全 文 ：·化学成分·
Studies on chemical constituents from Tetrastigma hypoglaucum
LIU Dong , JU Jian-hua, YAN G Jun-shan
*
( Institute of Medicina l Plant Development, CAM S& PUM C, Beijing 100094, China )
Abstract: Object　 To investiga te the chemical consti tuents in the aerial par t of Tetrast igma hypoglau-
cum Planch. nativ e to China in o rder to obtain a mo re comprehensiv e understanding of i ts effectiv e compo-
nents. Methods　 Compounds w ere isola ted by various column chromato graphies wi th silica g el. Thei r
st ructures w ere elucida ted by spect ral analy sis ( IR, M S,
1
HNMR,
13
CNM R) and chemical ev idence. Re-
sults　 Ten compounds w ere iso la ted f rom the ethano l ex t racts of T . hypoglaucum , w hich w ere identified
as β -sitosterol (Ⅰ ) , palmi tic acid (Ⅱ ) , pentacosane (Ⅲ ) , daucostero l (Ⅳ ) , resv erat rol (Ⅴ ) , gallic acid
(Ⅵ ) , ethy l gallate (Ⅶ ) , catechin (Ⅷ ) , 7-O-gallo ylca techin (Ⅸ ) , a nd 3, 3′-dimethoxy ellagic acid-4-O-β-
D-g lucopyranoside (Ⅹ ) . Conclusion　 All compounds w ere obtained from this plant for the f irst time.
Key words: Tetrastigma hypoglaucum Planch. ; ca techin; chemica l consti tuents
狭叶崖爬藤化学成分的研究
刘　东 ,鞠建华 ,杨峻山
(中国医学科学院 中国协和医科大学药用植物研究所 ,北京　 100094)
摘　要: 目的　研究我国特有药用植物狭叶崖爬藤 Tetrastigma hypoglaucum 的化学成分 ,为阐明其有效成分提供
依据。 方法　利用各种色谱技术进行分离 ,根据化合物的光谱数据 ( IR, M S, 1 HNM R, 13CNM R)和化学方法鉴定其
结构。 结果　从狭叶崖爬藤地上部分分离并鉴定了 10个化合物 ,分别为 β -谷甾醇 (β-sito stero l,Ⅰ ) ,棕榈酸
( palmitic acid,Ⅱ ) ,正二十五烷 ( pentacosane,Ⅲ ) ,胡萝卜苷 ( daucoste rol,Ⅳ ) ,白藜芦醇 ( resv erat rol,Ⅴ ) ,没食子酸
( g allic acid,Ⅵ ) ,没食子酸乙酯 ( ethy l g alla te,Ⅶ ) ,儿茶素 ( cat echin,Ⅷ ) , 7-氧 -没食子酰基 -儿茶素 ( 7-O-gallo ylca te-
chin,Ⅸ )和 3, 3′-二甲氧基鞣花酸 -4-氧 -葡萄糖苷 ( 3, 3′-dimethoxy ellagic acid-4-O-β -D-g lucopy rano side,Ⅹ )。结论
所有化合物均系首次从该植物中分离得到。
关键词: 狭叶崖爬藤 ;儿茶素 ;化学成分
中图分类号: R284. 1　　　文献标识码: A　　　文章编号: 0253 2670( 2003) 01 0004 03
　　 Tetrastigma hypoglaucum Planch. ( Vi taceae)
is widely dist rbuted in Sichuan and Yunnan
Provinces of China. As a Chinese fo lk herb
medicine, i ts rhizomes o r the w hole plants are used
fo r the t reatment of f racture, t raumatic injury ,
and sw elling pain due to pathogenic w ind-damp-
ness
[1 ] . In o rder to obtain a mo re comprehensive
understanding of it s ef fectiv e consti tuents, ten
compounds (Ⅰ -Ⅹ ) w ere isolated f rom the 95%
EtO H ex tracts o f this plant. These ten com-
pounds, namely, β -sitosterol (Ⅰ ) , palmi tic acid
(Ⅱ ) , pentacosane (Ⅲ ) , daucosterol (Ⅳ ) , resv er-
at ro l (Ⅴ ) , gallic acid (Ⅵ ) , ethy l g allate ( Ⅶ ) ,
ca techin (Ⅷ ) , 7-O-galloy lcatechin (Ⅸ ) , and 3,
3′-dimethoxy ellagic acid-4-O-β-D-g lucopy ranoside
(Ⅹ ) w ere identi fied by comparison wi th authentic
samples, and on the basis o f thei r physical con-
stants and spect ral da ta
[2～ 7 ] .
