Stable Isotope Labeling Derivatization Combined with Multiple-Mass Spectrometry Technologies to Monitor Metabolites of Tenuifoliside A Incubated with Intestinal Bacteria Incubation Model.

Aromatic carboxylic acids (ACAs), play important roles in preventive and therapeutic effects for some diseases. However, complex matrix effect and poor detection sensitivity make it difficult and even rare to detect ACAs in complex bio-samples. Herein, a stable isotope labeling derivatization (SILD) method based on one-pot synthesis of carboxylic amides by aniline (AN) and aniline-d5 (AN-d5) was firstly designed for quantitatively monitoring ACAs under mild conditions. The detection sensitivity was improved up to 500 folds. Importantly, when taking the trace tenuifoliside A (TA) containing p-hydroxyl-benzoyl- (HB) and 3, 4, 5-trimethoxylcinnamoyl- (TC) unit as a special example via intestinal bacteria incubation, the metabolites ACAs and whole metabolic profiles of TA were firstly accurately and systematically monitored by applying the SILD method combined with multiple-mass spectrometry (MMS) technologies. It provides a convenient, universal, high-sensitivity and high-recovery methodological tool for the systematically metabolic study of trace drugs in vitro and in vivo.