Rapid separation and identification of 96 main constituents in Huanglian Jiedu decoction via ultra‐high performance liquid chromatography–Orbitrap Fusion Tribrid mass spectrometer

Abstract

Huanglian Jiedu decoction is a widely used traditional Chinese medicine with a broad spectrum of therapeutic effects, including heat clearing, detoxification, and attenuation of inflammation. However, the composition of Huanglian Jiedu decoction is still unclear due to its complexity and limitations of analytical methods. In this study, we established a fast and reliable analytical method based on ultra-performance LC-Orbitrap Fusion Tribrid mass spectrometer for high-speed separation and structural identification of multiple compounds in Huanglian Jiedu decoction. The analysis was carried out using a Hypersil GOLD C18 column (2.1 Â 100 mm, 1.9 μm) with gradient elution coupled to a high-definition mass spectrometer system operating in both positive and negative ESI modes. According to the chromatographic retention time, precise molecular weight, fragment ion peaks, and published data, the main chromatographic peaks were attributed to specific molecules whose chemical structures were determined. In total, 96 components were identified, including 34 flavonoids and their glycosides, 23 alkaloids, 18 organic acids, 13 terpenoids, and 8 miscellaneous compounds. This study revealed the detailed chemical composition of Huanglian Jiedu decoction, which is of great importance for quality control and further pharmacological and mechanistic studies.

Screening of the active Ingredients in Huanglian Jiedu decoction through amide bond-Immobilized magnetic nanoparticle-assisted cell membrane chromatography

“…Identifying the active ingredients in TCM is essential for quality control and drug development. CM chromatography has attracted increasing attention due to its high screening efficiency and specificity in elucidating the active ingredients of TCMs (Liao et al, 2018a;Liao et al, 2018b;He et al, 2022a;He et al, 2022b). However, this method has several limitations.…”

“…Currently, there are two common strategies for immobilizing MBs and CMs. One of them is based on physical methods (Sherwood et al, 2019;Xu et al, 2019;Qi et al, 2020;He et al, 2022a;He et al, 2022b). Although this method is simple and the receptor activity on the CM is well preserved during its course, the separation of the CMs from the MBs is performed under a strong external magnetic field, which may result in instability.…”

“…The cells were then cultured in a cell incubator at 37 °C. The CMs were prepared according to previously reported methods (He et al, 2022a;He et al, 2022b). The amount of CMs was examined using the BCA protein assay.…”

“…Subsequently, 5 mL citrate-sodium citrate buffer (pH 4.0) was added, and the resultant solution was incubated for 30 min to dissociate the specific binding components from the CM. The dissociated solution was collected and fractionated by SPE, according to a previously reported method (He et al, 2022a;He et al, 2022b). This sample was used for mass spectrometry (MS) analysis.…”

“…Moreover, the high accumulation of reactive oxygen species is closely related to glutamate-induced neuronal damage (Pan et al, 2020;Tonsomboon et al, 2021). The Huanglian Jiedu decoction (HLJDD), which originates from "Arcane Essentials from the Imperial Library", is composed of Coptis chinensis Franch (Ranunculaceae; Coptidis Rhizoma), Scutellaria baicalensis Georgi (Lamiaceae; Scutellariae Radix), Phellodendron amurense Rupr (Rutaceae; Phellodendri Amurensis cortex), and Gardenia jasminoides Ellis (Rubiaceae; Gardeniae fructus) in a ratio of 3:2:2:3 (He et al, 2022a;He et al, 2022b;Liao et al, 2022). As a Chinese classical formula with heat-clearing and detoxicating effects, HLJDD has been proved to have the potential to treat AD by many modern pharmacological studies (Sun et al, 2018;Wu et al, 2020).…”