山螺的化學(xué)成分研究

山海市的山海市。ethook是桔?？贫嗄晟ь愔参锏囊环N又名土黨參和四葉參的植物。生于山坡、林緣河谷兩邊等較陰濕的地方,廣泛分布于我國(guó)東北及河北、山西、山東等地。以根入藥,性味甘、辛,平。具有消腫、解毒、排膿、祛痰、催乳的功效。目前,國(guó)內(nèi)外對(duì)其化學(xué)成分的研究還不夠全面。為了對(duì)山海螺資源進(jìn)行開發(fā)利用,筆者對(duì)其化學(xué)成分進(jìn)行系統(tǒng)研究,并從中分離鑒定了6個(gè)化合物,分別為:紫丁香苷(1)、莽草酸(2)、木栓酮(3)、α-菠菜甾醇(4)、豆甾醇(5)、Δ7-豆甾烯醇(6)。其中,化合物3～6為首次從該植物中分離得到。1儀器、試藥和儀器B-540型熔點(diǎn)測(cè)定儀(BüCHI公司,溫度計(jì)未校正),RE-52AA旋轉(zhuǎn)蒸發(fā)儀(上海亞榮生化儀器廠),Agilent1100型LC-MSDTrapSL型ESI-MS質(zhì)譜儀(美國(guó)Agilent公司),AM500型核磁共振儀(Brucker公司,TMS為內(nèi)標(biāo))。層析硅膠(100～200目、200～300目,青島海洋化工有限公司),凝膠SephadexLH-20(Pharmacia公司),其他試劑均為分析純。實(shí)驗(yàn)用山海螺藥材于2009年10月采自吉林吉安,經(jīng)筆者鑒定為桔?？泣h參屬植物山海螺CodonopsislanceolataBenth.etHook.的根。2萃取部位的確定山海螺干燥根10kg,粉碎后用95%乙醇加熱回流提取3次,每次2h,合并提取液,濃縮得浸膏1.9kg,加適量水混懸后依次用石油醚、乙酸乙酯、水飽和正丁醇萃取,萃取液濃縮后分別得石油醚部位(90g)、乙酸乙酯部位(160g)、水飽和正丁醇部位(320g)。取乙酸乙酯部位浸膏上硅膠柱(100～200目),氯仿-甲醇系統(tǒng)梯度洗脫,經(jīng)凝膠SephadexLH-20純化和重結(jié)晶得到化合物1～6。3基因表達(dá)的化合物化合物1:白色針晶(氯仿-甲醇),mp190～192℃。FeCl3反應(yīng)呈綠色,Molishi反應(yīng)呈棕紅色。FAB-MSm/z:395[M+Na]+。1H-NMR(CD3OD,300MHz)δ:6.72(d,J=2.0Hz,H-3,5),6.44(d,J=15.0Hz,H-7),6.34(d,J=15.0Hz,H-8),4.10(2H,t,H-9),4.81(d,J=9.0Hz,H-1′),3.19(m,2′-H),3.16(m,3″-H),3.11(m,H-4),3.02(m,H-5),3.57(m,6″-H),3.75(s,-OCH3×2);13C-NMR(CD3OD-d6,300MHz)δ:129.9(C-1),152.8(C-2),104.5(C-3),132.4(C-4),104.5(C-5),152.8(C-6),132.6(C-7),130.2(C-8),61.3(C-9),102.4(C-1′),74.1(C-2′),76.4(C-3′),70.1(C-4′),76.9(C-5′),61.1(C-6′),56.5(-OCH3×2)。以上數(shù)據(jù)與文獻(xiàn)對(duì)照基本一致,故確定化合物1為紫丁香苷。化合物2:白色晶體,mp184～187℃。IR(KBr)cm-1:3479,1684,1645,1401,1278,930,855。EI-MSm/z:174[M]+。1H-NMR(CD3OD,300MHz)δ:6.68(1H,m,2-H),4.29(1H,m,3-H),3.90(1H,t,4-H),3.65(1H,t,5-H),2.59(1H,m,6-Ha),2.12(1H,m,6-Hb);13C-NMR(CD3OD-d6,300MHz)δ:169.7(-COOH),131.2(C-1),138.5(C-2),71.1(C-3),66.6(C-4),65.7(C-5),30.4(C-6)。以上數(shù)據(jù)與文獻(xiàn)對(duì)照基本一致,故確定化合物2為莽草酸?；衔?:白色針晶(乙酸乙酯),mp260～262℃。Liebermann-Burchard反應(yīng)陽(yáng)性。EI-MSm/z:426[M]+,411,341,302,273,246,231,205,179,163,137,125,123,109,95,81,69(基峰)。