全 文 ：香橙化学成分的分离与鉴定
朴香兰
(中央民族大学 生命与环境科学学院 , 中国少数民族传统医学研究中心 , 北京 100081)
摘　要:　本文通过采用色谱分离技术 , 对香橙抗氧化作用的物质基础进行研究.通过色谱分离技术 ,从香橙
中分离得到 7个化学成分 , 包括 3个香豆素类 、2个柠檬苦素类和 2 个黄酮类化合物.
关键词:　香橙;香豆素;柠檬苷;黄酮醇
中图分类号:Q946　　文献标识码:A　　文章编号:1005-8036(2008)03-0036-04
收稿日期:2008-03-21
基金项目:中国博士后科学基金项目(No.20060390373);留学回国人员科研启动基金资助项目(教外司留[ 2007]
1108).
作者简介:朴香兰(1965-), 女(朝鲜族),吉林人 , 中央民族大学生命与环境科学学院 、中国少数民族传统医学研究
中心讲师 ,主要研究方向:抗氧化天然药物化学.
　　香橙(Citrus junos)为芸香科(Rutaceae)植物香橙 Citrus junos Sieb.的果实 , 是 C.inchangensis 和C.
reticulate var.austera 的杂交果实[ 1] .分布在中国 、韩国和日本等国.香橙在民间一直用于芳香类苦味健
胃 、祛痰 、止咳等.在韩国南海岸大量栽培 , 主要做香橙蜜及香橙汁.香橙含有柠檬苦素类化合物[ 2 ～ 3] ,
香豆素类化合物[ 4 ～ 5] , 以及黄酮类化合物[ 6 ～ 7] .这些化合物具有各种药理活性 , 如:抗癌[ 8] 、抗肿
瘤[ 9 ～ 10] 、抗高血压[ 11] 、降低胆固醇[ 12] , 以及抗血栓作用[ 13] .
本课题组在不同天然植物筛选抗氧化剂中[ 14 ～ 17] , 筛选出水果香橙的有效部位为其二氯甲烷萃取
物.运用现代分离技术 , 从香橙二氯甲烷萃取物中分离得到 7个化合物(化合物 1-7).其中有 3个香
豆素类化合物 , 2个柠檬苦素类化合物和 2个黄酮类化合物.这 7个化合物均是对二氯甲烷萃取物.
1　材料与方法
1.1　化学试剂与仪器
香橙购于韩国超市.所有化学试剂均为分析纯(北京化工厂).硅胶(青岛海洋化工厂分厂).硅胶
板(德国Merck公司).紫外吸收光谱仪UV Series 600(美国), 核磁共振仪 Avance 500(德国),质谱 Jeol
JMS-700(日本).
1.2　提取
取香橙 10 kg , 用甲醇加热回流3次 , 每次 3 h.溶剂减压浓缩回收 , 制得甲醇提取浸膏(980 g);用
1000 mL 的水混悬后 , 再用 800 mL二氯甲烷萃取得二氯甲烷萃取物 (12.8 g).水溶部分再用 700 mL水
饱和正丁醇萃取得正丁醇萃取物(66.7 g).
1.3　化学成分的分离
二氯甲烷萃取部分 10 g , 用硅胶柱色谱(50 g , 4×50 cm), 氯仿-甲醇梯度洗脱(100∶1※ 5∶1), 得
到7个组分.对组分 3再次柱色谱分离 , 得到化合物 1至 7.
2008年 8月
第 17卷　第 3期
中央民族大学学报(自然科学版)
Journal of the CUN(Natural Sciences Edition)
Aug., 2008
Vol.17　No.3
2　结果与讨论
香橙是一个即可用来观赏又可食用的水果.含有丰富的维生素 , 香豆素类 , 柠檬苦素类及黄酮类
成分.因此 , 本研究用体外与细胞系统 , 评价了香橙的自由离子基消除活性 , 并证实了它们的抗氧化
活性.对有效部位二氯甲烷萃取 , 用硅胶柱进行了分离 ,得到了化合物 1至 7的结构鉴定如下:
化合物 1:黄色结晶.EI MS m z:162([M] + , C9H6O3).1H-NMR(300MHz , CD3OD , δppm):7.73
(1H , d , J =9.51Hz , H-4), 7.34(1H , d , J =8.55 Hz , H-5), 6.69(1H , dd , J =8.55 , 2.19 Hz , H-
6), 6.60(1H , d , J =2.19 Hz , H-8), 6.74(1H , d , J =9.51Hz , H-3).13C-NMR(75MHz , CD3OD ,
δppm):162.0(C-7), 161.6(C-2), 156.0(C-9), 145.4(C-4), 130.4(C-5), 114.0(C-10),
111.8(C -3), 111.8 (C -6), 102.7 (C -8).以上数据与文献基本一致 , 故鉴定为伞形酮
(umbelliferone).
