全 文 ：天然产物研究与开发 NatProdResDev2009, 21:960-962, 983
文章编号:1001-6880(2009)06-0960-04
ReceivedJuly8, 2008;AcceptedAugust3, 2009
＊CorrespondingauthorTel:86-871-5223903;E-mail:xcheng@mail.
kib.ac.cn
菜蕨的化学成分研究
王紫娟 1, 2 ,赵勤实 3 ,彭丽艳 3 ,成 晓 1＊
1中国科学院昆明植物研究所 植物园;2中国科学院研究生院 ,北京 100049;
3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:采用硅胶柱层析和凝胶柱层析对菜蕨的丙酮提取物进行分离纯化 ,首次从中分离得到 10个化合物 , 通
过波谱学数据并和已知化合物数据比较 , 鉴定它们分别为 β-sitosterol(1), Stigmast-4-ene-6β-ol-3-one(2), Stig-
mast-4-ene-3, 6-dione(3), Benzeneaceticacid(4), 3β-Hydroxy-5α, 8α-epidioxyergosta-6, 22-diene(5), Stigmast-4-ene-
3β, 6β-diol(6), Stigmast-5-ene-3β, 7α-diol(7), Stigmast-4-ene-6α-ol-3-one(8), Glycerol-1, 3-dihexadecanoate(9)以及
Daucosterol(10)。
关键词:菜蕨;化学成分;豆甾类
中图分类号:R284.1;Q946.48 文献标识码:A
StudyontheChemicalconstituentsofCalipterisesculenta(Athyriaceae)
WANGZi-juan1, 2 , ZHAOQin-shi3 , PENGLi-yan3 , CHENGXiao1＊
1 KunmingInstituteofBotany, theChineseAcademyofSciences;2GraduateUniversityofChineseAcademy
ofSciences, Beijing100049 , China;3TheStateKeyLaboratoryofPhytochemistry
andPlantResourcesinWestChina, KunmingInstituteofBotany, theChineseAcademyofSciences, Kunming650204 , China
Abstract:TencompoundswerefirstlyisolatedfromtheacetoneextractionofthefernCallipterisesculenta(Retz.)J.Sm.
exMooreetHoulst..Thestructuresofthecompoundswereidentifiedasβ-sitosterol(1), Stigmast-4-ene-6β-ol-3-one
(2), Stigmast-4-ene-3, 6-dione(3), Benzeneaceticacid(4), 3β-Hydroxy-5α, 8α-epidioxyergosta-6, 22-diene(5), Stig-
mast-4-ene-3β, 6β-diol(6), Stigmast-5-ene-3β , 7α-diol(7), Stigmast-4-ene-6α-ol-3-one(8), Glycerol-1, 3-dihexade-
canoate(9)andDaucosterol(10)onthebasisofNMRspectroscopyandcomparisonwiththedataofknowncompounds.
Keywords:Callipterisesculenta;Chemicalconstituents;Stigmasterols
Introduction
Calipterisesculenta(Retz.)J.Sm.ex Moore et
Houlst., akindofbigevergreenhygrophilousterrestrial
leptosporangiatefern, belongstothefamilyAthyriaceae
widelydistributedinthetropical, subtropicalofAsia
andtheislandsofPolynesia.Theyusualyinhabitthe
valeysandravinesofshadyforests.Thetenderfronds
areofemplasticandtasty.However, thechemicalcon-
stituentsofthisplanthavenotbeenstudieduptonow.
Inthisstudy, wehaveisolatedandpurifiedtencom-
poundsfrom thefrondsofCalipterisesculenta.The
structureofthecompoundswereidentifiedasβ-sitos-
terol(1), Stigmast-4-ene-6β-ol-3-one(2)[ 1, 2] , Stig-
mast-4-ene-3, 6-dione(3)[ 1, 3, 4] , Benzeneaceticacid
(4)[ 5] , 3β-Hydroxy-5α, 8α-epidioxyergosta-6, 22-diene
(5)[ 6-9] , Stigmast-4-ene-3β, 6β-diol(6), Stigmast-5-
ene-3β, 7α-diol(7)[ 1, 10-12] , Stigmast-4-ene-6α-ol-3-one
(8)[ 1] , Glycerol-1, 3-dihexadecanoate(9)andDau-
costerol(10)mainlybyNMRspectradataandbycom-
parisonthedatawiththoseofknowncompounds.
Experimental
General
The1Hand13CNMRspectrawererecordedonBruker
AM-400 spectrometerswithTMSasinternalstandard.
