Studies On Chemical Constituents Of Astilboides Tabularis (Hemsl.) Engler

"Shan Heye", the whole plant of Astilboides tabularis (Hemsl.) Engler,is widely used for treatment of diarrhea. So far, there are not any researches in chemical constituents and the indexes of content determination about this plant, For the sake of the better use of this herb medicine, this paper described the extraction, isolation, identification of the chemical components and established a method to determine the content of one typical chemical component in the leaves of Astilboides tabularis (Hemsl.) Engler.Extraction , Separation and identification.1.1 ExtractionDried leaves 20kg of Astilboides tabularis (Hemsl.) Engler were extracted by heat reflux in Ethanol for three times (2h, 1.5h and 1h, respectively), the solvent volume were 30L, 25L and 20 L respectively, the ethanol was evaporated in vacuum and concentrated to cream(1160g).1.2 SeparationMix the cream with appropriate amount silica gel .and the mixture was subjected to silica gel column chromatography and eluted by different mobile phase to afford compound 1(112mg),compound 2(109mg),compound 3(30.7mg),compound 4(124mg),compound 5(90mg),compound 6(97mg),compound 7(32mg),compound 8 (83mg)。1.3 IdentificationCompounds 1～8 were identified respectively by MS and NMR as quercitrin ,quercetin-3-O-β-D-galactoside , kaempferol-3-O-α-L- rhamnoside , gallic acid ,bergenin, quercetin,β-sitosterol and daucosterol,The content determination1:Apparatus and ReagentApparatus:Agilent 1200 HPLC instrumentColumn :Agilent HC-C8 4.6×150mm ODS 5μmReagent:Methanol(AR), Purified Water2:Methods and Results 2.1:experimental conditionMobile phase: Methanol: water=35:65Flow rate: 1.0mL?min-1Wavelength: 360 nm2.2:Results2.2.1 Linear RangeThe linear range of quercitrin is 0.1μg～10.0μg.Y=16.17×102X-53.365 r=0.99992.2.2 Precision testQuercitrin solution was injected into HPLC for six times, calculated the RSD of peak area , RSD= 0.76 % (n=6).2.2.3 Repeatability testPrepare six sample solution for the repeatability tests, and each solution was injected into HPLC for two times, calculated the RSD of peak area, RSD = 1.63 % (n=6).2.2.4 DetectabilityWhen the concentration of compound 1 solution was descreased to 0.0002μg·mL-1 and determined by HPLC, S/N≥3, so the detectability was 0.001μg.2.2.5 StabilityThe test of stability showed the quercitrin solution is stable in ten hours. The RSD of the peak area was 0.64% .2.2.6 Recovery testThe average recovery of quercitrin was 102.0% and the RSD of the peak area was no more than 2%。2.2.7 The determination of the content of quercitrin.Prepare three sample solution for the repeatability tests, and each solution was injected into HPLC for two times, calculated the content of quercitrin.The results of this paper can provide a new index of content determination for establishment of quality standards of the leaves of Astilboides tabularis (Hemsl.) Engler, and also can offer science foundation for the development and research of this medicinal plant.