Experimental section
Genera l experimental procedures. Melting
points were determined on a Fisher-Johns appara-
tus and w ere uncorrected. UV spect ra w ere mea-
sured on a Philips PYE Unican PU 8800 spectrome-
ter. IR spect ra w ere recorded on a Perkin-Elmer
·4· 中草药　 Chinese T raditional and Herbal Drug s　第 34卷第 1期 2003年 1月
收稿日期: 2002-07-05基金项目:国家自然科学基金资助重点项目 ( No. 29732040)作者简介:刘　东 ,男 ,博士 ,助理研究员 ,主要从事天然药物化学方面的研究。
* 通讯作者　 Tel: ( 010) 62899707　　 Fax: ( 010) 62899739　　 E-mail: ju nshanyang@ hotmai l. com
983 G spect rometer. NM R spect ra ( 500 MHz fo r
1
HNM R and 125 M Hz for
13
CNMR) w ere mea-
sured on a Burker AM-500 spect rometer, using
TM S as internal standa rd. Coupling constants (J
va lues ) are giv en in Hz. V G ZAB-2F mass spec-
trometer w as used to reco rd the EI-M S, and an
Autospec-Ultima ETO F spect rometer w as used to
record the FAB-M S. Si g el and Si gel GF254 sheets
( 0. 20-0. 25 mm ) ( both f rom Qingdao M arine
Chemical Group Co. , Qingdao , Shandong
Province, China) w ere used fo r column chromatog-
raph y and T LC, respectiv ely. D101 micro reticular
resin was made by Tianjin Nankai Chemical Indus-
try. Sephadex L H-20 ( Pharmacia , 40μm) was
purchased from Sigma Chemicals.
Plant ma terial. The who le plant of T . hy-
poglaucum was collected in Li jiang, Yunnan
Province of China, in June, 1998, and identi fied by
Professo r Lian Wen-yan, from Insti tute of M edici-
nal Plant Development, Chinese Academy o f Medi-
cal Sciences & Peking Union M edical Co lleg e. A
voucher specimen ( HB-98-0609) is deposi ted at the
Institute o f Medicinal Plant Development , Chinese
Academy of Medical Sciences & Peking Union
Medical College.
Ex t raction and isola tion. The air-dried and
pulv erized aerial part o f T . hypoglaucum ( 11. 2
kg ) w as ex tracted th ree times wi th 95% EtO H for
tw o hours under ref lux. Af ter remova l of solv ent ,
the residue ( 1. 64 kg ) wa s suspended in wa ter
( 3 000 mL ) and parti tioned successiv ely wi th
pet ro leum ether ( 3 000 mL× 3) , and n-BuOH
( 3 000 mL× 3) . The pet roleum ether-so luble f rac-
tion ( 150 g ) w as isolated by repeated co lumn chro-
ma tog raphy wi th silica gel and Sephadex LH-20 to
af fo rd compoundsⅠ ( 200 mg ) , Ⅱ ( 40 mg ) , Ⅲ ( 68
mg ) and Ⅳ ( 40 mg ) . The ethy l acetate-soluble
f raction ( 100 g ) was iso lated by repea ted column
chroma tog raphy w ith silica g el and Sephadex LH-
20 to affo rd compoundsⅤ ( 10 mg ) , Ⅵ ( 400 mg ) ,
Ⅶ ( 34 mg ) , Ⅷ ( 200 mg ) and Ⅸ ( 25 mg ) . Com-
poundⅩ ( 22 mg ) w as obtained by repeated column
chroma tog raphy w ith D101 micro reticular resin,
si lica g el and Sephadex LH-20 from the n-butanol-
so luble part.
β-Si to sterol (Ⅰ ): colo rless needles, mp 137
℃-139℃ ( acetone ) ; IRνKBrmax cm- 1: 3 420 ( OH) ,
2 980, 2 960, 1 650 ( C= C) ; EI-M S m /z: 414
[M
+ ] ( 65) , 396 ( 68) , 381 ( 37) , 329 ( 35) , 303
( 40) , 273 ( 31) , 255 ( 65) , 233 ( 30) , 213 ( 55) ,
159 ( 55) , 81 ( 100) . It w as identified by compari-
son wi th an authentic sample.
Palmi tic acid (Ⅱ ): w hi te so lid, mp 56℃ -58
℃ ( acetone ) ; EI-M S m /z: 256 [M+ ] ( 1) , 99
( 12) , 85 ( 48) , 71 ( 65) , 57 ( 100) . It was identi-
fied by comparison w ith an authentic sample.
Pentacosane (Ⅲ ): whi te solid; mp 56℃ -58
℃ ( MeOH); EI-M Sm /z: 352[M+ ] ( 1) , 99 ( 12) ,
85 ( 48) , 71 ( 65) , 57 ( 100) . It was identified by
compa rison wi th an authentic sample.
