The Analysis of <i>Leontopodium leontopodioides</i> (Willd.) Beauv. Chemical Composition by GC/MS and UPLC-Q-Orbitrap MS

Chen, Yuanyuan; Dong, Yu; Song, Lin; Bai, Changxi; Wang, Buhechaolu; Sa, Chula

doi:https://doi.org/10.1155/2024/3525212

International Journal of Analytical Chemistry

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Abstract Introduction Experimental Results Discussion Conclusions Data Availability Additional Points Conflicts of Interest Authors’ Contributions Acknowledgments References Copyright Related Articles

Research Article | Open Access

Volume 2024 | Article ID 3525212 | https://doi.org/10.1155/2024/3525212

The Analysis of Leontopodium leontopodioides (Willd.) Beauv. Chemical Composition by GC/MS and UPLC-Q-Orbitrap MS

Yuanyuan Chen,¹Yu Dong,¹Lin Song,¹Changxi Bai,¹Buhechaolu Wang,¹and Chula Sa¹

Academic Editor: Chanbasha Basheer

Received08 Feb 2023

Revised06 Apr 2024

Accepted15 Apr 2024

Published07 May 2024

Abstract

Leontopodium leontopodioides (Willd.) Beauv. (L. leontopodioides.) has been used to treat lung diseases in traditional Chinese medicine (TCM). However, a systematic analysis of its chemical components has not been reported so far. In this study, UPLC-Q-Orbitrap MS and GC-MS were applied to investigate the chemical composition of the water extracts and essential oils of L. leontopodioides. UPLC-Q-Orbitrap MS adopts a heating electrospray ionization source, collecting primary and secondary mass spectrometry data in positive and negative ions, respectively, and uses Compound Discoverer 3.2 software to analyze the collected raw data. As a result, a total of 39 compounds were identified from their high-resolution mass spectra in both positive and negative ionization modes, including 13 flavonoids and their glycosides, 15 phenolic acids, 4 oligosaccharides and glycosides, 4 pentacyclic triterpenoids, and 3 other compounds. Among them, 18 chemical components have not been reported in L. leontopodioides. In the GC-MS section, two common organic solvents (n-hexane and diethyl ether) were used to extract essential oils, and the mass spectra were recorded at 70 eV (electron impact) and scanned in the range of 35∼450 m/z. Compounds were identified using NIST (version 2017), and the peak area normalization method was used to calculate their relative amounts. Finally, 17 components were identified in the volatile oil extracted with n-hexane, accounting for 80.38% of the total volatile oil, including monoterpenoids, phenylpropene, fatty acids, and aliphatic hydrocarbons. In the volatile oil extracted with diethyl ether, 16 components were identified, accounting for 73.50% of the total volatile oil, including phenylpropene, aliphatic hydrocarbons, monoterpenoids, fatty acids, and esters. This study was the first to conduct a comprehensive analysis of the chemical composition of the L. leontopodioides water extract and its essential oil, and a comprehensive chemical composition spectrum was constructed, to lay a foundation for its further pharmacodynamic material basis and quality evaluation.

