海南薄叶红厚壳叶的化学成分研究

海南薄叶红厚壳叶的化学成分研究

红杉是属的植物，世界上约80种，主要分布在亚洲的热带区，其次是美国和印度洋。我国有4种,其中海南有2种,分别为红厚壳(CalophylluminophyllumLinn.)和薄叶红厚壳(C.membranaceumGardn.etChamp.)。红厚壳属植物有很高的药用价值,民间常用作防腐剂、收敛剂、祛痰剂、利尿剂和催泻剂。种子和根的油对治疗伤口和疥疮有好处。国外学者从该属植物中分离出了香豆素类、黄酮类、三萜类和其它生物活性化合物。据报道从C.inophyllum中分离出的几种红厚壳素(inophyllums)和从C.lanigerum中分离出的calanolides具有抗HIV活性,Itoigawa等在筛选抗肿瘤催化剂的实验中还发现,异海棠果酸比红厚壳素(inophyllum-E)显示出更强的活性,而前者是后者的香豆素内酯开环所致。而海南薄叶红厚壳作为海南特有植物,其化学成分和药理作用研究至今未见报道,为了开发利用海南特有植物资源,本试验对薄叶红厚壳叶的化学成分进行了研究。1实验1.1核磁共振检测仪器5-X国产显微熔点测定仪,温度计未校正;VGZAB-HS质谱仪;INOVA-500核磁共振仪;FT-5DX红外光谱仪;薄层色谱用硅胶G(60型),柱色谱用硅胶(200～300目),所用试剂均为市售AR试剂。1.2木栓酮的合成将采自海南陵水县的薄叶红厚壳叶阴干,称取约8kg粉碎后,用95%乙醇在室温下浸提3次,每次15h,合并浸提液并减压浓缩,依次用石油醚、乙酸乙酯和甲醇浸提乙醇浸膏,合并乙酸乙酯相并浓缩得粗提物48g。粗提物以石油醚、乙酸乙酯、甲醇按极性递增进行硅胶柱层析,并按体积收集馏分。其中馏分1～6合并浓缩后,在室温下析出晶体,用石油醚重结晶后得白色针状晶体木栓酮(1)。其母液经过硅胶柱层析后再用石油醚重结晶后分别得白色针状晶体β-谷甾醇(2)和海棠果醇(3)。馏分18～22合并浓缩后经过硅胶柱层析,以EtOAc/MeOH为洗脱剂,重结晶后得到异海棠果酸(4),穗花杉双黄酮(5)和海棠果酸(6)。1.3化合物的物理和波谱数据1.3.1计算c.[m+1]3.3]白色针晶,mp260～262℃。IR(KBr)νmax(cm-1):2970,2927,2866,1715,1456,1387,1258;FAB-MS(m/z):427[M+1]+;1HNMR(CDCl3,TMS)δ:0.73和0.87(each3H,s,H-24,25),0.88(3H,d,J=6.5Hz,H-23),0.95(3H,s,H-30),1.00(3H,s,H-26),1.01(3H,s,H-29),1.05(3H,s,H-27),1.18(3H,s,H-28),1.20～1.77(m,23H),1.95(m,1H),2.22～2.32(m,2H)。化合物1的熔点、氢谱数据与文献报道的木栓酮数据一致,因此化合物1为木栓酮。1.3.2对照品-谷哌醇无色针状结晶(氯仿-甲醇),mp135～137℃,Liebeman-Burchard反应呈阳性,喷5%H2SO4溶液105℃加热显橙红色斑点。IR(KBr)νmax(cm-1):3430,2959,2938,2870,1640,1464,1379,1058;EI-MS(m/z):414(M+),396,330,329,273,255,231,213。以上数据与文献报道的β-谷甾醇一致,与对照品β-谷甾醇共TLC、共熔点证明系同一化合物。确定化合物2为β-谷甾醇。1.3.3cnmrcdcl3,tms无色针晶,mp279～281℃。IR(KBr)νmax(cm-1):3545,2931,2865,1706,1464,1388,1081;1HNMR(CDCl3,TMS)δ:0.72和0.86(each3H,s,H-24,25),0.88(3H,d,J=6.0Hz,H-23),0.91,0.98,0.99和1.12(each3H,s,H-27,26,29,30),2.50,2.20(3H,m,H-4,2α,2β),3.65(2H,s,H-28)。13CNMR(CDCl3,TMS)δ:22.3(C-1),41.5(C-2),212.7(C-3),58.3(C-4),42.1(C-5),41.3(C-6),18.3(C-7),52.5(C-8),37.5(C-9),59.6(C-10),35.5(C-11),30.1(C-12),39.4(C-13),38.2(C-14),31.4(C-15),29.1(C-16),35.2(C-17),39.5(C-18),34.5(C-19),28.1(C-20),31.5(C-21),33.4(C-22),6.8(C-23),14.7(C-24),18.1(C-25),19.1(C-26),19.2(C-27),68.0(C-28),34.3(C-29),32.8(C-30)。与文献对照可知化合物3为海棠果醇。