Simultaneous determination of harpagoside and cinnamic acid in rat plasma by high-performance liquid chromatography: application to a pharmacokinetic study |
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Authors: | Peifan Li Yunhui Zhang Li Xiao Xinghua Jin Kun Yang |
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Institution: | (1) School of Pharmaceutical Science and Technology, Tianjin University, No. 92, Weijin Road, 300072 Tianjin, People’s Republic of China;(2) Analysis and Testing Center, Tianjin Medical University, No. 22, Qixiangtai Road, 300070 Tianjin, People’s Republic of China |
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Abstract: | Radix Scrophulariae (Xuanshen) is one of the famous Chinese herbal medicines widely used to treat rheumatism, tussis, pharyngalgia, arthritis,
constipation, and conjunctival congestion. Harpagoside and cinnamic acid are the main bioactive components of Xuanshen. The
purpose of this study was to develop an HPLC–UV method for simultaneous determination of harpagoside and cinnamic acid in
rat plasma and investigate pharmacokinetic parameters of harpagoside and cinnamic acid after oral administration of Xuanshen
extract (760 mg kg−1). After addition of syringin as internal standard, the analytes were isolated from plasma by liquid–liquid extraction. Separation
was achieved on a Kromasil C18 column, and detection was by UV absorption at 272 nm. The described assay was validated in terms of linearity, accuracy,
precision, recovery, and limit of quantification according to the FDA validation guidelines. Calibration curves for both analytes
were linear with the coefficient of variation (r) for both was greater than 0.999. Accuracy for harpagoside and cinnamic acid ranged from 100.7–103.5% and 96.9–102.9%, respectively,
and precision for both analytes were less than 8.5%. The main pharmacokinetic parameters found for harpagoside and cinnamic
acid after oral infusion of Xuanshen extract were as follows: C
max 1488.7 ± 205.9 and 556.8 ± 94.2 ng mL−1, T
max 2.09 ± 0.31 and (1.48 ± 0.14 h, AUC0–24 10336.4 ± 1426.8 and 3653.1 ± 456.4 ng h mL−1, 11276.8 ± 1321.4 and 3704.5 ± 398.8 ng h mL−1, and t
1/2 4.9 ± 1.3 and 2.5 ± 0.9 h, respectively. These results indicated that the proposed method is simple, selective, and feasible
for pharmacokinetic study of Radix Scrophulariae extract in rats.
Figure Radix Scrophulariae |
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Keywords: | Radix Scrophulariae (Xuanshen) extract Harpagoside Cinnamic acid HPLC– UV Pharmacokinetics |
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