Study On The Primary Efficacy Material In Prepared Rhubarb On Promoting Blood Circulation And Removing Blood Stasis By Spectrum-Activity Relationship Analysis

This study conducted a systematic research on material basis of prepared rhubarb obtained from raw rhubarb.The entire study was undertook from chemical compounds differences between decoction of raw and prepared rhubarb and the spectrum-effect relationship of prepared rhubarb respectively to explore the primary efficacy material in prepared rhubarb on promoting blood circulation and removing blood stasis.First of all,10 batches of raw rhubarbs were collected from different origins all over the country.All these samples collected needed to undergo a strict quality evaluation according to the Chinese pharmacopoeia 2015 edition-part one and the best quality sample was selected to make prepared rhubarb according to the optimal processing techniques found from literature.The prepared rhubarbs obtained were used for further experiments.By using UPLC-Q-TOF-MS method coupled with automatic analysis software Peakview,decoctions of 18 batches of raw rhubarb and prepared rhubarb(9 batches for each)were detected,from which 45 chemical compounds were identified totally.All of the data were visualized using principal component analysis(PCA)and t-test to check for outliers and variation.16 compounds were screened out to be the main contributors for the significant differences between raw rhubarb and prepared rhubarb.The intensity of rhein in prepared rhubarb was found to be higher than in raw rhubarb.It was found that the therapeutic effects on promoting blood circulation and removing blood stasis of prepared rhubarb were superior to raw rhubarb by comparative study on the pharmacology.The result verified that prepared rhubarb shows quite a different curative effect from raw rhubarb after processing,and its effect of promoting blood circulation and removing blood stasis was reinforced.Basing on spectrum-effect relationship analysis method,the blood of rats in different groups were collected at different time points to detect pharmacodynamics indexes and plotted the efficacy-time curves.Besides,UPLC-Q-TOF-MS method was utilized to detect the rat plasma.19 compounds were eventually identified,among which 9 were the original form of compounds in prepared rhubarb and the other 10 were the metabolites.According to the results of UPLC-Q-TOF-MS detection,the intensity-time curves of all these 19 compounds were also plotted.Gray correlation analysis method was employed to calculate the correlation grades between efficacy-time curves and intensity-time curves of all 19 compounds to reflect the relationship between chemical compounds in vivo and drug efficacy.Rhein glucuronide was eventually found possessing the highest possibility to be the main active ingredient for treatment of noxious heat with blood stasis syndrome.