Study On Chemical Constituents And Bio-activities Of Tribulus Terrestris L.

Tribuli fructus is the dry fruit of Tribulus terrestris L.which are plants of Zygophylaceae family.It has many pharmacological functions,such as anti-aging,antibiosis,ease pain,regulating blood lipid,reducing blood sugar,anticancer,diureses,robustness,tyrosinase inhibition.It is often used to treat coronary heart disease,angina,cerebral thrombosis,diabetes,hyperlipemia and myocardial ischemia etc.In this paper,21 monomer compounds?JL-1^JL-21?were isolated from the water extract of Tribuli fructus and 19 of them were identified by 1D-NMR,2D-NMR,MS,IR spectral method combining with the physical and chemical properties,including 3 new compounds and 3 compounds obtained from Tribulus terrestris L.for the first time.The three new compounds are 26-O-?-D-glucopyranosyl-?25R?-5?-furostan-2?,3?,22?,26-tetraol-3-O-?-D-glucopyranosyl-?1?4?-?-D-galactopyranoside?JL-18?,26-O-?-D-glucopyranosyl-?25S?-5?-furostan-2?,3?,22?,26-tetraol-3-O-?-D-glucopyranosyl-?1?4?-?-D-galactopyranoside?JL-19?,26-O-?-D-glucopyranosyl-?25S?-5?-furostan-3?,22?,26-triol-3-O-?-L-rhamnopyranosyl-?1?2?-[?-D-glucopyranosyl-?1?4?]-?-D-galacto pyranoside?JL-21?.The three first isolated compounds are gitogenin-3-O-?-D-glucopyranosyl-?1?4?-?-D-galactopyranoside?JL-6?,manogenin-3-O-?-D-glucopyranosyl-?1?2?-O-?-D-glucopyranosyl-?1?4?-?-D-galactopyranoside?JL-1?,neohecogenin-3-O-?-L-rhamnopyranosyl-?1?2?-[?-D-glucopyranosyl-?1?4?]-?-D-galactopyranoside?JL-9?.On the basis of studying the chemical composition,the paper established a SPE-ELSD-HPLC method to determine 8 kinds of spirostanol saponins JL-1^JL-8.The eight compounds are manogenin-3-O-?-D-glucopyranosyl-?1?2?-O-?-D-glucopyranosyl-?1?4?-?-D-galactopyranoside?JL-1?,hecogenin-3-O-?-D-galactopyra nosyl-?1?2?-[?-D-xylopyranosy?1?3?]-?-D-glucopyranosyl-?1?4?-?-D-galactopyrano side?JL-2?,?25R,S?-hecogenin-3-O-?-D-galactopyranosyl-?1?2?-?-D-glucopyranosyl-?1?4?-?-D-galactopyranoside?JL-3?,hecogenin-3-O-?-D-glucopyranosyl-?1?4?-?-D-galactopyranoside?JL-4?,?25R,S?-gitogenin-3-O-?-D-galactopyranosyl-?1?2?-?-D-glu copyranosyl-?1?4?-?-D-galactopyranoside?JL-5?,gitogenin-3-O-?-D-glucopyranosyl-?1?4?-?-D-galactopyranoside?JL-6?,?25R,S?-Tigogenin-3-O-?-D-glucopyranosyl-?1?4?-[?-L-rhamnopyranosyl-?1?2?]-?-D-galactopyranoside?JL-7?and hecogenin?JL-8?.The optimal extraction method was determined by the extraction of solvent,extraction method,solvent dosage and extraction times,and then the methodology had been explored.Experimental results show that the compound JL-1,JL-2,JL-3,JL-4,JL-7had a good linear relationship with the ranges of 0.40²0.0?g and the compound JL-5,JL-6,JL-8 had a good linear relationship with the ranges of 0.20¹0.0?g.The compound JL-1^JL-8 had a good performance in precision,stability,repeatability and sample recovery rate with the RSD all less than 2%.The method established in this experiment has the advantages of high precision and accuracy.The content determination results indicate that tthe difference in the content of the compound JL-1^JL-8 is not obvious between different regions.Among them,the average contents of JL-1^JL-8 are showed as 0.049 mg·g^-1?JL-1?,0.105 mg·g^-1?JL-2?,0.125mg·g^-1?JL-3?,0.063 mg·g^-1?JL-4?,0.050 mg·g^-1?JL-5?,0.039 mg·g^-1?JL-6?,0.081mg·g^-1?JL-7?,0.013 mg·g^-1?JL-8?,respectively.In this paper,the steroidal saponins of Tribuli fructus was separated into total furostanol saponins and total spirostanol saponins by the method of macroprous adsorption resin.Then a comparative study of two kinds of total steroidal saponins on three bio-activities,including the ability to remove hydroxyl radicals,the ability to inhibit tyrosinase and the proliferation of lung cancer A549 cells,were investigsted.Experimental results show that total furostanol saponins and total spirostanol saponins all had the ability to remove hydroxyl radicals,inhibit tyrosinase and inhibit the proliferation of lung cancer A549 cells.But,theinhibitory ability of total spirostanol saponins was better than total furostanol saponins.From what has been discussed above,in this paper,21 monomer compounds were isolated from the water extract of Tribuli fructus and 19 of them were identified by nuclear magnetic resonance combining with the physical and chemical properties,including 3 new compounds and 3 known compounds obtained from Tribulus terrestris L.for the first time.The paper established a SPE-ELSD-HPLC method to determine 8 kinds of high-content spirostanol saponins JL-1^JL-8.Comparative study of the bioactivities of two steroidal saponins were finished.The results of this paper provide a new scientific basis for the study of the Tribulus terrestris L.chemical constituents,the improvement of the quality standard of Tribulus terrestris L.and the bio-activities of steroidal saponins in the Tribulus terrestris L..