目的 建立HPLC-UV-DPPH在线分析方法，比较不同生产企业的地黄药材在炮制前后抗氧化活性成分的变化，为中药炮制及质量控制提供有效的评价方法。方法 HPLC-UV-DPPH在线联用筛选系统中HPLC流动相为乙腈-0.1%醋酸水溶液，梯度洗脱，体积流量为1.0 mL/min，色谱柱为Aglient Extend C₁₈柱（250 mm×4.6 mm，5μm），柱温30℃，检测波长334 nm，DPPH溶液体积流量0.5 mL/min，检测波长517 nm。以毛蕊花糖苷为阳性对照，通过"量-效"研究思路评价样品的总抗氧化活性及各成分对总抗氧化活性的贡献率。采用HPLC-FT-MS对药材中主要的抗氧化活性成分进行鉴定并比较药材炮制前后的差异。结果 HPLC-UV-DPPH方法检测到地黄中主要含有9个抗氧化活性成分，而熟地黄含有13个，地黄和熟地黄有8个共有抗氧化活性成分，地黄在炮制前后抗氧化活性成分明显不同，不同生产企业的样品中各抗氧化活性成分的活性差异较大。抗氧化活性贡献率较高的成分分别为玉叶金花苷酸、海胆苷、焦地黄苯乙醇苷A1/A2、毛蕊花糖苷、异毛蕊花糖苷。结论 HPLC-UV-DPPH法稳定、灵敏、重现性好，适用于地黄和熟地黄中抗氧化活性成分的快速筛选和质量评价。

Objective To develop an HPLC-UV-DPPH method to compare anti-oxidants of Rehmanniae Radix and Rehmanniae Radix Praeparata sample from different manufactories and to provide an effective method for the processing and quality control of traditional Chinese medicine.Methods HPLC in HPLC-UV-DPPH system was performed on Aglient Extend C₁₈ (250 mm×4.6 mm, 5 μm) column with gradient elution of acetonitrile-0.1% acetic acid at the flow rate of 1.0 mL/min, and the detection wavelength was at 334 nm. The column temperature was 30℃. Flow rate of DPPH solution is 0.5 mL/min, and detection wavelength was set at 517 nm. The total activities of the samples and the contribution rate of each component to the total activity were evaluated by the "quantity-effect" research idea, and verbascoside was regarded as a positive reference. The main anti-oxidants in original and processed herbs were identified by HPLC-FT-MS and were compared.Results The detection of HPLC-UV-DPPH method showed that there were nine anti-oxidants in Rehmanniae Radix extract, while 13 anti-oxidants were found in Rehmanniae Radix Praeparata. There were eight common anti-oxidants in the two herbs. The anti-oxidants were obviously different before and after Rehmanniae Radix processed. The activities of the antioxidants in different samples were markedly different. Anti-oxidants with higher contributions were mussaenosidic acid, echinacoside, jionoside A1/A2, verbascoside, and isoverbascoside, respectively.Conclusion The HPLC-UV-DPPH method is stable, sensitive, reproducible, and suitable for rapid screening of anti-oxidants and quality evaluation of Rehmanniae Radix and Rehmannia Radix Praeparata.

国家自然科学基金项目（81403093）；湖北省食品药品监督管理局科研项目（2016010＋02）；中央高校基本科研业务费资助项目（175220006）