建立2D-HPLC同时检测钩藤碱、异钩藤碱、去氢钩藤碱、异去氢钩藤碱的方法

Development of a 2D-HPLC for simultaneous determination of rhynchophylline， isorhynchophylline， corynoxeine and isocorynoxeine

中文关键词:  二维高效液相法  钩藤碱  异钩藤碱  去氢钩藤碱  异去氢钩藤碱

英文关键词:2D-HPLC  Rhynchophylline  Isorhynchophylline  Corynoxeine  Isocorynoxeine

中文摘要:

      目的 建立二维高效液相色谱法（2D-HPLC）同时检测钩藤碱、异钩藤碱、去氢钩藤碱、异去氢钩藤碱四种钩藤生物碱含量的方法。方法 采用Aston SC2（3.5 mm×25 mm，5 μm）一维色谱柱，Aston SH C₁₈（3.5 mm×10 mm，5 μm）中间色谱柱，Aston SCB（4.6 mm×125 mm，5 μm）分析色谱柱，一维流动相为CAA-1，分析流动相为BPI-1∶MPI-1∶OPI=45∶14∶41，流速1.0 mL/min，柱温40℃，检测波长254 nm，进样量500 μL，分析时间9.5 min。结果 钩藤碱、异钩藤碱、去氢钩藤碱、异去氢钩藤碱线性范围分别为9.77~10 000.00 ng/mL（r=0.999 6）、10.74~11 000.00 ng/mL（r=0.999 7）、10.74~11 000.00 ng/mL（r=0.999 7）、10.74~11 000.00 ng/mL（r=0.999 6），精密度、稳定性和重复性相对标准偏差（RSD）<5.00%，准确度为95.20%~104.01%，方法回收率为93.63%~101.38%。结论 建立的2D-HPLC同时检测钩藤中四种生物碱含量的方法简单准确，可作为钩藤药材质量控制的新方法。

英文摘要:

      Objective To develop a two-dimensional high-performance liquid chromatography （2D-HPLC） for simultaneous determination of the contents of four kinds of Uncaria alkaloids： rhynchophylline， isorhynchophylline， corynoxeine and isocorynoxeine.Methods The 2D-HPLC apparatus was comprised of a first chromatographic column in version Aston SC2 （3.5 mm×25 mm， 5 μm）， an intermediate column in version Aston SH C₁₈ （3.5 mm×10 mm， 5 μm）， and an analytical column in version Aston SCB （4.6 mm×125 mm， 5 μm）. The mobile phase of the first and second liquid chromatography system were CAA-1 and mixed mobile phase （V _{BPI-1 basic mobile phase} ∶ V _{MPI-1 mobile phase} ∶ V _{OPI-1 organic mobile phase} = 45∶14∶41）. The chromatographic parameters included a flow rate of 1.0 mL/min， a column temperature of 40℃， a wavelength of 254 nm， an injection volume of 500 μL and a detection time of 9.5 min.Results The linear ranges of rhynchophylline， isorhynchophylline， corynoxeine and isocorynoxeine were 9.77~10 000.00 ng/mL （r=0.999 6）， 10.74~11 000.00 ng/mL （r=0.999 7）， 10.74~11 000.00 ng/mL （r=0.999 7）， 10.74~11 000.00 ng/mL（r=0.999 6）， respectively. The relative standard deviation （RSD） of precision， stability and repeatability were all less than 5.00%. The accuracy was 95.20%~104.01%， and the recovery rate was 93.63%~101.38%.Conclusion The 2D-HPLC developed for simultaneous determination of four kinds of alkaloids in Uncaria is simple and accurate， which can be used as a new method for quality control of Uncaria.