDs of precision,stability and reproducibility tests were lower than 3%;recoveries were96.19%-101.90%(RSD=2.19%,n=6),95.35%-101.16%(RSD=2.27%,n=6)and 95.92%-98.98%(RSD=1.33%,n=6),respectively.CONCLUSIONS:The method is rapid and accurate,and suitable for the simultaneous determination of chlorogenic acid,evodiamine,rutecarpine in different places of E. rutaecarpa.