1. Introduction

As a traditional Chinese medicine, Leontopodium leontopodioides (Willd.) Beauv. has the functions of clearing away the pulmonary heat, relieving cough, and expectorating phlegm and is generally used to treat lung diseases in TCM [1]. L. leontopodioides belongs to the Asteraceae family and is a perennial herb with a height of 5∼45 cm, and it is widely distributed in northeast, north, and northwest China and grows in arid grasslands, loess slopes, gravel, and mountain grasslands at an altitude of 100∼3200 m [2], as shown in Figure 1. Previous studies of L. leontopodioides were isolated by chromatographic methods, such as silica gel, ODS, Sephadex LH-20, and HPLC, and identified by chemical and physical methods, especially spectral analysis [3–5]. Modern pharmacological studies have shown that L. leontopodioides has anti-inflammatory, antibacterial, antioxidant, hypoglycemic, diuretic, and other effects [6–9]. Chen [10] et al. used chemical and spectroscopic methods to study a 70% EtOH extract of the whole plants of Leontopodium leontopodioides (Wild.) Beauv obtained leontoaerialosides A (1), B (2), C (3), D (4), and E (5). Zhao et al. [11] found that chlorogenic acid and ferulic acid are components with obvious antioxidant effects in L. leontopodioides, and most of the chemical components related to antioxidant activity are phenolic acids. Wu et al. [12] obtained an abundant higher monomer compound by silica gel column chromatography and preparative thin-layer chromatography in ethyl acetate parts of the alcohol extract; through analysis of ultraviolet, infrared, hydrogen, and carbon spectrum, it was presumed as para-hydroxyl-acetophenone. Gao et al. [13] reported the essential oil from the aerial parts of L. leontopodioides and found that it not only has low antioxidant activity but also possesses a potent antibacterial activity against S. aureus and B. subtilis. Although some chemical components, such as flavonoids, phenylpropanoids, phenolics [14], and essential oils, have been isolated from L. leontopodioides, systematic analysis of its chemical components has not been investigated. More importantly, L. leontopodioides has not been recorded in the Chinese Pharmacopoeia. Therefore, it is necessary to carry out a systematic and comprehensive study of the chemical composition in order to elucidate its pharmacodynamic material basis. At present, there is no research on the chemical composition of the water extract and essential oil of L. leontopodioides at the same time.

2. Experimental

2.1. Sample Preparation

The whole plant of L. leontopodioides was powdered. Pulverized samples of L. leontopodioides (50g) were accurately weighed, add 15 times of distilled water, decocting 3 times for 1 hour each time, combining the three filtrates, evaporating, drying and weighing to prepare the L. leontopodioides water extrac, and its yield was 27.63%. Fifty grams of crushed L. leontopodioides was precisely weighed, and the supercritical CO₂ extraction method was used for 5 h at a temperature of 45°C and a pressure of 18 MPa to obtain 1.20 mL of dark green essential oil of L. leontopodioides.

An appropriate amount of L. leontopodioides water extract was weighed, and 1 mL of 80% methanol was added to prepare a solution with a concentration of 10 mg·mL⁻¹, vortexed, ultrasonicated for 10 min, and centrifuged at 14000 rpm for 10 min. Then, 0.8 mL of the supernatant was placed in a centrifuge tube and centrifuged again, and the supernatant was placed into a sample bottle for analysis by UHPLC-MS.

2.2. Methods

In this study, water extracts and essential oils of L. leontopodioides were analyzed using UPLC-Q-Orbitrap MS and GC-MS techniques, respectively. Featuring high resolution, high sensitivity, and high speed, ultra-performance liquid chromatography quadrupole-Orbitrap mass spectrometry (UPLC-Q-Orbitrap MS), a cutting-edge molecular separation and determination technique, has been applied to the analysis of various complex samples [15, 16]. Based on UPLC-Q-Orbitrap MS technology, a rapid identification method was established for the chemical composition of the water extract of L. leontopodioides. According to the precise molecular mass and fragmentation information of the compounds, the main compounds were identified by means of databases and references, and their cracking laws were discussed, which provided a data basis for further elucidating their pharmacodynamic material basis. GC-MS a highly effective and versatile analytical technique is widely used in pharmaceutical industries for analytical research and development, quality control, and quality assurance [17]. The essential oil was extracted by supercritical carbon dioxide extraction and then dissolved in n-hexane and diethyl ether. The resulting fractions were analyzed by GC-MS, using NIST (version 2017) for similarity search, enabling identification of the components, while calculating their relative amounts using peak area normalization. The instruments and materials used in the experiment are listed in Table 1.

