Immobilized Magnetic Beads-Based Multi-Target Affinity Selection Coupled with HPLC-MS for Screening Active Compounds from Traditional Chinese Medicine and Natural Products

Abstract

Screening and identifying active compounds from traditional Chinese medicine (TCM) and other natural products plays an important role in drug discovery. Here, we describe a magnetic beads-based multi-target affinity selection-mass spectrometry approach for screening bioactive compounds from natural products. Key steps and parameters including activation of magnetic beads, enzyme/protein immobilization, characterization of functional magnetic beads, screening and identifying active compounds from a complex mixture by LC/MS, are illustrated. The proposed approach is rapid and efficient in screening and identification of bioactive compounds from complex natural products.

Development of a HPLC‐MS/MS Method to Determine 11 Bioactive Compounds in Tongmai Yangxin Pill and Application to a Pharmacokinetic Study in Rats

“…The pharmacological effects of TMYX are based on the diverse chemical composition. Chen reported 80 compounds were identified or surmised using HPLC-MS, including 23 flavones and their glucuronides, 6 phenethyl alcohol glycosides, 20 triterpene saponins, 15 lignans, and 18 other compounds [ 13 ]. Fan characterized 40 absorbed bioactive components after oral administration of TMYX in rat serum by UPLC/Q-TOF-MS [ 14 ].…”

<p>Multiple Components Rapidly Screened from Perilla Leaves Attenuate Asthma Airway Inflammation by Synergistic Targeting on Syk</p>

“…But there are lots of disadvantages in traditional methods for TCM study due to the complexity and diversity of components, length of time required, high screening cost, large sample demand and extremely easy loss of bioactive trace components. 5 Moreover, TCM works best with combinatory therapies of multi-component due to their synergistic interactions. 6 Sometimes an herbal product may be showing better effect than an equivalent dose of a single-isolated active ingredient, and its therapeutic efficacy may come from synergistic or additive actions of multiple bioactive ingredients within the herb.…”

“…Magnetic beads have been widely used for biological and chemical affinity isolation with the assistance of a magnetic field due to its excellent suspension stability, easy surface modification, and convenient solid-liquid separation. 5 Carboxyl-modified beads provide an excellent solid support for amide bonding with an enzyme's amino group, 16 and immobilization of enzymes on beads surface can be used to isolate bioactive components without inhibiting enzymatic activity. 17 Therefore, Syk-conjugated beads were employed to screen for Syk affinitive compounds from PLE, and HPLC/MS was used for compounds identification and semi-quantification.…”

Classification of Chinese herbs based on the cluster analysis of delayed luminescence

“…There are also many tools available for the study and characterization of TCMs. These include a variety of chromatography schemes, mass spectrometry, ultraviolet and infrared spectrophotometry, as well as nuclear magnetic resonance . However, these methods cannot fully reflect the cold and heat concepts of traditional Chinese medicine.…”