Microwave plasma torch desorption ionization mass spectrometry for chemical constituents of Pinellia ternata (Thunb.) Ten. ex Breitenb.

Microwave plasma torch desorption ionization mass spectrometry for chemical constituents of Pinellia ternata (Thunb.) Ten. ex Breitenb. | Authorea try { document.documentElement.classList.add('js'); } catch (e) { } var _gaq = _gaq || []; _gaq.push(['_setAccount', 'G-8VDV14Y67G']); _gaq.push(['_trackPageview']); (function() { var ga = document.createElement('script'); ga.type = 'text/javascript'; ga.async = true; ga.src = ('https:' == document.location.protocol ? 'https://ssl' : 'http://www') + '.google-analytics.com/ga.js'; var s = document.getElementsByTagName('script')[0]; s.parentNode.insertBefore(ga, s); })(); Skip to main content Preprints Collections Wiley Open Research IET Open Research Ecological Society of Japan All Collections About About Authorea FAQs Contact Us Quick Search anywhere Search for preprint articles, keywords, etc. Search Search ADVANCED SEARCH SCROLL This is a preprint and has not been peer reviewed. Data may be preliminary. 12 October 2025 V1 Latest version Share on Microwave plasma torch desorption ionization mass spectrometry for chemical constituents of Pinellia ternata (Thunb.) Ten. ex Breitenb. Author : Jiarui Gao 0009-0007-9001-2492 [email protected] Authors Info & Affiliations https://doi.org/10.22541/au.176029759.94104507/v1 Published Rapid Communications in Mass Spectrometry Version of record Peer review timeline 212 views 156 downloads Contents Abstract Information & Authors Metrics & Citations View Options References Figures Tables Media Share Abstract Rationale: Pinellia ternata (Thunb.) Ten. ex Breitenb. is a perennial herb that has a long history of use in traditional Chinese therapy due to its diverse pharmacological activities including antiemetic, antitussive, and anti-tumor activities. Characterization and quality control of P. ternata will be challenging due to its highly complex and diverse chemical composition. Methods: In this study, we utilized microwave plasma torch desorption ionization mass spectrometry (MPT-MS) for the first time to investigate P. ternata for the purpose of direct in situ analysis of individual samples. Chemical analysis of P. ternata was performed without pretreatment, while accurately identifying classes of chemical constituents, including alkaloids, organic acids, phenolic compounds, and fatty acids. Meanwhile, tandem MPT-MS was used to verification Compounds got in MPT-MS. Results: Results yielded reproducible, and unique, and generated chemical fingerprints reflected the overall chemical composition of P. ternata. And 41 compounds was detected, including alkaloids, organic acids, phenolic compounds, and fatty acids. Conclusion: MPT-MS facilitates rapid and direct characterization and quality assessment of this medicinal herb promoting better safety and reliability for clinical uses. Introduction P. ternata, the dried tuber of a plant belonging to Araceae family, has been prescribed as one of the most popular Traditional chinese medicines (TCM) 1 . Tubers are usually collected in summer and autumn, cleaned, the outer peel is removed and fibrous roots removed before sun-drying to be used medicinally 2 . It was reported that alkaloids, amino acids and organic acids, polysaccharides, volatile oils and nucleosides were found in P. ternate 2,3 . These various compositions are responsible for its wide spread pharmacological activities like antitussive, inhibition of glandular secretion, antiemetic, antifertility and anticancer-action 3 4 . However, it has been reported that unprocessed P. ternata, in contrast to its processed medicinal form, can also cause hepatotoxicity, irritant toxicity to oral, throat and gastrointestinal mucosa, vomiting and diarrhea 5,6 . The multiple components and fickle chemical composition lead to the requirement of effective analytical techniques for its characterization and quality control to ensure its safety and efficacy in clinical treatment. Thus far, numerous analytical methods have been employed to characterize