全 文 ：天然产物研究与开发 NatProdResDev2009, 21:435-437, 505
文章编号:1001-6880(2009)03-0435-04
　
　
　ReceivedApril13, 2007;AcceptedJune28, 2007
　FoundationItem:ThisworkwassupportedbytheNationalNaturalSci-
enceFoundationofChina(20672120).
＊CorrespondingauthorTel:86-871-5223263;E-mail:haoxj@mail.kib.
ac.cn
脉叶虎皮楠的生物碱成分研究
张　于 1, 2 ,何红平 1 ,邸迎彤 1 ,王跃虎 1 ,高　锁 1, 2 ,郝小江 1＊
1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;
2中国科学院研究生院 ,北京 100049
摘　要:从脉叶虎皮楠(Daphniphyllumpaxianum)的枝干中分离并鉴定了四个生物碱 , 分别为 daphnilactoneA
(1)、daphnicyclidinD(2)、daphniphylline(3)、daphnicyclidinH(4)。其中首次对化合物 1的碳谱数据进行了归
属。
关键词:脉叶虎皮楠;生物碱;化学成分;DaphnilactoneA
中图分类号:R284.1;Q946.91 文献标识码:A
AlkaloidsfromDaphniphylumpaxianum
ZHANGYu1, 2 , HEHong-ping1 , DIYing-tong1 , WANGYue-hu1 , GAOSuo1, 2 , HAOXiao-jiang1＊
1StateKeyLaboratoryofPhytochemistryandPlantResourcesinWestChina, KunmingInstituteofBotany, Chinese
AcademyofSciences, Kunming650204 , China;2GraduateUniversityofChineseAcademyofSciences, Beijing100049 , China
Abstract:Fouralkaloids, daphnilactoneA(1), daphnicyclidinD(2), daphniphylline(3)anddaphnicyclidinH(4),
wereisolatedfromthebranchesofDaphniphyllumpaxianumforthefirsttime.Theirstructureswereestablishedonthe
basisofspectroscopicanalysis.The13CNMRdataofthecompound1wereassignedforthefirsttime.
Keywords:Daphniphylumpaxianum;alkaloid;chemicalconstituent;DaphnilactoneA
Introduction
AlkaloidsfromthegenusDaphniphylumhaveatracted
greatinterestsforitshighlycomplexpolycyclicstruc-
turesfromabiogeneticandtotalsyntheticpointsof
view[ 1, 2] .DaphniphylumpaxianumRosenthisanative
evergreentreeinSouthernChina[ 3] .Thepreviousstud-
iesonthisspeciesresultedinseveralnovelDaph-
niphylumalkaloids[ 4, 5] .Herewereportedtheisolation
andstructureelucidationoffouralkaloids, daphnilac-
toneA(1), daphnicyclidinD(2), daphniphyline(3)
anddaphnicyclidinH(4).
ResultsandDiscussion
Compound1 hasamolecularformulaofC23 H35 NO2
providedbyESI-MSandNMRspectra.The1Hand13C
NMR(DEPT)spectrashowedsignalsoffivequaterna-
rycarbons, fourmethines, elevenmethylenesandthree
methyls.Amongthem, twomethylenes(δC 57.3, δH
2.64 and3.78;δC 65.8, δH 2.67 and2.87)andone
methine(δC 61.5, δH 2.71)weretypicalofnitrogenat-
edgroups, andoneestercarbonylatom(δC 172.9)
fromlactonering.Thesefeatureswereinaccordance
withdaphnilactoneA[ 6] , andtheHMBCcorrelations
werefurtherconfirmeditsstructure(Table1).The13C
NMRdataof1 wereassignedforthefirsttime.
Thestructuresofcompounds2-4 weresuccessivelyde-
terminedasdaphnicyclidinD(2), daphniphyline(3)
anddaphnicyclidinH(4)bycomparisonoftheirNMR
datawiththosereported[ 7, 8] .
