思茅豆腐柴中的黄酮类化学成分研究

思茅豆腐柴中的黄酮类化学成分研究 思茅豆腐柴中的黄酮类化学成分研究 李钦,沈月毛,李萍(1.中国药科大学,现代中药教育部重点实验室,南京210009;2.中 国科学院昆明植物研究所植物化学与西部植 物资源持续利用国家重点实验室,昆明650204) 摘要:目的研究民族药物思茅豆腐柴(PremnaszemaoensisPei)中的黄酮类化学成分. 利用SephadexLH一20及硅胶 等色谱技术进行分离纯化,根据理化性质,光谱数据进行结构鉴定.结果得到7个黄 酮类化合物,分别鉴定为5,4一二羟基. 3,7,3一三甲氧基黄酮(1),5一羟基'3,4,6,7-四甲氧基黄酮(2),5,4一二羟基_7一甲氧 基黄酮醇(3),5,3一二羟基_7,4一二甲 氧基黄酮醇(4),3,4,5一三羟基一3,7一二甲氧基黄酮(5),5,7一二羟基一4一甲氧 基黄酮(6),5一羟基_7,3,4一三甲氧基黄酮醇 (7).结论这7个化合物均为首次从思茅豆腐柴中分离得到. 关键词:思茅豆腐柴;黄酮;黄酮醇 中图分类号:R284.1文献标识码:A文章编号:1001—2494(2008)06—0417—03 StudyonFlavanoidsintheLeavesofPremnaszemaoensisPei LIQin,SHENYue— rnao,LIPing(1.KeyLaboratoryofModemChineseMediciandDepartmentofPharmacogno,China PharmaceuticalUniversity,Nang210009,China;2.StateKeyLaboratoryofPhytochemistryandPlantResourcesinWestChina, KunmingInstituteofBotany,ChineseAcademyofSciences,Kunming650204,China) ABSTRACT:OBJECTIVEToinvestigatethechemicalconstituentsofPremnaszemaoensisPei.METHODSThecompoundswere isolatedandpurifiedviasilieagelandSephadexLH一 20columnchromatographyrepeatedlyandtheirstructureswereelucidatedonthe basisofspectroscopicevidencesandphysicochemicalproperties.RESULTSSevenflavanoi dswereobtainedandidentifiedas5.4. dihydroxy-3,7,3一trimethoxynavone(1),5-hydroxy一3,4,6,7-tetramethoynavone(2),5,4'-dihydroxy-7一methoxyflavonol(3),5, 3dihydroxy-7,4一dimeth0xynavonol(4),3,4,5一trihydroxy-3,7-dimethoxynavone(5),5,7-dihydroxy一4'-methoxyflavone(6),5一 hydroxy-7,3,4一trimethoxvnavonol(7).CONCLUSIONAIlcompoundsareisolatedfromtheplantforthefir sttime. KEYW0RDS:tunaszemaoensisPei;flawme;flavonol 马鞭草科(Verbenaceae)豆腐柴属(PremnaL.) 植物全世界约有200种,主要分布在亚洲和非洲的 热带地区,我国有约44种1变种,主产于南部各省, 尤集中于云南地区….根据文献调查研究的结果, 豆腐柴属植物的化学成分及生物活性方面的研究都 还不多见,从已有的研究来看,本属植物主要含有倍 半萜,二萜,三萜,黄酮,酚酸及生物碱类成分,生物 活性方面主要具有抗菌及细胞毒活性引. 思茅豆腐柴(PremnaszemaoensisPei)为高大乔 木,高3,12m,主分布于云南南部的思茅,西双版 纳及澜沧等地区.由于在当地具有多种用途,遭到 了严重的砍伐,并且其自然繁殖能力较差,因此成年 植株大量减少,渐成濒危种.为了对该濒危植物的 利用及保护提供科学依据,我们对思茅豆腐柴进行 了化学成分研究.利用SephadexLH一20及硅胶等 柱色谱技术,从中分离得到了7个黄酮类化合物. 1仪器和试剂 BrukerAM一400及DRX一500核磁共振仪, TMS为内标;VG—Auto—Spec一3000及Thermo— FinniganLCQ—Advantage质谱仪;JascoDIP一370 digitalpolarmeter微量旋光仪;BiichiR一114旋转蒸 发仪;BSZ一100自动部分收集仪(上海青浦沪西仪 器厂);SHZ—D(?)