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目的建立HPLC-DPPH在线筛选法筛选中药提取物中抗氧化活性成分,对黄芩根、茎、叶提取物进行在线抗氧化活性筛选,比较10个不同产地黄芩根提取物中抗氧化活性成分差异。方法HPLC-DPPH在线筛选系统中HPLC流动相为乙腈-甲酸水(pH=3),梯度洗脱,体积流量为1mL/min,色谱柱为Spherigel C18柱,柱温25℃,检测波长为276nm;DPPH反应体系中NaHCO3体积流量为0.1mL/min,NaHCO3质量浓度为0.1μg/mL;DPPH体积流量为0.5mL/min,DPPH质量浓度为10μg/mL,检测波长517nm。结果黄芩根、茎、叶提取物均具有抗氧化活性,但所含抗氧化活性成分有较大差异。黄芩根提取物中抗氧化活性成分因产地不同而不同,道地药材河北承德黄芩提取物中含有7种抗氧化活性成分,其中有3种抗氧化活性成分是其他产地黄芩所不具有的,但10种产地黄芩提取物中都含黄芩素、黄芩苷、二氢黄芩苷、去甲基汉黄芩素、野黄芩苷这5种抗氧化活性成分。结论该法适用于中药提取物中抗氧化活性成分的筛选,具有快速、准确、直接、重现性好的特点。
OBJECTIVE To establish an on-line HPLC-DPPH screening method for screening anti-oxidative components of extracts from Chinese traditional medicine. On-line anti-oxidative activities of extracts from roots, stems and leaves of Scutellaria baicalensis Georgi were screened. The differences of antioxidant activity in extracts from Scutellariae Radix were compared. Methods The mobile phase of HPLC-DPPH on-line was acetonitrile-formic acid water (pH = 3) with gradient elution. The volume flow rate was 1 mL / min and the column was Spherigel C18 column with the detection wavelength of 276 nm at 25 ℃. The volume flow of NaHCO3 in the DPPH reaction system was 0.1 mL / min, the concentration of NaHCO3 was 0.1 μg / mL, the DPPH volume flow rate was 0.5 mL / min, the DPPH concentration was 10 μg / mL and the detection wavelength was 517 nm. Results Scutellaria root, stem and leaf extracts have antioxidant activity, but the antioxidant activity of the ingredients are quite different. Scutellaria root extract antioxidant activity of different ingredients due to different origin, authentic medicine Hebei Chengde skullcap extract contains seven kinds of antioxidant active ingredients, of which three kinds of antioxidant ingredients from other areas skullcap does not have, but 10 kinds of origin Scutellaria extract containing baicalein, baicalin, dihydro baicalin, to the baicalein, wild baicalin five antioxidant active ingredients. Conclusion This method is suitable for the screening of antioxidant active ingredients in Chinese herbal extracts, with fast, accurate, direct, reproducible characteristics.