Identification of in vitro and in vivo Chemical Constituents of Guipi Anmian Decoction Based on UPLC-Q Exactive Orbitrap HRMS

doi:10.19803/j.1672-8629.20250862

Abstract

Abstract: Objective To quickly identify the main chemical components of Guipi Anmian decoction and those absorbed into the bloodstream and brain based on ultra-high performance liquid chromatography-quadrupole-orbitrap mass spectrometry (UPLC-Q Exactive Orbitrap HRMS) technology. Methods The herbs were decocted in water before Amomum villosum was added and Panax ginseng decocted separately. The filtered residue was extracted with 50% ethanol using ultrasonication. The filtrates were combined to prepare the extract of Guipi Anmian decoction. Eight SPF male SD rats were randomly divided into the control group and administration group, with four rats in each. The rats in the administration group were given the extract of Guipi Anmian decoction by gavage (the crude drug concentration was 15 g·kg^-1), while those in the control group received an equal dose of normal saline. Blood and brain tissue samples were collected after drug administration. Chromatographic separation was performed on a Waters ACQUITY UPLC BEH Shield RP C₁₈column (2.1 mm×100 mm, 1.7 μm). The mobile phase composed of acetonitrile (A) and water containing 0.1% formic acid aqueous solution (B) was used under gradient elution. As mass spectrometry, a full-scan mode with positive/negative ion switching and data-dependent secondary mass spectrometry scanning (Full MS/dd MS²) was adopted to acquire data so that precise primary information on mass spectrometry and abundant secondary fragmental data on mass spectrometry were collected. Combined with the reference standard retention times, PubChem database entries, and related literature, the main components in the extract and the prototype components in the plasma or brain were identified. Results A total of 158 chemical constituents were identified from GuipiAnmian decoction, including 55 terpenes, 34 flavonoids, 15 tannins, 19 phenylpropanoids, 8 alkaloids, 5 phenolic acids, 5 amino-acids, 4 nucleotides and 13 other compounds. Twenty-five of the prototype constituents were detected in the plasma and 29 in the brain. Conclusion A method for rapid multicomponent identification of GuipiAnmian decoction in vitro and in vivo has been established in this study, which is expected to contribute to research on effective substances and quality control.

Key words: Guipi Anmian Decoction, Phyllanthus emblica, Panax ginseng, Constituents Absorbed into Plasma, Constituents Absorbed into the Brain, UPLC-Q Exactive Orbitrap HRMS, Rats

CLC Number:

ZHENG Haiyun, WANG Ruifen, XIONG Jie, ZHAO Fei, WANG Bin, YU Wen. Identification of in vitro and in vivo Chemical Constituents of Guipi Anmian Decoction Based on UPLC-Q Exactive Orbitrap HRMS[J]. Chinese Journal of Pharmacovigilance, 2026, 23(4): 414-419.