Table1　NMRdataandHMBCcorrelationsofcompound1a(CDCl3)
δH δC HMBC(H※C) δH δC HMBC(H※C)
1 2.71 (1H, m) 61.5, 61.5, 61.(d) 2, 7, 8, 13, 22 13a 1.44(1H, m) 24.5(t) 5, 8, 9, 14, 21, 22
2 1.26 (1H, m) 40.5(d) 1, 3, 19, 20 13b 2.18(1H, m)
3a 1.63 (1H, m) 18.9(t) 1, 2, 4, 5 14a 1.40(1H, m) 40.5(t) 13, 22
3b 1.81 (1H, m) 14b 1.92(1H, m)
4a 1.66 (1H, m) 37.4(t) 2, 3, 5, 21 15a 1.58(1H, m) 30.5(t) 9, 10, 16, 17, 22
4b 2.67 (1H, m) 15b 2.18(1H, m)
5 37.8(s) 16a 1.62(1H, m) 27.0(t) 9, 10, 15, 17
6 1.61 (1H, m) 46.4(d) 7, 8, 12, 21 16b 1.80(1H, m)
7a 2.64(1H, d, 14.9) 57.3(t) 1, 5, 11, 21, 23 17a 1.43(1H, m) 38.1(t) 9, 10, 11, 15, 16, 23
7b 3.78(1H, dd, 14.9, 6.1) 17b 1.80(1H, m)
8 39.5(s) 18 1.62(1H, m) 31.3 (d) 2
9 97.5(s) 19 0.90(3H, d, 6.7) 21.1(q) 18, 20
10 51.1(s) 20 0.93(3H, d, 6.5) 21.4(q) 18, 19
11a 1.56 (1H, m) 29.9(t) 6, 12, 17, 10, 23 21 1.05(3H, s) 28.1 (q) 4, 6, 8
11b 2.10 (1H, m) 22 172.9(s)
12a 1.26 (1H, m) 29.2(t) 5, 11 23a 2.67(1H, m) 65.8(t) 9, 11, 12, 17, 22
12b 2.69 (1H, m) 23b 2.87(1H, m)
1HNMRdatawereobtainedat400MHzand13CNMR
dataat100 MHz.2DNMRexperimentswerecarried
outat500 MHz.
MaterialsandMethods
Apparatusandplantmaterials
NMRspectraweremeasuredonaBrukerAM-400(1D
NMR)andBrukerDRX-500(2DNMR)spectrometer
withTMSasinternalstandard.ESI-MSspectrawere
carriedonFinniganMAT90.IRspectrawererecorded
onaBio-RadFTS-135 Spectrophotometer(KBr).Opti-
calrotationsweremeasuredonaPerkin-Elmer241au-
tomaticpolarimeter.C18 reverse-phasedsilica(Pharma-
cia), SephadexLH-20 (Pharmacia)andsilicagel
(300-400 mesh, QingdaoHaiyangChemicalCo.,
Ltd., China)wereusedforcolumnchromatography.
SilicagelGF254 plates(QingdaoHaiyangChemical
Plant)wereusedforTLC.
ThebranchesofD.paxianumwerecolectedfromSi-
chuanProvince, andauthenticatedbyProf.GongXun
atKunmingInstituteofBotany, ChineseAcademyof
Sciences.
Extractionandisolation
ThebranchesofD.paxiaum(20 kg)wereextracted
threetimeswith95% EtOH.Thecrudeextractwasad-
justedtopH≈ 4withtartaricacid.Theacidicmixture
waspartionedwithEtOAc.Theaqueouslayerwasad-
justedtopH≈ 10withsaturatedNa2CO3 andthenex-
tractedwithCHCl3 toobtaincrudealkaloids(20 g).
Thecrudealkaloidsweresubjectedtoasilicagelcol-
umn(CHCl3 /MeOH, 1∶0※ 20∶1※10∶1※5∶1※0∶1,
v/v)togivefractions1-5.Fraction1(120 mg)was
separatedoverasilicagelcolumn(CHCl3 /MeOH, 10
∶1※4∶1 , v/v)andthenpurifiedbySephadexLH-20
columnelutedwithmethanoltogive1 (15 mg,
0.075%), 2 (6 mg, 0.03%).Fraction3 wassubjec-
tedtoaRP-18 column(MeOH/H2O, 3∶7※7∶3 , v/v)
togettwofractions, theywerealseparatedoversilica
gelcolumnelutedwithCHCl3 -MeOH(8∶1※2∶1, v/
v)togive3(5mg, 0.025%)and4(8mg, 0.04%).
Identification
DaphnilactoneA(1)　C23 H35NO2 , colorlesssolid.
[ α] 20D-21.9°(c0.45, CHCl3 );IRυKBrmaxcm-1:1732;
436 NatProdResDev　　　　　　　　　　　　　　　　　　　　 　Vol.21
ESI-MSm/z:358[ M+H] +.NMR(CDCl3)datasee
Table1.
