Rapid discovery of active ingredients from complex matrices is one of great challenges for modern drug development. Traditional methods often require many sample treatment steps, including an extraction step with exclusively dedicated solvents followed by repeated separation and activities assessment. This present work described an integrated analytical setup for natural antioxidants discovery in which the online extraction (OLE) of a solid sample is directly coupled to its analysis by high performance liquid chromatography-diode array detector-quadrupole time-of-flight tandem mass spectrometry and 2,2'-Azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS) antioxidant assay (OLE-HPLC-DAD-QTOF-MS/MS-ABTS). This developed approach makes sample extraction, chromatographic separation and chemical detection, and antioxidant assay integrated into a single HPLC injection and was successfully applied for the rapid discovery of natural antioxidant bioactives from Polygonum viviparum. A total of 21 secondary metabolites were characterized according to their retention times, ultraviolet (UV) spectra, exact mass and fragmentation ions in MS/MS spectra, and 18 of them displayed antioxidant activity (response as negative peaks in antioxidant assay). This work describes a simple, green and efficient approach to minimize the sample consumption (only 0.4 mg was required) and eliminate complex sample treatment procedures. The developed OLE-HPLC-DAD-QTOF-MS/MS-ABTS system offers new perspectives for rapid chemical profiling of natural products and their antioxidants discovery.

从复杂基质中快速发现活性成分是现代药物开发的重大挑战之一。传统方法通常需要许多样品处理步骤，包括使用专用溶剂的萃取步骤，然后进行重复分离和活性评估。本工作描述了一种用于天然抗氧化剂发现的综合分析装置，其中固体样品的在线提取（OLE）直接通过高效液相色谱-二极管阵列检测器-四极杆飞行时间串联质谱和2,2'-叠氮双-（3-乙基苯并噻唑啉-6-磺酸盐）（ABTS）抗氧化剂测定（OLE-HPLC-DAD-QTOF-MS / MS-ABTS）。这种先进的方法可以进行样品提取，色谱分离和化学检测，虎杖。根据其保留时间，紫外（UV）光谱，MS / MS光谱中的精确质量数和碎片离子，对总共21种次级代谢产物进行了表征，其中18种具有抗氧化活性（在抗氧化剂测定中为负峰）。这项工作描述了一种简单，绿色和有效的方法，以最大程度地减少样品消耗（仅需要0.4 mg）并消除复杂的样品处理程序。所开发的OLE-HPLC-DAD-QTOF-MS / MS-ABTS系统为天然产物的快速化学分析及其抗氧化剂的发现提供了新的视角。