化合物 2:黄色针晶 (C8H10 N2O2).EI MS m z:232 [M] +.1H-NMR(300 MHz , DMSO-d6)(:
10.07(1H , s , 9-OH), 8.15(1H ,d , J =9.75Hz , H-5), 8.03(1H , d , J =2.19Hz , H-2), 7.28(1H ,
d , J =2.19 Hz , H-3), 6.30 (1H , d , J =9.75 H-6), 4.08 (3H , s , 4-OCH3).13 C-NMR(75 MHz ,
DMSO-d6)(:146.61(C-2), 105.78(C-3), 115.23(C-3a), 145.90(C-4), 107.51(C-4a), 140.28
(C-5), 112.81(C-6), 160.35(C-7), 141.61(C-8a), 125.87(C-9), 147.31(C -9a), 61.51(4-
OCH3).以上数据与文献基本一致 , 故鉴定为 9-羟基-4-甲氧补骨脂素 (9-hydroxy-4-methoxyl
psoralen).
化合物3:白色片晶.EI MS m z:298([M] + , C19H22O3).1H-NMR(300MHz , CDCl3 , δppm):7.61
(1H , d , J =9.51Hz , H-4), 6.22(1H , d , J =9.51Hz , H-3), 7.33(1H , d , J =8.55Hz , H-5), 6.82
(1H , dd , J =8.55 , 2.43Hz , H-6), 6.80(1H , d , J =2.43Hz , H-8), 4.58(2H , d , J =6.6Hz , H-1′),
5.44(1H , t , J =6.6Hz , H -2′), 2.09(2H , m , H-4′), 2.09(2H , m , H -5′), 5.05(1H , m , H -6′),
1.74(3H , s , H -3′-CH3), 1.64(3H , s , H -8′), 1.58(3H , s , H -7′-CH3).13C -NMR(75 MHz ,
CDCl3 , δppm):161.26(C-2), 112.95(C-3), 143.41(C-4), 128.64(C-5), 113.22(C-6), 162.13(C
-7), 101.58(C-8), 155.85(C-9), 112.40(C-10), 65.48(C-1′), 118.39(C-2′), 142.34(C-3′),
39.49(C-4′), 26.21(C-5′), 123.59(C-6′), 131.94(C-7′), 25.63(C-8′), 17.69(7′-CH3), 16.74
(3′-CH3).以上数据与文献基本一致 , 故鉴定为橙皮油素(auraptene).
化合物4:白色针晶.C19H38O4 .FAB MS m z:471 [M+H] +.1H-NMR(DMSO-d6 , δppm):7.71
(1H , s , H-21), 7.65(1H , t , J =1.7Hz , H-23), 6.50(1H , d , J =1.7Hz , H-22), 5.46(1H , s , H-
17), 4.91(1H , d , J =13.0Hz , H-19), 4.47(1H , d , J =13.0Hz , H-19), 4.10(1H , s , H-15), 4.10
(1H , s , H-1), 3.11(1H , t , J =15.3Hz , H-5), 2.76(1H , d , J =16.3Hz , H-2), 2.61(1H , dd , J =
16.3 , 4.0Hz , H-2), 2.55(1H , dd , J =12.4 , 3.1Hz , H-9), 2.44(1H , dd , J =15.0 , 3.2Hz , H-6),
2.26(1H , dd , J =15.0 , 3.2Hz , H-6), 1.81(1H , m , H-11β), 1.71(1H , m , H-11α), 1.70(1H , m , H
-12β), 1.22(1H , m , H-12α), 1.17(3H , s , 4α-CH3), 1.01(3H , s , 4β - CH3), 0.98(3H , s , 8-
CH3), 1.09(3H , s , 13-CH3), 13C-NMR(DMSO-d6 , δppm):78.51(C-1), 35.79(C-2), 170.44(C
-3), 79.64(C-4), 58.01(C-5), 36.31(C-6), 208.21(C-7), 50.37(C-8), 46.55(C-9), 45.34(C
-10), 17.62(C-11), 29.83(C-12), 37.70(C-13), 66.78(C-14), 53.79(C-15), 167.46(C-16),
77.52(C-17), 64.90(C-19), 120.31(C-20), 141.84(C-21), 110.32(C-22), 143.48(C-23), 29.83
(4α-CH3), 21.52(4β-CH3), 19.77(17-CH3), 17.10(8-CH3).以上数据与文献基本一致 , 故鉴定
为柠檬苦素(limonin).