and analyze natural products derived from plants, like ultraviolet-visible (UV-Vis) spectroscopy, infrared spectroscopy (IR), nuclear magnetic resonance spectroscopy (NMR) 7 , chromatography (GC), capillary electrophoresis (CE) and liquid chromatography (LC) with mass spectrometry (MS) 8-13 , hybrid tandem mass spectrometry liquid chromatography high-resolution mass spectrometry (LC-HRMS) 14,15 , gas chromatography time-of-flight mass spectrometry (GC-TOF-MS) 16 , matrix-assisted laser desorption/ionization tandem time-of-fight mass spectrometry (MALDI‑TOF/TOF-MS) 17 . Although these methods are effective for the separation and identification of natural compounds in P. ternata, they still present notable limitations in that they can require intricate sample pretreatment protocols, as well as take relatively long periods of time for analysis 12 , which renders them unsuitable for analyzing the high-throughput, as well as for distinguishing large quantities of P. ternata from related species. Therefore, analytical methods that provide complete molecular information about P. ternata components, and real-time, in situ analytics, will provide new goals for the case study. The microwave plasma torch (MPT) represents a new cite of ambient ionization source that utilizes microwave plasma oscillating at one specific frequency, 2450 MHz. The MPT produces a stable, visible flame-like plasma at atmospheric pressure 18 . The plasma is composed of high energy electrons, argon ions and reactive species, which are ideal for facilitating desorption and ionization of analytes, and therefore, favorable in direct mass spectrometric analysis of complex samples. Recently, the microwave plasma torch desorption ionization mass spectrometry (MPT-MS) is widely used in natural products in situ analysis. For instance, differentiation Navel Oranges 19 , in Situ Analysis of Tobacco Leaves 20 , chemical characterization of aromatic secondary organic aerosol 21 . In this study, we utilized an MPT-MS-based method for the rapid in situ characterization of P. ternata. By generating reproducible chemical fingerprints and providing evidence of representative constituents, our data demonstrates the capability of the MPT-MS method as a rapid chemical characterization and quality control system for TCM. 2 Experimental Section 2.1 Instruments The MPT ion source was designed by Pan’s laboratory 22 and it comprises three copper tubes nested in a central tube (CT), an intermediate tube (IT), and an outer tube (OT) as shown in figure 1 and figure S1. The working gas utilized in the CT was argon, which had an adjustable flow rate ranging from 0 to 2 mL/min with the microwave generator is used to generate plasma from 0 to 100 W at 2450 MHz. The MPT-MS analysis through a nebulizer spray. The whole device is mounted on a multi-axis platform used to adjust the distance between the front of the MPT and the MS inlet. We used a Orbitrap Exploris 120 Mass Spectrometer (Thermo Scientific, San Jose, CA) for real-time detection of chemicals. The mass spectrometer settings for data collection were: capillary temperature 320 ̵ͦ C; Lens voltage 80 V. For standard sample experiments we analyze in positive electrospray ionization modes (ESI); systems with the ion source voltage set at 3.5 kV. The MS has a resolution of 30000 and the MS data was collected using Thermo Fisher Scientific Xcalibur 2.2 SP1 software. Fig. 1 Schematic diagram of in-situ analysis of P. ternata by MPT-MS 2.2 Compound Identification Theoretical data for the identification of compounds were gathered by compiling reported chemical components of P. ternata and relevant taxa from the previous literature 23,24 . The molecular formulas of these components were drawn using ChemDraw 23.1.1, and theoretical exact mass was calculated. Acquired experimental mass spectra by MPT-MS were then compared to the theoretical calculated values. The compounds that had a close measured and theoretical mass and were considered to have an acceptable score values were deemed reliably identified. This methodology confirmed the presence of representatives of various constituents including alkoloids, organic acids, phenolics and fatty acids. 