海南薄葉紅厚殼葉的化學(xué)成分研究

海南薄葉紅厚殼葉的化學(xué)成分研究

紅杉是屬的植物，世界上約80種，主要分布在亞洲的熱帶區(qū)，其次是美國和印度洋。我國有4種,其中海南有2種,分別為紅厚殼(CalophylluminophyllumLinn.)和薄葉紅厚殼(C.membranaceumGardn.etChamp.)。紅厚殼屬植物有很高的藥用價值,民間常用作防腐劑、收斂劑、祛痰劑、利尿劑和催瀉劑。種子和根的油對治療傷口和疥瘡有好處。國外學(xué)者從該屬植物中分離出了香豆素類、黃酮類、三萜類和其它生物活性化合物。據(jù)報道從C.inophyllum中分離出的幾種紅厚殼素(inophyllums)和從C.lanigerum中分離出的calanolides具有抗HIV活性,Itoigawa等在篩選抗腫瘤催化劑的實(shí)驗(yàn)中還發(fā)現(xiàn),異海棠果酸比紅厚殼素(inophyllum-E)顯示出更強(qiáng)的活性,而前者是后者的香豆素內(nèi)酯開環(huán)所致。而海南薄葉紅厚殼作為海南特有植物,其化學(xué)成分和藥理作用研究至今未見報道,為了開發(fā)利用海南特有植物資源,本試驗(yàn)對薄葉紅厚殼葉的化學(xué)成分進(jìn)行了研究。1實(shí)驗(yàn)1.1核磁共振檢測儀器5-X國產(chǎn)顯微熔點(diǎn)測定儀,溫度計未校正;VGZAB-HS質(zhì)譜儀;INOVA-500核磁共振儀;FT-5DX紅外光譜儀;薄層色譜用硅膠G(60型),柱色譜用硅膠(200～300目),所用試劑均為市售AR試劑。1.2木栓酮的合成將采自海南陵水縣的薄葉紅厚殼葉陰干,稱取約8kg粉碎后,用95%乙醇在室溫下浸提3次,每次15h,合并浸提液并減壓濃縮,依次用石油醚、乙酸乙酯和甲醇浸提乙醇浸膏,合并乙酸乙酯相并濃縮得粗提物48g。粗提物以石油醚、乙酸乙酯、甲醇按極性遞增進(jìn)行硅膠柱層析,并按體積收集餾分。其中餾分1～6合并濃縮后,在室溫下析出晶體,用石油醚重結(jié)晶后得白色針狀晶體木栓酮(1)。其母液經(jīng)過硅膠柱層析后再用石油醚重結(jié)晶后分別得白色針狀晶體β-谷甾醇(2)和海棠果醇(3)。餾分18～22合并濃縮后經(jīng)過硅膠柱層析,以EtOAc/MeOH為洗脫劑,重結(jié)晶后得到異海棠果酸(4),穗花杉雙黃酮(5)和海棠果酸(6)。1.3化合物的物理和波譜數(shù)據(jù)1.3.1計算c.[m+1]3.3]白色針晶,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的熔點(diǎn)、氫譜數(shù)據(jù)與文獻(xiàn)報道的木栓酮數(shù)據(jù)一致,因此化合物1為木栓酮。1.3.2對照品-谷哌醇無色針狀結(jié)晶(氯仿-甲醇),mp135～137℃,Liebeman-Burchard反應(yīng)呈陽性,噴5%H2SO4溶液105℃加熱顯橙紅色斑點(diǎn)。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。以上數(shù)據(jù)與文獻(xiàn)報道的β-谷甾醇一致,與對照品β-谷甾醇共TLC、共熔點(diǎn)證明系同一化合物。確定化合物2為β-谷甾醇。1.3.3cnmrcdcl3,tms無色針晶,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)。與文獻(xiàn)對照可知化合物3為海棠果醇。1.3.4cnmrcd.3,tms黃色針晶,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-單晶衍射證明化合物4為異海棠果酸。同時也與文獻(xiàn)報道的一致。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’,黃色針晶(甲醇),mp246～248℃,紫外光下呈黃色,噴AlCl3溶液后出現(xiàn)黃色熒光,Zn-濃HCl顯色反應(yīng)呈陽性。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’’’)。以上數(shù)據(jù)與文獻(xiàn)報道的一致,化合物5即為穗花杉雙黃酮。1.3.6cnmrdmso,tms無色針晶,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)。與文獻(xiàn)對照得知化合物6為海棠果酸。2結(jié)構(gòu)鑒定結(jié)果從采自海南陵水縣的薄葉紅厚殼葉的乙醇浸膏的乙酸乙酯部分得到6個化合物,即木栓酮(1),β-谷甾醇(2),海棠果醇(3),異海棠果酸(4),穗花杉雙黃酮(5)和海棠果酸(6)。經(jīng)理