DaphnicyclidinD(2)　C23H27NO4 , lightyelowsol-
id;[ α] 24D-85°(c0.43, CH3OH);IRυKBrmaxcm-1:3427,
2935, 1679, 1558;ESI-MSm/z:382 [ M +H] +;1H
NMR(CDCl3)δ:2.60(1H, brs, H-2), 2.27 (1H, d,
15.9Hz, H-3a), 2.43 (1H, m, H-3b), 3.31 (1H, m,
H-4), 2.64(1H, m, H-6), 2.60 (1H, m, H-7a), 4.20
(1H, m, H-7b), 2.65(1H, m, H-11a), 2.91(1H, dt,
11.9Hz, 17.2 Hz, H-11b), 1.63 (1H, m, H-12a),
2.44 (1H, m, H-12b), 3.10 (1H, dd, 4.0 Hz, 16.5
Hz, H-16a), 3.29 (1H, dd, 3.2 Hz, 16.5 Hz, H-
16b), 4.22 (1H, m, H-17a), 4.75 (1H, dd, 5.2 Hz,
10.7Hz, H-17b), 2.46 (1H, m, H-18), 3.24 (2H,
m, H-19), 1.36(3H, d, 7.2 Hz, H-20), 1.53(3H, s,
H-21), 3.9 (3H, s, H-22-OMe);13CNMR(CDCl3)
δ:197.3(C-1), 47.6 (C-2), 17.1 (C-3), 65.4 (C-
4), 51.3 (C-5), 47.7 (C-6), 59.3 (C-7), 137.1
(C-8), 121.0 (C-9), 179.8 (C-10), 31.2 (C-11),
30.2 (C-12), 134.8 (C-13), 123.4 (C-14), 129.8
(C-15), 22.6 (C-16), 69.1 (C-17), 28.1 (C-18),
52.6(C-19), 18.1(C-20), 33.4 (C-21), 166.9 (C-
22), 51.7 (C-22-OMe).TheESI-MS, NMRspectral
datawereinconsistentwiththosereported[ 7] .
Daphniphyline(3)　C32 H49 NO5 , colorlesssolid;
[ α] 24D +31°(c0.9, CHCl3);IRυKBrmaxcm-1:1742, 1672;
ESI-MSm/z:528 [ M+H] +;1HNMR(CDCl3)δ:
3.38 (1H, d, 12.6 Hz, H-1), 1.64 (1H, m, H-2),
1.88 (1H, m, H-3a), 2.04 (1H, m, H-3b), 1.62
(1H, m, H-4a), 1.91 (1H, m, H-4b), 1.90 (1H, m,
H-6), 3.4 (2H, m, H-7), 2.61 (1H, d, 18.3 Hz, H-
9), 1.62 (1H, m, H-11a), 1.92 (1H, m, H-11b),
1.55 (2H, m, H-12), 1.47 (1H, m, H-13a), 2.36
(1H, m, H-13b), 5.62 (1H, dd, 3.0 , 12.5, H-14),
1.45(1H, m, H-15a), 2.18 (1H, m, H-15b), 1.34
(1H, m, H-16a), 1.89 (1H, m, H-16b), 1.84 (1H,
m, H-17a), 2.32 (1H, m, H-17b), 2.20 (1H, m),
0.90(3H, d, 6.5 Hz, H-19), 1.04 (3H, d, 6.0 Hz,
H-20), 0.98 (3H, s, H-21), 0.89 (3H, s, H-24),
3.69(1H, d, 14.8 Hz, H-25a), 4.50 (1H, dd, 2.0
Hz, 14.8 Hz), 4.60 (1H, d, 6.6 Hz, H-26), 1.91
(1H, m, H-27a), 2.04 (1H, m, H-27b), 1.89 (1H,
m, H-28a), 2.08 (1H, m, H-28b), 1.47 (3H, s, H-
30), 2.10 (3H, s, H-32);13CNMR(CDCl3)δ:62.4
(C-1), 37.8 (C-2), 21.7 (C-3), 39.8 (C-4), 37.1
(C-5), 37.8 (C-6), 46.2 (C-7), 47.5 (C-8), 52.9
(C-9), 77.3 (C-10), 25.2 (C-11), 28.4 (C-12),
30.2 (C-13), 73.5 (C-14), 31.0 (C-15), 25.1 (C-
16), 36.1 (C-17), 30.5 (C-18), 20.8 (C-19), 20.9
(C-20), 23.8 (C-21), 212.6 (C-22), 50.5 (C-23),
18.8 (C-24), 65.2 (C-25), 82.1 (C-26), 25.1 (C-
27), 33.7 (C-28), 105.3 (C-29), 25.1 (C-30),
170.4 (C-31), 21.7(C-32).ItsNMRdatawereiden-
ticalwiththosereported[ 8] .