2.2.1. Chromatography and Mass Spectrometry Conditions

The UPLC-Q-Orbitrap LC/MS system used a Waters ACQUITY UPLC HSS T3 C₁₈ column (2.1 mm × 100 mm, 1.8 μm; Waters Corporation, USA); the column temperature was 35°C, and the flow rate was 0.2 mL·min⁻¹. The injection volume was 5 μL; mobile phase was 0.1% formic acid acetonitrile (A)-0.1% formic acid water (B), gradient elution: 0∼10 min, 100% B; 10∼20 min, 100%∼70% B; 20∼25 min, 70%∼60% B; 25∼30 min, 60%∼50% B; 30∼40 min, 50%∼30% B; 40∼45 min, 30%∼0% B; 45∼60 min, 0% B; 60∼60.1 min, 0%∼100% B; and 60.1∼70 min, 100% B. The wavelength of DAD was set as a full scan in the range of 190∼400 nm.

The heating electrospray ionization source (HESI) was used as the ion source to detect the positive and negative ion modes, the positive spray voltage was 3.2 kV, the negative spray voltage was 3.0 kV, the detection method was full MS/dd-MS2, the sheath gas flow was 40 arb, the auxiliary gas flow rate was 15 arb, the capillary temperature was 320°C, and the auxiliary gas heater temperature was 350°C. The resolution of MS was 70000, the resolution of MS/MS was 17500, and the mass spectrum was recorded with a positive ion spectrum scan range of m/z 100∼500. Unknown compounds were identified using Compound Discoverer 3.2 software, and mzCloud (https://www.mzcloud.org/) and mzVault (self-built database) were used to identify compounds.

2.2.2. GC-MS Conditions

Inject 1.0 μL essential oils dissolved in n-hexane/diethyl ether each in splitless mode. An HP-5 ms fused silica capillary column (30 m, inner diameter 0.25 mm, and film thickness 0.25 μm) was used with helium as the carrier gas, and the oven temperature was increased from 80°C to 90°C at a rate of 3°C/min (maintained for 2 min), 95°C to 140°C at a rate of 3°C/min, 155°C to 185°C at a rate of 2°C/min, and finally 195°C at a rate of 5°C/min (hold for 8 min). The total run time was 54.5 min, and the ion source temperature was set at 250°C. The GC interface temperature was 270°C. The mass spectra were recorded at 70 eV (EI) and were scanned in the range 35∼450 m/z. Compounds were identified using the NIST Chemistry WebBook (https://webbook.nist.gov).

3. Results

3.1. Compounds Confirmation of L. leontopodioides Water Extract by UPLC-Q-Orbitrap MS

First, samples were injected according to the chromatographic and mass spectrometry conditions, and Compound Discoverer 3.2 software was used to search for the target compound peaks on the collected raw data and screen for compounds with a score greater than 80, and after matching, compounds were obtained, and the secondary fragmentation fragment ion information was analyzed to further accurately identify chemical components. As a result, a total of 39 chemical components were identified from the water extract of L. leontopodioides, mainly including flavonoids, phenolic acids, pentacyclic triterpenes, oligosaccharides, and glycosides. The total ion chromatogram is shown in Figure 2, and the chemical composition identification results are shown in Table 2.

(a)

(b)

3.1.1. Identification of Flavonoids and Their Glycosides

In this study, a total of 13 flavonoids and their glycosides (peaks 4, 5, 6, 7, 8, 9, 28, 31, 32, 35, 36, 37, and 38) were identified from the extract of L. leontopodioides. The excimer ion of peak 35 with a retention time of 32.827 min and a molecular formula of C₁₅H₁₀O₆ given in negative ion mode was m/z 285.0403 [M − H]⁻, and it loses a molecule of carbon dioxide forming m/z 241.0501 [M − H − CO₂]⁻. At the same time, 1, 3 cracking could occur to generate fragment ions m/z 151.0035 [M − H]⁻ and m/z 133.0294 [M − H]⁻. Among them, m/z 133.0294 consisted of residues on the B ring and C ring, and its intensity was larger than that of m/z 151.0035. Combined with the databases, peak 35 was identified as luteolin. Its MS/MS spectrum and the fragmentation pathway are shown in Figure 3.

