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Determination of Daphnetin and its 8-O-Methylated Metabolite in Rat Plasma by UFLC-MS/MS: Application to a Pharmacokinetic Study

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Abstract

Daphnetin, which has been developed as a drug against obliterative vasculitis, can be rapidly and stereoselectively metabolized to an active 8-O-methylated metabolite, namely daphnetin 8-methyl ether (daphnetin-Me). Herein, a rapid, sensitive and reliable ultrafast liquid chromatography tandem mass spectrometry (UFLC-MS/MS) method was developed and validated to simultaneously determine daphnetin and daphnetin-Me in rat plasma after intragastric administration. The MS quantification for the two analytes and 3-aminocoumarin (internal standard, IS) was carried out on a triple quadrupole mass spectrometer using an ESI source in positive multiple reaction monitoring mode (daphnetin: m/z 179.15 → 51.10; daphnetin-Me: m/z 193.30 → 150.05; IS: m/z 162.00 → 106.20). The method exhibited a broad linear range of 1–2000 ng mL−1. The intra- and inter- assay precisions (RSD%) were ≤ 8.29% with the accuracies (RME%) within ± 5.95%. This newly developed method was successfully applied to a pharmacokinetic study of daphnetin after a single dose of 20 mg kg−1 in rats. Daphnetin and daphnetin-Me peaked almost at the same time. Compared with that of daphnetin, a 2.1-fold higher area under the concentration–time curve (AUC) for daphnetin-Me were observed. These results would be beneficial in facilitating further investigation of pharmacological mechanisms, as well as the rational application of daphnetin and daphnetin-containing herb preparations.

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Abbreviations

COMT:

Catechol-O-methyltransferase

IS:

Internal standard

ESI:

Electrospray ionization

UFLC:

Ultra-fast liquid chromatography

NMR:

Nuclear magnetic resonance

LLOQ:

Lower limits of quantification

MRM:

Multiple reaction monitoring

PK:

Pharmacokinetic

C max :

Maximum concentration

T max :

Time to Cmax

T 1/2 :

Terminal elimination half-life

AUC:

Area under curve

MRT:

Mean residence times

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Acknowledgements

This work was supported by the National Natural Science Foundation (81672458, 82003850), the Grants from the Science and Technology Planning Project of Sichuan Province (2019JDPT0010, 2021JDTD0003), the China Postdoctoral Science Foundation (2020M673290), the Sichuan Province Post Doctoral Fund Special Assistance Program (202027), and the Joint Fund of Luzhou City and Southwest Medical University (2018LZXNYD-ZK18, 2018LZXNYD-ZK28). The authors gratefully acknowledge Prof. Jianming Wu for valuable suggestions, and Mr. Linjie Zhu for research assistance. We thank Dr. Jiangwei Zhang for improving the manuscript.

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  1. Zhongqiong Wang and Chengyi Wang contributed equally to this work.

Authors and Affiliations

  1. Department of Gastroenterology, The Affiliated Hospital of Southwest Medical University, Luzhou, 646000, China

    Zhongqiong Wang, Chengyi Wang, Wei Zhang, Li Liu, Mingming Deng, Muhan Lü & Sicheng Liang

  2. Department of Dermatology, The Affiliated Hospital of Southwest Medical University, Luzhou, 646000, China

    Chengyi Wang & Xiaoyi Qi

  3. The Public Platform of Advanced Detecting Instruments, Public Center of Experimental Technology, Southwest Medical University, Luzhou, 646000, China

    Bing He & Sicheng Liang

  4. The Pharmacy School of Southwest Medical University, Luzhou, 646000, China

    Bing He

  5. Human Microecology and Precision Diagnosis and Treatment of Luzhou Key Laboratory, Luzhou, 646000, China

    Muhan Lü & Sicheng Liang

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  1. Zhongqiong Wang
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Wang, Z., Wang, C., He, B. et al. Determination of Daphnetin and its 8-O-Methylated Metabolite in Rat Plasma by UFLC-MS/MS: Application to a Pharmacokinetic Study. Chromatographia 85, 333–341 (2022). https://doi.org/10.1007/s10337-022-04131-5

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