DaphnicyclidinH(4)　C23H29NO5 , lightyelowsol-
id;[ α] 20D -108°(c0.58, CH3OH);IRυKBrmaxcm-1:3430,
2930, 1678, 1566;ESI-MSm/z:400 [ M+H] +;1H
NMR(CDCl3)δ:2.32 (1H, m, H-2), 2.26 (2H, m,
H-3), 3.63 (1H, br, H-4), 2.56 (1H, m, H-6), 2.67
(1H, m, H-7a), 3.76 (1H, m, H-7b), 2.36 (1H, m,
H-11a), 2.60 (1H, m, H-11b), 1.76 (1H, m, H-
12a), 1.88 (1H, m, H-12b), 2.64 (1H, m, H-16a),
2.88 (1H, m, H-16b), 3.64 (2H, m, H-17), 2.00
(1H, m, H-18), 3.19 (1H, br, H-19a), 3.87(1H, m,
H-19b), 1.25 (3H, d, 6.9 Hz, H-20), 1.34 (3H, s,
H-21), 3.68 (3H, s, H-22-OMe);13CNMR(CDCl3)
δ:195.7 (C-1), 47.8 (C-2), 17.7 (C-3), 66.9 (C-
4), 49.7 (C-5), 47.9 (C-6), 59.9 (C-7), 131.4
(C-8), 125.9 (C-9), 203.0 (C-10), 38.8 (C-11),
29.4 (C-12), 122.9 (C-13), 121.2(C-14), 130.2
(C-15), 29.7 (C-16), 64.9 (C-17), 28.2 (C-18),
54.3 (C-19), 17.4(C-20), 34.8(C-21), 171.9(C-
22), 51.8 (C-22-OMe).TheESI-MS, NMRdatawere
inconsistentwiththosereported[ 7] .
Acknowledgment　Wearegratefultoalthemembers
oftheanalyticalgroupforrecordingspectruminthe
StateKeyLaboratoryofPhytochemistryandPlantRe-
sourcesinWestChinaKunmingInstituteofBotony
CAS.ThisworkwassupportedbytheNationalNatural
ScienceFoundationofChina(20672120 ).
References
1　KobayashiJ, MoritaH.TheAlkaloids.Cordell, GAED;New
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(下转第 505页)
437Vol.21 　　　　　　ZHANGYu, etal:AlkaloidsfromDaphniphyllumpaxianum 　
峰为吡喃糖 β型 OH变角振动特征峰 , 1635 cm-1处
νC=O的伸缩振动反映的是乙酰氨基 ,其吸收强弱与
脱乙酰度有直接的关系。
图 4　壳聚糖红外光谱图
Fig.4　IRspectrumofchitosan
3.5.3　壳聚糖水分的测定
壳聚糖水分含量为 8.96%,符合商品壳聚糖的
标准。
3.5.4　壳聚糖的含量(纯度)的测定
采用溴甲酚绿显色比色法测得壳聚糖的含量为
90.2%。
通过从拟康氏木霉菌丝中提取壳聚糖研究 ,获
得了产率高 、提取不同脱乙酰度和不同分子量壳聚
糖的提取条件 ,这些条件为同时获得不同脱乙酰度
和分子量的壳聚糖提供了可能 。另外本研究用拟康
氏木霉提取壳聚糖产率可达 14.4%,高于其它菌丝
体来源的壳聚糖产率 。与虾蟹来源的壳聚糖相比 ,
具有耗能少 ,提取工艺相对简单 ,无需脱色等优点 ,
由于拟康氏木霉还是兰迪多邦多功能生物制剂的副
产品 ,对其开发利用可节约资源 ,减少污染 ,具有较
大的经济效益。
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505Vol.21 　　　　　　何圣红等:从拟康氏木霉中提取壳聚糖的初步研究