TheChemicalshiftswerereportedinppmusingCDCl3
assolvent.Silicagel(200-300 mesh)forcolumnchro-
matographyandGF254(10-40μ)forTLCweresupplied
bytheQingdaoMarineChemicalInc.China.Spotson
DOI :10.16333/j.1001-6880.2009.06.036
TLCweremonitoredunderUVlampandbyheatingsil-
icagelplatessprayedwith10%H2SO4 inEtOH.Seph-
adexLH-20(25-100 μm, pharmcia)wereusedforgel
chromatographyandMCI(MCI-gelCHP-20P)forde-
colorizing.
Plantmaterial
ThefrondsofCalipterisesculentawerecolectedfrom
DaWeiShanMountain, HeKouinJune, 2006.A
voucherspecimenCheng20060620 wasdepositedin
theKUN.
ExtractionandIsolation
ThedriedandpowderedfrondsofCalipterisesculenta
(8.0 kg)wereextractedwithacetone(10 L×3, each
oneday)atroomtemperature.Thesolventwasremoved
undervacuum, andaresidue(134 g)wasobtained.Af-
terdecolorizedonMCIcolumnelutedwith 95%
EtOH, theresiduewassubjectedtosilicagelcolumn
(φ8 ×L21 cm)elutedwithpetroleumether:acetone
(10:0, 9:1, 7:3, 6:4)togivefractions1-4.Through
recrystalizationfrompetroleumether, compound1(5.5
g)waspurifiedfromfraction2 andcompound2(4 g)
fromfraction3.Thefraction3 wasfurtherisolatedand
purifiedthroughsilicagelcolumnchromatographyand
gelchromatographyrepeatedly, togivethecompound3
(3 mg), 4(240 mg), 5(8 mg), 6(26 mg), 7(118
mg), 8(49 mg)andCompound9(190 mg).compound
10(about1 g)waspurifiedfromfraction4.
Identification
Compound1 and10 wereidentifiedasβ-sitosteroland
Daucosterolbycomparingwithauthenticsampleson
TLC.
Stigmast-4-ene-6β-ol-3-one(2) Colorlessplates
(acetone);1HNMR(CDCl3 , 400 MHz)δ:5.80(1H,
s, H-4), 4.34(1H, t, J=7.5 Hz, H-6), 0.73(3H, s,
H-18), 1.37(3H, s, H-19), 0.92(3H, d, J=6.5 Hz,
H-21), 0.84(3H, d, J=7.3 Hz, H-26), 0.82(3H, d,
J=6.8 Hz, H-27), 0.85(3H, t, J=7.6 Hz, H-29);13
CNMR(CDCl3 , 100 MHz)δ:37.03(t, C-1), 34.23(t,
C-2), 200.56(s, C-3), 126.24(d, C-4), 168.68(s, C-
5), 73.17(d, C-6), 38.51(t, C-7), 29.68(d, C-8),
53.57(d, C-9), 37.96(s, C-10), 20.93(t, C-11),
39.55(t, C-12), 42.46(s, C-13), 55.84(d, C-14),
24.12(t, C-15), 28.16(t, C-16), 56.00(d, C-17),
11.94(q, C-18), 19.47(q, C-19), 36.08(d, C-20),
18.70(q, C-21), 33.84(t, C-22), 26.00(t, C-23),
45.76(d, C-24), 29.08(d, C-25), 19.80(q, C-26),
18.99(q, C-27), 23.02(t, C-28), 11.99(q, C-29).
Stigmast-4-ene-3, 6-dione(3) Colorlesssolids;1H
NMR(CDCl3 , 400 MHz)δ:6.17(1H, s, H-4), 0.72
(3H, s, H-18), 1.17(3H, s, H-19), 0.93(3H, d, J=
6.4Hz, H-21), 0.83(3H, d, J=7.1 Hz, H-26), 0.81
(3H, d, J=7.0 Hz, H-27), 0.84(3H, t, J=6.8 Hz,
H-29);13 CNMR(CDCl3 , 100 MHz)δ:35.52(t, C-
1), 33.98(t, C-2), 199.57(s, C-3), 125.45(d, C-
4), 161.10(s, C-5), 202.43(s, C-6), 46.83(t, C-7),
34.20(d, C-8), 50.96(d, C-9), 39.80(s, C-10),
20.86(t, C-11), 39.11(t, C-12), 42.51(s, C-13),
55.82(d, C-14), 23.97(t, C-15), 28.02(t, C-16),
56.53(d, C-17), 11.97(q, C-18), 17.51(q, C-19),
36.03(d, C-20), 18.70(q, C-21), 33.80(t, C-22),
25.98(t, C-23), 45.77(d, C-24), 29.09(d, C-25),
19.83(q, C-26), 18.99(q, C-27), 23.04(t, C-28),
11.89(q, C-29).Theabovespectradatawereinac-
cordancewithreportofshenetal[ 3] ;whileinthere-
portsofGrecaetal[ 1] andGasparetal[ 4] , the13 C
NMRchemicalshiftsofC-8, C-10 andC-12 were
39.1, 34.2and39.8ppmrespectively, theasignments
ofthethreecarbons signalsshouldbeinterchanged.