37　第 3期 朴香兰:香橙化学成分的分离与鉴定
化合物5:白色粉末.C26H32O8 , FAB MS m z:473 [M+H] +.1H-NMR(DMSO-d6 , δppm):7.71
(1H , s , H-21), 7.65(1H , t , J =1.7Hz , H-23), 6.49(1H , d , J =1.7Hz , H-22), 5.42(1H , s , H-
17), 5.39(1H , d , J =5.4Hz , 1-OH), 3.81(1H , s , H-15), 3.63(1H , t , J =6.0Hz , H-1), 3.01(1H ,
t , J =14.4Hz , H-5), 2.65(2H , m , H-2), 2.59(1H , d , J =11.0Hz , H-9), 2.42(1H , dd , J =14.2 ,
3.7Hz , H-6), 2.28(1H , dd , J =14.2 , 3.7Hz , H-6), 1.70(2H , m , H-11), 1.37(1H , m , H-12β),
1.17(1H , m , H-12α), 1.45(3H , s , 4α-CH3), 1.26(3H , s , 4β -CH3), 1.14(3H , s , 10-CH3), 1.09
(3H , s , 8-CH3), 0.98(3H , s , 13-CH3).13C-NMR(DMSO-d6 , δppm):68.39(C-1), 39.09(C-
2), 170.85(C-3), 83.84(C-4), 49.44(C-5), 38.88(C-6), 208.77(C-7), 52.04(C-8), 43.75(C-
9), 44.25(C-10), 16.74(C-11), 31.32(C -12), 36.85(C -13), 65.79(C -14), 52.69(C-15),
167.24(C-16), 77.56(C -17), 120.20(C-20), 141.62(C-21), 110.26(C -22), 143.38(C -23),
23.10(4α-CH3), 33.03(4β -CH3), 20.22(13-CH3), 16.11(8-CH3), 15.95(10-CH3).以上数据与文
献基本一致 , 故鉴定为脱乙酰诺米林(deacetylnomilin).
化合物 6:黄色结晶.C16H12O6.EI MS m z:300 [M] +.1H-NMR(300MHz , DMSO-d6 , δppm):
13.06(1H , s , 5-OH), 10.37(1H , s , 4′-OH), 7.91(2H , d , J =8.79 Hz , H-2′,6′), 6.91(2H , d , J
=8.79 Hz , H-3′,5′), 6.76(1H , s , H-8), 6.58(1H , s , H-3), 3.27(3H , s , 6-OCH3).13C-NMR
(DMSO-d6 , δppm):182.6 (C-4), 164.2 (C-2), 161.6 (C-4′), 157.7 (C-7), 153.2(C -5),
152.8(C-9), 131.8(C-6), 128.9(C-2′, 6′), 121.6(C-1′), 116.4(C-3′, 5′), 104.5(C-10),
102.8(C-3), 94.7 (C-8), 60.4 (6-OCH3).以上数据与文献基本一致 , 故鉴定为粗毛豚草素
(hispidulin).