型循环水式多用真空泵(巩义 市予华仪器有限公司).SephadexLH一20(Amers— han公司),D101大孔树脂(天津农药厂),硅胶G (青岛海洋化工厂),所用试剂均为纯. 思茅豆腐柴样品采集于西双版纳热带植物园, 经西双版纳热带植物园的兰芹英研究员鉴定为 PremnaszemaoensisPei,凭证标本存于昆明植物所. 2提取与分离 思茅豆腐柴粗粉(干燥叶片)1.7kg,用体积分 数为90%乙醇热回流提取3次,每次4h,合并乙醇 提取液后减压浓缩,浓缩液依次用石油醚,乙酸乙酯 萃取.取乙酸乙酯部分198g,经D101大孔树脂分 作者简介:李钦,男,硕士研究生通讯作者:李萍,女,教授,博t生导师Tel:(025)85322256E—mail:lipingli@public1.ptt.jS.CB 孬丽丽—Chin—Pha—rmJ,2008Marc—h,Vol,43No,6'' 段脱色,用乙醇.水(0:1,5L;20:80,8L;30:70,20 L:50:50.32l;70:30,23L;80:20,16L;1:0,3 L);每份1L系统洗脱,得到A,,A7个部分,合并 A及A部位,称重共有33g,上一60g的硅胶柱,石 油醚.丙酮100:1到10:1梯度洗脱,得到B,,B3 个部分,其中B,部分共8.3g,经过反复的硅胶柱色 谱及SephadexLH一20柱分离纯化,得到化合物l (42mg),2(38mg),3(13mg),4(9mg),5(21 mg),6(33mg),7(7mg). 3结构鉴定 化合物1:淡黄色粉末,mp:177,178,盐 酸.镁粉反应阳性.ESI—MS:m/z343[M—H]一,结 合tt.NMR,C.NMII谱数据推断分子式为 C18Hl607.H.NMII(500MHz,DMSO—d6)6:7.66 (1H,d,J=2.0Hz,H一2),7.63(1H,dd,J=2.0,8.4 Hz.It-6),6.96(1H,d,J=8.4Hz,H-5),6.77(1H, d,J=2.2ttz,H.8),6.36(1H,d,J=2.2Hz,1-t-6), 3.85(6tt,s,.OCH),3.80(3H,s,.OCH).C-NMR (125MHz,DMSO.d)6:178.0(C-4),165.1(C-7), 160.8(C-5),156.2(C.9),1557(C-2),149.9(C一 4),147.4(C.3),137.9(C-3),122.3(C-6),120.6 (C.1),l15.6(C-5),112.0(C-2),97.7(C-6), 92.4(C.8),120.0(C.6),59.7(,OCH),56.0(一 OCH),55.7(一OCH).与文献对照,鉴定其为 5,4.二羟基_3,7,3.三甲氧基黄酮. 化合物2:淡黄色粉末,mp:197,199?,盐酸-镁 粉反应阳性.ESI.MS:m/z357[M—H]一,结合H- NMR13CNMR谱数据推断分子式为C9l-I,8O.tt-NMR (500MH_z,DMSOd)6:7.74(1H,dd,J=2.0,8.6Hz,H- 6),7.59(1H,d,J=2.0Hz,H-6),7.14(1H,d,J=8.6 Hz,H-5),7.04(1tt,d,J:2.2ttz,H.8),6.98(IH,d, J=2.2Hz,H-6),3.93(3H,S,一OCH),3.88(3H,s, 一 OCH),3.85(3H,S,一OCH3),3.72(3It,S, 一 OCtt).C.NMR(125MHz,DMSO-d)6:182.3(C- 4),163.6(C-2),158.7(C一7),152.7(C-5),157.2(C-9), 152.0(C-4),149.0(C.3),131.9(C-6),122.8(C-l), 120.1(C.6),1l1.7(C-5),109.4(C-2),105.1(C-10), 103.6(C一3),91.7(C一8),60.0(一OCH),56.5 (一OCH),55.9(一OCH,),55.8(一OCH).与文 献对照,鉴定其为5.羟基..3,4,6,7.四甲氧基黄酮. 化合物3:淡黄色粉末,mp:221,223?,盐 酸.镁粉反应阳性.ESI.MS:m/z299[M—H,结 合H.NMII,C—NMR谱数据推断分子式为cHO. H.NMR(500MHz,DMSO.d)6:813(2H,d, ? 418?Chi nPharmJ.2008March.Z43No6 J=8.4Hz,H.2,6),7.10(2H,d,J=8.4Hz,H-3, 5).6.34(1tt,d,J=2.2ttz,H-8),6.18(1tt,d, J=2.2Hz,It-6),3.82(3H,s,一OCH3)."C-NMR (125MHz,DMSO.d)6:175.9(C4),163.9(C-7), 160.6(C-5),160.4(C-4),156.1(C-9),145.2(C- 2),135.9(C-3),129.2(C.2,6),123.2(C-1), 113.9(C-3,5),103.0(C一10),98.1(C-6),93.4(C- 8),55.3(一OCH).