钩藤药材中异钩藤碱含量的超高效液相色谱和高效液相色谱法测定

Comparison of UPLC and HPLC for the Determination of Isorhychophyline in Uuncaria rhynchophylla

目的比较钩藤中异钩藤碱含量测定方法。方法采用高效液相色谱(HPLC)方法进行转换并优化为超高效液相色谱(UPLC)方法。高效液相色谱方法色谱柱为Alltech prevail C18柱(4.6mm×250 mm,5μm),流动相:乙腈-水(含三乙胺,冰醋酸调pH7.5),流速:1 ml.min-1,检测波长:246 nm,柱温:25℃。超高效液相色谱方法色谱柱为Waters AC-QUITY UPLCTMT3 C18柱(2.1 mm×50 mm,1.7μm),流动相:乙腈-水(含三乙胺,冰醋酸调pH 8.0),流速:0.6 ml.min-1,检测波长:246 nm,柱温:50℃。结果两种方法所得含量测定结果一致。结论超高效液相色谱方法能够替代高效液相色谱分析方法测定钩藤药材中异钩藤碱含量,既加快了分析速度,达到了样品分析的高通量,减少有机溶剂的使用,又得到更高的分析灵敏度。

Objective To compare the methods of ultra-performance liquid chromatography(UPLC) and HPLC for the determination of isorhychophyline in Uncaria rhynchophylla.Methods Alltech prevail C18 column(4.6 mm×250 mm,5 μm) was applied.With acetonitrile-water(triethylamine,acetic acid,pH7.5)as mobile phase at a flow rate of 1.0 ml/min,the detection wavelength was set at 246 nm and coulumn terperature was 25℃ in HPLC.A Waters ACQUITY UPLCTM BEH C18 column(2.1 mm×50 mm,1.7 μm)was applied,with acetonitrile-water(triethylamine,acetic acid,pH 8.0) as mobile phase at a flow rate of 0.6ml/min,the detection wavelength was set at 246 nm and column temperature was 50℃ in UPLC.Results Both of the two methods for the determination had the same results.Conclusion UPLC can replace HPLC for the determination of isorhychophyline in U.rhynchophylla.UPLC can increase separation speed and efficiency,and the major advantages of UPLC with 1.7 μm particles,over HPLC are the speed of analysis,the narrower peaks(giving increased signal –to-noise ratio).