Original articleRapid discovery and identification of 68 compounds in the active fraction from Xiao–Xu–Ming decoction (XXMD) by HPLC–HRMS and MTSF technique
Graphical abstract
Using MTSF technique, 50 related chemical components (red indication, 1–50) of Xiao–Xu–Ming decoction was discovered and identified based on the 14 templated compounds (green target recognition, R1–R14). These results showed that this technology has the advantage of rapidly identifying components of Chinese prescription. Notably, the unrelated peak is the compounds whose structures exhibited significant difference from the 14 templated compounds (such as orange indication, N1–N4). Such information will contribute to the research on targeted phytochemical separation.
Introduction
Chinese prescription is the fundamental method of traditional Chinese clinical medication and therefore the essence of traditional Chinese medicine (TCM). In modern science, the chemical components of Chinese prescriptions are responsible for their medicinal effects. Therefore, identifying the chemical components of Chinese prescriptions is important to understand their functional mechanisms. However, Chinese prescriptions contain complicated components, and identification and analysis of these multiple components can be difficult. With the development of modern analytical techniques, a variety of tandem high-resolution mass spectrometers (HRMS), such as Q-TOF, IT-TOF, LTQ-FTICR-MS, and LTQ-Orbitrap, have been widely used for the detection and identification of compounds in TCM [1], [2], [3], [4]. With advantages such as excellent resolution, high mass accuracy, superior sensitivity, fast scan rate, and multiple-stage mass spectrometry scanning (MSn), these mass spectrometers can provide HRMS and MSn data for nearly all compounds present in TCM using a simple data acquisition method [5], [6]. However, automatically acquired mass spectrometric data contain numerous endogenous components. Some challenges that need to be addressed include discovering relevant chemical components from the data automatically collected by the equipment and selecting a method that quickly identifies the structures of the components using the obtained mass spectrometric data.
Recently, mass spectral tree similarity has been introduced and applied to the identification of unknown compounds based on a library of known compounds [7], [8], [9], [10]. The reported methods [7], [8], [9], [10] of mass spectral tree similarity have presented practical values for the identification of chemical compounds, especially for the identification of metabolites in metabolomics. In our previous study [11], a novel mass spectral tree similarity filter (MTSF) technique was developed to discover and identify related metabolites based on the similarity of high resolution and multiple-stage mass spectrometric data between the detected compounds and templated library compounds.
This work is the first to apply MTSF technique to study the chemical components of the active fraction from “Xiao–Xu–Ming decoction (XXMD)”. XXMD was a traditional Chinese prescription and first recorded in “Bei Ji Qian Jin Yao Fang”. The formula consists of 12 crude drugs including Saposhnikovia divaricata (Turcz.) Schischk., Scutellaria baicalensis Georgi, Paeonia lactiflora Pall., Glycyrrhiza uralensis Fisch., Zingiber officinale Rosc., Stephania tetrandra S. Moore, Panax ginseng C. A. Mey., Cinnamomum cassia Presl, Prunus armeniaca L. var. ansu Maxim., Ephedra sinica Staph, Ligusticum chuanxiong Hort., and Aconitum carmichaeli Debx. in a ratio of 3:3:3:3:3:3:6:6:6:9:9:9 on a dry weight basis. It has been widely used to treat theoplegia and the effects of theoplegia in China. Through this study, we increased the application range of the MTSF technique and obtained satisfactory results.
Section snippets
Materials
Paeoniflorin (R1), prim-O-glucosylcimifugin (R2), cimifugin (R4), 4′-O-β-d-glucosyl-5-O-methylvisamminol (R5), baicalin (R6), baicalein (R9), glycyrrhizic acid (R10), wogonin (R11), chrysin (R12), and glycyrrhetinic acid (R14) were purchased from the National Institute for Control of Pharmaceutical and Biological Products (Beijing, China). Liquiritigenin (R3) was obtained from the Dalian Fusheng Pharmaceutical Co., Ltd. Wogonoside (R8) was ordered from the Shanghai Usea Biotech Co., Ltd.
Results and discussion
Optimized chromatographic conditions were adjusted based on the previous fingerprint analysis method [13]. Such adjustments included a change to a small size of the column (250 mm × 4.6 mm, 5 μm changed to 150 mm × 2.1 mm, 3 μm) and optimization of the elution gradient of the mobile phase. A suitable simple data acquisition method was mainly developed through the maximum development of LTQ FT acquisition capability. By optimizing injection time (200 ms for FT, 50 ms for LTQ), micro scan (1), and resolution
Conclusion
A total of 68 compounds were identified in the active fraction of XXMD. However, in our previous study [13], we just identified 14 major compounds of XXMT using manual mining approach. Briefly, the results demonstrated that MTSF technique should be useful to rapid discovery and identification of compounds in Chinese prescription. Aside from showing the advantages of this technology in terms of rapid discovery and identification of the chemical components of Chinese prescription, this study also
Acknowledgment
The authors would like to thank the Natural Science Foundation of Beijing (No. 7133252) for financial support of this work.
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Both these authors contributed equally to this work.