HPLC波长切换技术同时测定板蓝根中4种活性成分含量

Simultaneous Determination of Four Active Components in Radix Isatidis by HPLC with Double-wavelength Switch

目的:建立反相高效液相色谱法同时测定板蓝根药材中尿苷、腺苷、鸟苷和表告依春4种活性成分含量的方法,并比较不同时间采收的板蓝根中上述各成分的含量情况。方法:采用Kromasil C18色谱柱(4.6 mm×250 mm,5μm),流动相为甲醇-0.1%磷酸溶液,梯度洗脱,流速0.7 mL.min-1;双波长切换时间序列采样:0～30 min为254 nm,30～45 min为245 nm;柱温25℃。结果:在上述色谱条件下,测得尿苷、腺苷、鸟苷和表告依春分别在14.80～1 184 ng(r=0.999 9),14.55～1 164ng(r=0.999 9),13.75～1 100 ng(r=0.999 9),12.25～980 ng(r=0.999 9)与峰面积呈良好的线性关系;平均加样回收率:尿苷99.5%(RSD 1.2%),腺苷98.3%(RSD 1.4%),鸟苷98.1%(RSD 1.2%),表告依春99.0%(RSD 0.9%)。不同时间采收板蓝根中4种活性成分的含量是动态变化的。结论:方法操作简便、准确,重复性良好,可用于板蓝根药材的质量控制。

Objective： To develop an HPLC method for simultaneous determination of uridine, adenosine, guanosine and epigoitrin in Radix Isatidis, and compare the content variation of the four components of herbs harvested at different time. Method： Four components were separated by Kromasil C18 column （4.6 mm × 250 mm, 5 μm） with gradient mobile phase of methanol-0. 1% phosphoric acid at the flow rate of 0.7 mL · min ^（-1） The detection wavelength was set at 254 nm （0-30 min） and 245 nm （30-45 min）. The column temperature was maintained at 25 ℃. Result： The calibration curves of uridine, adenosine, guanosine and epigoitrin were in good linearity over the range of 14. 80-1 184 ng （r =0.999 9）, 14. 55-1 164 ng （r=0.999 9）, 13.75-1 100 ng （r= 0. 999 9） , 12.25-980 ng （r = 0. 999 9） , respectively. The average recoveries of the four components were 99.5% （RSD 1.2%）,98.3% （RSD 1.4%）,98.1% （RSD 1.2%）,99.0% （RSD0.9%）, respectively. The content of four components in Radix Isatidis varied with different harvest time. Conclusion： The method is simple, accurate and reproducible for quality control of Radix Isatidis.