花茵陈总黄酮提取纯化工艺优化及其抗氧化药效研究

Optimization of the extraction and purification process for total flavonoids from Artemisia capillaris Thunb. （flowering stage）and investigation of their antioxidant efficacy

关键词:  茵陈  总黄酮  大孔吸附树脂  黄嘌呤氧化酶  抗氧化

Key words:  Artemisia capillaris Thunb.  total flavonoids  macroporous resin  xanthine oxidase  antioxidant activity

摘要:

[目的] 优化花茵陈总黄酮（TFHC）的提取纯化工艺，评价其体外抗氧化活性，为TFHC产业化制备与药理应用研究提供实验依据。[方法] 采用超声辅助热浸法，结合正交实验-响应面法优化TFHC提取工艺；通过6种大孔树脂（NKA-9、HPD-600、AB-8、DM-130、D-101、D-4020）筛选与动态纯化制备精制TFHC；采用高效液相色谱（HPLC）法、紫外分光光度法系统考察纯化产物的2，2-二苯基-1-苦基肼（DPPH）自由基清除能力及对黄嘌呤氧化酶（XOD）的抑制能力。[结果] 优化筛选出TFHC最佳提取工艺参数：液料比51 mL/g、乙醇体积分数38.2%、超声时间30 min、热浸温度70℃及热浸时间1 h；静态吸附和解析能力较好的为D-101树脂；确定最佳纯化工艺为：上样浓度1.5 mg/mL，上样流速7 BV/h，树脂柱径高比1∶12，上样量2.35 BV，除杂洗脱液用量2.94 BV，洗脱剂70%乙醇，洗脱流速10.6 BV/h，洗脱剂用量2.35 BV。经最佳纯化工艺处理后，TFHC纯度由38.33%上升至92.00%，纯化效果明显。纯化后的TFHC对XOD的半数抑制浓度（IC₅₀）由5.061 mg/mL下降至1.905 mg/mL，其对DPPH自由基清除能力的IC₅₀值由0.070 mg/mL下降至0.033 mg/mL。[结论] TFHC最佳提取纯化工艺显著提升其得率与纯度，纯化后产物体外DPPH自由基清除能力及XOD抑制能力增强，抗氧化活性显著。

Abstract:

[Objective] To optimize the extraction and purification process of total flavonoids from Artemisia capillaris Thunb（. flowering stage）（TFHC）and evaluate their in vitro antioxidant activity，providing an experimental basis for the industrial preparation and pharmacological application of TFHC. [Methods] The extraction process of TFHC was optimized by ultrasound-assisted hot maceration combined with orthogonal experiment and response surface methodology. Six macroporous resins（NKA-9，HPD-600，AB-8，DM-130，D-101，D-4020）were screened for dynamic purification to obtain refined TFHC. High-performance liquid chromatography（HPLC） and UV spectrophotometry were used to systematically evaluate the DPPH radical scavenging capacity and xanthine oxidase （XOD）inhibitory activity of the purified product. [Results] The optimal extraction parameters were determined as follows：liquid-to-solid ratio of 51 mL/g，ethanol volume fraction of 38.2%，ultrasonication time of 30 min，hot maceration temperature of 70 ℃，and hot maceration time of 1 h. D-101 resin exhibited the best static adsorption and desorption capacity. The optimal purification process was：loading concentration of 1.5 mg/mL，loading flow rate of 7 BV/h，resin column diameter-to-height ratio of 1∶12，loading volume of 2.35 BV，impurity elution volume of 2.94 BV，eluent of 70% ethanol，elution flow rate of 10.6 BV/h，and eluent volume of 2.35 BV. After the optimal purification process，the purity of TFHC increased from 38.33% to 92.00%. The half-maximal inhibitory concentration（IC₅₀）of purified TFHC against XOD decreased from 5.061 mg/mL to 1.905 mg/mL，and the IC₅₀ for DPPH radical scavenging capacity decreased from 0.070 mg/mL to 0.033 mg/mL. [Conclusion] The optimized extraction and purification process significantly improved the yield and purity of TFHC. The purified product showed enhanced DPPH radical scavenging capacity and XOD inhibitory activity，demonstrating potent antioxidant activity.