| Investigation on the principle combination of couple herbs is the key to elucidate the combing foundation of complex prescription, and also the key researching area supported in the 12th five years proposed project by Chinese governments. Pharmacological effects method, high throughput screening technologies and the generalized methods in clinic are all the valid ways to study the combing rules of couple herbs in previous publications. The above methods also contributed much to elucidating foundational substance of the rules. However, up to date, the investigation on combing rules is still a hot topic of the above researching area. In this dissertation, the couple herbs of Salvia milltiorrhiza and borneol were selected as the example. Moreover, the construction of the researching strategy on combing rules of couple herbs was processed too, based on using "Jun-Shi compound" as the molecular probes, in this dissertation. Those researches would provide method and strategy to elucidate the combing rules of couple rules. The contents of this dissertation consist of five chapters, and the important contributions of the author are as follows:1. A novel high performance liquid chromatography-tandem mass spectrometry method for the determination of caffeic acid in rat plasma and tissue was established and applied in its pharmacokinetic study and tissue distribution after administration of Salvia milltiorrhiza. The results showed that the detection limits of caffeic acid in plasma and tissue were 0.12 and 0.05 ng·mL-1. The calibration curves of caffeic acid in plasma and tissue had good linearity over the range concentration of 0.01～10.0μg·mL-1 and 0.005～5.0μg·mL-1. With regards to recovery, precision, and stability, the results of the methodological evaluation showed the relative standard deviation meet the requirements of FDA and SFDA biological samples determination. The proposed method was also applied in the investigation on pharcokinetic and tissue distribution of caffeic acid in rat. The statistical results indicated that the plasma concentration-time course of caffeic acid in rat confirmed to a 2-compartment open model and the its tissue distribution was of ckidney>cheart>cliver>clung>cbrain>cspleen, demonstrating that the method can be applied in studying the clinical and non-clinical pharmacokinetic of caffeic acid.2. The effects of borneol on the pharmacokinetic and tissue distribution of caffeic acid after admistrated with couple herbs of Salvia milltiorrhiza and borneol was carried out by the above method. The results showed the pharmacokinetic parameters of t1/2α, t1/2βV1/F, AUC(0-t), AUC(0-∞) and Ka in rat increased significantly, while the parameters of CL/F, K10, K12, K21 and t1/2Ka decreased obviously as compared with administration of Salvia milltiorrhiza. Those results indicated that caffeic acid had longer therapeutic time and stronger transfer capacity from blood into tissue in rat. The study of tissue distribution showed more caffeic acid distributed in heart and brain. Borneol enhanced the targeting power and affinity of caffeic acid to heart and brain, which was consisted with the illustration and definition of shi herb in Chinese medical works.3. The "Jun-shi compound"—bornyl caffeiate was designed and synthesed with using the bioactive compound of Salvia milltiorrhiza and borneol as the materials. Then, its pharmacokinetic and tissue distribution were performed. Compared with the same dose of Salvia milltiorrhiza and caffeic acid, bornyl caffeinate increased AUC(0-t) and AUC(0-∞) of its metabolic product(caffeic acid) greatly, which indicated more caffeic acid could be absorbed into blood. Compared with administration of caffeic acid combined with borneol, bornyl caffeinate could increase the bioavailability of caffeic acid. Caffeic acid concentrated in heart and brain reflected the interacting characteristics of Jun herb (Salvia milltiorrhiza) and Shi herb (borneol) in rat. All those results indicate that bornyl caffeinate can be selected as the molecular probe to investigate the principle combination of couple herbs, and provide the method and strategy for the research of this area.
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