2D-HPLC结合捕集柱同时测定黄连药材中6个生物碱

目的： 建立二维高效液相色谱法（2D-HPLC）结合捕集柱同时测定黄连药材中非洲防己碱、药根碱、表小檗碱、黄连碱、巴马汀和小檗碱的含量。 方法： 第一维液相色谱系统色谱柱为Kromasil-C₁₈（100 mm×4.6 mm，5 μm），流动相为100 mmol·L^-1磷酸二氢铵溶液（pH为5.30）-乙腈（50：50）；第二维液相色谱系统色谱柱为Ultimate XB-Phenyl柱（250 mm×4.6 mm，5 μm），以100 mmol·L^-1磷酸二氢铵溶液（用磷酸调节pH为3.30）（A）-乙腈（B）为流动相进行梯度洗脱（1～2 min，36%B→30%B；2～10 min，30%B→34%B；10～16 min，34%B→64%B）。采用"中心切割"模式及捕集柱转移目标物，检测波长345 nm，柱温40℃。 结果： 不同产地黄连药材中非洲防己碱、药根碱、表小檗碱、黄连碱、巴马汀和小檗碱在19 min内完成分析，含量分别为3.8～4.4、3.3～4.0、10.1～11.2、18.9～20.8、14.4～16.6、 55.6～61.3 mg·g^-1。6个生物碱的平均回收率为95.5%～104.2%，RSD为0.23%～1.4%。 结论： 该方法能同时测定黄连药材中非洲防己碱、药根碱、表小檗碱、黄连碱、巴马汀和小檗碱的含量，可用于黄连药材的质量评价。

Objective: To establish a method for simultaneous determination of columbamine,jatrorrhizine,epiberberine,coptisine,palmatine and berberine in Coptidis Rhizoma using two-dimensional liquid chromatography(2D-HPLC)coupled with trapping column. Methods: The applied column was Kromasil-C₁₈(100 mm×4.6 mm,5 μm)and the mobile phase was 100 mmol·L^-1 ammonium dihydrogen phosphate solution(pH=5.30)-acetonitrile(50:50)in the first dimensional liquid chromatographic system;for the second dimensional liquid chromatographic system,the column was Ultimate XB-Phenyl(250 mm×4.6 mm,5 μm)and the mobile phase was 100 mmol·L^-1 ammonium dihydrogen phosphate solution(pH adjusted to 3.30 with phosphoric acid)(A)-acetonitrile(B)with gradient elution(1-2 min,36%B→30%B;2-10 min,30%B→34%B;10-16 min,34%B→64%B).Targets were transferred through a "central cut" mode with the trapping column.The detection wavelength was 345 nm.The column temperature was 40℃. Results: Six alkaloids of Coptidis Rhizoma were separated and analyzed in 19 min.And the content of the six alkaloids,i.e.,columbamine,jatrorrhizine,epiberberine,coptisine,palmatine,berberine,was 3.8-4.4,3.3-4.0,10.1-11.2,18.9-20.8,14.4-16.6,55.6-61.3 mg·g^-1.The average recovery of the six alkaloids was 95.5%-104.2%,RSD was 0.23%-1.4%. Conclusion: This method can simultaneously determine columbamine,jatrorrhizine,epiberberine,coptisine,palmatine and berberine in Coptidis Rhizoma and can be used to evaluate the quality of Coptidis Rhizoma.