全 文 ：应用与环境生物学报　 2006, 12(4):487～ 495 　 　
Chin J A pp lEnviron B io l=ISSN 1006-687X 　 2006-08-25
　Chem ical Study on the Stem ofLonicera landeolata
MA Chunhui
1, 2 , LI Bogang1 , XU Q ing1＆ ZHANG Guolin1*
(1Chengdu In stitu te of B io logy, 2Chengdu In st itu te ofO rgan ic Chem istry, Ch inese Academ y of S cien ces, C hengdu 610041, C hina)
Abstract　Tw en ty com poundsw ere obta ined fo r the first tim e from the 95% - e thano l ex trac t o f the stem s o fLonicera landeola-
taW al.l In Roxb. They w ere identified as(+)-cyclooliv il(1), ( +)-frax iresinol(2), prinsep io l(3), pa lm itic ac id (4),
4-me thy lpheno l(5), m ethy l 2, 4-dihydroxy-3, 6-dime thy lbenzoa te (6), apigenin (7), luteo lin (8), 5, 7, 4′-trihydroxy fla-
vonol 7-O-β-D-g lucopyranoside (9), querce tin (10), gen iste in (11), prune tin (12), geniste in 5-O-β-D-g lucopy ranoside
(13), prune tin 5-O-β-D-g lucopyranoside (14), 5, 7, 4′-trihydroxy flavanone 7-O-β-D-g lucopy ranoside (15), succinic acid
(16), ( +)-ca techin(17), ( -)-epica techin(18), β-sito ste ro l(19) and β-daucostero l(20). The ir structure sw ere dete r-
m ined by spectroscop icm e thods or comparison w ith au then tic sam ples. Among them , compounds 10, 17 and 18 w ere theACE I
active constituents. T ab 4, Re f 63
Keywords　Lonicera landeola ta;ang iotensin-converting enzym e;flavono id;lignin
CLC　Q949. 781. 206
柳叶忍冬的化学成分研究
马春辉 1, 2　李伯刚 1　许 庆 1　张国林 1*
(1中国科学院成都生物研究所;2中国科学院成都有机化学研究所　成都　610041)
摘　要　从柳叶忍冬 95%乙醇提取物中首次分离并鉴定了 20个化合物:(+)-cyclooliv il(1)、 (+)-frax iresinol(2)、
p rinsepio l(3)、 软脂酸(4)、对甲基苯酚(5)、 2, 4-二羟基-3, 6-二甲基苯甲酸甲酯 (6)、洋芹素(7)、 木犀草素(8)、 5,
7, 4′-三羟基黄酮醇 7-O-β-D-吡喃葡萄糖苷(9)、 槲皮素(10)、染料木素(11)、 5, 4′-二羟基-7-甲氧基异黄酮 (12)、 染
料木素 5-O-β-D-吡喃葡萄糖苷(13)、 5, 4′-二羟基-7-甲氧基异黄酮 5-O-β-D-吡喃葡萄糖苷(14)、 5, 7, 4′-三羟基双氢黄
酮 7-O-β-D-吡喃葡萄糖苷(15)、 丁二酸 (16)、 (+)-儿茶素 (17)、 ( -)-表儿茶素 (18)、 谷甾醇 (19)和胡萝卜苷
(20).应用波谱方法及与已知品对照进行结构鉴定.其中 , 化合物 10、 17和 18是抑制血管紧张素转化酶的活性成分.
表 4参 63
关键词　柳叶忍冬;血管紧张素转换酶;黄酮;木质素
CLC　Q949. 781. 206
　　The genus Lonicera (Caprifo liaceae) consists o f about 200
spec ie s in thew o rld. Abou t 100 spec ie s of them are found in Chi-
na[ 1] . The p lants of th is genus are rich in flavono ids[ 2, 3] , organ ic
acids[ 4] and seco iridoid g ly co sides[ 5 , 6] .
Lonicera landeolata W al.l In Roxb. m a in ly occu rs in T ibe t,
Nepal and Bhutan[ 7] . Its chem ical constituents we re no t reported
befo re.
Tw en ty compounds w ere ob ta ined from the 95% - e thano l ex-
tract ofL. landeolata. On the basis o f spec tra l ev idence, they w ere
identified as(+)-cycloo liv il(1), ( +)-fraxiresino l(2), prin-
sep io l(3), pa lm itic acid (4), 4-m e thy l-pheno l(5), m e thy l 2,
4-dihydroxy-3, 6-dim e thy lbenzoa te (6), ap igenin (7), lu teo lin
(8), 5, 7, 4′-trihydroxy flavono l 7-O-β-D-g lucopy ranoside (9),
que rce tin (10), gen iste in (11), prune tin(12), geniste in 5-O-β-
D-g lucopy rano side ( 13), prunetin 5-O-β-D-g luco-py rano side
( 14), 5, 7, 4′-trihydroxy flavanone 7-O-β-D-g lucopy rano side
(15), succ inic ac id (16), ( +)-ca techin (17), ( -)-epica te-
chin(18), β-sito stero l(19) and β-dauco ste ro l(20).
Received:2005-07-13　　A ccepted:2005-09-20
*C orresponding au thor(E-m ail:zhanggl@ cib. ac. cn)
1　Experiment Section
1. 1　General expermi ent procedures
M elting po ints w ere de term ined using a XRC-1 m elting po in t
appa ra tus(Science Instrum ents Factory, S ichuan Unive rsity) and
unco rrected. IR spectra and op tical ro ta tions we re obtained on a
Pe rkin E lm er spec trum one FT-IR spec trom eter (KBr disc) and a
Pe rkin E lm er 341 au tom atic polarim eter, re spec tively. M ass spec-
tra we re ob tained on F inngen-LCQDECA m ass spectrom e te r (ES-
IM S). NM R spectra we re recorded on B ruker Avance 600 spec-
trom e te rs w ith TM S as in te rnal standard. Silica ge lH (160 ～ 200
and 200 ～ 300 m esh, QingdaoH aiyang Chem ica lCo. , L td. ), S il-
ica ge l 60(230 ～ 400 m esh, M erck) and RP -18 silica ge l(P re-
pex 40 ～ 63 μm , Phenom enex) we re used fo r column ch rom a tog ra-
phy. P recoated plates ( silica ge l GF
254
, 0 ～ 40 μm , Q ingdao
H aiyang Chem ical Co. , L td. ), activa ted a t 110 ℃ fo r 2 h, w ere
used fo r TLC ana ly sis. PTLC was v isualized by spraying aniline-
phtha lic acid(0. 93 g an iline and 1. 66 g ph thalic ac id in 100 mL
n -bu tano l) reagent. The sam e reagent toge therw ith 10% (w )
e thano l so lution o f phosphomo lybdic ac id and 5% (w) FeC l
3
aque-
ous so lution w ith sm a ll am oun t o f hydrochloride acid w ere used fo r
the v isualization o f TLC. A ll the so lvents we re distilled prior to
use. Petro leum e ther:60 ～ 90℃.
1. 2　P lantm aterial
The stem s o f L. landeolata w ere co llec ted in April 2003 from
Yanb ian County, Sichuan, China. The p lant m a te rial w as identi-
fied by P ro.f FU Fading of Chengdu Institute o f B io logy, the Chi-
nese Academ y o f Sciences (CAS). A vouche r specim en (A-35)
w as deposited at the H e rba rium o f Chengdu Institu te o f B io logy,
CAS.
