Studies On Chemical Constituents Of Sinopodophyllum Emodi Wall

Studies on Chemical Constituents of Sinopodophyllum emodi Wall.Sinopodophyllum emodi Wall. named Taoerqi and Jisutai as well,is a rare perennial herbaceous plant, whose roots and fruits are used as a medicine in traditional Chinese medicine. A series of chemical investigations revealed that Sinopodophyllum emodi contains lignans (podophyllotoxin is the major composition ). In addition,there are flavonols, saponins, polyose,tannins as well as amio acids in it,Many pharmacological activities such as antitumor, expelling phlegm to arrest coughing,antiinflammatory, antivirus were evidenced by pharmacological studies.Isolation and structure elucidation of chemical constituents from Sinopodophyllum emodi Wall..were performanced, As a result, 10 compounds were isolatsted and identified by various spectral methods as Podophyllotoxin(T1), 4'-demethyl-podophyllotoxin(T2),Kaempferol(T3),Kaempferol-3-O-β-D-glucoside (T4),L-podophillotoxin-4-O-β-D-glucopyranoside(T5),Quercetin(T6),Podorhizol1-O-β-D-glucopyranoside(T7),Isopicropodophyllone(T8),Deoxypodophy-llotoxin(T9), Podophyllotoxone (T10)。Extraction,Separation and identificationDried samples (3kg) were extracted by heat reflux in ethyl acetate (18L, 15L,and 12L respectively) for three times (1.5h, 1.0h and1.0h, respectively). The combined extract was evaporated to dryness under reduced pressure to yield a greenish viscous extract. 120g extracts had been weighed and then subjected to silica gel column chromatography using ethyl acetate-petroleum ether as mobile phase to afford compounds T1(2.6g), T2(466mg), T3(92mg), T4(110mg),T5 (113mg), T6(75mg), T7(60mg), T8(273mg), T9(30mg), T10(5mg). All the compounds was identified respectively by 1DNMR and 2DNMR as Podophyllotoxin(T1),4'- demethylpodophyllotoxin(T2),Kaempferol(T3),Kaempferol3-O-β-D-glucoside(T4),L-podophillotoxin-4-O-β-D-glucopyranoside(T5),Quercetin(T6),Podorhizol1-O-β-D- glucopyranoside(T7),Isopicropodophyllone(T8),Deoxypodophyllotoxin(T9), Podophyllotoxone(T10). Contents determination of lignans in the Sinopodophyllum emodi Wall.In this paper we present a more sensitive quantification method by HPLC for analyzing the main lignans (Ⅰ~Ⅳ) in the traditional chinese medicine of Sinopodophyl- lum emodi Wall.HPLC was performed on Agilent 1100 HPLC instrument and ZORBAX SB C18 coloum (250mm×4.6mm, 5μm), using a mixture of A (acetonitrile) and B(water) as the mobile phase A linear solvent gradient was used, 0~13min: 0% to 25%A, 13~20min: 25% to 30%A, 20~30min: 30% to 32%A, 30~32min: 32% to 50%A. The flow rate was 1.2 mL·min-1 at 25℃, the wavelength for measurement was 290 nm.ResultsI The linearity was in the range of 0.05~5.0μg(n=7)for compoundsⅠ~Ⅳ, and the correlation coefficients were more than 0.9999. Y=3.300×102X+0.24 r=1.0000 Y=4.974×102X-7.84 r=0.9999 Y=4.897×102X+1.00 r=1.0000 Y=5.654×102X+1.69 r=1.0000II Solutions of CompoundsⅠ~Ⅳwere injected into HPLC for six times,the values of RSD as the results of precision tests were 0.97%,0.97%,1.43%,1.10% respectively.III The repeatability tests for six samples showed their RSD were 1.89%, 1.26%, 1.59%, 0.44% respectively. IV The stability of compoundsⅠ~Ⅳwere good in twelve hours ,because the RSD were 1.41%,1.92%,0.98%,1.01% ,respectively.V The average recoveries (n=6) forⅠ~Ⅳcompounds were 101.2% (RSD=1.62%), 100.6%(RSD=1.99%), 99.8%(RSD=0.80%), 101.2%(RSD=1.36%),respectivelyVI The samples of Podophyllum emodi Wall. from different place were determined, the average results show the compoundsⅠ~Ⅳare 0.85%,0.46%,3.31%,0.39% (from Beijing),0.28%,0.20%,2.59%,0.23%(from Bozhou of Anhui province) 0.15%,0.20%,1.01%,0.03% (from Anguo of Hebei province) respectively. The results of this paper can offer new index of content determination for quality standard establishment of Sinopodophyllum emodi Wall. and its agents. Also, the results can offer science foundations for the development and utilization of Sinoodophyllum emodi Wall.