Identification of bioactive compounds in Shaoyao‐Gancao decoction using β2‐adrenoceptor affinity chromatography

Abstract

Shaoyao-Gancao decoction, a Chinese herbal formula, is composed of Paeoniae Radix alba and Glycyrrhiza Radix et rhizoma. It has been widely used to treat muscle spasms and asthma. However, little is known about the bioactive components of Shaoyao-Gancao decoction. In the present study, the bioactive compounds in water-extract of Shaoyao-Gancao decoction were separated by the immobilized β -adrenoceptor affinity column and identified using quadrupole time-of-flight mass spectrometry. The affinity constants of the separated compounds that bind to β -adrenoceptor were determined by frontal analysis. Compound bioactivity was tested in a rat tracheal smooth muscle relaxation assay. We identified the bioactive compounds in the water extract of Shaoyao-Gancao decoction that bound to the β -adrenoceptor as paeoniflorin and liquiritin. Paeoniflorin and liquiritin had only one binding site on the immobilized β -adrenoceptor, and the affinity constants were (2.16 ± 0.10) × 10 M and (2.95 ± 0.15) × 10 M , respectively. Both compounds induced a concentration-dependent relaxation of tracheal smooth muscle following K -stimulated contraction, and the relaxation effects were abrogated by the β -adrenoceptor antagonist, ICI 118551. Therefore, paeoniflorin and liquiritin are bioactive compounds in Shaoyao-Gancao decoction and the β -adrenoceptor affinity chromatography is a useful tool for identifying potential β -adrenoceptor ligands in natural products used in traditional Chinese medicine.

Rapid characterization of chemical constituents of Shaoyao Gancao decoction using UHPLC coupled with Fourier transform ion cyclotron resonance mass spectrometry

“…The inferences of each ingredient were carried out with the help of the molecular formulas and fragmentation pathways, followed by additional conrmation with reference to the previous literatures. [8][9][10][11][12][13][14][15][16] Table 1 lists the retention time, formula, molecular weight, calculated m/z, detected m/z, error value and MS/MS data of ingredients. A concrete illustration of the ingredients' characterization was performed as hereunder.…”

“…Some of these chemical components have been reported to be the active ingredients in SGD. 8,10,13,17,18 For example, paeoniorin was reported to have anti-inammatory, hepatoprotective and neuroprotective effects. 19,20 Albiorin was shown to be both anti-inammatory and antioxidant.…”

Comparative pharmacokinetics and metabolites study of seven major bioactive components of Shaoyao‐Gancao decoction in normal and polycystic ovary syndrome rats by ultra high pressure liquid chromatography with tandem mass spectrometry

“…Shaoyao‐Gancao decoction (SGD) is a TCM formula that consists of only two herbs: Radix Paeoniae Alba ( Paeonia lactiflora Pall, root ) (RP) and Glycyrrhizae uralensis ( Glycyrrhiza uralensis Fisch., root and rhizome, honeyed ) (GU) with an equal proportion . The prescription was first recorded in Treatise on Febrile Diseases, a classical work of TCM written by the famous Chinese physician Zhongjing Zhang in Han dynasty of Chinese history .…”

Rapid screening and identification of bioactive compounds specifically binding to beta 2-adrenoceptor from San-ao decoction using affinity magnetic fine particles coupled with high-performance liquid chromatography–mass spectrometry

“…More recently, developed functional assays and phenotypic screens, such as cellular membrane affinity chromatography and computer-aided virtual screening technologies have received increasing attention [ 11 – 15 ]. In our previous work, beta 2-adrenoceptor ( β 2 -AR) chromatography was established for the identification of bioactive compounds of the receptor [ 16 ]. This strategy has limitations of time-intensity and labor-intensity in preparation of chromatographic columns.…”