1. 3　Extraction and separation
The air-dried and pow de red stem s of L. landeolata (4. 0 kg)
w ere soaked w ith 95% e thano l(25 L ×3, 5 day s each) a t room
tem pe ra ture. The ethanolw as evapo ra ted unde r reduced pre ssure to
g ive 170 g residue, which w as suspended in H2O (2. 5 L) and
fractionated w ith petroleum ethe r(2. 5 L×4), E tOAc (2. 5 L×
4) and n-bu tano l(2. 5 L×4) to give co rresponding frac tions A (5
g), B(30 g) and C(48 g). The aqueous phase was concentrated
in vacuo to g ive frac tion D (85 g).
F rac tion A w as subjected to silica ge l co lum n(150 g, 160 ～
200 m esh, ф 50 mm ×L 150 mm). G rad ient e lution using pe tro-
leum ethe r-ace tone (20 ∶ 1, 10 ∶ 1, each 2 000 m L) y ie lded
A1 (4. 1 g, non-po lar constituents, 400 ～ 2 000 m L), 6 (22 m g,
2 600 ～ 3 250 mL) and 19 (34 m g, 3 400 ～ 3 900 m L).
F rac tion B w as divided into five sub frac tions B1 ～ B5 by silica
ge l co lum n(1 200 g, 160 ～ 200 m esh, ф 80 mm ×L 430 mm),
using CHC l
3
-CH
3
OH (60∶ 1, 40 ∶ 1, 15∶ 1, 5∶ 1, each
5 000 mL) and m e thano l(5 000 mL) as so lvents. B4 (3. 2 g)
and B5 (1. 5 g) w ere added to C due to overlapping o f com po-
nen ts. B1 (1. 5 g) w as sepa ra ted ove r silica ge l co lum n (50 g,
200 ～ 300 m esh, ф 30 mm ×L 160 mm) w ith pe tro leum ethe r-
ace tone (5 ∶ 1, 2 200 m L) to g ive 11 (110 mg, 550 ～ 1 700
mL) and 2(15 mg, 1 750 ～ 2 100 m L). Separa tion o f B2(13. 2
g) by silica ge l co lum n (540 g, 200 ～ 300 m esh, ф 50 mm ×L
450 mm) w ith pe tro leum e ther - acetone (3 ∶ 1, 3 800 m L) af-
488 　　　　　　　　应 用与 环 境生 物学 报　　Chin J App lEnviron B io l　　　　　　　　　　　　　　 　　　12卷
forded B2-1 (5. 4 g, 650 ～ 1600 mL), B2-2 (4. 8 g, 1 700 ～
2 900 m L) and 16(8 mg, 3 050 ～ 3 700 m L). B2-1 w as purified
ove r silica gel 60 co lumn (240 g, ф 50 mm ×L 320 mm) e luted
w ith petro leum e ther-E tOAc (2. 5∶ 1, 1 800 m L) to give 5(60
m g, 550～ 1 750 m L). Compounds 3(5 mg, 450 ～ 750 mL) and
4 (40 m g, 850～ 1 650 mL)w ere ob ta ined from B2-2 by silica ge l
60 co lum n(240 g, ф 50 mm ×L 320 mm), e lu ted w ith petro le-
um e ther-E tOAc (2 ∶ 1, 1 700 m L). B3 w as separated by silica
ge l column (400 g, 200 ～ 300 m esh, ф 50 mm ×L 370 mm) e-
lu ted w ith CHC l3-CH3OH (20 ∶ 1, 2 800 mL) to affo rd 1 (20
m g, 450～ 900 mL), B3-1 (6. 8 g, 950 ～ 2100 m L) and 7 (10
m g, 2 250 ～ 2 750 m L). The sepa ra tion o f B3-1 by silica ge l 60
co lumn (280 g, ф 50 mm ×L 380 mm) e lu ted w ith pe tro leum e-
the r-ace tone (2 ∶ 1, 2 000 m L) y ielded 8 (50 mg, 450 ～ 1 200
mL) and 10 (15 m g, 1 600 ～ 1 900 m L).
F rac tion C (52. 7 g) w as first sub jec ted to m acroporous re sin
co lumn (D101 , pore size 13 ～ 14 nm , 26 ～ 60 m esh, ф 90 mm ×
L 500 mm) to rem ove suga r by CH3CH2OH -H2O (0∶ 1, 1∶
0, each 5 000 mL) and fraction C′(10. 3 g) w as y ie lded. C′w as
div ided into subfrac tions C1 ～ C4 by silica ge l co lum n(600 g, 160
～ 200 m esh, ф 80 mm ×L 270 mm) e luted w ith CHC l3 - CH3
OH (8 ∶ 1, 5 ∶ 1, 3 ∶ 1 and me thano l, each 6 000 m L).
Com pounds 20(35 m g, 450 ～ 1 050 mL), 15 (15 mg, 1 200 ～
1 900 m L) and C1-1 (2. 5 g, 2 000 ～ 4 100 m L) w ere iso lated
from C1 (4. 8 g) by silica ge l co lum n(180 g, 200 ～ 300m esh, ф
40 mm ×L 250 mm) w ith CHC l
3
- CH
3
OH (10 ∶ 1, 4 200
mL). Sepa ra tion o f C1-1 by silica g el 60 co lum n (120 g, ф 40
mm ×L 255 mm) w ith CHC l3 -CH3OH -H2O (9 ∶ 1 ∶ 0. 1,
2 000 m L) affo rded 12 (120 mg, 550 ～ 1 100 mL) and 14 (200
m g, 1 150 ～ 1 950 mL). C2(3. 2 g) w as sepa ra ted over R-18 sil-
ica gel column (50 g, 40～ 63 μm , ф 40 mm ×L 80 mm) e luted
w ith CH3OH -H2O (3. 5 ∶ 6. 5, 3 000 m L) to g ive 18 (18 m g,
650 ～ 1 300 mL), 13(30 m g, 1 500 ～ 2 300 mL) and 17(7 m g,
2 400 ～ 2 900 mL). C3 (1. 4 g) w as sepa ra ted ove r R-18 silica
ge l column (50 g, 40 ～ 63 μm , ф 40 mm ×L 80 mm) e luted
w ith CH3OH -H2O (3. 5∶ 6. 5, 3 000 mL) to g ive 9 (25 m g,
400 ～ 750 mL). F raction D (85 g)w as no t iso lated because itw as
inac tive to ACE.
1. 4　Identification of sugars in 9, 13, 14 and 15
The samp le (5 m g) was disso lved in 5 m L 95% e thano l, and
5 mL 2 m ol /L HC lw as then added to the m ix tu re. Unde r the at-
m osphere of N2 , them ix turew as refluxed fo r2 hours and the e tha-
nol was evaporated under reduced p ressure. The aqueous phase
w as ex trac ted w ith organic so lvents(CHC l3 o rE tOAc). The suga rs
w ere le ft in the aqueous phase.
The sugars we re identified by com pa ring the ir behav iou rw ith
tha t of authentic sam ples on silica g el TLC (S1:CHC l3∶ CH3OH
∶ H2O =18∶ 10 ∶ 1) and PTLC (S2:EtOAc∶ py ridine∶
w ate r=2∶ 1 ∶ 2;S3:n-butano l∶ ace tic acid∶ w a te r=4∶
1 ∶ 5, uppe r laye r;S4:n-bu tano l∶ benzene∶ py ridine∶ wa-
ter=5 ∶ 1∶ 3∶ 3). The configurations of the sugars we re de-
term ined by the ir optica l ro tations.
1. 5　Angiotensin-converting enzym e assay
ACE ac tivity w as de te rm ined by the m e thod of C arm el[ 8] .