Benzeneaceticacid(4) Colorlessplates(petroleum
ether);1HNMR(CDCl3 , 400MHz)δ:7.25-7.35(5H,
m), δ3.65(2H, s);13 CNMR(CDCl3 , 100 MHz)δ:
178.02(s, C-2′), 133.18(s, C-1), 129.36(d, C-2),
129.36(d, C-6), 128.63(d, C-3), 128.63(d, C-5),
127.35(d, C-4), 41.03(t, C-1′).Thestructureestab-
lishmentofcompound4wasmainlybycomparisondata
withthoseofesterofBenzeneaceticacid[ 5].
3β -Hydroxy-5α, 8α-epidioxyergosta-6, 22-diene(5)
Colorlessneedles(MeOH);1HNMR(CDCl3 , 400
MHz)δ:6.51(1H, d, J=8.5 Hz, H-6), 6.25(1H, d, J
=8.5 Hz, H-7), 5.20(1H, dd, J=7.4, 15.10 Hz, H-
22), 5.16(1H, dd, J=8.1, 15.04 Hz, H-23), 3.96
(1H, m, H-3);13 CNMR(CDCl3 , 100 MHz)δ:34.65
(t, C-1), 30.07(t, C-2), 66.45(d, C-3), 36.92(t, C-
4), 82.14(s, C-5), 135.38(d, C-6), 130.72(d, C-
7), 79.40(s, C-8), 51.02(d, C-9), 36.88(s, C-10),
23.37(t, C-11), 39.29(t, C-12), 44.52(s, C-13),
961Vol.21 WANGZi-juan, etal:StudyontheChemicalconstituentsofCalipterisesculenta(Athyriaceae)
51.64(d, C-14), 20.60(t, C-15), 28.64(t, C-16),
56.14(d, C-17), 12.84(q, C-18), 18.16(q, C-19),
39.73(d, C-20), 20.85(q, C-21), 135.17(d, C-22),
132.26(d, C-23), 42.73(d, C-24), 33.03(d, C-25),
19.93(q, C-26), 19.62(q, C-27), 17.53(q, C-28).
Stigmast-4-ene-3β, 6β-diol(6) Colorlesscrystals
(MeOH);1HNMR(CDCl3 , 500MHz)δ:5.74(1H, d,
J=3.9 Hz, H-4), 4.25(2H, m, H-3, H-6);13CNMR
(CDCl3 , 100 MHz)δ:36.70(t, C-1), 31.41(t, C-2),
71.35(d, C-3), 120.70(d, C-4), 146.10(s, C-5),
73.91(d, C-6), 42.27(t, C-7), 37.14(d, C-8),
42.69(d, C-9), 37.41(s, C-10), 20.77(t, C-11),
39.00(t, C-12), 42.05(s, C-13), 49.03(d, C-14),
24.25(t, C-15), 28.24(t, C-16), 55.62(d, C-17),
11.45(q, C-18), 18.24(q, C-19), 36.12(d, C-20),
18.79(q, C-21), 33.90(t, C-22), 25.90(t, C-23),
45.79(d, C-24), 29.09(d, C-25), 19.82(q, C-26),
18.99(q, C-27), 23.03(t, C-28), 11.98(q, C-29).
Stigmast-5-ene-3β, 7α-diol(7) Colorlessneedles
(MeOH);1HNMR(CDCl3 , 500 MHz)δ:5.61(1H,
dd, J=5.4, 1.33 Hz, H-6), 3.85(1H, brs, H-7),
3.55(1H, m, H-3), 0.68(3H, s, H-18), 0.99(3H, s,
H-19), 0.93(3H, d, J=6.5 Hz, H-21), 0.84(3H, d,
J=6.9 Hz, H-26), 0.82(3H, d, J=6.8 Hz, H-27),
0.86(3H, t, J=7.5Hz, H-29);13CNMR(CDCl3 , 125
MHz)δ:36.99(t, C-1), 31.35(t, C-2), 71.31(d, C-
3), 41.99(t, C-4), 146.22(s, C-5), 123.84(d, C-
6), 65.33(d, C-7), 37.50(d, C-8), 42.25(d, C-9),
37.39(s, C-10), 20.68(t, C-11), 39.15(t, C-12),
42.12(s, C-13), 49.40(d, C-14), 24.27(t, C-15),
28.24(t, C-16), 55.70(d, C-17), 11.60(q, C-18),
18.22(q, C-19), 36.07(d, C-20), 18.78(q, C-21),
33.90(t, C-22), 25.92(t, C-23), 45.81(d, C-24),
29.12(d, C-25), 19.76(q, C-26), 19.00(q, C-27),
23.04(t, C-28), 11.96(q, C-29).Thedatawerere-
sembletothoseofliteratures[ 10, 12] , whileinthelitera-
tures[ 1, 11] , the13 CNMRchemicalshiftsofC-5(δ
143.88), C-15(δ25.90), andC-16(δ29.22), C-23
(δ28.88)werediferent, theyshouldbecorected.
