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KMID : 1146920200500020159
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Journal of Pharmaceutical Investigation
2020 Volume.50 No. 2 p.159 ~ p.172
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A sensitive UPLC?ESI?MS/MS method for the quantification of cinnamic acid in vivo and in vitro: application to pharmacokinetic and protein binding study in human plasma
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Jeong Seung-Hyun
Jang Ji-Hun
Cho Hea-Young
Oh In-Joon
Lee Yong-Bok
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Abstract
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Purpose: The purpose of this study was to develop a sensitive method for quantifying cinnamic acid in human plasma using UPLC?ESI?MS/MS.
Methods: Cinnamic acid was separated using water containing 0.005% (v/v) formic acid and acetonitrile as a mobile phase by gradient elution at a flow rate 0.2 mL/min, equipped with a HALO-C18 column (2.1?×?100 mm, 2.7 μm). Quantitation of this analysis was performed on a triple quadrupole mass spectrometer employing electrospray ionization technique, operating in multiple reaction monitoring negative ion mode. The mass transitions were m/z 146.8?→?103.0 for cinnamic acid, and 432.9?→?225.0 for geniposide as internal standard. Liquid?liquid extraction and protein precipitation with ethyl acetate?methanol-1.2% acetic acid (4/16/1, v/v/v) were used in the sample extraction.
Results: The chromatograms showed high resolution, sensitivity, and selectivity with no interference with plasma constituents. The calibration curves for cinnamic acid in human plasma were 0.1?500 ng/mL and displayed excellent linearity with correlation coefficients (r) greater than 0.99. Both the intra- and inter-day precisions (CV%) were within 3.88% for human plasma. The accuracy was 99.34?106.69% for human plasma. In vitro plasma protein binding of cinnamic acid was 64.26?±?1.89 and 65.50?±?1.78% for the spiked human plasma concentrations of 100 and 1000 ng/mL, respectively.
Conclusion: The developed analytical method satisfied the criteria of international guidance and could be successfully applied to the pharmacokinetic study of cinnamic acid after oral administration of Socheongryong-tang tablets to humans.
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KEYWORD
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Cinnamic acid, UPLC?ESI?MS/MS, Socheongryong-tang, Pharmacokinetics, Plasma protein binding
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