Brie fly, 10 μL of ACE ex trac t(containing 10 μg of tota l prote in)
w as incubated w ith 10 μL o f sam ples o f va rious concen tra tions fo r
30 m in at 37℃. Then, 120 μL o fHHL w as added and incubated
for 15 m in a t 37 ℃. A fte rw ard, 40 μL o f 1 m o l /L NaOH and 10
μL of 20 g /L O-ph thaldialdehyde we re added, and the reac tion
w as term inated afte r 10 m in by add ition o f 20 L of 3 mo l /L HC l
(aq). The fluo rescence intensity o f the produc t H is - Leu w as
m easured at 405 nm (excita tion) and 535 nm (em ission) w ith a
fluore scence spec tropho tome ter(W allac V ic to r2). F or each a ssay,
the b lank contro l, negative control and positive control we re pre-
pared. If the lette rs N, S and B represent the m easured fluores-
cence intensity va lues o f the reacting sy stem s o f negative contro l,
de tec ted samp le and b lank contro l, re spective ly, theACE inhibito-
ry ra te(%)=100% ×(N -S) /(N -B). A t least three sepa-
ra te de te rm ina tions we re conduc ted fo r each sam ple.
1. 6　Prelmi inary screen ing
95% E tOH extrac t o f L. landeolata disp layed ang io tensin-
conve rting enzym e inh ib itory (ACE I) ac tivity. The ACE I ac tivities
o f different fractions a re show n in Table 1.
Tab le 1　ACEI ac tiv ities of d ifferen t frac tions
of 95% E tOH extrac t ofL. landeola ta (λ/%) a, b
Testing
con cen tration
(ρ/m g m L- 1)
10 5 1 0. 1
FractionA 102. 11 103. 65 22. 72 Inactive
F raction B 96. 05 102. 51 95. 09 51. 53
F raction C 94. 62 100. 93 103. 0 51. 68
W ater fraction Inactive Inactive Inactive Inactive
a FractionsA , B and C are acqu ired by extracting the E tOH extract suspen-
ded in w aterw ith petroleum ether, E tOA c andn-B uOH. bExperim en tsw ere
perform ed for at least three t im es w ith con sistent and repeatab le resu lts
2　Identification
(+)-Cyc loolivil(1):W hite pow de r, mp 173 ～ 174 ℃;
( li.t [ 9] 175 ～ 176 ℃);[ α] 20D + 56. 1°(c 0. 122, CH3OH);R f
0. 50 (CHC l3 -CH3OH , 6 ∶ 1);ESIMS (negative mode)m /z:
375 [ M-H] -;UV (CH3OH) λm ax nm ( logε) 214 (4. 46), 278
(4. 23); IRνm ax cm - 1:3 411, 2 934, 2 842, 1 604, 1 514,
1 449, 1 430, 1 371, 1 275, 1 254, 1 149, 1 123, 1 059,
489　 4期 MA Chunhu i, et a l. :C hem ical S tudy on th e S tem ofLon icera land eo lata 　　
1 028, 883, 860, 762, 640;1H NMR (600MH z, DMSO-d6):δ
8. 82 (1H , s, OH-4), 8. 40 (1H , s, OH-4′), 6. 71 (1H , d, J
=8. 0 H z, H-5), 6. 65 (1H, s, H-2), 6. 55 (1H , s, H-2′),
6. 53 (1H , d, J =8. 0 H z, H-6), 6. 06 (1H , s, H-5′), 4. 97
(1H , s, OH-9), 4. 74 (1H , s, OH-9′), 4. 33 (1H , s, OH-
8′), 3. 85 (1H , d, J=11. 6 H z, H-7), 3. 69 (6H, s, CH
3
O-3
and 3′), 3. 60 (2H , m , H-9a and 9′a), 3. 37 (2H , m , H-9b
and 9′b), 3. 08 (1H , d, J=16. 5 H z, H-7′a), 2. 43(1H , d, J
=16. 5 H z, H-5), 1. 88(1H, d, J =11. 2 H z, H-8);13C NM R
(150MH z, DMSO-d6) δ147. 9 (C-3), 146. 2 (C-3′), 145. 2
(C-4), 144. 4 (C-4′), 137. 4 (C-1), 132. 6 (C-1′), 125. 6
(C-6′), 122. 3 (C-6), 116. 6 (C-5′), 115. 8 (C-5), 113. 9
(C-2), 112. 9 (C-2′), 73. 5 (C-8′), 68. 5 (C-9′), 59. 5 (C-
9), 56. 2(CH
3
O-3), 56. 0(CH
3
O-3′), 46. 1(C-8), 43. 6(C-
7), 39. 3 (H-7′);The optica l ro ta tion, IR, UV andNMR da ta
w ere in acco rdancew ith those o f(+)-cyclooliv il[ 10] .
(+)-Fraxiresinol(2):White pow de r, mp 189 ～ 190 ℃;
[ α] 20D +28. 5°(c 0. 312, CHC l3);R f 0. 40(CHC l3 -CH3OH ,
20 ∶ 1);ESIMS (nega tive mode) m /z:403 [ M-H ] -;UV
(CH
3
OH) λ
m ax
nm ( logε) 280 (4. 08), 234 (4. 51), 214
(4. 72); IRνm ax cm - 1:3 433, 2 924, 2 852, 1 614, 1 518,
1 463, 1 428, 1 383, 1 317, 1 274, 1 213, 1 158, 1 033, 854,
798, 750;1H NMR (600MH z, DM SO-d6):δ6. 97(1H , s, H-
2″), 6. 80 (1H , d, J=7. 8 H z, H-6″), 6. 75 (1H , d, J =7. 8
H z, H-5″), 6. 64 (2H , s, H-2′and 6′), 4. 74 (1H , d, J=4. 8
H z, H-6), 4. 52 (1H , s, H-2), 4. 36 (1H , t, J=8. 6 H z, H-
4α), 3. 97(1H, d, J=9. 1 H z, H-8β), 3. 71(1H , d, J =9. 1
H z, H-8α), 3. 76 (3H, s, CH3O-3″), 3. 74 (6H , s, CH3O-3′
and 5′), 3. 62 (1H , dd, J=8. 4, 6. 5 H z, H-4β), 2. 90 (1H ,
d t, J=6. 8, 6. 0 H z, H-5);13C NMR (150MH z, DM SO-d
6
)δ
147. 9(C-3″), 147. 8 (C-3′and 5′), 146. 5(C-4″), 135. 5(C-
4′), 132. 8(C-1″), 127. 6(C-1′), 119. 4 (C-6″), 115. 7 (C-
5″), 111. 3 (C-2″), 105. 9 (C-2′and 6′), 91. 6 (C-1), 87. 8
(C-2), 85. 9 (C-6), 75. 1 (C-8), 70. 8 (C-4), 61. 2 (C-5),
56. 5 (CH3 O-3′and 5′), 56. 1 (CH3 O-3″);1H NMR (600
MH z, CDC l3):δ6. 99 (1H , s, H-2″), 6. 90 (2H , s, H-5″and
6″), 6. 65 (2H , s, H-2′and 6′), 4. 87(1H , d, J=4. 7 H z, H-
6), 4. 83 (1H , s, H-2), 4. 55 (1H, t, J =8. 6, 8. 6H z, H-
4α), 4. 06(1H, d, H-8β), 3. 91 (1H , d, H-8α), 3. 91 (6H ,
s, CH
3
O-3′and 5′), 3. 90 (3H , s, CH
3
O-3″), 3. 84(1H , dd,
J =9. 2, 6. 3 H z, H-4β), 3. 13 (1H , td, J=6. 7 H z, 6. 3 H z,
H-5);13C NMR (150 MH z, CDC l3):δ147. 4 (C-3′and 5′),
146. 8 (C-3″), 145. 5 (C-4″), 135. 1 (C-4′), 132. 4 (C-1″),
126. 2 (C-1′), 119. 7 (C-6″), 114. 4 (C-5″), 109. 1 (C-2″),
103. 5(C-2′and 6′), 91. 6 (C-1), 88. 0 (C-2), 85. 9 (C-6),
74. 6 (C-8), 71. 7 (C-4), 60. 0 (C-5), 56. 4 (CH3O-3′and
5′), 55. 9 (CH3O-3″);The optica l ro tation, IR, UV andNM R
data in DM SO-d
6
we re in accordance w ith tho se o f(+)-frax ires-
ino l[ 11] .
