| Chapter one, literature study.A synthetical review was given to the ancient and present research of Danggui-Baishao herb couple from home and abroad.Chapter two, study of chemical characteristics of Danggui-Baishao compatibilityMultiple techniques of UPLC-Q-TOF/MS and pattern recognition were applied to analyze the chemical characteristics of Danggui-Baishao herb couple of anterior-posterior compatibility and identify the main components in this herb couple in the case of the lack of reference substances to explore the main active ingredient variation before and after compatibility. By comparison with the reference and MS, we identified a total of 15 major chemical constituents in Danggui-Baishao herb couple. The OPLS-DA result showed that catechin, caffeic acid, gallic acid, paeoniflorin and benzoylpaeoniflorin were found to be the differentiating components between before and after compatibility. The compatibility of the two herbs could increase the content of catechin, caffeic acid, and benzoylpaeoniflorin and decrease the content of gallic acid and paeoniflorin. There were no new components detected after compatibility.Chapter three, the study of the characteristics of efficacy preference of the compatibility of Danggui-Baishao herb couple.Section one, the compatibility study of the blood nourishing efficacy of Danggui-Baishao herb couple.Based on the blood nourishing efficacy preference of Danggui-Baishao herb couple, the blood-deficiency mouse model induced by cyclophosphamide and acetylphenyhydrazine was established to study the blood nourishing efficacy under different proportion of Danggui-Baishao herb couple. We designed compatibility proportion by increase-decrease design method of pxperiment with base line geometric proportion. Periphera hemogram, immune organ index and BRC ATPase activity were made statistics based on PCA. The scores of PCA showed that 1:3-3:1 proportions of Danggui-Baishao hern couple were closer to the normol group than other proportions.Section two, the mechanism of blood nourishing efficacy of Danggui-Baishao herb couple based on the metabolomics study.UPLC-Q-TOF/MS and MarkerLynx software based on multivariate statistics were applied to explore the endogenous metabolism of blood-deficiency mouse after intragastric administration of Danggui-Baishao herb couple. Ten bio-markers such as phenylalanine, tryptophan, cholic acid, LysoPC and so on were identified and could be considered as the potential markers of the blood-deficiency model. The result of IPA analysed showed that Danggui-Baishao achieve the blood nourishing efficacy mainly by regulating the amino acid metabolism, energy metabolism and lipid metabolism.Chapter four, the study of the physiological disposition of the compatibility of Danggui-Baishao herb couple.Section one, the pharmacokinetics study of major components in rats after intragastric administration of different combination of Danggui-Baishao herb couple.In our study, ferulic acid, caffeic acid, vanillic acid, ligustilide, peoniflorin, albiflorin and oxypaeoniflorin were selected as detection index. We established a method of simultaneous determination of these seven ingredients rat plasma by UPLC-MS/MS and applied to the pharmacokinetic comparative study in the rats after intragastric administration of Radix Angelicae Sinensis (RAS), Radix Paeoniae Alba (RPA) and their herb couple (RR) and provide some scientific basis of the compatibility of Danggui-Baishao herb couple. The result showed that phenolic acids such as ferulic acid, caffeic acid and vanillic acid and ligustilide had different pharmacokinetic characteristics in RAS and RR herb couple group. RAS combined with RPA could decrease extent of absorption of ferulic acid and caffeic acid, delay the elimination of vanillic acid and promote the elimination of ligustilide in the body. Meanwhile, RAS combined with RPA could increase the absorption rate of peoniflorin, albiflorin and oxypaeoniflorin.Section two, the tissue distribution study of major components in rats after intragastric administration of different combination of Danggui-Baishao herb couple.UPLC-TQ/MS with TargetLynx software were applied to analyze the influence on the tissues distribution of ferulic acid, vanillic acid, peoniflorin, albiflorin and oxypaeoniflorin in different compatibility of Danggui-Baishao herb couple. The result showed that ferulic acid and vanillic acid distributed mainly in the kidney and liver in rat body. The distribution content of ferulic acid in rat heart, liver, lung and kidney was significantly increased at 5 min of herb couple compared with the RAS group. The distribution content of vanillic acid in rat liver, lung and kidney at 5 min and 60 min was significantly increased in the RR herb couple group compared with the RAS group. Peoniflorin, albiflorin and oxypaeoniflorin distributed mainly in liver, lung and kidney. The distribution content of these three components was significantly decreased in liver and kidney after 20 min oral administration of RR herb couple group compared with the RPA single group. Accordingly, the compatibility of RAS and RPA could promote the tissue distribution of active constituent in RAS.Section three, the metabolism study of major components in rats after intragastric administration of Danggui-Baishao herb couple.UPLC-Q-TOF/MS and MetaboLynx software based on mass defect filter (MDF) were applied to analyze the metabolites in the urine, plasma, bile and feces of normal rats after intragastric administration of different combination of Danggui-Baishao herb couple. Eight metabolites of ferulic acid such as ferulic acid sulfated, ferulic glucuronated and so on; four metabolites of vanillic acid such as vanillic acid sulfated,2×Methylation metabolites and so on. Peoniflorin and albiflorin existed in parent most and mainly eliminated from feces. Compared with the RAS group, the peak area of each metabolites of ferulic acid and vanillic acid in the herb couple group was significantly lower than the RAS group. Similarly, the peak area of each metabolite of peoniflorin and albiflorin in the herb couple group was significantly lower than the RPA group. |
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