Abstract
Quality-marker (Q-marker) is an emerging concept to ensure the quality and batch-to-batch consistency of Chinese medicine (CM). However, significant difficulties remain in the identification of Q-markers due to the unclear relationship between complex chemical compositions and the pharmacological efficacy of CM. In the present study, we proposed a novel strategy to identify the potential Q-marker of danhong injection (DHI) by an in vivo zebrafish thrombosis model. The anti-thrombotic effects of DHI and its major constituents were evaluated by the zebrafish model of arachidonic acid (AA)-induced thrombosis. The results indicated that DHI can attenuate tail venous thrombus and recover the decrease of heart red blood cell (RBC) intensity in a dose-dependent manner. The result that DHI prevented the formulation of thrombosis in zebrafish was also validated in the zebrafish thrombosis model with green fluorescence protein (GFP)-labeled hemoglobin. The major components of DHI, namely danshen (DS) and honghua (HH), as well as the major chemical constituents of DHI, also exerted anti-thrombotic effects, among which rosmarinic acid (RA) and p-coumaric acid (pCA) showed moderate anti-thrombotic effects. This is the first time that pCA from HH has been found as an active compound exerting an anti-thrombotic effect in a dose-dependent manner, whose IC50 value is approximately 147 μg/mL. By analyzing 10 batches of normal DHI samples and five abnormal samples by high-performance liquid chromatography (HPLC), we found the contents of pCA and RA can be positively correlated to the anti-thrombotic effect of DHI, suggesting that pCA and RA could be potential Q-markers of DHI to ensure batch-to-batch consistency. Our findings illustrated that discovering major active compounds from CM by in vivo pharmacological models can be a useful approach to identifying Q-markers of CM, and in vivo pharmacological models can be a potential tool to evaluate batch-to-batch consistency of CMs.
Highlights
Quality assessment of Chinese medicines (CMs) is an important issue in the modernization of traditional Chinese medicine (TCM), for CMs have been widely applied in clinical therapy in China and other developing countries [1,2]
Xiang [8] combined a metabolomics study with quality control based on gas chromatography-mass spectrometry (GC-MS) and successfully distinguished the samples from different species and ecotypes
In spite of great achievements, current approaches for CM quality assessments have always concentrated on the method of content analysis, which might be unsuitable for CM since CM is a complex multi-component system [10]
Summary
Quality assessment of Chinese medicines (CMs) is an important issue in the modernization of traditional Chinese medicine (TCM), for CMs have been widely applied in clinical therapy in China and other developing countries [1,2]. Various analytical methods have been adopted to ensure the quality. Xie et al [6] developed an approach to evaluate the quality consistency of liuwei dihuang pills produced by different manufacturers based on HPLC-fingerprint and chemometric methods. Xiang [8] combined a metabolomics study with quality control based on gas chromatography-mass spectrometry (GC-MS) and successfully distinguished the samples from different species and ecotypes. Liu [9] proposed that the intestinal permeability assay can be utilized for quality and safety control since intestinal absorption is one of the most important issues for the pharmacokinetic properties of CMs
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