Peak 9, with a retention time of 29.949 min and a molecular formula of C₂₁H₂₀O₁₁, combined with the databases was identified as cynaroside. It responds well in positive ion mode, and the excimer ion given in positive ion mode was m/z 449.1077 [M + H]⁺. In secondary mass spectrometry, luteoloside lost a glucose to form aglycone ion m/z 287.0549 [M + H − Glc]⁺, and the aglycone was further cleaved by RDA to form fragment ions m/z 153.0183 [^1,3A]⁻ and m/z 135.0441 [^1,3B]⁻. Its MS/MS spectrum and the fragmentation pathway are shown in Figure 4.

The excimer ion of peak 31 in negative ion mode was m/z 447.0922 [M − H]⁻. The excimer ion peaks were cracked and lost the fragment groups of C₆H₁₀O₅ and C₆H₁₁O₅, respectively, and fragment ions of m/z 285.0393 [M − H − C₆H₁₀O₅]⁻ and m/z 284.0324 [M − H − C₆H₁₁O₅]⁻ were obtained, respectively. Subsequently, the fragment ion of m/z 284.0324 continued to fragment, losing 1 neutral CO molecule, and producing a fragment ion of m/z 257.0424 [M − H − C₆H₁₁O₅ − CO]⁻. At the same time, the fragment ion of m/z 285.0393 can continue to be fragmented, and after losing one neutral CO molecule, it rearranges and removes 2 H atoms, and a fragment ion of m/z 255.0298 [M − H − C₆H₁₁O₅ − CO − 2H]⁻ was produced. Finally, the fragment ion continues to fragment and loses the CO molecule, producing a fragment ion of m/z 227.0349 [M − H − C₆H₁₁O₅ − CO − 2H − CO]⁻. Compared with the databases, peak 31 was identified as astragalin. Its MS/MS spectrum and the fragmentation pathway are shown in Figure 5.

3.1.2. Identification of Phenolic Acids

A total of 15 phenolic acid compounds (peaks 2, 3, 14, 17, 18, 21, 22, 23, 24, 26, 27, 29, 30, 33, and 34) were detected in the extract of L. leontopodioides, and these compounds responded better in negative ion mode. The excimer ion of peak 21 with a retention time of 19.948 min and a molecular formula of C₇H₆O₄ in negative ion mode was m/z 153.0192 [M − H]⁻. Through the loss of CO₂, the secondary spectrum generates fragment ion peaks at m/z 109.0293 [M − H − CO₂]⁻. Compared with the databases, it was identified as protocatechuic acid. Its MS/MS spectrum and the fragmentation pathway are shown in Figure 6.

The excimer ion of peak 24 with a retention time of 25.764 min and a molecular formula of C₇H₁₀O₅ in negative ion mode was m/z 173.0452 [M − H]⁻. The excimer ion peak lost one molecule of H₂O to generate m/z 155.0348 [M − H − H₂O]⁻ and also lost one molecule of H₂O to generate m/z 137.0240 [M − H − 2H₂O]⁻. Finally, this fragment ion continues to fragment and loses the COOH molecule, producing a fragment ion of m/z 93.0344 [M − H − 2H₂O − COOH]⁻. Compared with the databases, peak 24 was identified as shikimic acid. Its MS/MS spectrum and the fragmentation pathway are shown in Figure 7.

3.1.3. Identification of Pentacyclic Triterpenoids

A total of four pentacyclic triterpenoids (peaks 10, 12, 13, and 39) were identified in this study. Ursolic acid is a pentacyclic triterpenoid with a retention time of 48.962 min and a molecular formula of C₃₀H₄₈O₃. The excimer ion given in the positive ion mode was m/z 457.3670 [M + H]⁺. After it lost the neutral molecule H₂O, a fragment ion of m/z 439.3575 was generated, and at the same time, the COOH molecule was lost to obtain a fragment ion of m/z 411.3622, and this fragment further lost H₂O to produce the fragment ion of m/z 393.3508. Based on a comprehensive database, this structure was speculated as ursolic acid, and its MS/MS spectrum and the fragmentation pathway are shown in Figure 8.