2.3 Preparation samples Plant Samples Plant materials used in this study were the tubers of P. ternata, Arisaema amurense Maxim, Pinellia cordata N. E. Brown, and Arisaema heterophyllum Blume. Tubers, both fresh and dried, were collected, cleaned, and cut into half as hemispherical or flake like. The sample was then used for MPT-MS analysis without the use of any pre-treatment of the plant material. An advantage with collecting samples in situ, including dried samples, allowed for rapid acquisition of mass spectra. 3 Results and Discussion 3.1 Optimization of MPT-MS condition The ionization efficiency of MPT-MS is influenced by multiple factors, including the distance of the plasma torch to the inlet of the MS, the power of the plasma, the flow rate of the carrier gas, and the incident angle. These parameters had been systematically optimized in previous work from our group 20 , using anatabine as the model compound to assess the ionization efficiencies. The ion intensities yielded by the optimization studies were maximized at the following settings: distance from torch to inlet: 10 mm; power of plasma: 100 W; flow rate of argon: 1.0 L/min; plasma direct towards MS inlet. In this work, we merely adopted the values that were previously optimized for the analysis of P. ternata, rather than performing new optimization experiments to provide reproducible ionization efficiencies, as well as a reproducible number of scans for obtaining spectral data to provide a robust basis for assessing the subsequent in situ detection of chemical constituents. 3.2 Chemical Profile of P. ternate Fig. 2 Mass spectra of high intensity components in P. ternate The main chemical constituents of P. ternata are organic acids, alkaloids, volatile oils, sterols, and amino acids 25,26 , which were identified using MPT-MS. The results are shown in Figure 2 (Figure S2-S8) and table 1. There are 41 compounds were detected, including Organic acids: 3-phenylpropionic acid([M+H] + m/z 151.0759), caffeic acid([M+H] + m/z 181.0499), palmitelaidic acid ([M+H] + m/z 255.2324), which are the main bioactive components of P. ternata. The alkaloids are a class of basic organic compounds found in nitrogen-containing plants 27 , and they are also important secondary metabolites with significant bioactivity and physiological functions. They are also identified during the experiments, like trigonelline([M+H] + m/z 138.0553), N-ethylaniline([M+H] + m/z 122.0968), N-benzylidenemethylamine([M+H] + m/z 120.081). However, in the analysis, the most compounds detected were Volatile oils, terpenes & aromatics, which are liquid extract of aromatic plants 28 , like D-limonene([M+H] + m/z 137.1329), α-pinene([M+H] + m/z 137.1328), (+)-pulegone([M+H] + m/z 153.1278), terpinyl acetate([M+H] + m/z 197.1538), cis-anethol([M+H] + m/z 149.0964), O-cymene([M+H] + m/z 135.1172), butylbenzene([M+H] + m/z 135.1172), m-xylene([M+H] + m/z 107.0858), 1-octen-3-ol([M+H] + m/z 129.1278), 1-penten-3-ol([M+H] + m/z 87.0806), 6-methyl-5-hepten-2-one([M+H] + m/z 127.1121) and so on. Table 1 . Natural products detected in P. ternata 1 5-methyl-2,3-dihydrofuran 84.118 C₅H₈O 85.0648 85.0651 3.53 2 gamma butyrolactone 86.09 C₄H₆O₂ 87.0441 87.0443 2.3 3 1-penten-3-ol 86.134 C₅H₁₀O 87.0805 87.0806 1.15 4 2-methylpyrazine 94.117 C₅H₆N₂ 95.0604 95.0606 2.1 5 2,4-dimethylfuran 96.129 C₆H₈O 97.0648 97.065 2.06 6 acetylacetone 100.117 C₅H₈O₂ 101.0598 101.06 1.98 7 pivalaldehyde oxime 101.149 C₅H₁₁NO 102.0914 102.0917 2.94 8 m-xylene 106.168 C₈H₁₀ 107.0856 107.0858 1.87 9 2,5-dimethyl pyrazine 108.144 C₆H₈N₂ 109.0761 109.076 -0.92 10 hydroquinone 110.112 C₆H₆O₂ 111.0441 111.0443 1.8 11 Catechol 110.112 C₆H₆O₂ 111.0441 111.0443 1.8 12 3,4-dihydro-2-methoxy-2H-pyran 114.144 C₆H₁₀O₂ 115.0754 115.0757 2.61 13 benzeneacetonitrile 117.151 C₈H₇N 118.0652 118.0654 1.69 14 2,1-benzisoxazole 119.123 C₇H₅NO 120.0444 120.0446 1.67 15 N-benzylidenemethylamine 119.167 C₈H₉N 120.0808 120.081 1.67 16 phenylacetaldehyde 120.151 C₈H₈O 121.0648 121.0652 3.3 17 