Study on the Chemical Components of Jiangxi Specialty Traditional Chinese Medicine Chaxiong Based on UPLC-Q-TOF-MS/MS Technology
Chaxiong comes from the dried rhizomes of Ligusticum sinense Oliv. cv. Chaxiong Mss., a plant belonging to the Umbelliferae family. It has a warm nature, a pungent taste, and is beneficial to the liver and heart meridians. It has the effects of promoting blood circulation, regulating qi, dispelling wind, and relieving pain. In recent years, studies have found that Rhizoma Ligustici Texiong also has anti-inflammatory, analgesic, antithrombotic, vasodilator and other effects, and has certain therapeutic effects in radiation stomatitis, stroke, thrombotic diseases, novel coronavirus pneumonia and other aspects. Chaxiong is one of the specialty Chinese medicinal herbs in Jiangxi Province, mainly produced in counties and cities such as Wuning, Ruichang, and De’an in the Jiujiang area. It is an important traditional Chinese medicine resource and economic crop in the local area. Cha Xiong has a long history of medicinal use. In ancient times, it was called “Fu Xiong”, and together with Chuanxiong and other herbs, it is called “Xiong Qi”. It was first seen in the “Shennong Bencao Jing” and is classified as a top-grade herb. After the Song Dynasty, there was a differentiation in the name of “Fu Xiong”, which was recorded in works such as the “Compendium of Materia Medica”, “Compendium of Materia Medica”, and “Compendium of Materia Medica” in the Qing Dynasty. It is now included in the 2014 edition of the “Jiangxi Province Traditional Chinese Medicine Standards”.
At present, there are relatively few research reports on the chemical components of Camellia oleifera. Existing literature mainly involves the isolation and identification of chemical components such as phthalates, organic phenolic acids, alkaloids, and polysaccharides. However, these research cycles are long, the efficiency is low, and most of the literature is early, lacking comprehensiveness and innovation. In order to improve the separation efficiency and purity of the chemical components of Chaxiong, Gong et al. established a fingerprint of Chaxiong using high-performance liquid chromatography (HPLC), and identified and determined its main components such as chlorogenic acid and ferulic acid. However, related pharmacological studies have mostly focused on crude extracts, and the pharmacological substance basis and mechanism of action of Chaxiong have not been thoroughly explored, which has limited its clinical application. Therefore, clarifying the material basis of Chaxiong is of great significance for conducting relevant pharmacological research, further application, and development of Chaxiong.
The ultra-high performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS) analysis technology has the characteristics of fast, high efficiency, and high sensitivity, which can achieve rapid separation and accurate identification of compounds. It has been widely used in traditional Chinese medicine research. Previous studies have found that in positive ion mode, the response of phthalein, amino acid, and alkaloid components is stronger, while in negative ion mode, the response of organic phenolic acid components is better. Therefore, this study is based on UPLC-Q-TOF-MS/MS technology and uses both positive and negative ion ionization modes for rapid analysis of the chemical components of Camellia oleifera. It further clarifies the composition of various chemical components in Camellia oleifera, such as phthalates, organic phenolic acids, alkaloids, etc., providing scientific basis for the quality control and pharmacological substance research of Camellia oleifera.

This study used two ionization modes, positive ion and negative ion, to detect and identify compounds in Camellia oleifera through retention time, precise relative molecular weight, and secondary mass spectrometry fragmentation. As a result, a total of 72 compounds were identified from Camellia oleifera, including 33 phthalein compounds, 32 organic acid components (including 11 amino acids and 21 organic phenolic acid components), 5 alkaloid components, and 2 other components. Analysis of the results revealed that there are the most types of phthalein and organic acid components in Camellia oleifera, which is consistent with the results reported in existing literature.
Phthalide compounds are a class of organic compounds with a phthalein structure and are one of the characteristic components of plants in the Umbelliferae family. They are widely present in the Ligusticum chuanxiong plant and the local plant Angelica sinensis. Han summarized existing research results and found that more than 70 phthalide compounds have been isolated and identified in Ligusticum chuanxiong medicinal materials; Zhang et al. identified 55 phthalein compounds from Angelica sinensis using UPLC-Q-TOF/MS technology. Modern research has found that benzophenone compounds have various biological activities such as prevention and treatment of cardiovascular and cerebrovascular diseases, antibacterial, anti-tumor, anti-inflammatory, analgesic, and insecticidal effects. For example, components such as n-butyl phthalein, ligustilide, and ligustilide A can protect brain neurons from ischemic damage through different mechanisms.
Organic acid compounds are a type of compound with carboxyl groups in their molecular structure, widely distributed in medicinal plants and mainly enriched in the root and stem parts of plants, such as Ligusticum chuanxiong and Angelica sinensis in the Umbelliferae family. They are also present in high concentrations in the leaves and fruits of plants. Organic acid components can regulate the acid-base balance in the body, promote metabolism, enhance immunity, antioxidant, anti-inflammatory, antibacterial, etc. Among them, amino acid components are one of the essential nutrients for the human body, which can promote metabolism and improve immunity. For example, arginine has the effect of improving cardiovascular and cerebrovascular diseases and enhancing immune function; Organic phenolic acids have antioxidant, antiplatelet aggregation, anti atherosclerosis, immune regulation and other effects. For example, ferulic acid can prolong the coagulation time in mice, reduce the dry and wet weight of thrombosis in arteriovenous bypass model rats, and significantly inhibit thrombosis.
Therefore, the UPLC-Q-TOF-MS/MS method established in this study can quickly, accurately, and comprehensively analyze and identify the chemical components in Chaxiong, providing a scientific basis for the quality control and efficacy evaluation of Chaxiong, and laying the foundation for the discovery and identification of other unknown components in Chaxiong. However, this study also has some limitations, such as inaccurate structural inference of compounds, lack of quantitative analysis of characteristic components, and failure to consider the interactions and synergistic effects between compounds. Based on this, future research directions can be explored from the following aspects: optimizing the analysis conditions and parameters of UPLC-Q-TOF-MS/MS technology to improve the detection efficiency and accuracy of compounds in tea leaves; Using other analytical methods, such as nuclear magnetic resonance, infrared spectroscopy, ultraviolet spectroscopy, etc., to verify and supplement the structure of compounds in Camellia oleifera, in order to improve the reliability and integrity of compound identification in Camellia oleifera; Quantitative analysis of characteristic components in Camellia oleifera, establishment of quality standards and evaluation system for Camellia oleifera, reflecting the differences in content and relative contributions of various compounds in Camellia oleifera; By combining pharmacological and biological methods, explore the mechanism of action and interactions of compounds in Camellia oleifera, elucidate the pharmacological substance basis and composition of its active ingredients.