3.1.4. Identification of Oligosaccharides and Glycosides

In the negative ion mode, the primary mass spectrometry mainly exists in the form of quasimolecular ion peak [M − H]⁻. Under the high-energy collision of mass spectrometry, the cleavage of the glycosidic bond mainly occurs and loses the glycosyl group. A total of four oligosaccharide and glycoside compounds (oligosaccharides: peaks 15, 16, and 19; glycosides: peak 20) were identified in this experiment. The excimer ion of peak 15 with a retention time of 2.189 min and a molecular formula of C₁₂H₂₂O₁₁ in negative ion mode was m/z 341.1085 [M − H]⁻. Its negative ion mode of MS² spectra revealed 179.0560 [M − H − Glc] and 161.0454 [M − H − Glc − H₂O]. Based on a comprehensive database, it was tentatively identified as sucrose, and its MS/MS spectrum and the fragmentation pathway are shown in Figure 9.

3.1.5. Other Compounds

In addition, two alkaloids (peaks 1 and 11) and one coumarin (peak 25) were identified in positive ion mode from the water extract of L. leontopodioides. The excimer ion of peak 11 with a retention time of 40.018 min and a molecular formula of C₁₇H₁₉NO₃ in positive ion mode was m/z 286.1435 [M + H]⁺. The most abundant fragment of m/z 201.0545 was formed by the cleavage of the amide bond, and loss of piperidine ring (–C₅H₁₁N), in the process of further cracking, will be obtained the characteristic ion m/z 171.0439 [M + H − C₅H₁₁N − CH₂O]⁺ with molecular formula C₁₁H₇O₂. This ion continues to lose CO to obtain the ion m/z 143.0492 [M + H − C₅H₁₁N − CH₂O − CO]⁺ and m/z 115.0542 [M + H − C₅H₁₁N − CH₂O − CO − CO]⁺. Compared with the database, this compound was identified as piperine. Its MS/MS spectrum and the fragmentation pathway are shown in Figure 10.

3.2. Compound Confirmation of L. leontopodioides Essential Oil by GC-MS

Samples were injected according to the GC-MS conditions and obtained a total ion chromatogram of volatile components in L. leontopodioides, and the obtained data were searched and matched by the mass spectrometry database of the National Institute of Standards and Technology (NIST 2017), and compounds with similarity scores above 80% were taken into account. After comparing the chemical composition of essential oils extracted with n-hexane and diethyl ether, a total of 33 volatile compounds (there were seven identical components) were identified, as shown in Figure 11 and Tables 3 and 4. The relative content of each component was estimated by the peak area normalization method. It can be seen from Table 3 that the main components of the volatiles of L. leontopodioides extracted with n-hexane mainly included phenylpropene (64.52%), monoterpenes (10.96%), fatty acids (10.03%), and contained some aliphatic hydrocarbons. Among them, the components with higher content were methylconiferylaldehyde (14.77%), (E)-2,6-dimethoxy-4-(prop-1-en-1-yl) phenol (12.04%), and eugenol (11.51%). As shown in Table 4, the main components included fatty acids (28.99%), phenylpropene (28.37%), aliphatic hydrocarbons, and some esters. Among them, the components with higher content were methylconiferylaldehyde (9.61%), pentadecanoic acid (9.25%), and 8-methylnonanoic acid (8.63%). Furthermore, terpinolene, terpinen-4-ol, γ-terpinene, methyleugenol, methylconiferylaldehyde, tetradecanoic acid, and n-hexadecanoic acid were identical components.

