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Abstract
The Flos Lonicerae-Fructus Forsythiae herb couple is the basic components of Chinese herbal preparations (Shuang-Huang-Lian tablet, Yin-Qiao-Jie-Du tablet and Fufang Qin-Lan oral liquid), and its pharmacological effects were significantly higher than that in Flos Lonicerae or Fructus Forsythiae, but the reasons remained unknown. In the present study, pattern recognition analysis (hierarchical cluster analysis (HCA) and principal component analysis (PCA)) combined with UHPLC-ESI/LTQ-Orbitrap MS system were performed to study the chemical constitution difference between co-decoction and mixed decoction in the term of chemistry. Besides, the pharmacokinetics in vivo and intestinal absorption in vitro combined with pattern recognition analysis were used to reveal the discrepancy between herb couple and single herbs in the view of biology. The observation from the chemical view in vitro showed that there was significant difference in quantity between co-decoction and mixed decoction by HCA, and the exposure level of isoforsythoside and 3, 5-dicaffeoylquinic acid in co-decoction, higher than that in mixed decoction, directly resulted in the discrepancy between co-decoction and mixed decoction using both PCA and HCA. The observation from the pharmacokinetics displayed that the exposure level in vivo of neochlorogenic acid, 3, 4-dicaffeoylquinic acid, isoforsythoside and forsythoside A, higher than that in single herbs, was the main factor contributing to the difference by both PCA and HCA, interestingly consistent with the results obtained from Caco-2 cells in vitro, which indicated that it was because of intestinal absorption improvement of neochlorogenic acid, 3, 4-dicaffeoylquinic acid, isoforsythoside and forsythoside A that resulted in a better efficacy of herb couple than that of single herbs from the perspective of biology. The results above illustrated that caffeic acid derivatives in Flos Lonicerae-Fructus Forsythiae herb couple could be considered as chemical markers for quality control of its preparations.
Highlights
Herbs used together in couples (Yaodui in Chinese) are the basic composition units of Chinese herbal formulas and have special clinical significance in Traditional Chinese Medicine (TCM)
It was shown (Fig. 4a) that there was significant difference in quantity between mixed decoction and co-decoction analyzed by hierarchical cluster analysis (HCA), and we found (Fig. 4b, c) that the most possible ingredients resulted in the discrepancy between co-decoction and mixed decoction were isoforsythoside and 3, 5-dicaffeoylquinic acid analyzed by principal component analysis (PCA) combined with HCA
The systemic biopharmaceutics combined with HCA and PCA were firstly performed to rapidly elucidate the compatibility foundation of FLJ-Fructus Forsythiae (FF) herb couple, and it was found consistently (Fig. 4 & 7) that the caffeic acid derivatives including isoforsythoside, 3, 5-dicaffeoylquinic acid, neochlorogenic acid, 3, 4-dicaffeoylquinic acid and forsythoside A were the most important ingredients resulting in difference between herb couple and single herbs, and interestingly, their exposure levels in the views of chemistry in vitro and biology in vivo were all significantly higher than that in single herbs, highly consistent with the data that the pharmacological activities such as antiinflammatory and antipyretic effects in FLJ-FF herb couple were significantly higher than that in FLJ or FF [17]
Summary
Herbs used together in couples (Yaodui in Chinese) are the basic composition units of Chinese herbal formulas and have special clinical significance in Traditional Chinese Medicine (TCM). There are magnitudes of methods to study the compatibility rule and principles of herb couples to some extent including literature research [2], extraction and separation [3], pharmacological effects [4,5,6], pharmacokinetics in vivo of active components [7] and serum pharmaco-chemistry methods [8], etc. The two herbs are the basic components of Chinese herbal preparations such as Shuang-Huang-Lian oral liquid, Yin-Qiao-Jie-Du tablet, Qin-Re-Jie-Du oral liquid and Fufang Qin-Lan oral liquid, which are extensively used in clinical practice [11], and it was shown by Lin (2008) [12] that the pharmacological effects such as anti-inflammatory and antipyretic effects in FLJ-FF herb couples were significantly stronger than that in FLJ or FF, but the reasons were unknown.
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