Recent advances of affinity ultrafiltration-liquid chromatography-mass spectrometry in screening natural active substances

Medicinal plants serve as a valuable source of naturally occurring bioactive compounds， which have played a vital role in promoting human health， preventing and treating various diseases for thousands of years. However， the phytochemical profiles of medicinal plants， including traditional Chinese herbal medicines （TCHMs）， are notably complex and highly diverse， posing a tough challenge for the rapid screening and identification of their potential bioactive constituents. This challenge is central to pharmacological research and drug development efforts in medicinal plants including TCHMs. Therefore， the rapid and efficient screening of bioactive components from complex natural products has become a key research direction in academia. To address this issue， affinity ultrafiltration coupled with liquid chromatography-mass spectrometry （AUF-LC-MS） has emerged as an efficient technique that integrates affinity-based capture， ultrafiltration， and liquid chromatographic-mass spectrometric analysis to enable rapid screening and characterization of potential small-molecule bioactive compounds. Known for its low sample consumption， high specificity， exceptional sensitivity， operational simplicity and high efficiency， this approach leverages specific interactions between target molecules and chemical extracts to identify bioactive ligands from complex mixtures， making it particularly suitable for studying intricate matrices such as medicinal plant extracts and TCHM formulations. This study summarizes the principles， characteristics， and recent applications of AUF-LC-MS technology in the screening of potential active components in medicinal plants including TCHMs over the past decade （2015-2025）. Based on different levels of AUF-LC-MS screening， including compound level， single TCHM level， and TCHM formula level， a comprehensive summary is provided. AUF-LC-MS screening of single TCHM level is further analyzed and summarized based on the number of targets， including single target screening， dual-targets screening， and multi-targets screening. At the same time， comparisons were made between other different natural active substance screening approaches and the AUF-LC-MS method. In addition， it outlines prospective research directions for the further development of AUF-LC-MS with the aim of providing new insights for the study of the pharmacological substances of TCHMs and the development of new drugs.