Pr in sep iol(3):W hite pow de r;mp 189 ～ 190 ℃;( li.t [ 17]
191 ～ 192 ℃);[ α] 20D -19. 2°(c 0. 127, CHC l3);R f 0. 30
(CHC l3 -CH3OH , 20 ∶ 1);ESIMS (negative mode)m /z:373
[ M-H] -;UV (CH3 OH) λmax nm ( logε) 281 (3. 86), 234
(3. 91); IRνm ax cm - 1:3 200, 1 601, 1 503, 1 451, 1 423,
1 363, 1 341, 1 245, 1 201, 1 151, 1 119, 1 080, 1 023, 987,
956, 936, 905, 870, 837;1H NMR (600MH z, CDC l
3
):δ6. 94
(2H , s, H-2 and 2′), 6. 74 (2H , dd, J =8. 0 H z, 1. 5 H z, H-6
and 6′), 6. 69(2H, d, J=8. 0 H z, H-5 and 5′), 4. 86(2H , s,
H-7 and 7′), 3. 96 (2H , d, J =9. 2 H z, H-9a and 9′a), 3. 78
(2H , d, J=9. 2 H z, H-9b and 9′b), 3. 73 (6H , s, CH3-10 and
10′);13C NM R (150MH z, CDC l
3
) ∶ δ147. 6 (C-4 and 4′),
146. 6 (C-3 and 3′), 129. 1(C-1 and 1′), 120. 8(C-6 and 6′),
115. 2(C-5 and 5′), 112. 9(C-2 and 2′), 88. 2(C-8 and 8′),
87. 7 (C-7 and 7′), 75. 2 (C-9 and 9′), 56. 2 (C-10 and 10′);
The optica l ro tation, IR, UV andNMR data in DMSO-d6 we re in a-
g reem ent w ith those of pr insepiol[ 12] .
Palm itic ac id (4):W hite amo rphou s so lid (CH3OH);m p
64 ～ 65 ℃ ( li.t [ 13] 63 ～ 64 ℃);R f 0. 60 (CHC l3 -CH3OH , 10
∶ 1);ESIM S (negative mode) m /z:255 [ M-H] - and 511
[ 2M-H] -;IRνmax cm -1:2 960, 2 915, 1 703, 1 640, 1 463,
1 405, 1 301;1H NMR (600MH z, CDC l3):δ2. 25(2H, t, J =
7. 8 H z, -CH2CH2COOH), 1. 61 (2H , m, -CH2 CH2 COOH),
1. 311(24H , brs, CH3(CH2)12CH2CH2COOH), 0. 92 (3H, t,
J =7. 2 H z, CH3);13C NMR (150MH z, CDC l3):δ179. 5 (C
=O), 36. 4 (-CH2CH2COOH), 33. 1 (-CH2CH2COOH), 30. 8
～ 23. 6 (12C , CH3(CH2)12CH2CH2COOH), 14. 5 (CH3);The
IR data w ere in ag reem ent w ith tho se of pa lm itic acid[ 13] .
4-M ethylphenol (5):W hite powder; mp 36 ～ 37 ℃
( li.t [ 14] 35 ～ 37 ℃);R f 0. 65 (CHC l3 -CH3OH , 10 ∶ 1);ES-
IM S (nega tivem ode) m /z:215 [ 2M-H] -;IRν
m ax
cm - 1:3 367,
2 964, 2 929, 1 611, 1 598, 1 509, 1 445, 1 362, 1 237,
1 217, 1 176, 826;1H NMR (600MH z, CDC l3):δ11. 3 (1H ,
s, OH), 7. 28 (2H , dd, J =8. 4, 2. 1 H z, H-3 and 5), 7. 13
(2H , dd, J =8. 4, 2. 1 H z, H-2 and 6), 1. 69 (3H , s, CH3);
13C NMR (150MH z, CDC l
3
):δ156. 4 (C-1), 141. 9 (C-4),
128. 1(C-3 and 5), 115. 5 (C-2 and 6), 31. 2 (CH3);The
NM R data we re no t repo rted.
Methyl 2, 4-d ihydroxy-3, 6-d im ethylbenzoate (6):Co lo r-
less needles(CHC l3);mp 138 ～ 139℃ ( li.t [ 15] 139 ～ 141 ℃);
R f 0. 30 (pe tro leum e ther-acetone, 10 ∶ 1);ESIM S (negative
m ode)m /z:195 [M-H] -;UV (CH3OH) λmax nm ( logε) 270
(4. 17), 223 (4. 01); IRνm ax cm -1:3 409, 2 925, 2 854,
1 627, 1 500, 1 447, 1 368, 1 314, 1 273, 1 197, 1 161,
1 110, 1 032, 992, 799;1H NM R (600MH z, CDC l3):δ12. 05
(1H, s, OH-2), 6. 22 (1H , s, H-5), 5. 79 (1H , s, OH-4),
3. 92 (3H , s, -COOCH3), 2. 45 (3H , s, CH3-6), 2. 11 (3H ,
s, CH3-3);13C NMR (150 MH z, CDC l3):δ172. 6 (C =O),
163. 1 (C-4), 158. 3 (C-2), 140. 1 (C-6), 110. 6 (C-5),
108. 6(C-3), 105. 1(C-1), 51. 8 (OCH3), 24. 05 (CH3-6),
7. 64 (CH
3
-3);The IR and NMR da ta we re c lo sely resem ble to
490 　　　　　　　　应 用与 环 境生 物学 报　　Chin J App lEnviron B io l　　　　　　　　　　　　　　 　　　12卷
those reported[ 15] .
Apigen in (7):Pa le ye llow needles (CH3OH);m p >300
℃ ( li.t [ 16] 347 ～ 348 ℃);R
f
0. 20 (CHC l
3
- CH
3
OH , 10 ∶
1);ES IM S(nega tivem ode)m /z:269 [ M-H] -;UV (CH3OH)
λm ax nm ( logε) 335 (4. 30), 296 (4. 28), 268 (4. 41);IRνm ax
cm - 1:3 250, 1 652, 1 607, 1 502, 1 351, 1 243, 1 180,
1 032, 910, 825;1H NM R (600 MH z, DMSO-d6 ):δ12. 94
(1H , s, OH-5), 10. 45 (1H , s, OH-7), 7. 91(2H , d, J=8. 6
H z, H-2′and 6′), 6. 91 (2H , d, J =8. 6 H z, H-3′and 5′),
6. 76 (1H , s, H-3), 6. 47 (1H , d, J =1. 6 H z, H-8), 6. 18
(1H , d, J =1. 5 H z, H-6);13C NMR (150MH z, DM SO-d6)
data see Table 2;The IR, UV and 1H NMR da ta w ere in accord-
ance w ith those o f apig enin[ 16 ～ 18] .