(a)

(b)

4. Discussion

4.1. UPLC-Q-Orbitrap MS Section

Flavonoids mainly exist in natural plants in the form of free or combined with sugar to form glycosides or in the form of carbon sugars, and they have anti-inflammatory, antioxidant, antibacterial, antidiabetic, antihypertensive, and other pharmacological activities [18]. The mass spectrometry fragmentation characteristics of flavonoid aglycones were mainly the loss of CO, COO, and CH₃ groups, or the loss of neutral molecules such as H₂O and the occurrence of reverse Diels–Alder reaction (RDA) fragmentation to form a series of characteristic ion peaks. Flavonoid glycosides first lose the glycosyl group to form the corresponding aglycone and then further cleave [19, 20]. In this study, taking luteolin, cynaroside, and astragalin as examples, the cracking rules of flavonoids and their glycosides were described, and it was found that the cracking rules of the three were consistent with those reported in the literature [21–23]. Among the 13 flavonoids and their glycosides obtained from the analysis, isoquercitrin, kaempferol, kaempferol-7-O-β-D-glucopyranoside, cymaroside, hyperoside, apigenin-7-O-β-D-glucoside, luteolin, quercetin, and apigenin compounds with previous reports [24–28] on the chemical composition of L. leontopodioides, scutellarein, morin, diosmetin, and astragalin have not been reported.

Phenolic acids mainly contain carbonyl, carboxyl, and hydroxyl groups, so neutral fragments of CO, H₂O, and CO₂ were easily lost in mass spectrometry collisions. The cracking rules of protocatechuic acid and shikimic acid obtained by database analysis are consistent with those reported in the literature [29, 30]. Chlorogenic acid, quinic acid, protocatechuic acid, protocatechualdehyde, cryptochlorogenic acid, caffeic acid, p-coumaric acid, isochlorogenic acid B, isochlorogenic acid C, and salicylic acid have been reported [31, 32]. Moreover, basic research on pharmacodynamics found that protocatechuic acid, protocatechuic aldehyde, chlorogenic acid, and caffeic acid in L. leontopodioides can resist acute inflammation [33]. Cinnamic acid, citric acid, fumaric acid, and shikimic acid have not been reported in L. leontopodioides in previous research.

The mass spectrometry fragmentation of pentacyclic triterpenoids was mainly loss of neutral molecules, such as H₂O and CO, and the occurrence of Diels–Alder reaction, and the oligosaccharide and glycoside mass spectrometry was relatively simple. The cracking rules of ursolic acid and sucrose were consistent with literature reports [34, 35]. In previous research, pentacyclic triterpenoids, oligosaccharides, and glycosides have not been reported in L. leontopodioides. Newly discovered pentacyclic triterpenoids have a wide range of pharmacological effects and important biological activities, including anti-inflammatory, antibacterial, antiviral, immunomodulatory, blood sugar regulation, blood pressure lowering, and antitumor activities [36]. In particular, ursolic acid has the same inhibitory effect on glycosidase in vivo and in vitro and has an obvious hypoglycemic effect [37]. Oligosaccharides possess various bio-activities, including immune regulation, antitumor, antioxidation, and anti-infection, and modulate the gut microflora [38].

In addition, there has been no research on the chemical composition of L. leontopodioides using UPLC-Q-Orbitrap MS technology at present. UPLC-Q-Orbitrap MS technology adopts full MS/dd-MS2 mode, which greatly shortens the analysis time and can quickly detect multiple chemical components, with its advantages of high separation, high resolution, and high sensitivity, and it can provide accurate mass, elemental composition, mass spectrometry fragments, and other information required for the structural characterization of compounds without the need for reference substances. Then, the possible structure of the compound can be speculated for rapid qualitative analysis. This study collects data in both positive and negative ion modes to obtain more complete mass spectrometry data. Therefore, this method was used to analyze the water extract of L. leontopodioides in this study. Compared with previous studies, not only flavonoids and phenolic acids, but also pentacyclic triterpenes, oligosaccharides, and glycosides, which have never been reported before, were obtained using UPLC-Q-Orbitrap MS. The above research results show that the anti-inflammatory, antibacterial, antioxidant, and hypoglycemic effects of L. leontopodioides may be derived from the presence of chemical components, such as flavonoids, phenolic acids, pentacyclic triterpenes, oligosaccharides, and glycosides.