1H-NMR(CDCl3)δ:0.73(3H,s,24-CH3),0.88(3H,s,25-CH3),0.89(3H,d,J=6.9Hz,23-CH3),0.94(3H,s,30-CH3),1.01(3H,s,29-CH3),1.04(3H,s,26-CH3),1.05(3H,s,27-CH3),1.19(3H,s,28-CH),2.37～2.39(1H,m,H-4),2.21～2.33(2H,m,2-H);13C-NMR(CDCl3)δ:213.4(C=O),59.5(C-10),58.2(C-4),52.5(C-8),43.2(C-18),42.2(C-5),41.3(C-2),41.2(C-6),39.5(C-22),39.1(C-13),38.7(C-14),37.3(C-9),36.0(C-16),35.9(C-19),35.2(C-11),34.8(C-29),32.8(C-21),32.3(C-17),32.0(C-30),31.6(C-28),30.5(C-15),29.5(C-20),25.9(C-12),22.0(C-1),20.0(C-27),18.4(C-7),18.0(C-25),17.5(C-26),14.6(C-23),7.0(C-24)。以上數(shù)據(jù)與文獻(xiàn)對(duì)照基本一致,故確定化合物3為木栓酮?；衔?:白色結(jié)晶(丙酮),mp164～165℃。Liebermann-Burchard反應(yīng)陽(yáng)性。IR(KBr)cm-1:3400,2940,2860,1650(C=C),1442。EI-MSm/z:412[M]+,397,369,271,81,55,43。1H-NMR(CDCl3,400MHz)δ:5.12(1H,s,7-H),5.23(1H,d,22-H),5.31(1H,s,23-H),3.53(1H,m,3-H),1.55(1H,s,OH),1.03(3H,d,J=6.5Hz),0.88(3H,t,J=9.0Hz),0.81(6H,d,J=8.0Hz),0.82(3H,s),0.51(3H,s,C-CH3);13C-NMR(125MHz,CDCl3)δ:36.1(C-1),32.3(C-2),71.3(C-3),38.1(C-4),40.8(C-5),29.6(C-6),116.5(C-7),138.5(C-8),47.5(C-9),34.0(C-10),22.5(C-11),39.8(C-12),43.8(C-13),55.5(C-14),23.1(C-15),28.8(C-16),55.9(C-17),12.3(C-18),13.6(C-19),41.2(C-20),22.2(C-21),139.0(C-22),129.8(C-23),51.9(C-24),32.6(C-25),19.4(C-26),21.9(C-27),26.9(C-28),13.7(C-29)。以上數(shù)據(jù)與文獻(xiàn)報(bào)道數(shù)據(jù)一致,故確定化合物4為α-菠菜甾醇?；衔?:無(wú)色針晶,mp168～169℃。EI-MSm/z:412[M]+。Libermann-Burchard反應(yīng)陽(yáng)性。IR(KBr)cm-1:3400,1664,1630。1H-NMR(CDCl3)δ:0.7～2.5(m,43H),3.47(br,OH),3.52(1H,m,H-3),5.29(1H,m,H-6),5.18(1H,dd,H-22),5.10(1H,dd,H-23);13C-NMR(CDCl3)δ:37.2(C-1),31.6(C-2),71.6(C-3),42.2(C-4),140.7(C-5),121.6(C-6),31.8(C-7),31.9(C-8),50.0(C-9),36.5(C-10),21.2(C-11),39.5(C-12),42.0(C-13),56.9(C-14),24.3(C-15),28.9(C-16),56.1(C-17),12.1(C-18),19.2(C-19),40.4(C-20),21.3(C-21),138.3(C-22),129.2(C-23),51.2(C-24),31.8(C-25),19.0(C-26),21.2(C-27),25.4(C-28),12.0(C-29)。以上數(shù)據(jù)與文獻(xiàn)對(duì)照基本一致,故確定該化合物為豆甾醇。化合物6:白色針晶,mp153～154℃。Libermann-Burchard反應(yīng)陽(yáng)性。1H-NMR(CDCl3)δ:0.53(3H,s,CH3-18),0.77～0.85(12H,s,CH3-19,26,27,29),0.90(3H,d,CH3-21),3.60(1H,m,H-3),5.16(1H,m,H-7);13C-NMR(CDCl3)δ:37.3(C-1),31.6(C-2),71.1(C-3),38.7(C-4),40.1(C-5),29.8(C-6),117.7(C-7),140.9(C-8),50.2(C-9),34.8(C-10),22.9(C-11),40.3(C-12),43.9(C-13),5