Quality evaluation of Angelica Sinensis Radix dispensing granules by integrating microvascular activity and chemical analysis

As a new form of crude slices of traditional Chinese medicine (TCM), traditional Chinese medicine dispensing granules (TCMDGs) have been used for clinical formula. It is necessary to evaluate whether the chemical composition and biological activity are consistent among the different batches. Angelica Sinensis Radix (ASR), the root of Angelica sinensis (Oliv.) Diels, is one of the most frequently used medicinal materials in gynecology, senile and cardiovascular diseases. In this paper, the quality of TCMDGs is examined taking the Angelica Sinensis Radix dispensing granules (ASRDGs) as a typical case.In this study, integrating bioequivalence and chemical analysis was used to evaluate the quality of dispensing granules taking ASRDGs as a typical case.According to the clinical efficacy of ASR, the intestinal absorption liquid of ASRDGs (IAL-ASRDGs) in 15 batches prepared by the everted gut sac (EGS) method was used to evaluate its effects of vasodilatation on isolated vascular rings. Then, the chemical profiling analysis of IAL-ASRDGs from the 15 batches was carried out by ultra-high performance liquid chromatography coupled with quadrupole-time of flight mass spectrometry (UPLC-Q-TOF-MS). The components in IAL-ASRDGs were identified using mass spectrometry data and the obtained reference standards. Pearson correlation analysis was further performed for the selection of quality control markers based on the extracted ion chromatograms of the identified compounds and vasodilatory activities of different batches of IAL-ASRDGs. Moreover, the selected chemical components in ASRDGs were further verified by vasodilatory activities and quantitatively analyzed by ultra-high performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-QQQ-MS).The IAL-ASRDGs showed favorable vasodilatory activities. There were significant differences among the 15 batches. The ranges of maximum vasodilation rate (%) and EC50 were 69.33 ± 7.16 to 19.52 ± 5.05 and 0.07-25.58 g raw materials/mL, respectively. A total of 46 compounds in IAL-ASRDGs were identified based on accurate mass measurements, fragmentation behavior and the reference standards. Among them, 8 compounds including butylidenephthalide, butylphthalide and senkyunolide A showed positive correlation with vasodilatory effect of IAL-ASRDGs. The 8 compounds were further verified, showing obvious vasodilatory activities. The content of the 8 compounds also showed some differences in 15 batches of ASRDGs. Among them, the content of ferulic acid, senkyunolide I and senkyunolide H varied the most in different batches of ASRDGs. By Pearson correlation analysis, the total content of senkyunolides (senkyunolide A, senkyunolide I and senkyunolide H) in ASRDGs was found to reflect the vasodilatory activity of ASRDGs mostly.This study provides new strategies for the quality assessment by bioequivalence and explore the chemical quality control markers for ASRDGs.