Determination of Multiple Neurotransmitters through LC-MS/MS to Confirm the Therapeutic Effects of <i>Althaea rosea</i> Flower on TTX-Intoxicated Rats
Tetrodotoxin (TTX) inhibits neurotransmission in animals, and there is no specific antidote. In clinical practice in China, <i>Althaea rosea</i> (<i>A. rosea</i> flower) extract has been used to treat TTX poisoning. In this work, the efficacy of the ethyl acetate fraction ext...
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MDPI AG
2023-05-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/28/10/4158 |
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| author | Yichen Wang Renjin Zheng Pingping Wu Youjia Wu Lingyi Huang Liying Huang |
| author_facet | Yichen Wang Renjin Zheng Pingping Wu Youjia Wu Lingyi Huang Liying Huang |
| author_sort | Yichen Wang |
| collection | DOAJ |
| description | Tetrodotoxin (TTX) inhibits neurotransmission in animals, and there is no specific antidote. In clinical practice in China, <i>Althaea rosea</i> (<i>A. rosea</i> flower) extract has been used to treat TTX poisoning. In this work, the efficacy of the ethyl acetate fraction extract of <i>A. rosea</i> flower in treating TTX poisoning in rats was investigated. A high-performance liquid chromatography–tandem mass spectrometry (LC-MS/MS) method was developed to determine nine neurotransmitters in rat brain tissue, including γ-aminobutyric acid (GABA), dopamine (DA), 5-hydroxytryptamine (5-HT), noradrenaline (NE), 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), 5-hydroxyindole-3-acetic acid (5-HIAA), epinephrine (E), and tyramine (Tyn). The detoxifying effect of <i>A. rosea</i> flower was verified by comparing the changes in neurotransmitters’ content in brain tissue before and after poisoning in rats. The assay was performed in multiple reaction monitoring mode. The quantification method was performed by plotting an internal-standard working curve with good linearity (R<sup>2</sup> > 0.9941) and sensitivity. Analyte recoveries were 94.04–107.53% (RSD < 4.21%). Results indicated that the levels of 5-HT, DA, E, and NE in the brains of TTX-intoxicated rats decreased, whereas the levels of GABA, Tyn, and 5-HIAA showed an opposite trend, and HVA and DOPAC were not detected. The levels of all seven neurotransmitters returned to normal after the gavage administration of ethyl acetate extract of <i>A. rosea</i> flower to prove that the ethyl acetate extract of <i>A. rosea</i> flower had a therapeutic effect on TTX poisoning. The work provided new ideas for studies on TTX detoxification. |
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| language | English |
| last_indexed | 2024-03-11T03:27:10Z |
| publishDate | 2023-05-01 |
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| spelling | doaj.art-2ced7e3259a0493ba6ff9a17dba82a792023-11-18T02:40:06ZengMDPI AGMolecules1420-30492023-05-012810415810.3390/molecules28104158Determination of Multiple Neurotransmitters through LC-MS/MS to Confirm the Therapeutic Effects of <i>Althaea rosea</i> Flower on TTX-Intoxicated RatsYichen Wang0Renjin Zheng1Pingping Wu2Youjia Wu3Lingyi Huang4Liying Huang5School of Pharmacy, Fujian Medical University, Fuzhou 350122, ChinaSchool of Pharmacy, Fujian Medical University, Fuzhou 350122, ChinaSchool of Pharmacy, Fujian Medical University, Fuzhou 350122, ChinaSchool of Pharmacy, Fujian Medical University, Fuzhou 350122, ChinaSchool of Pharmacy, Fujian Medical University, Fuzhou 350122, ChinaSchool of Pharmacy, Fujian Medical University, Fuzhou 350122, ChinaTetrodotoxin (TTX) inhibits neurotransmission in animals, and there is no specific antidote. In clinical practice in China, <i>Althaea rosea</i> (<i>A. rosea</i> flower) extract has been used to treat TTX poisoning. In this work, the efficacy of the ethyl acetate fraction extract of <i>A. rosea</i> flower in treating TTX poisoning in rats was investigated. A high-performance liquid chromatography–tandem mass spectrometry (LC-MS/MS) method was developed to determine nine neurotransmitters in rat brain tissue, including γ-aminobutyric acid (GABA), dopamine (DA), 5-hydroxytryptamine (5-HT), noradrenaline (NE), 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), 5-hydroxyindole-3-acetic acid (5-HIAA), epinephrine (E), and tyramine (Tyn). The detoxifying effect of <i>A. rosea</i> flower was verified by comparing the changes in neurotransmitters’ content in brain tissue before and after poisoning in rats. The assay was performed in multiple reaction monitoring mode. The quantification method was performed by plotting an internal-standard working curve with good linearity (R<sup>2</sup> > 0.9941) and sensitivity. Analyte recoveries were 94.04–107.53% (RSD < 4.21%). Results indicated that the levels of 5-HT, DA, E, and NE in the brains of TTX-intoxicated rats decreased, whereas the levels of GABA, Tyn, and 5-HIAA showed an opposite trend, and HVA and DOPAC were not detected. The levels of all seven neurotransmitters returned to normal after the gavage administration of ethyl acetate extract of <i>A. rosea</i> flower to prove that the ethyl acetate extract of <i>A. rosea</i> flower had a therapeutic effect on TTX poisoning. The work provided new ideas for studies on TTX detoxification.https://www.mdpi.com/1420-3049/28/10/4158tetrodotoxinLC-MS/MS<i>Althaea rosea</i> flowerneurotransmittersdetoxification |
| spellingShingle | Yichen Wang Renjin Zheng Pingping Wu Youjia Wu Lingyi Huang Liying Huang Determination of Multiple Neurotransmitters through LC-MS/MS to Confirm the Therapeutic Effects of <i>Althaea rosea</i> Flower on TTX-Intoxicated Rats Molecules tetrodotoxin LC-MS/MS <i>Althaea rosea</i> flower neurotransmitters detoxification |
| title | Determination of Multiple Neurotransmitters through LC-MS/MS to Confirm the Therapeutic Effects of <i>Althaea rosea</i> Flower on TTX-Intoxicated Rats |
| title_full | Determination of Multiple Neurotransmitters through LC-MS/MS to Confirm the Therapeutic Effects of <i>Althaea rosea</i> Flower on TTX-Intoxicated Rats |
| title_fullStr | Determination of Multiple Neurotransmitters through LC-MS/MS to Confirm the Therapeutic Effects of <i>Althaea rosea</i> Flower on TTX-Intoxicated Rats |
| title_full_unstemmed | Determination of Multiple Neurotransmitters through LC-MS/MS to Confirm the Therapeutic Effects of <i>Althaea rosea</i> Flower on TTX-Intoxicated Rats |
| title_short | Determination of Multiple Neurotransmitters through LC-MS/MS to Confirm the Therapeutic Effects of <i>Althaea rosea</i> Flower on TTX-Intoxicated Rats |
| title_sort | determination of multiple neurotransmitters through lc ms ms to confirm the therapeutic effects of i althaea rosea i flower on ttx intoxicated rats |
| topic | tetrodotoxin LC-MS/MS <i>Althaea rosea</i> flower neurotransmitters detoxification |
| url | https://www.mdpi.com/1420-3049/28/10/4158 |
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