Method for rapidly discovering active components in Yupingfeng granules by UPLC‐ESI‐Q‐TOF‐MS

Abstract

Yupingfeng granules (YPFG) were isolated from a traditional Chinese medicine (TCM) formulation composed of three herbs (Astragali Radix, Atractylodis Macrocephalae Rhizoma, and Saposhnikoviae Radix). This formulation is used in TCM to tonify qi, and it can help strengthen exterior and reduce sweating. Nevertheless, the active components of YPFG remain unclear. In this study, the chemical constituents of YPFG were systematically characterized by ultra-performance liquid chromatography coupled with electrospray ionization/ quadrupole time-of-flight mass spectrometry (UPLC-ESI-Q-TOF-MS). Fifty-eight compounds, namely, 20 flavonoids, 19 saponins, nine organic acids, four volatile coumarins, three lactones, one alkaloid, and two other components, were identified. In addition, the constituents of YPFG with the potential for in vivo bioactivities following oral administration were investigated in Sprague-Dawley rats. Thirteen compounds, namely, 11 flavonoid-related and 2 saponin-related components, were detected in rat plasma. After enriching flavonoids and saponins in YPFG by extraction, the extracts and YPFG were administrated to immunosuppressed rats, respectively. Plasma samples were analyzed by UPLC-ESI-Q-TOF-MS, and principal component analysis (PCA) confirmed that the extracts had similar effects to YPFG. This method could discover active ingredients in YPFG quickly and provide a scientific basis for quality control and mechanism research. K E Y W O R D S absorbed components in rat plasma, active components, chemical profiling, principal component analysis, UPLC-ESI-Q-TOF-MS, Yupingfeng granules 1 | INTRODUCTION Yupingfeng (YPF) has been used as an herbal medicine in China for centuries. YPF was first recorded in Dan xi xin fa, 1 which is an ancient book written by Zhenheng Zhu during the Yuan dynasty. YPF, composed of Astragali Radix, Atractylodis Macrocephalae Rhizoma, and Saposhnikoviae Radix, shows excellent activities including in tonifying qi, strengthening exterior, and reducing sweating. Clinically, YPF is often used to treat recurrent common cold, 2 asthmatic bronchitis, 3 allergic rhinitis, 4 and chronic urticaria, 5 all of which are often caused by immune disorders. Recent studies have shown that YPF granules (YPFG) can improve low thymus and spleen function caused by dexamethasone (Dex). Treatment with YPFG restored lymphocytic activity in Dex-treated mice back to the normal level and inhibited Dex-induced apoptosis of lymphocytes. YPFG mitigated the low levels of CD4(+)/CD8(+) caused by Dex and upregulated Dex

Identification of Chemical Components of Qi-Fu-Yin and Its Prototype Components and Metabolites in Rat Plasma and Cerebrospinal Fluid via UPLC-Q-TOF-MS

“…In recent years, LC-MS technology has been widely used in the analysis of components of TCM, combining the high separation ability of liquid chromatography with the high sensitivity of mass spectrometry [ 47 , 48 ]. Up to now, the only research on the identification of components in Qi-Fu-Yin was based on UPLC-Q-TOF-MS in vitro [ 10 ].…”

Comprehensive characterization and identification of chemical constituents in Yangwei decoction using ultra high performance liquid chromatography coupled with electrospray ionization quadrupole time‐of‐flight tandem mass spectrometry

“…Based on the cleavage rule of reference standards and reference data from literature [40,41], lactones could easily eliminate the small neutral fragments such as H 2 O (18 Da), CO (28 Da), CO 2 (44 Da), CH 3 (15 Da), MeOH (32 Da), and/or the combination of these fragments. These fragmentation mechanisms concluded the characterization of F I G U R E 6 Characteristic MS/MS spectra and possible fragmentation pathways of chlorogenic acid, 6-gingerol, atractylenolide-III lactones in YWD, and a total of eight lactones including three lactones (labeled as P167, P196, P205), two limonins (labeled as P123, P142), and three coumarins (labeled as P17, P111, P178) were identified in YWD (Table S3).…”

Simultaneous quantitation of 15 bioactive components in Yupingfeng granules by LC–MS/MS

“…Our group characterized 58 constituents in YPFG and detected 13 metabolites in the plasma of rats administered with YPFG. In addition, it was observed that the enriched flavonoids, chromones, and saponins extract in YPFG could significantly improve rats with immunosuppression and the extract had similar efficacy to YPFG 9 . It has been found that these components may regulate the metabolism of bile acids by enhancing the expression of the Farnesoid X receptor (FXR) and liver X receptor (LXR) to achieve immune regulation by integrating metabolomics and network pharmacology 10 …”

“…The current study aimed to develop a method based on liquid chromatography–tandem mass spectrometry (LC–MS/MS) to simultaneously detect and quantify 15 bioactive components (prim‐O‐glucosylcimifugin, calycosin 7‐O‐glucoside, cimifugin, 5‐O‐methylvisammioside, ononin, sec‐O‐glucosylhamaudol, calycosin, astragaloside IV, astragaloside III, formononetin, astragaloside II, isoastragaloside II, cyclocephaloside II, astragaloside I, and isoastragaloside I) (Figure 1) in YPFG. These 15 compounds were prototypes of the metabolites of YPFG in plasma and have been found to have therapeutic effects on immunosuppressive rats induced by cyclophosphamide 9,10 . Furthermore, the content measured by LC–MS/MS was compared with that measured by HPLC, which provided a reference for further research on the material basis of YPFG and the quality evaluation of YPFG and other Yupingfeng preparations.…”

“…9,10 Furthermore, the content measured by LC-MS/MS was compared with that measured by HPLC, which provided a reference for further research on the material basis of YPFG and the quality evaluation of YPFG and other Yupingfeng preparations.…”