Abstract
Hawthorn leaves, officially listed in the 2015 Chinese Pharmacopoeia, comes from Crataegus pinnatifida Bge. var. major N. E. Br. and Crataegus pinnatifida Bge. It has the effect of invigorating blood and dissolving stasis, regulating qi and dredging veins, turbid, and lipid-lowering. This work was designed to explore the spectrum–effect relationship between high-performance liquid chromatography (HPLC) fingerprint and the invigorating blood and dissolving stasis effect of hawthorn leaves. Four extracts from hawthorn leaves, 50% ethanol extract (EE), macroporous resin extract (MRE), ethyl acetate extract (EAE), and n-butanol extract (BAE) were administered to rats at three doses of 0.4 g·kg−1, 0.6 g·kg−1, and 0.8 g·kg−1 (crude medicine weight/rat weight), respectively. Twenty-seven indexes were obtained and analyzed by principal component analysis (PCA) and one-way ANOVA. The spectrum–effect relationship was constructed by multiple linear regression (MLR) analysis. With the above separation method, HPLC fingerprints of 12 batches (three concentrations of four extracts) of hawthorn leaves extracts were obtained, and 32 common peaks were marked by similarity analysis. Additionally, 9 indexes of fibrinogen (FIB), erythrocyte total (ET), platelet count (PC), mean platelet volume (MPV), plateletcrit (PCT), whole blood viscosity-WBV(LSR-20S−1), WBV(LSR-60S−1), WBV(LSR-150S−1), and plasma viscosity (PV) were selected as the relevant indexes with pertinence and statistical significance (p < 0.05) on invigorating blood and dissolving stasis, and taken for the subsequent spectrum–effect experiments. Finally, the results of spectrum–effect relationships between 32 common peaks and 9 indexes showed the main peaks responsible for invigorating blood and dissolving stasis were 1–6, 8–11, 13–15, 17, 19, 21, 23, and 27–32. Compared with the standards and other references, peaks 3, 14, 17, and 19 were, respectively, identified as chlorogenic acid, vitexin glycoside, vitexin, and rutin. This work was helpful to the discovery of the active substance and quality control of hawthorn leaves.
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Acknowledgements
This work were supported by the Key Project of Natural Science Research Program of Chengde Medical University (Grant Number 201811), the Key Discipline Construction Projects of Higher School (Grant Number 2013.4), and Science and Technology Research Key Project of Higher School in Hebei Province, China (Grant Numbers ZD2015097 and QN2017005). The funding bodies played no role in study design, data acquisition, analysis and interpretation, or manuscript preparation and submission.
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Xu, B., Gao, J., Zhao, S. et al. The Spectrum–Effect Relationship Between HPLC Fingerprint and the Invigorating Blood and Dissolving Stasis Effect of Hawthorn Leaves. Chromatographia 83, 409–421 (2020). https://doi.org/10.1007/s10337-020-03861-8
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DOI: https://doi.org/10.1007/s10337-020-03861-8