Add to citation manager EndNote|Ris|BibTeX

URL: https://zgywjj.magtechjournal.com/EN/10.19803/j.1672-8629.20250862

https://zgywjj.magtechjournal.com/EN/Y2026/V23/I4/414

References

[1] YUAN Y, NIU D, PANG XX, et al.Research Progress on Analytical Techniques for the Material Basis of Efficacy in Traditional Chinese Medicine (TCM) Compound Prescriptions[J]. China Pharmacy(中国药房), 2025, 36(21): 2738-2741.
[2] ZHAO LR, LI J, ZHANG YJ, et al.Research Progress on Methods for Investigating the Material Basis of Efficacy in Traditional Chinese Medicine Based on in vivo Processes[J]. China Medicine and Pharmacy(中国医药科学), 2022, 12(23): 39-42.
[3] OUYANG LF, ZHU KX, SHAO X, et al.Research Progress on Intracranial Small Molecule Detection Technology and Ginsenoside Brain Entry[J]. World Science and Technology-Modernization of Traditional Chinese Medicine(世界科学技术-中医药现代化), 2016, 18(9): 1567-1574.
[4] MENG H, XU LT, LI PP, et al. Exploring the Active Substances and Mechanism of Physalis Calyx Seu Fructus Against Inflammation-related Respiratory Diseases via Integrating Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry, Network Pharmacological Analysis,Experimental Verifications[J/OL]. Chem Biodiver, (2025-11-25)[2025-12-12]. https://doi.org/10.1002/cbdv.202500496.
[5] ZHOU J, WANG R, HOU J, et al.Investigation of Antithrombotic Activity and in vivo Effective Forms of Kaempferitrin Using FeCl(3)-Induced Rat Arterial Thrombosis and UHPLC-Q-Exactive Orbitrap MS[J]. Molecules, 2025, 30(22): 4434.
[6] LI Y, LI Y, YANG K, et al. Screening of ERbeta-Targeted Antipos-tmenopausal Osteoporosis Chemical Constituents from Cornus Officinalis Based on Affinity Ultrafiltration and UHPLC-Q-Exactive Orbitrap MS[J/OL]. Biomed Chromatogr, (2025-11-28)[ 2025-12-12]. https://doi.org/10.1002/bmc.70270.
[7] KONG YM, LIANG LY, CAO JL, et al. Research Progress on Modern Applications and Pharmacological Effects of the Classical Formula Guipi Decoction, with Predictive Analysis of Its Potential Quality Markers[J/OL]. Chinese Journal of Traditional Chinese Medicine and Pharmacy(中华中医药学刊), (2025-07-17)[2025-12-12]. https://link.cnki.net/urlid/21.1546.R.20250716.1649.014.
[8] ZHENG HY, WANG B, ZHAI YS, et al.Study on the Sleep-Improving Function of a Phyllanthus emblica and Lycium barbarum Compound Formula in Mice[J]. Journal of Food Safety and Quality(食品安全质量检测学报), 2025, 16(3): 208-213.
[9] LIU JB, ZHANG XY, YANG SS, et al.Rapid Identification of Characteristic Chemical Constituents of Panax ginseng, Panax quinquefolius, and Panax japonicus Using UPLC-Q-TOF/MS[J]. J Anal Methods Chem, 2022: 6463770.
[10] GUAN P, LIU W, CAO Y, et al.Full Collision Energy Ramp-MS² Spectrum in Structural Analysis Relying on MS/MS[J]. Anal Chem, 2021, 93(46): 15381-15389.
[11] BALKRISHNA A, VERMA S, MPR, et al. Comprehensive Phytochemical Profiling of Phyllanthus emblica L. Flowers on UHPLC/MS Quadrupole Time of Flight, HPTLC, HPLC, and NMR Analytical Platforms Reveals Functional Metabolites with Potent Anti-Inflammatory Effects in Human (THP-1) Macrophages[J]. Phytochem Anal, 2025, 36(1): 218-233.
[12] WANG H, TANG L, HU S, et al.Chemical Profiling of Shengmai Injection, Tissue Distribution and Pharmacokinetic Characteristics of Ginsenosides after Intravenous Dosing Shengmai Injection in Rats with Cerebral Ischemia[J]. J Ethnopharmacol, 2024, 319(Pt1):117119.
[13] CHEN XW, SNEED KB, PAN SY, et al.Herb-Drug Interactions and Mechanistic and Clinical Considerations[J]. Curr Drug Metab, 2012, 13(5): 640-651.
[14] MOINUDDIN G, DEVI K, KUMAR KHAJURIA D.Evaluation of the Anti-Depressant Activity of Myristica Fragrans (Nutmeg) in Male Rats[J]. Avicenna J Phytomed, 2012, 2(2): 72-78.
[15] YANG J, XU Y, HU P, et al.Exploring the Mechanism of Action of Huoermai Essential Oil for Plateau Insomnia Based on the CAMP/CREB/BDNF/Gabaergic Pathway[J]. J Ethnopharmacol, 2025, 338(Pt2): 119092.
[16] DONG JJ, WANG X, GONG JY.Experimental Study on the Sleep-Promoting Effects of Ginseng[J]. Biotechnology World(生物技术世界), 2015, (6): 154.
[17] LU ZH, ZHENG QY, ZHANG X, et al.Research Progress on the Regulation of Neurotransmitter Metabolism by Panax ginseng[J]. Chinese Traditional and Herbal Drugs(中草药), 2023, 54(21): 7260-7272.
[18] LI J, WANG X, ZHANG Y, et al.Ginsenoside Rg₁ Alleviates PCPA-Induced Insomnia by Inhibiting NLRP3 Inflammasome Activation and Pyroptosis through the Nrf2/HO-1 Pathway in Mice[J]. Psychop-harmacology(Berl), 2025, 242(12): 2739-2755.
[19] JIN C, QIAO J, GONG H, et al.Ginsenoside Re Binds Drp1 Leu94 across Species to Restore Mitochondrial Homeostasis via Atg1/ULK1-Dependent Fission-Mitophagy Coupling in Age-Related Degeneration[J]. Pharmacol Res, 2025, 222: 108043.
[20] LEE HS, YANG EJ, LEE T, et al.Laccase Fermentation of Clove Extract Increases Content of Dehydrodieugenol, Which Has Neuropro-tective Activity against Glutamate Toxicity in HT22 Cells[J]. J Microbiol Biotechnol, 2018, 28(2): 246-254.