N-ethylaniline 121.183 C₈H₁₁N 122.0965 122.0968 2.46 18 1,3-benzodioxole 122.123 C₇H₆O₂ 123.0441 123.0444 2.44 19 phenethyl alcohol 122.167 C₈H₁₀O 123.0805 123.0808 2.44 20 3,4-dimethylphenol 122.167 C₈H₁₀O 123.0805 123.0808 2.44 21 5-methyl-2-acetylfuran 124.139 C₇H₈O₂ 125.0598 125.0601 2.4 22 6-methyl-5-hepten-2-one 126.199 C₈H₁₄O 127.1118 127.1121 2.36 23 1-octen-3-ol 128.215 C₈H₁₆O 129.1274 129.1278 3.1 24 2-comaranone 134.134 C₈H₆O₂ 135.0441 135.0444 2.22 25 butylbenzene 134.222 C₁₀H₁₄ 135.1169 135.1172 2.22 26 O-cymene 134.222 C₁₀H₁₄ 135.1169 135.1172 2.22 27 D-limonene 136.238 C₁₀H₁₆ 137.1325 137.1329 2.92 28 alpha-pinene 136.238 C₁₀H₁₆ 137.1325 137.1328 2.19 29 trigonelline 137.138 C₇H₇NO₂ 138.055 138.0553 2.17 30 2-pentylfuran 138.21 C₉H₁₄O 139.1118 139.1119 0.72 31 2,2,6-trimethylcyclohexanone 140.226 C₉H₁₆O 141.1274 141.1272 -1.42 32 2,6-dimethyl-5-heptenal 140.226 C₉H₁₆O 141.1274 141.1272 -1.42 33 cis-anethol 148.205 C₁₀H₁₂O 149.0961 149.0964 2.01 34 3-phenylpropionic acid 150.177 C₉H₁₀O₂ 151.0754 151.0759 3.31 35 (+)-pulegone 152.237 C₁₀H₁₆O 153.1274 153.1278 2.61 36 allyl 2-ethylbutyrate 156.225 C₉H₁₆O₂ 157.1224 157.1228 2.55 37 5-pentyl-2H-pyran-2-one 164.204 C₁₀H₁₂O₂ 167.1067 167.1071 2.39 38 methyl eugenol 178.231 C₁₁H₁₄O₂ 179.1067 179.1071 2.23 39 caffeic acid 180.159 C₉H₈O₄ 181.0496 181.0499 1.66 40 terpinyl acetate 196.29 C₁₂H₂₀O₂ 197.1537 197.1538 0.51 41 palmitelaidic acid 254.414 C₁₆H₃₀O₂ 255.2319 255.2324 1.96 3.3 MPT-MS/MS-Based Verification of Compounds Figure 3. MPT-MS/MS fragments of 5-methyl-2,3-dihydrofuran MS/MS databases is currently the fastest approach for confident compound annotations in small molecule analysis including metabolomics, lipidomics, food, and environmental sciences 29 . Thus, to further evaluate the reliability of the compounds identified by MPT-MS, the representative ions were carried out tandem MPT-MS (MPT-MS/MS) analysis to gain their structural information. And utilize MS-FINDER (version 3.60) to generated theoretical fragmentation patterns and compared them with experimental data (shown in Fig.3 and Fig.S9-S16). Each candidate compounds were based on match between observed and predicted one 30 . The results are shown in table 2. Multiple primary constituents of P. ternata, including alkaloids, organic acids and volatile oils, indicated a high score in the analysis, showing a strong agreement between available experimental data in MS/MS and the theoretical data in the fragments. For instance, active ingredients are detected such as 5-methyl-2,3-dihydrofuran (score=7.8), palmitelaidic acid (score=7.19) and N-ethylaniline (score=6.87). The outcomes of this study further substantiate the accuracy and reliability of MPT-MS with MS/MS verification in profiling the chemical components of P. ternata, while also demonstrating the capacity of MPT-MS for more comprehensive characterization and quality control of TCM overall. Table 2. the score each compound obtained in tandem MS (MS/MS) analysis 5-methyl-2,3-dihydrofuran 85.0648 85.0651 7.8 gamma butyrolactone 87.0441 87.0443 7.6 1-penten-3-ol 87.0805 87.0806 6.76 2-methylpyrazine 95.0604 95.0606 6.28 2,4-dimethylfuran 97.0648 97.065 7.23 acetylacetone 101.0598 101.06 7.4 2,5-dimethyl pyrazine 109.0761 109.076 7.19 hydroquinone 111.0441 111.0443 7.36 catechol 111.0441 111.0443 7.58 3,4-dihydro-2-methoxy-2H-pyran 115.0754 115.0757 7.14 benzeneacetonitrile 118.0652 118.0654 6.3 2,1-benzisoxazole 120.0444 120.0446 5.77 N-benzylidenemethylamine 120.0808 120.081 6.1 phenylacetaldehyde 121.0648 121.0652 7.64 N-ethylaniline 122.0965 122.0968 6.87 1,3-benzodioxole 123.0441 123.0444 6.9 phenethyl alcohol 123.0805 123.0808 7.64 3,4-dimethylphenol 123.0805 123.0808 7.3 5-methyl-2-acetylfuran 125.0598 125.0601 7.63 2-coumaranone 135.0441 135.0444 6.69 butylbenzene 135.1169 135.1172 7.23 O-cymene 135.1169 135.1172 7.27 D-limonene 137.1325 137.1329 7.61 α-pinene 137.1325 137.1328 7.51 trigonelline 138.055 138.0553 7.12 2,2,6-trimethylcyclohexanone 141.1274 141.1272 6.94 2,6-dimethyl-5-heptenal 141.1274 141.1272 7.43 cis-anethole 149.0961 149.0964 7.46 3-phenylpropionic