Screening of bioactive components from traditional Chinese medicine by immobilized β2 adrenergic receptor coupled with high performance liquid chromatography/mass spectrometry

Abstract

Traditional Chinese medicine (TCM) represents a valuable resource for lead compounds discovery. Given the complexity of TCM components, analytical methods play a key role in novel drug development. In our study, we established a high specific and reliable bio-active components screen system, where β2 adrenergic receptor (β2-AR) was immobilized on silica by non-covalent bonds and packed into a stainless steel column (4.6 × 50 mm, 7 μm) to form β2-AR chromatography column. The column was further coupled with high performance liquid chromatography-time of flight tandem mass spectrometry (TOF-MS/MS). By utilizing this strategy, we successfully identified four β2-AR-targeting compounds: tetrahydroberberine, tetrahydrocolumbamine, fumarine and corydaline from Corydalis Rhizome. The association constants between β2-AR and tetrahydroberberine (9.04 × 104/M) as well as fumarine (4.30 × 104/M) were determined by frontal chromatography. We also found that these two compounds shared the identical binding site on immobilized β2-AR with corresponding concentrations of 6.67 × 10−4 M and 5.88 × 10−4 M, respectively. The newly established method represents an efficient tool to identify the target specific natural compounds.