Luteolin (8):Pa le ye llow powder;m p >300 ℃ ( li.t [ 19]
326 ～ 328 ℃);ESIM S (nega tive mode) m /z:285 [ M-H] -;
IRν
m ax
cm - 1:3 403, 1 652, 1 603, 1 579, 1 502, 1 442, 1 258,
831, 742;1H NMR (600 MH z, DM SO-d6):δ12. 97 (1H , s,
OH-5), 10. 82 (1H , s, OH-7), 7. 40 (1H , d, J =8. 4 H z, H-
5′), 7. 39 (1H , s, OH-2′), 6. 89(1H , d, J =8. 4 H z, H-6′),
6. 66 (1H , s, H-3), 6. 44 (1H , d, J =1. 3 H z, H-6), 6. 18
(1H , d, J =1. 3 H z, H-8);13C NMR (150MH z, DM SO-d6)
da ta see Tab le 2;The IR, NMR data and the TLC behav io r unde r
d iffe rent so lvent system s(E tOAc, R f 0. 75;CHC l3∶ CH3OH =10∶ 1, R
f
0. 30;CHC l
3
∶ (CH
3
)
2
CHOH =8 ∶ 1, R
f
0. 75;
CHC l
3
∶ CH
3
COCH
3
=1 ∶ 1, R
f
0. 65) we re the sam e as those
o f lu teo lin[ 19 ～ 21] . The m elting po in t o f 8 w as the sam e as that of
the authentic sam ple.
Tab le 2　13C NMR data of com pounds 7 ～ 9 (150MHz in DM SO-d6)
C-atom 7 8 9 C-atom 7 8 9
2 164. 9 164. 6 148. 0 10 104. 3 104. 4 105. 2
3 103. 5 103. 5 136. 5 1′ 121. 8 119. 6 122. 0
4 182. 4 182. 3 176. 6 2′ 129. 1 114. 0 130. 1
5 157. 9 157. 9 160. 8 3′ 116. 6 146. 3 115. 9
6 99. 5 99. 5 99. 3 4′ 162. 1 150. 4 159. 8
7 164. 4 164. 8 163. 2 5′ 116. 6 116. 7 115. 9
8 94. 6 94. 5 94. 8 6′ 129. 1 122. 2 130. 1
9 161. 8 162. 1 156. 2
　 　 5, 7, 4′-Tr ihydroxyflavonol 7-O-β-D-glucopyranoside
(9):Pa le ye llow pow de r;m p 263. 8 ～ 264. 5 ℃, dec. ( li.t [ 22]
265 ～ 267 ℃, dec. );[ α] 20D -119. 5°(c 0. 223, CH3 OH);R f
0. 30 (CHC l3 - CH3OH , 4 ∶ 1);ESIMS (negative mode)m /z:
447 [M-H] - and 895 [ 2M-H] -;UV (CH3OH)λmax nm ( logε)
366(4. 04), 261(4. 34);UV (CH3ONa) λm ax nm ( logε) 437
(4. 09), 262 (4. 39);UV (A lC l3) λm ax nm ( logε) 425
(4. 18), 261 (4. 39);UV (A lC l
3
+HC l) λ
max
nm ( logε) 425
(4. 12), 351 (3. 79), 262 (4. 39);UV (NaOA c) λm ax nm
( logε) 369 (4. 05), 261(4. 43);UV (NaOAc +H3BO3)λm ax
nm ( logε) 367 (4. 10), 261 (4. 39); IRνmax cm - 1:3 407,
1 655, 1 636, 1 607, 1 505, 1 372, 1 326, 1 179, 1 080, 984,
831;D-g lucose w as ob tained by hydro ly sing 9 in 2 m ol /L HC l
(aq. )-95% E tOH and identified by TLC (S1:R f =0. 30) and
PTLC (S2:R f =0. 28;S3:R f =0. 18;S4:Rf=0. 24) behaviour
w ith tha t o f authentic sam ple and optica l ro tation ([ α] 20D +52. 7
(c 0. 108, H2O));1H NMR (600MH z, DMSO-d6):δ12. 51
(1H , s, OH-5), 10. 19 (1H , s, OH-3), 9. 58 (1H , s, OH-
4′), 8. 08 (2H , d, J=8. 4 H z, H-2′and 6′), 6. 95 (2H , d, J
=8. 4 H z, H-3′and 5′), 6. 81 (1H , d, J =1. 8 H z, H-8),
6. 43 (1H , d, J=1. 8 H z, H-6), 5. 08 (1H , d, J=7. 5 H z, H-
G lc1);13C NM R (150MH z, DMSO-d6):δ100. 4(C-1″), 77. 6
(C-5″), 76. 9 (C-3″), 73. 6 (C-2″), 70. 1 (C-4″), 61. 1 (C-
6″), othe r data see Table 2;The IR, UV and 13C NMR da ta o f 9
w ere in acco rdancew ith those reported[ 23] .
Querce tin (10):Ye llow powder;mp 309 ～ 310℃ ( li.t [ 24]
312 ～ 315℃);UV (CH
3
OH)λ
m ax
nm ( logε) 376 (4. 40), 302
(3. 92), 257 (4. 38); IRνm ax cm -1:3 320, 1 661, 1 610,
1 561, 1 521, 1 453, 1 406, 1 383, 1 317, 1 260, 1 211,
1 191, 1 169, 1 130, 1 093, 940;ES IM S(nega tive mode)m /z:
301 [ M-H] -;The TLC behavior under d iffe rent so lvent system s
(E tOAc, R f 0. 80;CHC l3∶ CH3OH =10 ∶ 1, R f 0. 30;CHC l3
∶ (CH
3
)
2
CHOH =8 ∶ 1, R
f
0. 65;CHC l
3
∶ CH
3
COCH
3
=1
∶ 1, R f 0. 70) w as the sam e as tho se o f quercetin. The m e lting
point o f 10 w as the sam e as tha t o f the authentic sam ple.
Gen iste in (11):Co lo rless need le s (CH3 OH);m p 286 ～
287 ℃ ( li.t [ 25] 288 ～ 289 ℃);R f 0. 80(CHC l3 - CH3OH , 10∶
1);ESIM S(nega tivem ode)m /z:269 [ M-H] -;UV (CH3OH)
λm ax nm ( logε) 332 (3. 85), 262(4. 43), IRνm ax cm - 1:3 408,
1 652, 1 614, 1 565, 1 517, 1 503, 1 308, 1 044, 790;1H
NM R (600MH z, DMSO-d6):δ12. 96 (1H , s, OH-5), 10. 98
(1H , s, OH-7), 9. 63 (1H, s, OH-4′), 8. 32 (1H, s, H-2),
7. 37 (2H , d, J =8. 4 H z, H-2′and 6′), 6. 82 (2H , d, J=8. 4
H z, H-3′and 5′), 6. 39(1H , d, J=1. 9 H z, H-8), 6. 23 (1H ,
d, J =1. 9 H z, H-6);13C NM R (150MH z, DMSO-d
6
) da ta see
Table 3;The IR, UV and NMR da taw ere in ag reem ent w ith those
o f geniste in[ 25, 26] .
