Exploring the changes in anticoagulant active ingredients before and after bovine bile fermentation of the traditional Chinese medicine Yaomu via an aggregation-induced emission fluorescence sensor

Abstract

Fermentation is a significant and specialized processing technology that primarily serves to enhance efficacy and reduce toxicity. Yaomu (YM) was fermented in a cellar using bovine bile as the fermentation matrix. However, the alterations in its active ingredients before and after processing remain poorly understood. An aggregation-induced emission fluorescence sensor (NITPA-S) was developed that employed S-2238 (S) to encapsulate the NITPA-Py-603 (NITPA) probe with the help of glutaraldehyde, creating an “ON” state. Thrombin hydrolyzed the sensor, releasing the probe in the “OFF” state, while thrombin inhibitors reversed this effect. We combined an HPLC fully automatic partial fraction collector and UHPLC-Q-TOF/MS for the separation, screening and identification of YM active compounds, which were used as indicators of anticoagulant activity to explore the changes in the content of YM before and after processing by using UHPLC-MS/MS and the sensor. A total of 211 compounds were collected and identified. Notably, karakoline, talatisamine, neoline, chasmanine and schaftoside exhibited the highest inhibition rates in YM. Furthermore, these compounds generally increased in concentration after processing (p < 0.05). The anticoagulant inhibition rates of the mixture of these five compounds were 86.48% and 83.44% when compared with those of unfermented YM and YM, respectively. Consequently, the fluorescent sensor could completely screen out the anticoagulant active ingredients, providing a novel analytical approach for exploring and screening changes in the active components before and after TCM processing.