与文献对照,鉴定为5,4-二 羟基-7.甲氧基黄酮醇. 化合物4:淡黄色粉末,mp:230,232?,盐 酸.镁粉反应阳性.ESI.MS:m/z329[M—H]一,结 合H.NMll,"C.NMR谱数据推断分子式为C17HO7. H.NMR(500MHz,DMSO.d^)6:7.77(1H,d, J=2.0Hz,H.2),7.74(1H,dd,J=2.0,8.4Hz,H- 6),6.94(1H,d,J=8.4Hz,H-5'),6.77(1H,d,J= 2.2Hz,tt.8),6.34(1tt,d,J=2.2Hz,tt-6),3.85 (3H,s,.OCH3),3.84(3tt,s,一OCH3).C-NMR (125Mttz,DMSO.d)6:176.0(C-4),165.0(C-7), 160.4(C.5),156.1(C-9),149.0(C-4),147.5(C. 2),147.1(C-3),136.1(C-3),121.9(C一1),121.0 (C-6),l15.6(C-2),111.9(C-5),104.0(C-10), 97.5(C-6),92.1(C.8),56.1(,OCH),55.9 (一OCtt).与文献对照,鉴定其为5,3-二羟基- 7,4.二甲氧基黄酮醇. 化合物5:淡黄色粉末,mp:235,237?,盐 酸.镁粉反应阳性.ESI-MS:m/z329[M—H],结 合H.NMtl,C.NMR谱数据推断分子式为CH.O7. tt.NMR(500MHz,DMSO.d6)6:7.74(1H,d, J=2.0Hz,tt.2),7.66(1H,dd,J:2.0,8.4Hz,H一 6),6.99(1H,d,',=8.4Hz,H-5),6.67(1H,d,J: 2.2I-lz,tt-8),6.30(1tt,d,J=2.2ttz,tt-6),3.88 (3H,S,一OCH),3.86(3H,S,,OCH)."C-NMR (125MHz,DMSO.d)6:179.1(C4),166.1(C-7), 162.3(C-5),157.3((2-2),156.7((2-9),151.0(C- 4),148.4(C.3),138.9(C-3),123.1(C-1),121.8 (C.6),116.3(C一2),112.9(C-5),106.1(C一10), 98.2(C-6),92.7(C-8),60.0(一0CH),56.3 (一0cH).与文献对照,鉴定为3,4,5-三羟 基.3,7-二甲氧基黄酮. 化合物6:淡黄色粉末,mp:263,265?,盐 酸.镁粉反应阳性.EsI.Ms:m283[M—H],结 合H.NMR,"c.NMR谱数据推断分子式为CH,2 0.H.NMR(500MHz,DMS0-d6)6:8.05(2H,d, J=8.4Hz,H_2,6),7.13(2H,d,J=8.4Hz,H_3, 中国药学杂志2o08年3月第43卷第6期 5),6.88(1H,s,H一3),6.51(1H,d,J=2.2Hz,H一 8),6.20(1H,d,J=2.2Hz,H.6),3.87(3H,s, 一 OCH3)."C—NMR(125MHz,DMSO—d6)6:181.8(C一 4),164.2(C-7),163.3(C一2),162.3(C-9),161.8 (C4),157.3(C-5),128.3(C一2,6),122.8(C一 1),114.6(C一3,5),103.7(C一10),103.5(C一3), 98.9(C),94.0(C一8),55.5(一OCH).与文献 对照,鉴定其为5,7一二羟基一4一甲氧基黄酮. 化合物7:淡黄色粉末,mp:189,192cC,盐 酸一镁粉反应阳性.ESI—MS:m/z343[M—H]一,结 合H—NMR,"C—NMR谱数据推断分子式为c,H, 07.H—NMR(500MHz,DMSO—d6)6:7.79(1H,d, J:2.0Hz,H一2),7.76(1H,dd,J=2.0,8.4Hz,H一 6),6.95(1H,d,J=8.4Hz,H一5),6.79(1H,d,J= 2.2Hz,H一8),6.35(1H,d,J=2.2Hz,H一6),3.86 (6H,s,一OCH3),3.84(3H,s,一OCH3).C—NMR(125 MHz,DMSO—d)6:175.8(C4),164.5(C-7),160.4 (C一5),156.0(C-9),150.7(C4),148.5(C一3), 145.3(C一2),138.1(C一3),123.4(C一1),121.7(C一 6),112.3(C-2),112.0(C-5),103.7(C一10),97.5 (C),91.8(C.8),56.3(一OCH),55.9(一 OCH.),55.8(一OCH.).与参考文献对照,鉴定 其为5一羟基-7,3,4一三甲氧基黄酮醇. 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