Prunetin (12):Co lo rle ss needles(CH3OH);m p 240～ 241
℃ ( li.t [ 27] 241 ～ 242 ℃);R f 0. 40 (CHC l3 - CH3OH , 4 ∶ 1);
ES IM S(po sitivem ode) m /z:285 [M +H] +;UV (CH3OH)
λm ax nm ( logε) 262(4. 51);UV (CH3ON a)λm ax nm ( logε) 273
(4. 49);UV (A lC l
3
) λ
m ax
nm ( logε) 376 ( 3. 61), 273
(4. 51);UV (A lC l3 +HC l)λm ax nm ( logε) 376 (3. 60), 273
491　 4期 MA Chunhu i, et a l. :C hem ical S tudy on th e S tem ofLon icera land eo lata 　　
(4. 51);UV (NaOAc) λm ax nm ( logε) 263 (4. 50);UV
(NaOAc +H3BO 3)λm ax nm ( logε) 263 (4. 54);IRνmax cm - 1:
3 380, 1 667, 1 614, 1 569, 1 518, 1 505, 1 431, 1 356,
1 315, 1 273, 1 257, 1 220, 1 191, 1 155, 1 052, 952, 824,
785;1H NMR (600MH z, DMSO-d6):δ12. 94 (1H , s, OH-
5), 9. 63 (1H, s, OH-7), 8. 37 (1H , s, H-2), 7. 38 (2H , d,
J =8. 4 H z, H-2′and 6′), 6. 83(2H , d, J=8. 4 H z, H-3′and
5′), 6. 61 (1H , d, J =2. 1 H z, H-8), 6. 38 (1H , d, J =2. 1
H z, H-6), 3. 85 (3H, s, -OCH3);13C NM R (150 MH z, DM-
SO-d6) data see Table 3;The IR, UV and 1H NMR data we re in
acco rdance w ith those o f prune tin[ 27] . 13C NMR data w ere no t re-
ported befo re.
Gen iste in 5-O-β-D-glucopyranoside (13):W hite powder;
m p 174 ～ 175℃;[ α] 20
D
-158. 2°(c 0. 387, CH
3
OH);R
f
0. 40
(CHC l3 -CH3OH , 3∶ 1);ESIMS (nega tive m ode) m /z:431
[ M-H] -;UV (CH3 OH) λm ax nm ( logε) 255 (4. 66);UV
(CH3ONa)λm ax nm ( logε) 261 (4. 66), UV (A lC l3) λmax nm
( logε) 255 (4. 59), UV (A lC l3 + HC l) λm ax nm ( logε) 255
(4. 58), UV (NaOAc) λm ax nm ( logε) 261 (4. 69), UV
(NaOAc +H3BO 3)λm ax nm ( logε) 255 (4. 66);IRνmax cm - 1:
3 253, 1 627, 1 609, 1 581, 1 508, 1 456, 1 309, 1 246,
1 213, 1 085, 1 046, 843;D-g luco sew as obtained by hydro ly sing
14 in 2 m o l /L HC l( aq. ) - 95% E tOH and identified by TLC
(S1:R f =0. 30) and PTLC (S2:R f =0. 28;S3:R f =0. 18;S4:
R
f
=0. 24) w ith authentic sam ple and optica l ro tation ([ α] 20
D
+
52. 9°(c 0. 122, H2O));1H NM R (600MH z, DM SO-d6):δ
10. 97(1H, s, OH-7), 9. 56 (1H , s, OH-4′), 8. 19 (1H, s,
H-2), 7. 31 (2H , d, J =8. 4 H z, H-2′and 6′), 6. 80(2H , d,
J =8. 4 H z, H-3′and 5′), 6. 78 (1H , d, J =2. 1 H z, H-6),
6. 59 (1H , d, J=2. 1 H z, H-8), 4. 77 (1H , d, J=7. 5 H z, H-
G lc1);13C NM R (150MH z, DMSO-d6):δ104. 1(C-1″), 78. 0
(C-5″), 76. 1 (C-3″), 74. 1 (C-2″), 70. 1 (C-4″), 61. 2 (C-
6″), othe r data see Table 3;The UV and NM R data of 13 w ere in
ag reem ent w ith those repo rted[ 28] . The m elting point and optica l
ro tation we re no t repo rted.
Grunetin 5-O-β-D-glucopyranoside(14):White powder;mp
240～ 241℃ ( li.t [ 29] 237 ～ 239 ℃);[ α] 20
D
-129. 4°(c 0. 257, CH
3
OH);R f 0. 40 (CHC l3 -CH3OH, 4∶ 1, w ater saturated);ESIMS
(positivem ode)m /z:469 [M +Na] + and 915 [ 2M +Na] +;UV
(CH3OH)λm ax nm ( logε) 254 (4. 63);UV (CH3ONa) λm ax nm
( logε) 254(4. 55), UV (A lCl3) λm ax nm ( logε) 255(4. 63), UV
(A lC l3 +HCl)λm ax nm (logε) 255(4. 61), UV (NaOAc)λm ax nm
( logε) 255(4. 66), UV (NaOAc +H3BO3) λm ax nm ( logε) 255
(4. 65);IRνmax cm - 1:3 330, 3 146, 1 634, 1 622, 1 597, 1 515,
1 433, 1 263, 1 068, 1 026, 841;D-glucosew as obtained by hydro l-
ysing 14 in 2 mo l /L HCl(aq. ) -95% EtOH and identified by TLC
(S1:R f =0. 30) and PTLC (S2:R f =0. 28;S3:R f =0. 18;S4:R f =
0. 24) w ith authentic sample and optical ro ta tion ([ α] 20
D
+52. 8 (c
0. 125, H2O));1H NMR (600MH z, DMSO-d6):δ9. 58(1H, s,
OH-4′), 8. 27(1H , s, H-2), 7. 32 (2H , d, J=8. 4 H z, H-2′and
6′), 6. 89(1H, d, J=2. 2H z, H-6), 6. 87(1H, d, J =2. 2 H z, H-
8), 6. 80 (2H , d, J=8. 4 Hz, H-3′and 5′), 4. 82(1H , d, J=7. 5
H z, H-1″);13C NMR(150MH z, DMSO-d6):δ103. 1(C-1″), 78. 1
(C-5″), 76. 3 (C-3″), 74. 0 (C-2″), 70. 4 (C-4″), 61. 4 (C-6″),
o ther data see Tab le 3;The UV and NMR data of 14 we re in acco rd-
ance w ith those reported[ 29] .
