Highly selective screening of the bioactive compounds in Huoxue capsule using immobilized β2-adrenoceptor affinity chromatography
Section snippets
Materials and reagents
Huoxue capsule was acquired from the Xi’an Datang Pharmaceutical Group (Xi’an, China). Bambuterol, clorprenaline, phenylephrine, acetylcholine, Triton X-100, and ICI 118551 were purchased from Sigma (St. Louis, MO, USA). Naringin, hydroxysafflor yellow A (HSYA), and ferulic acid (purity > 98%) were purchased from the National Institute for the Control of Pharmaceutical and Biological Products of China (lot nos. 110722-200610, 110722-200610, and 0773-9910, respectively). HPLC-grade solvents and
Purification of the β2-AR
The purity of the obtained β2-AR was 95.3%, as determined by analysis with sodium dodecyl sulfate polyacrylamide gel electrophoresis and high-performance size exclusion chromatography.
Selectivity and system suitability of β2-AR column
The results of the positive control experiment revealed that the tR values of bambuterol and clorprenaline that bound to the β2-AR on the stationary phase were 5.51 and 6.86 min, respectively (Figs. 2B). However, the tR values of prazosin and terazosin, which target the α-AR and were used as negative control drugs,
Conclusion
In this study, a method using porcine recombinant β2-AR affinity chromatography on-line with LC/MS was established to screen for β2-AR ligands from complicated systems such as natural plant extracts and traditional Chinese medicines. Using this method, ferulic acid, HSYA, and naringin were identified. This method can simultaneously recognize, separate, and identify multiple bioactive compounds and will be useful in drug discovery efforts using natural medicinal herbs to identify potential β2-AR
Acknowledgments
This study was supported by grants from the National Natural Science Foundation of China (Major International [Regional] Joint Research Project, 81120108002; General Program, 81071765), the Specialized Research Fund for the Doctoral Program of Higher Education (20130201130008), the Program for Changjiang Scholars and Innovative Research Team in the University of China (IRT1174), the Program for Innovative Research Team of Shaanxi Province (2013KCT-24), and the Scientific Research Plan Projects
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Advances in receptor chromatography for drug discovery and drug–receptor interaction studies
2023, Drug Discovery TodayModern research thoughts and methods on bio-active components of TCM formulae
2022, Chinese Journal of Natural MedicinesG protein-coupled receptor-in-paper, a versatile chromatographic platform to study receptor-drug interaction
2021, Journal of Chromatography ACitation Excerpt :These stationary phases have been evaluated by lead compound screening and receptor-drug interaction analysis [1–5]. The HPAC method has at least two merits: i) it combines the separation capacity of chromatography and the specificity of receptor-ligand bindings, thus making the targeted screening of lead compounds and receptor-drug interaction analysis in a reliable, fast and high throughput manner; ii) the stability of the receptors is highly improved after immobilization, thus the affinity column can give a continuously stable and effective readout of the injected analytes within one month [7–10]. However, it still has some drawbacks to overcome: i) the cost of the microspheres (particle size < 7.0 μm) is relatively high (30 dollars or higher per gram) and it is difficult to recover the microspheres when the immobilized receptor is inactive; ii) the affinity columns are usually packed under a pressure of 400 bar and works in a high-pressure environment (i.e. 20 bar or above).
Screening of bioactive components from traditional Chinese medicine by immobilized β<inf>2</inf> adrenergic receptor coupled with high performance liquid chromatography/mass spectrometry
2019, Journal of Chromatography B: Analytical Technologies in the Biomedical and Life SciencesCitation Excerpt :In our previous work [9], the affinity chromatography stationary phase was obtained by binding β2-AR to macropore silica gel by non-covalent bonding and then affinity chromatographic column was constructed. Immobilized β2-AR still maintained biological activity and selectivity that could study the interaction between receptor and ligands and receptor conformation changes by ligands, temperature, pH and so on [10]. In this research we constructed β2-AR column and used to screen the bioactive compounds of Rhizome Corydalis which is one of the well-known TCM [11].
Recent applications of immobilized biomaterials in herbal analysis
2019, Journal of Chromatography ARecent advances in bio-affinity chromatography for screening bioactive compounds from natural products
2019, Journal of Pharmaceutical and Biomedical AnalysisCitation Excerpt :Compared with an offline system, the online coupling system could significantly simplify the operation procedure and increase the screening efficiency [8,100,147]. So far, several BAC-online-HPLC-MS systems have been used for screening programs [8,54,57]. He’s group developed a CMC-online-HPLC-MS system using an eight-port switching valve and two capture loops [100].