Daucostero l (Ⅳ ): white amo rphous powder,
mp 290℃ -295 ℃ ( MeOH) ; EI-M S m /z: 576
[M
+ ] , ( 2) , 414 ( 28) , 396 ( 100) , 381 ( 45) , 392
( 14) , 303 ( 14) , 255 ( 32) , 231 ( 12) , 213 ( 24) . It
w as identi fied by comparison wi th an authentic
sample.
Resvera tro l (Ⅴ ) [3 ]: co lorless needles, mp 258
℃-260℃ ( M eOH); IRνKBrmax cm- 1: 3 240, 1 600,
1 583, 1 510, 1 460, 1 440, 1 380, 1 260, 1 145,
1 010, 860, 805. EI-M S m /z: 228 [M
+ ] ( 100) , 211
( 8) , 199 ( 6) , 181 ( 20) ; 1HNM R( CD3OD)δ6. 10
( 1H, t ,J= 2. 0 Hz, H-4′) ; 6. 39( 2H, d, J= 2. 0 Hz,
H-2′and H-6′) , 6. 70( 2H, dd, J= 7. 0, 2. 0 Hz, H-3
and H-5) , 6. 73 ( 1H, d, J = 16. 0 Hz, H-β ) , 6. 89
( 1H, d, J= 16. 0 Hz, H-α) , 7. 29( 2H, dd, J= 7. 0,
2. 0 Hz, H-2 and H-6) ;
13
CNM R( CD3OD)δ102. 6
( C-4′) , 105. 7( C-2′and C-6′) , 116. 0( C-3 and C-
5 ) , 127. 0 ( C-1) , 128. 8 ( C-2 and C-6) , 129. 4 ( C-
1′) , 130. 4( C-β ) , 141. 3( C-α) , 158. 3( C-4) , 159. 6
( C-3′and C-5′) .
Gal lic acid (Ⅵ ) [2 ]: white needles, mp 238℃-
240℃ ( MeOH) ; EI-M S m /z: 170 [M+ ] ( 100) ,
152 [M-H2O ]
+ ( 90) , 135 ( 22) , 125 ( 30) , 107( 16) ,
79 ( 28) . 1HNMR( CD3OD)δ7. 00 ( 2H, s, H-3 and
H-7) .
13
CNM R( CD3OD)δ110. 3 ( C-3 and C-7) ,
122. 0 ( C-2) , 139. 6 ( C-5) , 146. 4 ( C-4 and C-6) ,
170. 4 ( C-1) .
Ethy l g allate (Ⅶ ) [4 ]: whi te needles, mp 146
·5·中草药　 Chinese T raditional and Herbal Drug s　第 34卷第 1期 2003年 1月
℃ -148℃ ( MeOH); IRνKBrmax cm- 1: 3 400, 2 980,
1 700, 1 610, 1 530, 1 450, 1 370, 1 230, 1 100,
1 020, 870, 760. EI-M S m /z: 198 [M+ ] ( 52) , 170
( 20) , 153 ( 100) , 125 ( 18) .
1
HNM R( CD3OD)δ1. 27
( 3H, t , J= 7. 0 Hz, H-9) , 4. 20( 2H, q, J = 7. 0 Hz,
H-8 ) , 6. 99 ( 2H, s, H-3 and H-7 ) . 13CNMR
( CD3OD)δ14. 6( C-9) , 61. 7( C-8) , 109. 9( C-3 and
C-7) , 121. 7( C-2) , 139. 6( C-5) , 146. 4( C-4 and C-
6) , 168. 5( C-1) .
Catechin (Ⅷ ) [5, 6 ]: w hi te needles, mp 154℃ -
156℃ ( MeOH); UV λmax ( nm ): 278 ( MeOH);
294 ( NaOM e) ; 278, 314 ( sh ) , 360 ( AlCl3 ) ; 278,
314, 360 ( AlCl3 + HCl ) ; 278 ( NaO Ac ) ; 278
( H3 BO3 ) . IRνKBrmax cm- 1: 3 380, 1 623, 1 520, 1 465,
1 185, 1 146, 1 025, 820; EI-M S m /z: 290 [ M
+ ]
( 45) , 272 ( 8) , 152 ( 60) , 139 ( 100) , 123 ( 63) .