4.2. GC-MS Section

The essential oil of L. leontopodioides was extracted by supercritical carbon dioxide (SC-CO₂) extraction technology. As a new advanced “green” separation technology, SC-CO₂ is easy to operate and can not only extract and separate the desired substances quickly and efficiently but also the yield and purity of the obtained substances are higher than those of traditional methods [39].

The composition of L. leontopodioides essential oil was analyzed by GC-MS, and the chemical composition of essential oils extracted with different organic solvents was compared, and it was found that extracting essential oils with n-hexane can obtain a large amount of phenylpropene compounds, such as eugenol. The pharmacological effects of eugenol include antibacterial, anticancer, antioxidant, and other effects [40]; using diethyl ether to extract essential oils can obtain a large amount of fatty acids. Aparna et al. [41] through research suggested that the n-hexadecanoic acid might function as an anti-inflammatory agent. Analysis of L. leontopodioides essential oil by GC-MS found that the high content of fatty acid components and phenylpropene components may be an essential ingredient for its medicinal effect.

Gao et al. [13] extracted essential oil from aerial parts of Leontopodium leontopodioides (Willd.) Beauv. by water distillation and analyzed it by GC-FID and GC-MS. The main components in the essential oil were identified as palmitic acid (11.6%), n-pentadecanal (5.7%), linalool (3.8%), β-ionone (3.3%), hexahydrofarnesyl acetone (3.2%), bisabolone (3.2%), and β-caryophyllene (3.2%).

Compared with this, the results of this study are quite different, but the composition types are roughly the same, which may be related to the origin of L. leontopodioides, the extraction methods of volatile oil, and the extraction of volatile oil with different solvents.

5. Conclusions

In this study, UPLC-Q-Orbitrap MS and GC-MS analytical methods were established to comprehensively characterize the chemical composition of L. leontopodioides and provide a good research basis for the formulation compatibility and pharmacological mechanism of L. leontopodioides. However, the analysis of this study mainly focused on the identification and analysis of chemical components and did not carry out basic research on blood components and pharmacodynamic substances. Therefore, in the future, this analytical technique should be used to further improve the pharmacodynamic material basis of L. leontopodioides. At the same time, the mechanism of action of L. leontopodioides should be further elucidated by combining serum medicinal chemistry, network pharmacology, metabolomics, and other technologies.

Data Availability

The data used to support the findings of this study are included within the article.

Additional Points

Highlights. (1) In this study, we divided the water extract and volatile oil of L. leontopodioides to reveal its chemical constituents by UPLC-Q-Orbitrap MS and GC-MS for the first time. In addition, the constituents of volatile oil dissolved in two different solvents were investigated. (2) This is not only a comprehensive and systematic composition analysis of L. leontopodioides but also provides a basis for its development and utilization.

Conflicts of Interest

The authors declare that they have no conflicts of interest.

Authors’ Contributions

All authors contributed to the study’s conception and design. Chula Sa, Yu Dong, and Changxi Bai designed project development and protocol. Yuanyuan Chen, Lin Song, and Buhechaolu Wang performed material preparation, data collection, and analysis. Yuanyuan Chen wrote the first draft of the manuscript, and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Acknowledgments

This work was supported by the Inner Mongolia Medical University Mongolian Pharmacy “First-Class Discipline” Young Teachers Innovation Ability Improvement Project (No. myxylxkky2019-01), the Inner Mongolia Medical University Mongolian Pharmacy “First-class Discipline” scientific research project (No. myxylxk2021-11), Scientific and Technologically Innovative Research Team for Inner Mongolia Medical University of Bioanalysis of Mongolian medicine (YKD2022TD037), the General Program of Inner Mongolia Medical University (YKD2021MS040), and University Youth Science and Technology Talent Program (No. NJYT23135).

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