Stigmast-4-ene-6α-ol-3-one(8) Colorlesscrystals
(MeOH);1HNMR(CDCl3 , 500MHz)δ:6.17(1H, d,
J=1.8 Hz, H-4), 4.33(1H, m, H-6), 0.72(3H, s, H-
18), 1.19(3H, s, H-19), 0.82(3H, d, J=6.1 Hz, H-
27), 0.85(3H, d, J=6.0 Hz, H-26), 0.93(3H, d, J
=6.5 Hz, H-21), 0.86(3H, t, J=6.8 Hz, H-29);13C
NMR(CDCl3 , 100 MHz)δ:36.23(t, C-1), 34.11(t,
C-2), 199.52(s, C-3), 119.62(d, C-4), 171.54(s, C-
5), 68.69(d, C-6), 41.46(t, C-7), 33.79(d, C-8),
53.71(d, C-9), 39.01(s, C-10), 20.99(t, C-11),
39.40(t, C-12), 42.11(s, C-13), 55.51(d, C-14),
24.15(t, C-15), 28.10(t, C-16), 55.94(d, C-17),
11.91(q, C-18), 18.25(q, C-19), 36.07(d, C-20),
18.66(q, C-21), 33.57(t, C-22), 25.96(t, C-23),
45.75(d, C-24), 29.07(d, C-25), 19.81(q, C-26),
18.98(q, C-27), 23.01(t, C-28), 11.95(q, C-29).
Glycerol-1, 3-dihexadecanoate(9) Whitepow-
der;1HNMR(CDCl3 , 400 MHz)δ:0.88(6H, t, J=
6.4 Hz, H-16′), 1.26(44H, overlap, H-4′-H-14′),
1.36(4H, m, H-15′), 1.62(4H, m, H-3′), 2.35(4H,
t, J=7.5 Hz, H-2′), 2.49(1H, brs, OH), 4.08-4.21
(5H, m, H-1 , 2, 3);13CNMR(CDCl3 , 500 MHz)δ:
173.91(s, C-1′), 68.38(d, C-2), 65.02(t, C-1, 3),
34.09(t, C-2′), 31.91(t, C-14′), 29.67-29.11(t, C-
4′-C-13′), 24.88(t, C-3′), 22.68(t, C-15′), 14.10
(q, C-16′).
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(下转第 983页)
962 NatProdResDev Vol.21
品 、熏肉 、烟熏 、坚果等食用香精 。已报道在天然依
兰 、肉豆蔻 、丁香 、水仙 、晚香玉 、香石竹等精油中存
在。在香根油中检出该物质也属首次发现。
3.2.2 香兰素(组分 11)Vanilin
分子式为 C8H8O3 ,相对质量 152.14。 FEMA编
号为 3107, FDA182.60, COE107, 中国 GB2760-
1996批准为允许使用的食品香料 。香兰素是重要
的香荚兰的香料之一 ,作粉底香料 ,几乎用于所有香
型 ,如紫罗兰 、兰草 、葵花 、玫瑰 、茉莉等 。香兰素在
食品烟酒中应用也很广泛 ,在香子兰 、巧克力 、太妃
香型中是必不可少的香料 。天然存在于秘鲁香脂 、
丁子香兰油 、香子兰 、咖啡 、葡萄 、白兰地 、威士忌中。
在香根油中检出该物质属首次发现 。
4 问题与研究方向
香根油化学成分研究与提取工艺 、香根草种植
有密切关系 ,不同提取工艺 、种植时间可能合成的化
学物质不同 ,本研究将计划测定传统蒸馏法提取的
香根油化学成分 ,并与浸提法比较 ,分析香根油成分
与提取工艺之间的关系 。
对已发现的新香料成分的分离 、化学性质 、应
用 、合成等研究工作需要进一步进行 ,工业化生产及
应用有待进一步研究。
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983Vol.21 王飞生等:香根草油香气成分分析