Tab le 3　13C NMR data of compounds 11 ～ 14(150MHz, DMSO-d6)
C-atom 11 12 13 14 C-atom 11 12 13 14
2 154. 4 154. 8 152. 0 152. 2 10 104. 9 105. 9 109. 2 110. 2
3 121. 7 121. 5 125. 0 125. 3 1′ 122. 8 122. 9 122. 9 122. 7
4 180. 7 180. 9 175. 6 175. 5 2′ 130. 6 130. 6 130. 8 130. 8
5 158. 1 157. 9 159. 6 159. 2 3′ 115. 5 115. 6 115. 3 115. 4
6 99. 4 98. 5 104. 4 96. 3 4′ 162. 5 162. 2 157. 6 157. 7
7 164. 8 165. 7 163. 0 163. 9 5′ 115. 5 115. 6 115. 3 115. 4
8 84. 1 92. 8 98. 0 104. 1 6′ 130. 6 130. 6 130. 8 130. 8
9 157. 9 157. 9 159. 0 159. 1 CH3O
- 56. 5 56. 5
　 　 5, 7, 4′-Tr ihydroxyflavanone 7-O-β-D-glucopyranoside
(15):Pa le ye llow needles(CH3OH);m p 226 ～ 227 ℃ ( li.t [ 30]
228 ～ 230 ℃);[ α] 20D -171. 5°(c 0. 337, CH3OH);R f 0. 60
(CHC l3 -CH3OH , 3∶ 1);ESIMS (nega tive m ode) m /z:433
[ M-H] -;UV (CH
3
OH) λ
m ax
nm ( logε) 279 (4. 21), 261
(4. 28);UV (CH3 ONa) λm ax nm ( logε) 363 (3. 59), 281
(4. 20), 261 (4. 29);UV (A lC l3 ) λmax nm ( logε) 380
(3. 38), 305 (4. 11), 261(4. 18);UV (A lC l3 +HC l)λmax nm
( logε) 378 (3. 45), 301 (4. 05), 261 (4. 18);UV (NaOAc)
λm ax nm ( logε) 279 (4. 24), 261 (4. 29);UV (NaOAc +H3
BO
3
) λ
max
nm ( logε) 279 (4. 25), 261 (4. 30); IRν
m ax
cm -1:
3 429, 2 924, 2 854, 1 641, 1 457, 1 376, 1 296, 1 171,
1 065, 834;D-g lucose w as obtained by hydro ly sing 15 in 2 mo l /L
HC l(aq. ) - 95% E tOH and identified by TLC (S1:R f =0. 30)
and PTLC (S2:R f =0. 28;S3:R f =0. 18;S4:R f =0. 24) w ith
au then tic samp le and optica l ro ta tion ([ α] 20D +52. 2°(c 0. 120,
H2O));1H NMR (600MH z, DMSO-d6):δ12. 55(1H, s, OH-
5), 7. 50 (2H , d, J=8. 4 H z, H-2′and 6′), 7. 21 (2H , d, J
=8. 4 H z, H-3′and 5′), 6. 63 (1H , d, J =2. 3 H z, H-8),
5. 74 (1H , d, J=7. 8 H z, H-G lc1), 5. 47 (1H , dd, J =13. 2,
2. 7 H z, H-2 ), 3. 25 (1H, dd, J =16. 8, 13. 5 H z, H-3a),
2. 87 (1H , dd, J=16. 8, 2. 7 H z, H-3b);13C NMR (150MH z,
DM SO-d6):δ198. 2 (C =O), 167. 4 (C-7), 165. 3 (C-5),
164. 5 (C-9), 160. 5 (C-4′), 130. 4 (C-1′), 129. 7 (C-2′and
6′), 117. 3 (C-3′and 5′), 105. 1 (C-10), 102. 3(C-1′′), 98.
5 (C-6), 97. 2(C-8), 80. 6 (C-2), 79. 9 (C-3′′), 79. 2 (C-
492 　　　　　　　　应 用与 环 境生 物学 报　　Chin J App lEnviron B io l　　　　　　　　　　　　　　 　　　12卷
5′′), 75. 5 (C-2′′), 71. 8 (C-4′′), 63. 0 (C-6′′), 44. 1 (C-
3);The IR, UV, optica l ro tation and NMR da ta of 15 w ere in ac-
co rdancew ith those reported[ 30 , 31] .
Succ in ic ac id (16):W hite powder;m p 180 ～ 182 ℃
( li.t [ 32] 181～ 182 ℃);ESIMS (nega tive mode)m /z:117 [M-
H] -;The TLC behavio r under d iffe rent so lvent system s (E tOA c
∶ CH
3
OH =15 ∶ 1, R
f
0. 65;CHC l
3
∶ CH
3
OH =10 ∶ 1, R
f
0. 50;CHC l3∶ (CH3)2CHOH =8∶ 1, R f 0. 65;CHC l3∶ CH3
COCH
3
=1 ∶ 1, R
f
0. 55) we re the sam e as those o f succ in ic
acid. The me lting po int o f 16 w as the same as tha t o f the authentic
sam ple.
(+)-Catech in (17):Wh ite powder;m p 169 ～ 170 ℃
( li.t [ 33] 172 ～ 174℃);[ α] 20
D
+11. 2°(c 0. 517, CH
3
OH);UV
(CH3OH) λm ax nm ( logε);IRνm ax cm -1:3 350, 1 617, 1 539,
1 511, 1 467, 1 353, 1 228, 1 186, 1 144, 1 085, 954, 825;
ES IMS (nega tivem ode) m /z:289 [ M-H] -;The op tical ro ta tion
and the TLC behavio r unde r differen t so lvent system s(E tOA c, R f
0. 75;CHC l3∶ CH3OH =3 ∶ 1, R f 0. 55;CHC l3∶ (CH3)2
CHOH =8 ∶ 1, R f 0. 30;CHC l3∶ CH3 COCH3 =1 ∶ 1, R f
0. 45) w ere in accordance w ith those of(+)-ca techin. The me lt-
ing po int of 17 w as the same as that o f the au then tic samp le.
( -)-Epicatech in (18):Wh ite powder;mp 248 ～ 249 ℃
( li.t [ 34] 249 ℃);[ α] 20D -60. 2°(c 0. 226, CH3OH);UV (CH3
OH)λm ax nm ( logε) 287(3. 56), 281(3. 69), 227(4. 30);ES-
IM S(negativem ode)m /z:289 [ M-H] -;The optica l ro ta tion and
the TLC behavior unde r different so lvent sy stems(EtOAc, R f 0. 75;
CHC l3∶ CH3OH (w ater satu rated)=4 ∶ 1, R f 0. 35;CHC l3∶
(CH3)2CHOH =8 ∶ 1, R f 0. 30;CHC l3∶ CH3COCH3 =1 ∶ 1,
R
f
0. 45) w ere in accordance w ith those of ( -)-epicatechin. The
m elting po int o f 18 was the same as that of the au then tic sam ple.
3　D iscussion
Com pounds 17 and 18 show ed w eak ACE ( angio tensin-con-
ve rting enzyme) inhib itory ac tiv ity w ith IC
50
1. 593 and 1. 781
mm o l /L, re spective ly[ 35 , 36] . A t a concen tra tion of 1 m g /m L,
compounds 17 and 18 produced about 40% and 45 % ACE inh ibi-
tion of a homogenate of rabbit lung in v ivo, respective ly[ 37] . Com-
pound 10 could inh ib itACE w ith inh ib itory ra tio o f 31. 32%, when
88. 7 μmo l /kg was adm inistrated in vivo, w hich w as sim ila r to tha t
o f captop ril[ 38, 39] . Thus, it could be concluded that compounds
10, 17 and 18 we re the ACEI active compounds in th is plan.t
This plan t is rich in flavone, iso flavone and flavono l(0. 6%
o f thew eight of the plan t) that possessm any v ital ac tivities. H ow-
eve r, because of its distribution at an altitude of 3 000 m and diffi-
cu lty in colle cting, it is unsuitab le used fo r flavono ids resorces.
The po ssib le re sources fo r com pounds 7, 8, 10 and 11 a re summ a-
rized in Tab le 4.
Tab le 4　Act ivitie s of compounds 7, 8, 10 and 11 and their or igin
Com pounds Activities Orig in
A pigen in 7
Inh ib ition of locom otor behaviou r[ 40] , an tib acterial[ 41] , va-
sorelaxant ef fect[ 42] , antitumor[ 43] , steroid horm one activi-
ty[ 44] and rad ical scavenging activity[ 45]
Ma tricaria cham om illa l[ 46] , Spirodela polyr-
rth iza (L. ) S ch leid[ 47]
Luteolin 8
An titum or[ 48] , inh ib ition of p roliferation and col lagen ex-
p ression of hepatic stellate cells (HSC)[ 49] , b lood coagu-
lant ef fect[ 50] and ant ivirus[ 51]
Ajuga lupu lina var. M ajor[ 52] , Lon icera japon-
eca Thum b[ 53] , Nelum bo nucifera G aertn[ 54]
Quercetin 10 Inh ib ition of p latelet agg regat ion[ 55] and ACE I[ 11 , 12] A llium cepa L.