1.3.4cnmrcd.3,tms黄色针晶,mp182～183℃,FAB-MS(m/z):421[M+1]+,405,375,347;1HNMR(CD3COCD3,TMS)δ:0.96(3H,s,H-19),1.16(3H,d,J=7Hz,H-22),1.23(3H,s,H-20),1.44(3H,d,J=6Hz,H-21),2.69(1H,dq,J=7.5,3.5Hz,H-11),4.75(1H,dq,J=6.5,3Hz,H-10),5.56(1H,d,J=9.5Hz,H-7),6.45(1H,s,H-3),6.56(1H,d,J=9.5Hz,H-8),7.34(3H,m,H-15,16,17),7.42(2H,m,H-14,18),10.38(1H,br,OH),12.53(1H,s,OH);13CNMR(CD3COCD3,TMS)δ:166.3(C-2),121.4(C-3),148.4(C-4),109.3(C-4a),159.4(C-4b),77.5(C-6),127.8(C-7),116.2(C-8),101.6(C-8a),161.9(C-8b),78.8(C-10),44.8(C-11),201.9(C-12),102.1(C-12a),157.0(C-12b),141.8(C-13),127.0(C-14),129.1(C-15,17),129.5(C-16),127.8(C-18),28.5(C-19),28.1(C-20),16.5(C-21),9.7(C-22)。X-单晶衍射证明化合物4为异海棠果酸。同时也与文献报道的一致。1.3.5h,6.35s,1h,h-3’,5’5’,5’2’,5’2’,5’2’,5’2’,5’2’,5’2’,5’2’,5’3’,5’2’,5’2’,5’3’,5’2’,5’2’,5’2h,2h,2h,3h,3h,3h,3h,3h,3h,3h,3h,3h,3h,3h,3h,3h,3h,3h,3h,3h,3h,3h,3h,3h,3h,3h,3h,3h,3h,3h,3h,3h,3h,3h,3,3’,3,3’,3,3’,3,3’,3,3,3,3,3,3,3,3,3’,3,3’,3,3’,3,3’,3,3’,3,3’,3,3,3,3,3,3,3,3,3’,3,3’,3,3’,3,3’,3,3’,3,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,3’,黄色针晶(甲醇),mp246～248℃,紫外光下呈黄色,喷AlCl3溶液后出现黄色荧光,Zn-浓HCl显色反应呈阳性。FAB-MS(m/z):539[M+1]+,460,341,307;IR(KBr)νmax(cm-1):3500～3250(br,OH),1653,1608,1574,1493,1350,1240,1160,1105,1030,900,840,630;1HNMR(CD3COCD3,TMS)δ:6.21(d,2.5Hz,1H,H-6),6.35(s,1H,H-6’’),6.44(d,1.5Hz,1H,H-8),6.62(s,1H,H-3’’),6.70(s,1H,H-3),6.77(dd,2,6.5Hz,2H,H-3’’’,5’’’),7.15(d,9Hz,1H,H-5’),7.68(dd,2,6.5Hz,1H,H-2’’’,6’’’),7.96(dd,3,9Hz,1H,H-6’),8.21(d,2.5Hz,1H,H-2’),13.06(s,1H,OH-5),13.19(s,1H,OH-5’);13CNMR(CD3COCD3,TMS)δ:165.0(C-2),103.6(C-3),183.1(C-4),162.3(C-5),99.7(C-6),163.3(C-7),94.8(C-8),158.8(C-9),104.7(C-10),121.9(C-1’),128.1(C-2’),122.5(C-3’),161.7(C-4’),116.7(C-5’),132.6(C-6’),164.7(C-2’’),103.8(CH),182.2(C-4’’),161.7(C-5’’),101.1(C-6’’),162.6(C-7’’),105.2(C-8’’),156.1(C-9’’),104.7(C-10’’),122.5(C-1’’’),129.1(C-1’’’),116.7(C-3’’’),162.3(C-4’’’),118.7(C-5’’’),129.1(C-6’’’)。以上数据与文献报道的一致,化合物5即为穗花杉双黄酮。1.3.6cnmrdmso,tms无色针晶,mp310～312℃.IR(KBr)νmax(cm-1):3510(OH),1690(CO2H),1210;1HNMR(DMSO,TMS)δ:0.74(s,3H,H-24),0.79(s,3H,H-25),0.82(d,7Hz,3H,H-23),0.88(s,3H,H-27),0.90(s,3H,H-26),0.95(s,3H,H-29),0.99(s,3H,H-30),12.06(s,1H28-COOH).13CNMR(DMSO,TMS)δ:15.4(C-1),34.3(C-2),69.8(C-3),48.7(C-4),37.3(C-5),41.2(C-6),16.8(C-7),52.4(C-8),37.2(C-9),60.6(C-10),35.1(C-11),30.6(C-12),38.3(C-13),40.0(C-14),32.2(C-15),34.9(C-16),43.6(C-17),37.1(C-18),35.2(C-19),28.0(C-20),32.1(C-21),29.1(C-22),11.6(C-23),16.0(C-24),17.4(C-25),18.0(C-26),19.8(C-27),179.4(C-28),29.6(C-29),34.2(C-30)。与文献对照得知化合物6为海棠果酸。2结构鉴定结果从采自海南陵水县的薄叶红厚壳叶的乙醇浸膏的乙酸乙酯部分得到6个化合物,即木栓酮(1),β-谷甾醇(2),海棠果醇(3),异海棠果酸(4),穗花杉双黄酮(5)和海棠果酸(6)。经理