目的 设计一种反应性顶空系统，包括辅助加热装置和内插式反应池的反应性顶空瓶，用于中药中挥发油抗氧化活性研究，建立挥发油抗氧化活性评估方法。方法 辅助加热装置加热反应性顶空瓶底部，红外热成像仪研究反应性顶空瓶温度分布；挥发油进入顶空并与内插式反应池中自由基发生清除反应，分光光度法测定自由基吸光度变化；固相微萃取针从顶空反应池上方原位取样，气相色谱-质谱联用（GC-MS）测定顶空挥发油清除反应前后成分变化，表征抗氧化活性成分。结果 改进的金属浴辅助加热装置实现样品区及顶空温差大于50℃，使挥发油快速释放进入顶空的同时避免自由基热分解；金花清感颗粒（Jinhua Qinggan Granules，JQG）用量为1.50 g，样品温度90℃，反应30 min时，对2,2'-联氮-双(3-乙基苯并噻唑啉-6-磺酸)［2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonate)，ABTS］自由基清除率达88.95%（n＝6，RSD＝1.91%）；建立的抗氧化活性评估方法的精密度和重复性RSD（n＝8）分别为2.50%、1.24%；经GC-MS分析鉴定出57种挥发油，包括蒎烯、青蒿酮等，其中清除反应前后峰面积减少达到30%以上的有35种，占总挥发性油的61.40%，据此推断具有抗氧化活性的成分主要为萜类和醛酮、酚酸等化合物，主要包括罗勒烯、α-细辛脑等。结论 JQG挥发油具有良好的抗氧化活性；设计制作的反应性顶空系统可在一个小型密闭反应瓶中原位一步完成挥发油的释放-清除自由基-顶空微萃取进样3个过程；建立的方法具有快速、高效、重现性好等特点，可用于中药挥发油的抗氧化活性评估以及活性成分鉴定。

Objective A reactive headspace system was designed, including a reactive headspace bottle of an plug-in reaction tank and an auxiliary heating device, which was used to study the antioxidant activity of volatile oil in traditional Chinese medicine, the methods of antioxidant activity evaluation and component analysis of volatile oil were established.Methods The auxiliary heating device heats the bottom of the reactive headspace bottle and the infrared thermal imager studies the temperature distribution of the reactive headspace bottle; the volatile oil enters the headspace and scavenges the free radical in the interpolation reaction tank; the change of absorbance of free radical is measured by spectrophotometry; the changes of components before and after headspace volatile oil removal reaction were determined by SPME-GC-MS to characterize the antioxidant components. Results The improved metal bath auxiliary heating device realizes the temperature difference between the sample area and the headspace more than 50℃, so that the volatile oil can be quickly released into the headspace and avoid the thermal decomposition of free radicals. When the dosage of Jinhua Qinggan Granule (JQG) was 1.50 g, the sample temperature was 90℃, and the reaction time was 30 min, the free radical scavenging rate of JQG's volatile oil to the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) was 88.95% (n=6, RSD=1.91%); The RSD (n=8) of the precision and repeatability of the established antioxidant activity evaluation method were 2.50% and 1.24%; the solid-phase microextraction needle was sampled in situ from above the headspace reaction tank, and 57 kinds of volatile oils were identified by GC-MS analysis, including pinene, artemisinin, etc. among them, 35 kinds had a peak area reduction of more than 30% before and after the removal reaction, accounting for 61.40% of the total volatile oils. Therefore, it is inferred that the components with antioxidant activity are mainly terpenoids, aldehydes, ketones, phenolic acids and other compounds, mainly including basilene α-asarone, etc. Conclusion The volatile oil of JQG has good antioxidant activity; the reactive headspace system can complete the three reaction processes in a small closed reaction bottle in one step, including release, removal with free radical and headspace microextraction injection of volatile oil; this method is rapid, efficient and reproducible, and it can be used to evaluate the antioxidant activity and identify the active components of the volatile oil of traditional Chinese medicine.

R283.6

国家自然科学基金资助项目(21367025);云南省科技计划项目(2017FD118)