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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 establish a method for the simultaneous determination of uridine, adenosine, guanosine and epigallocatechin gallate in Radix isatidis by RP-HPLC and to compare the contents of these four components in Radix Isatidis Happening. Methods: Kromasil C18 column (4.6 mm × 250 mm, 5 μm) was used. The mobile phase consisted of methanol-0.1% phosphoric acid solution with gradient elution at a flow rate of 0.7 mL · min-1. 254 nm at 30-45 min and column temperature at 25 ° C. Results: Under the above chromatographic conditions, uridine, adenosine, guanosine and epirubicin were measured at 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) and 12.25 ~ 980 ng (r = 0.999 9) showed a good linear relationship with the peak area. The average recoveries were 99.5% (RSD 1.2%), adenosine 98.3% RSD 1.4%), guanosine 98.1% (RSD 1.2%), according to the report 99.0% Yichun (RSD 0.9%). The content of 4 active ingredients in Radix isatidis changed dynamically at different times. Conclusion: The method is simple, accurate and reproducible. It can be used for quality control of Radix isatidis.