Study On Hemostatic Activity And Chemical Constituents Of Ethyl Acetate Part Of Toddalia Asiatica Root Bark

Objective: To conduct the pharmacognosy identification and hemostatic activity screen of the root bark of Toddalia asiatica of the Rutaceae family, study the chemical constituents and qualitative analysis of acetic ether extraction of impregnation of the 95% ethanol extracts. Methods : 1. Through making horizontal slices of full root of Toddalia asiatica, permeabilizing slides, staining slides of the root bark and the xylem respectively, the tissue section characteristics of full root of Toddalia asiatica was perfected in traditional means of pharmacognostic identification, fine powder microscopic characteristics between the root bark and the root cores of Toddalia asiatica was also investigated. 2. For hemostatic activity of Toddalia asiatica root bark, bleeding time(BT), amount of bleeding(BA) and clotting time(CT) were chose as indexes, the different polarity fractions were studied by the typical mouse tail-cutting metho‘d, glass capillary tube method. 3. Different chromatographic methods and technologies were applied to the isolation and purification of acetic ether extraction of the root bark of Toddalia asiatica, and the structures were elucidated by their physical and chemical properties and spectroscopic techniques. 4. LC-MS/MS were used to analyze the compounds of acetic ether extraction. Rusults:1. Toddalia asiatica root bark contain stone cells, calcium oxalate crystals, cork cells, oil cells, et al. Meanwhile, the xylem respectively have wood fibers, calcium oxalate crystals, textured pit catheters, et al. 2. The ethyl acetate fraction showed the best hemostatic activity than petroleum ether fraction of cold maceration extract. 3. Six compounds were isolated from the acetic ether extraction of the root bark of Toddalia asiatica and elucidated as pimpinellin(1) 、chelerythrine(2) 、isopimpinellin(3)、skimmianine(4)、toddaculine(6)、toddalosin(7). 4. Spectral data of 30 compounds are choosed and the structure of 13 compounds were confired from mass of data. Conclusions:1. Stone cells, oil cells were identify characteristics of Toddalia asiatica root bark, which could be applied as indexes topharmacognostic identification of root bark and full root of Toddalia asiatica. 2. Toddalia asiatica root bark obtained excellent hemostatic and coagulation effects, and ethyl acetate fraction was the best. 3. Six compounds were isolated from the root bark of Toddalia asiatica, conducted compounds of coumarin and alkaloid, which lay the foundation for the deep studies on hemostatic activity of the root bark of Toddalia asiatica. 4. Structure of compounds are confirmed intuitively and accurately by LC-MS/MS data, which provide a great convenience for looking for the active ingredients.