An enhanced analytical strategy integrating offline two‐dimensional liquid chromatography with high‐resolution accurate mass spectrometry and molecular networking: Comprehensive characterization of HuangLian JieDu Decoction as a case study

Abstract

Comprehensive ingredient research is of great significance for understanding the effective material basis of herbal medicines, but due to the diversity and complexity of their phytochemicals, such research is challenging. Here, a multifaceted strategy was proposed to analyze and identify the composition of HuangLian JieDu Decoction based on offline two‐dimensional liquid chromatography combined with ultraviolet detection and high‐resolution mass spectrometry. Multiple components were separated by two‐dimensional liquid chromatography, which consisted of hydrophilic interaction chromatography and reversed‐phase liquid chromatography, and then further characterized by high‐resolution mass spectrometry with a full mass spectrometry/precursor ion list/data‐dependent secondary scan data acquisition method. For data processing, database screening and molecular networking were used to identify the components in HuangLian JieDu Decoction. The offline two‐dimensional liquid chromatography combined with ultraviolet detection and a high‐resolution mass spectrometry system showed good orthogonality of 76.35% and a high peak capacity of 5175, effectively separating multiple components. Finally, 527 compounds, including 164 alkaloids, 133 terpenoids, 88 flavonoids, 60 phenylpropanoids, 38 organic acids, and 44 other compounds, were characterized. This integrated approach is suitable for the comprehensive characterization of herbal medicines and other complex chemical systems.

Gain deeper insights into traditional Chinese medicines using multidimensional chromatography combined with chemometric approaches

“… 1 D: C 18 HCE (4.6 mm × 250 mm, 5 μm) UV Dai et al, 2023 2 D: HILIC Safflower (Honghua): flowers 1 D: TAC column ESI-MS Qiao et al, 2021 2 D: Shim-pack VP-ODS (10 mm × 2.0 mm, 5 μm) Homoisoflavonoids from Ophiopogon japonicus (L. f.) Ker Gawl. (Maidong): roots 1 D: HSCCC 2 D: SunFire™ C 18 (4.6 mm × 250 mm, 5 μm) DAD; Q-TOF-MS Deng, Xu, Tong, Shi, & Shi, 2020 Qingfei Paidu Decoction 1 D: C 18 YE column (50 mm × 250 mm, 10 μm) UV; Q-TOF-MS Shen et al, 2022 2 D: BEH Amide (2.1 mm × 150 mm, 1.7 μm) C. chinensis (Tusizi): seeds 1 D: Waters XBridge Amide (4.6 mm × 150 mm, 3.5 μm) Q-TOF-MS Wang et al, 2022c 2 D: Agilent Zorbax SB-Aq (2.1 mm × 100 mm, 1.8 μm) Compound Danshen Dripping Pill 1 D: XBridge Amide (4.6 mm × 150 mm, 3.5 μm) Q-TOF-MS Wang et al, 2022a 2 D: HSS T3 (2.1 mm × 100 mm, 1.8 μm) Huanglian Jiedu Decoction 1 D: Waters Xbridge HILIC column Q-Exactive Orbitrap-MS Li, et al, 2022b 2 D: Waters Acquity HSS T3 column O. indicum 1 D: Oasis HLB (2.1 mm × 20 mm, 25 μm) DAD; ESI-MS Cheng, Kong, Li, & Li, 2022 2 D: Kinetex C 18 (4.60 mm × 100 mm, 2.6 μm) …”

Rapid identification of the compounds of Bushen Huoxue Prescription based on offline two‐dimensional liquid chromatography with high‐resolution mass spectrometry and molecular network technology

“…When the similarity between two mass spectra is higher than a threshold, they are connected in a MN [22]. This method can infer unknown compounds from known compounds according to the similarity relationship between compounds in the network, thus offsetting the shortcomings of conventional identification methods [11,23]. To improve the quality of an MN, we used Mzmine software to generate a FBMN.…”

Qualitative identification and quantitative comparison of Physochlainae Radix from different regions based on chemometric methods

“…Chromatographic separation is a prerequisite to comprehensively identifying the multi-components in a complicated system like herbal extract. The columns with different bonding technologies or bonding groups may exert significantly different resolutions for the constituents [26][27][28][29]. Five different chromatographic columns on the separation of PR were evaluated by comparing the overall resolution and the number of the resolvable peaks eluted.…”

“…In R2, batches of samples 4 and 11 were also collected from Shaanxi province. In R3, batches of samples 13,14,15,16,17,18,19,20,21,22,23,24,25,26,27,28,29, and 30 were collected from Shanxi, Henan, and Guangxi provinces. HCA results enable the primary and non-primary producing areas, as well as superior and inferior products to be distinguished.…”