acid 151.0754 151.0759 7.43 methyl eugenol 179.1067 179.1071 7.69 palmitelaidic acid 255.2319 255.2324 7.19 3.4 Advantages and Limitations of MPT-MS The recent study demonstrated that MPT-MS has several significant advantages for the chemical analysis of P. ternata. At first, it can rapidly perform in situ detection without complicated pretreatment 31 ,which is an extremely valuable gain in terms of time efficiency on traditional methodologies such as GC-MS and LC-MS. Secondly, the method provides broader chemical coverage, allowing various metabolites (organic acids, alkaloids, etc.) to be detected in a single run. Thirdly, reproducible and characteristic chemical fingerprints could be developed to aid in quality check and standardization of traditional Chinese medicines. Although this approach has many benefits, MPT-MS does have limitations. High molecular weight compounds (ex. polysaccharides and glycosides) are less efficiently ionized in the current plasma environment, limiting the identification. In the future, optimizations in the plasma conditions, for instance, reducing the plasma power to limit excessive fragmentations, may provide better access to larger or labile molecules. Furthermore, combining artificial intelligence to interpret the spectra may enhance compound coverage and analytical reliability and broaden the use of MPT-MS to herbal quality control. 4 Conclusion In this study, MPT-MS was conducted for rapid in situ chemical analysis of P. ternata, and our results show the apparent ability of MPT-MS to analyze many different classes of components, including organic acids, alkaloids, and volatile compounds, and offer a robust representation of the chemical complexity of the phytochemical profile of this example of a TCM. By measuring accurate masses along with considering an assessment of MPT-MS/MS data analysis with MS-FINDER, our results also provided more certainty in our proposed tentative assignments of the major compounds which strongly suggests that MPT-MS may be a useful and reliable method for pants analysis. Overall, the results of this study suggest that MPT-MS may provide a rapid chemical characterization of herbal medicines. Author Contributions Jiarui Gao：Conceptualization, Methodology, Validation, Formal analysis, Investigation, Resources, Data curation, Writing – original draft, Writing – review & editing, Visualization, Supervision, Project administration Acknowledgements The authors acknowledge the department of Chemistry, Zhejiang University, for providing laboratory facilities and technical support during this research. Data availability All the relevant data are available from the authors upon reasonable request Conflicts of Interest The authors declare no conflicts of interest. Reference 1. Yi TS, Li H, Li DZ. Chromosome variation in the genus (Araceae) in China and Japan. Bot J Linn Soc . Apr 2005;147(4):449-455. doi:DOI 10.1111/j.1095-8339.2005.00381.x2. Zou T, Wang J, Wu X, et al. A review of the research progress on Pinellia ternata (Thunb.) Breit.: Botany, traditional uses, phytochemistry, pharmacology, toxicity and quality control. Heliyon . 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Oct 2017;14(10)doi:ARTN 112910.3390/ijerph14101129 Information & Authors Information Version history V1 Version 1 12 October 2025 Peer review timeline Published Rapid Communications in Mass Spectrometry Version of Record 15 Jan 2026 Published Copyright This work is licensed under a Non Exclusive No Reuse License. Keywords ion sources mass spectrometry microwave plasma torch pinellia ternata traditional chinese medicine Authors Affiliations Jiarui Gao 0009-0007-9001-2492 [email protected] Universiti Sains Malaysia View all articles by this author Metrics & Citations Metrics Article Usage 212 views 156 downloads .FvxKWukQNSOunydq8rnd { width: 100px; } Citations Download citation Jiarui Gao. Microwave plasma torch desorption ionization mass spectrometry for chemical constituents of Pinellia ternata (Thunb.) Ten. ex Breitenb.. Authorea . 12 October 2025. 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