Studies On Chemical Constituents Of Spirostanol Saponin Form The Fruits Of Tribulus Terrestris L.

Tribulus terrestris L. is one of the well-known traditional Chinese medicine, the fruit of Tribulus terrestris L. has been used for prevention cardiovascular and cerebrovascular diseases,antibacterial and anti-Aging, it was largely cultivated in most province of. china. The Effective chemical constituents of this plant is Saponins, Flavonoids, Alkaloids and Polysaccharide as reported.This paper describes the isolation, structural determinations and the analysis of the spirostanol saponin from the dried fruit of Tribulus terrestris L. Extraction and SeparationAir-dried and powdered fruits of Tribulus terrestris L.(20 kg) were extracted with 85%EtOH, The EtOH extract was then concentrated, and the residue obtained was suspended in 10 times volume of water. The subsequent suspension was subjected to AB-8 resin column using H2O,45% and 95% EtOH. The solvents of the fraction eluted by 95% EtOH were removed in vacuo. The residue obtained(280g) was subjected to column chromatography on silical gel eluted with EtOH- EtOAc-H2O (2.5:8:0.5) to give 3 frs.(A,B,C). Fr.B was chromatographed on silica gel eluted with EtOH- EtOAc- Petroleum ether- H2O (2.5:5:1:0.5) into frs B-1—B-3, Fr.B-1 was chromatographed on silica gel and eluted with EtOH- EtOAc- Petroleum ether- H2O (2:4:1.5:0.2) to give JA(600mg).and the remaining to give JD(400mg) and JG(30mg) by CHCl3 -MeOH-H2O (4:1:0.1).Fr.B-2 was chromatographed on ODS silica gel using MeOH-H2O (6.5:3.5) into frs B-2-1—B-2-3. Fr.B2-2-2 was chromatographed on silica gel with CHCl3 -MeOH-H2O (3.5:1:0.15) to give JC(150mg) and JI(60mg). Fr.B2-2-3 was chromatographed on silica gel with CHCl3-MeOH-H2O (3:1:0.1) to give JB(300mg),JF(180mg),JE(30mg) and JH(20mg)Compounds JA～JI were identified respectively by IR, MS, and 1DNMR as Terrestrosin D, Tribulosin, Terrestrosin E, Terrestrosin C, Terrestrosin B, Gitonin, Hecogenin 3-O-β-D)-glucopyranosyl(1→4)-β-D-galactopyranoside, Terrestrosin A, F-gitonin.Determination of Terrestrosin D (compounds JA) in the fruits of Tribulus terrestris L. by HPLC-ELSD Instruments and Conditions:HPLC was performed on an Agilent 1200 HPLC instrument and COSMOSIL C18-MS-Ⅱcolumn (4.6×250 mm,5μm) using methanol,Acetonitrile and water as mobile phase in the ratio of 40:10:50 with the flow rate of 0.8 ml/min at 25℃, The gas(air) pressure of ELSD was 2.15 Bar and the temperature of drift tube for measurement was 91℃ResultsⅠ.linearity:The range of the linearity was0.40μg～20.00μg for compound JA, and the correlation coefficients was 0.9991. The regressive equation is y=1.5543x+0.4066 ,r=0.9991.Ⅱ.Precision:Solutions of compound JA was injected into HPLC for six times,the values of RSD as the results of precision tests was 1.11%.(n=6)Ⅲ.Repeatability:The repeatability tests for six samples showed their RSD was 0.68%.(n=6)Ⅳ.Stability:The stability of compound JA was good in ten hours ,because the RSD was 1.08%.Ⅴ.Recovery:The average recovery (n=6) for compound JA was 98.5%, RSD=1.07%(n=6)ⅥContents determination:The samples of. Tribulus terrestris L. from jilin were determined, the average results show the content of compound JA is 0.0310%.The results of this paper can offer new index of content determination for quality standard establishment of Tribulus terrestris L., the results can offer science foundations for the development and utilization of Tribulus terrestris L.