1
HNM R( CD3OD)δ2. 46( 1H, dd, J = 7. 5, 16. 0 Hz,
H-4a) , 2. 79( 1H, dd, J= 5. 0, 16. 0 Hz, H-4b ) , 3. 92
( 1H, dd, J= 7. 5, 8. 0 Hz, H-3) , 4. 50( 1H, d, J= 7. 5
Hz, H-2) , 5. 86( 1H, d, J= 1. 5 Hz, H-8) , 5. 93( 1H,
d, J= 1. 5 Hz, H-6) , 6. 66( 1H, dd, J = 8. 0 Hz, H-
6′) , 6. 70( 1H, d, J= 8. 0 Hz, H-5′) , 6. 78( 1H, s, H-
2′) ; 13 CNM R( CD3OD)δ28. 5 ( C-4) , 68. 8 ( C-3) ,
82. 8 ( C-2) , 96. 2( C-6) , 95. 4( C-8) , 100. 8( C-10) ,
115. 2( C-2′) , 116. 0( C-5′) , 120. 0( C-6′) , 132. 0( C-
1′) , 146. 2( C-3′and C-4′) , 156. 9( C-7) , 157. 6( C-
9) , 157. 8( C-5) .
7-O-Gallo ylca techin (Ⅸ ) [ 6]: white needles,
mp 262℃-264℃ ( M eOH); UV λmax ( nm ): 278
( MeOH); 294 ( NaOMe ) ; 278, 314 ( sh ) , 360
( AlCl3 ) ; 278, 314, 360 ( AlCl3 + HCl ) ; 278
( NaOAc ) ; 278 ( H3 BO3 ) . IRνKBrmax cm- 1: 3 330,
1 700, 1 623, 1 530, 1 465, 1 350, 1 285, 1 203,
1 140, 1 025, 860, 760. EI-M S m /z: 290 ( 45) , 272
( 8) , 152 ( 60) , 139 ( 100 ) , 123 ( 63) ; 1 HNMR
( CD3OD)δ2. 43( 1H, dd, J= 7. 5, 15. 5 Hz, H-4a ) ,
2. 69 ( 1H, dd, J = 5. 5, 15. 5 Hz, H-4b) , 3. 94( 1H,
dt , J= 7. 5, 5. 5 Hz, H-3) , 4. 50( 1H, d, J= 7. 0 Hz,
H-2) , 6. 17( 1H, d, J = 2. 0 Hz, H-8) , 6. 20 ( 1H, d,
J= 2. 0 Hz, H-6) , 6. 66( 1H, dd, J= 7. 5, 2. 0 Hz, H-
6′) , 6. 71 ( 1H, d, J = 7. 5 Hz, H-5′) , 6. 78( 1H, d,
J= 2. 0 Hz, H-2′) , 7. 13( 2H, d, J= 1. 0 Hz, H-3″) .
13
CNM R( CD3OD)δ28. 6( C-4) , 68. 1( C-3) , 83. 0( C-
2 ) , 101. 6( C-10) , 103. 5( C-8) , 106. 4( C-6) , 110. 5
( C-3″and C-7″) , 115. 1( C-2′) , 116. 1( C-5′) , 120. 0
( C-2″) , 131. 7( C-1′) , 140. 6( C-4″and C-6″) , 146. 2
( C-4′) , 146. 3 ( C-3′) , 146. 7 ( C-5″) , 151. 7 ( C-7) ,
157. 0( C-9) , 158. 1( C-5) , 166. 5( C-1″) .
3, 3′-Dimethoxy ellagic acid-4-O-β -D-glucopy-
rano side (Ⅹ ) [7 ]: whi te amorphous powder, mp
285℃ -287℃ ( M eOH); posi tive to M olish test;
Po si tiv e-ion FAB-M S m /z 493 [M+ H ]
+
; m /z:
330 [M - 162 ]
+ ( 100) , 315 ( 40) ; 1HNM R( DM SO-
d6 )δ5. 16( 1H, d, J= 8. 0 Hz, Glu-H-1) , 3. 22-3. 71
[ m, Glu-H-( 2-6 ) ] , 4. 09 ( 1H, s, 3-OCH3 ) , 4. 05
( 1H, s, 3′-OCH3 ) , 7. 81 ( 1H, s, H-5) , 7. 53 ( 1H, s,
H-5′) . 13 CNMR( CD3OD)δ114. 8 ( C-1) , 142. 4( C-
2) , 142. 4 ( C-3) , 153. 5 ( C-4) , 112. 6 ( C-5) , 112. 5
( C-6) , 159. 1 ( C-7) , 111. 8 ( C-1′) , 141. 6 ( C-2′) ,
140. 8( C-3′) , 152. 3( C-4′) , 113. 5( C-5′) , 112. 3( C-
6′) , 159. 1 ( C-7′) , 62. 4 ( 3-OCH3 ) , 102. 0 ( C-1″) ,
74. 0( C-2″) , 77. 2( C-3″) , 70. 2( C-4″) , 78. 0( C-5″) ,
61. 2( C-6″) .
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保　护　环　境　　保　护　植　被
·6· 中草药　 Chinese T raditional and Herbal Drug s　第 34卷第 1期 2003年 1月