化合物 7:黄色粉末.C17H14O 6.EI MS m z:314 [M] +.1H-NMR(C5D5N , δppm):13.67(1H , s , 5
-OH), 12.74(1H , s , 4′-OH), 7.96(1H , d , J =8.8Hz , H-2′), 7.27(1H , d , J =8.8Hz , H-3′), 7.27
(1H , d , J =8.8Hz , H-5′), 7.96(1H , d , J =8.8Hz , H-6′), 6.96(1H , s , H-3), 6.80(1H , s , H-8),
3.99(3H , s , 6-OCH3), 3.87(3H , s , 7-OCH3).13C-NMR(C5D5N , δppm):164.7(C-2), 103.7(C-
3), 183.1(C-4), 153.6(C-5), 133.0(C-6), 159.3(C-7), 91.5(C-8), 153.4(C-9), 106.3(C-
10), 122.1(C-1′), 128.9(C-2′), 116.8(C-3′), 162.8(C-4′), 116.8(C-5′), 128.9(C-6′), 60.5(6
-OCH3), 56.3(7-OCH3).以上数据与文献基本一致 , 故鉴定为蓟黄素 (cirsimaritin).
图 1　香橙中分离的化合物结构
Fig.1　Structures of isolated compounds from Citrus junos
从香橙的二氯甲烷萃取物中分离得到 7个化合物(图 1).根据它们的理化性质与光谱数据 , 鉴定
出3个香豆素类化合物 , 伞形酮(umbelliferone), 9-羟基-4-甲氧补骨脂素 (9-hydroxy-4-methoxyl
38 中央民族大学学报(自然科学版) 第 17卷　
psoralen)和橙皮油素 (auraptene);2 个柠檬苦素类化合物 , 柠檬苦素 (limonin)和脱乙酰诺米林
(deacetylnomilin);2个黄酮类化合物 , 粗毛豚草素 (hispidulin)和蓟黄素 (cirsimaritin).
伞形酮 , 9-羟基-4-甲氧补骨脂素和橙皮油素等香豆素具有抗氧化作用[ 14] , 柠檬苦素和脱乙酰
诺米林均有抗癌作用[ 18] , 粗毛豚草素具有抗肿瘤作用[ 19] .蓟黄素具有抗幽门螺杆菌作用[ 20] .香橙资源
丰富 , 尚未得到有效的利用.近年来 , 有关香橙有效成分和药理作用的研究已经引起一定的关注 , 有
必要进一步研究活性成分的跟踪 , 本研究的结果对于促进香橙资源的利用有一定的参考价值.
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39　第 3期 朴香兰:香橙化学成分的分离与鉴定
[ 5 ] 　李宁 ,陈彬.基于数据库的海量GDF 导航电子地图数据处理方法[ J] .测绘科学 , 2007 , 32(2):157-159.
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Second Edition.The MIT Press , 2001.
Simplification of Forbidden Roads of A Graph
ZHOUMeng ,XU Ci-wen , E Li-li
(College of Science , Central University for Nationalities , Beijing 100081, China)
Abstract:In this paper ,we discuss the method and principle of finding the small set of forbidden roads.
We classified the roads as pass roads , forbidden roads and other roads.Get the small set by split road and
compare them.We try to offer a algorithm:1 Find the pass roads and forbidden roads , use them as the
precondition;2 Split every road , find the parts that could be used under the precondition , and record
those roads the part could replace;3 Choose the best parts , insert into a new set.
Key words:GDF;navigation;no entry;Optimizing
[责任编辑:杨　玉]
(上接第 39页)
Isolation and Identification of Chemical Constituents from Citrus junos
PIAO Xiang-lan
(Chinese Minority Traditional Medical Center , College of Life and Environmental Sciences ,
Central University for Nationalities , Beijing 100081 , China)
Abstract:The antioxidative constituents were studied.Seven components , including three coumarins ,
two limonolids and two flavonolids were isolated from Citrus junos using column chromatography.They
were identified as umbelliferone , 9-hydroxy-4-methoxyl psoralen , auraptene , limonin , deacetylnomilin ,
hispidulin and cirsimaritin , respectively.Their structures were characterized by various spectroscopic
techniques including IR , MS , 1D and 2D 1H-NMR and 13C-NMR.
Key words:Citrus junos;coumarin;limonolid;flavonolid
[责任编辑:白　玲]
56 中央民族大学学报(自然科学版) 第 17卷