目的 建立制备型高效液相色谱（PHPLC）分离竹叶黄酮中异荭草苷和荭草苷的方法。方法 竹叶黄酮经AB-8树脂吸附洗脱纯化后，采用PHPLC分离异荭草苷和荭草苷，制备色谱条件：流动相为甲醇-0.3%乙酸水溶液（32：68），体积流量为5 mL/min，柱温为20 ℃，检测波长330 nm，进样体积为400 μL，上样质量浓度为15.3 mg/mL。结果 HPLC检测表明，异荭草苷的质量分数>99%，荭草苷的质量分数>93%，加样回收率分别为（92.7±2.92）%和（85.1±3.45）%。单体结构通过核磁共振碳谱和氢谱鉴定，确定为异荭草苷和荭草苷。结论 该法分离效果较好，低毒，回收率高，可用于竹叶黄酮中2种黄酮苷的分离制备。

Objective To establish a method for preparation of orientin and isoorientin from bamboo leaf flavones (BLF) using preparative high-performance liquid chromatography (PHPLC). Methods After BLF was purified by AB-8 resin-based column chromatography, PHPLC was employed to isolate orientin and isoorientin, using a mobile phase consisting of methyl alcohol-water with 0.3% acetic acid (32:68). The flow rate was 5 mL/min while the column temperature was maintained at 20 ℃. The injection volume was 400 μL and the concentration of sample was 15.3 mg/mL. Signal was monitored at 330 nm with the diode array detector. Results HPLC analysis showed that the purities of isoorientin and orientin were > 99% and > 93%, respectively, and the recovery rates were (92.7±2.92)% and (85.1±3.45)%, respectively. The confirmation of chemical structures was performed by ¹H-NMR and ¹³C-NMR. Conclusion The method is simple, with low toxicity but high recovery rate, and can be applied to the separation and preparation of isoorientin and orientin.

浙江省重点科技创新团队项目（2010R50032）