Studies On The Quality Control Of Polygalae Radix And Antidepressant Activity And Pharmacokinetics Of Acvlated Oligosaccharides

Polygalae Radix (PR) has been prescribed for the treatment of insomnia, palpitations with anxiety, restlessness and disorientation for a long time. In the present study, multiple research modes and morden instrument analytical techniques were used to evaluate the quality of PR and the antidepressant effect and pharmacokinetics of its active components. The main innovation and academic contributions of this dissertation are as follow:1. A regioselective approach for preparation of tenuifoliside B (TFSB, a 3’,6-mixed diester of sucrose) using commercial available starting materials was developed, which was successively applied into the synthesis of main 3’-mono ester and 3’,6-diester of sucrose, such as sibiricose A5 (SA5), sibiricose A6 (SA6), tenuifoliside A (TFSA) and tenuifoliside C (TFSC).3’,6-disinapoyl sucrose (DISS) was extracted and prepared by cylic column chromatographic extraction (CCCE) and reversed phase flash chromatography (RPFC). Intergated extraction-adsorption method followed by semi-preparative high performance liquid chromatography (SPHPLC) was used for the preparation of three main acylated pentasaccharides from PR. Finally,9 Polygalae Radix acylated oligosaccharides (PRAOs) were prepared by the methods mentioned above.2. An HPLC-UV method with good linearity, precision and sensitivity was established and applied to the simultaneous quantitation of 8 PRAOs in PR. It could provide much more quality information of PRAOs than any other reported quantitative method.3. An HPLC-UV fingerprint was established, and prediction models for the quantifition of 8 PRAOs in PR were developed by HPLC-UV fingerprint analysis coupled with chemometrics method. The PLSR models were established with correlation coefficient for calibration higher than 0.95 and relative standard error for prediction lower than 5.1%. The proposed method extends the usage of HPLC fingerprint in the quality control of PR.4. A SMLR mathematical model for the prediction of the DPPH elimination activities of PR was established by HPLC-UV fingerprint analysis coupled with chemometrics method with a relative standard error of 3.96%. A rapid and efficient method using DPPH-HPLC/ESI-Q-TOF-MS/MS was used to screen antioxidants in PR for the first time. The identity confirmation was achieved by HPLC-ESI-Q-TOF-MS/MS. With this method, eight constituents were proposed possessing potential antioxidant activity. DISS was proposed to be the best one. The antioxidant activity of DISS was evaluated by DPPH, ABTS radical scavenging assay and ferric-reducing antioxidant power (FRAP) assay in vitro. The results demonstrated that DISS possesses moderate DPPH, ABTS radical scavenging capacity and ferric-reducing antioxidant activity.5. Forced swimming and yohimbine toxicity potentiation test in mice were used to assess the the effects of total acylated oligosaccharide of Polygalae Radix (TAOPR) rich fraction used alone or in combination with TGGR. The results indicated that TAOPR can significantly increase the death rate of mice in yuhimbine potentiation model and decrease the duration of immobility in forced swim test in mice. Concomitant administration may induce a more pronounced antidepressant effect than monotherapy. A serumal metabonomic method based on GC-MS was used to evaluate the antidepressant effect of TAOPR in chronic unpredictable mild stress (CUMS) induced depressed rats. The results obtained suggested that TAOPR may have therapeutic actions on depression-like behavior induced by CUMS in rats possibly by partially regulating the perturbed metabolic pathways of glycolysis metabolism.6. The metabolism of TFSA, TFSB and DISS in rat urine after intraperitoneal admination were investigated by HPLC-ESI-Q-TOF-MS/MS. SA6, an active component isolated from PR, was proved be to the main metalolite of DISS in rat urine after intraperitoneal admination. Further more, an HPLC-UV method was developed for the determination of SA6 in rat plasma. It was successively applied into the pharmacokinetic study of SA6. The nasal bioavailability (4.63%) was 16.8 times higher than oral adimination (0.26%). The present research may provide some references for the rational use of PR in clinic.