RESEARCH ARTICLE
Simultaneous determination of three flavonoids and one
coumarin by LC–MS/MS: Application to a comparative
pharmacokinetic study in normal and arthritic rats after oral
administration of Daphne genkwa extract
Luyi Zhou
|
Jing Li
|
Chong Yan
School of Pharmacy, Guangdong Medical
University, Dongguan, China
Correspondence
Luyi Zhou, School of Pharmacy, Guangdong
Medical University, Dongguan, China.
Email: zly235235@outlook.com
Funding information
National Nature Science Foundation of China,
Grant/Award Number: 81303156; Program
for Outstanding Young Teachers in Higher
Education Commission of Guangdong Prov-
ince of China, Grant/Award Number:
Yq2013085; Social Development Project of
Dongguan City, Grant/Award Number:
2014108101056
Abstract
A selective and sensitive liquid chromatography–tandem mass spectrometry method
was developed and validated for investigating the pharmacokinetics of umbelliferone,
apigenin, genkwanin and hydroxygenkwanin after oral administration of Daphne
genkwa extract. Plasma samples were treated by protein precipitation with acetoni-
trile. Analytes were detected by triple‐quadrupole MS/MS with an ESI source in
negative selection reaction monitoring mode. The transitions of m/z 161 → 133 for
umbelliferone, m/z 269 → 117 for apigenin, m/z 283 → 268 for genkwanin and m/z
299 → 284 for hydroxygenkwanin were confirmed for quantification. Chromato-
graphic separation was conducted using an Eclipse XDB‐C
18
column, and the applied
isocratic elution program allowed for simultaneous determination of the four analytes
for a total run time of 2.5 min. The linearity was validated over the plasma concentra-
tion ranges of 1.421–1421 ng/mL for umbelliferone, 0.845–845 ng/mL for apigenin,
1.025–1025 ng/mL for genkwanin and 0.845–845 ng/mL for hydroxygenkwanin.
The extraction recovery rate was >82.7% for each analyte. No apparent matrix effect
was observed during the bioanalysis. After full validation, the proposed method was
successfully applied to compare the pharmacokinetics of these analytes between
normal and arthritic rats.
KEYWORDS
comparative pharmacokinetics, Daphne genkwa,LC–MS/MS, rat plasma
1
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INTRODUCTION
The dried flowers of Daphne genkwa, known as Yuanhua in traditional
Chinese medicine, have been used for treatment of edema, asthma,
cough and cancer in China for thousands of years (Jiangsu New
Medical College, 1985; State Commission of Chinese Pharmacopoeia,
2015). Pharmacological studies on rheumatoid arthritis have shown
that the extract of D. genkwa significantly decreased arthritis in a
rat model through antioxidant, anti‐inflammatory and
hemorheological modulatory mechanisms (Jiang et al., 2014; Zhang
et al., 2014). The chemical components of D. genkwa are coumarins,
flavonoids, lignans and diterpenes (Hong, Chung, Lee, Park, & Lee,
2011; Huang et al., 2012). In particular, flavones, such as apigenin,
genkwanin and hydroxygenkwanin, are the main effective compo-
nents of this plant (Kim et al., 2014; Uyangaa et al., 2015). The phar-
macological activities of these three flavones have been reported.
Apigenin possesses anti‐inflammatory, anti‐oxidant and anticancer
activities (Gaballah, Gaber, & Mohamed, 2017; Patil et al., 2016; Xu
et al., 2016). Genkwanin shows significant anti‐tumor activity by
enhancing host immunity and decreasing inflammatory cytokine levels
Abbreviations: CFA, complete Freund's adjuvant.
Received: 20 July 2017 Revised: 12 January 2018 Accepted: 22 February 2018
DOI: 10.1002/bmc.4233
Biomedical Chromatography. 2018;32:e4233.
https://doi.org/10.1002/bmc.4233
Copyright © 2018 John Wiley & Sons, Ltd.wileyonlinelibrary.com/journal/bmc 1of10
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