[ 56] , App le[ 57] , Moru s alba
L. [ 58] , Sophora japon ica L. [ 59]
G en is tein 11
Protein tyrosine k inase (PTK) inh ib itor[ 60] and glycine re-
ceptor b locker[ 61] Soybean
[ 62] , Trifolium pratense L. [ 63]
R eferences
1　 In stitutum B otanicum Kunm ingen se A cadem iae S inicae (中国科学院云
南植物研究所). Flora Yunnanica (Vo l5). B eijing (北京):S cien ce
P ress (科学出版社), 1991. 402
2　X ing JB (邢俊波), L iH J(李会军), Li P (李萍), Xu L (许力 ).
S tudies on the quality cont ro l ofFosLon icera e-determ ination of total fla-
vonoids. Chin J MAP (中国现代应用药学), 2002, 19 (3):169～
170
3　Yan CY (颜承云), Gu JW (谷继伟). The dyn ast ic analysis on th e
conten t of total f lavone in fresh fru its ofLon icera edu lis. Ch in W ild P lan t
Resour (中国野生植物资源), 2004, 23 (2):51～ 52
4　Zhang YY (张艳焱), Zhang TL (张天伦 ), Liang GY (梁光义 ),
C ao PX (曹佩雪), Ruan JH (阮婧华). Determ ination of ch lorogen ic
acid in d ifferent processed crude d rugs of Lon icera fu lvotom en tosa by
HPLC. Ch in Trad＆Herba lDrug s(中草药), 2003, 34 (8):696～
697
5　K akuda R, Im aiM , Yaoita Y, M ach idaK , Kikuch iM. S eco iridoid g ly-
cosides from the flow er bud s of Lon icera japon ica. Phy tochem istry,
2000, 55 (8):879～ 881
6　Kum ar S, S atiOP, Semw alVD , Nau tiyalM , Sati S, Takeda Y. Iridoid
glycosides from Lon icera qu inqu elocu laris. Phytochem istry, 2000, 53
(4):499 ～ 501
7　 In stitutum Botanicum Kunm ingen se A cadem iae S inicae (中国科学院云
南植物研究所). Flora Yunn anica (Vo l5). B eijing (北京):S cien ce
Press(科学出版社), 1991. 414 ～ 416
8　C arme lS, YaronA. An in tram olecu larly quenched fluorescen t tripep tide
as a flu orogenic sub strate of angioten sin-I-converting enzym e and of bac-
terial d ipeptidyl carboxypeptidase. Eur J B ioch em , 1978, 87:265 ～
273
9　Zhang DM (张冬梅), Hu LH (胡立宏), YeWC (叶文才), Zhao
SX (赵守训). S tudies on the chem ica l cons tituen ts ofFraxinus chinen-
sis Roxb. Ch in J NatM ed (中国天然药物), 2003, 1 (2):79～ 81
493　 4期 MA Chunhu i, et a l. :C hem ical S tudy on th e S tem ofLon icera land eo lata 　　
10　Zheng XH (郑训海), Kong LY (孔令义). S tudies on chem ica l con-
sti tuen ts ofEp im ed ium koreanum. Ch in Trad＆ Herba l D rugs (中草
药), 2002, 33 (11):964～ 967
11　Tsukam oto H , H isada S, N ish ibe S. L ignans from bark of Fra xinus
m and shurica var. japon ica and F. japon ica. Chem ＆ Pha rm Bu ll,
1984, 32 (11):4482 ～ 4489
12　K ilidhar SB, Parthasarathy MR, Sharm a P. Prinsep iol, a lignan from
stem s ofPrinsepia u tilis. Phytochem istry, 1982, 21 (3):796～ 797
13　Gu ZP (顾志平), Li TM (李彤梅), X iao PG (肖培根), C hen JM
(陈建民). S tudies on the ch em ical constituen ts ofS trychnos nit ida G.
Don. Ch in J ChinMa tM ed (中国中药杂志), 1997, 22 (1):40～
41, 62
14　封士兰 , 潘宣 , 张晓光. 兔耳草的化学成分研究. 中草药 , 2000,
31 (7):500～ 501
15　Zheng WP (郑卫平), Tang YP (唐于平), Lou FC (楼凤昌), Zhi
F (植飞). S tudies on the constituen ts ofD end robium chryseum Rolfe.
J Ch ina Pha rm Un iv (中国药科大学学报), 2000, 31 (1):5～ 7
16　Lin LD (林立东), Q in GW (秦国伟), Xu RS (徐任生). S tudies
on chem ical consti tuen ts of Ilex cen troch inensis. Acta Bo tS in (植物学
报), 1994, 36 (5):393～ 397
17　W ang AG (王爱国), LuY (吕扬), Feng XZ (冯孝章). S tudies on
the chem ical consti tuen ts ofA juga forrestii D ie ls. Acta P harm S in (药
学学报), 1994, 29 (12):899～ 904
18　于德泉 , 杨俊山主编. 分析化学手册第七分册. 第 2版. 北京:化
学工业出版社 , 1999. 817
19　Liang QL (梁侨丽 ), D ing LS (丁林生 ). C hem ical study on th e
f low er ofD olichos lablab L. J Ch ina P harm Un iv (中国药科大学学
报), 1996, 27 (4):205～ 207
20　Shen YX (沈一行), Quan LH (权丽辉), Guan L (关玲), C hen
JM (陈建民 ). S tud ies on th e flavono ids from D endran th em a l
avandulifolium. Acta Pharm S in (药学学报), 1997, 32 (6):451～
454
21　于德泉 , 杨俊山主编. 分析化学手册第七分册. 第 2版. 北京:化
学工业出版社 , 1999. 818
22　Pan Q (潘勤), Yang PQ (杨培全), C heng GH (陈桂红). S tudies
on the chem ical con stitu ents of trad itional Tibetan m ed icinal herb
S hengdeng (Rhamnella gi lg itica) (I). Ch in Trad＆Herba lD rugs (中
草药), 1998, 29 (2):76～ 79
23　M arkh am KR, Ternai B, S tanley R, Geiger H , Mabry TJ. C arbon-13
NM R s tudies of flavonoid s-Ⅲ Natural ly occurring f lavonoid glycosides
and their acy lated derivatives. Tetrahedron, 1978, 34:1389～ 1397
24　C hen JJ(陈纪军), Chen JS (陈金素), C hen SY (陈泗英), Zhou J
(周俊). The ch em ical cons tituen ts ofRhod iola a tun tsuen sis. Acta B ot
Yunnan ica (云南植物研究), 1999, 21 (4):525～ 530
25　S ang YS (桑已曙), M in ZD (闵知大 ). S tud ies on the con stituen ts
ofP ueraria om eiensisTang etW ang. J Ch in PharmU niv (中国药科大
学学报), 2000, 31 (6):408～ 410
26　K in jo JE, Fu ru saw a JT, Baba J. S tud ies on the constituen ts ofP uerar-
ia loba ta Ⅲ:isoflavonoids and related com pounds in the roots and th e
vo lub le stem s. Chem ＆P harm Bu ll, 1987, 35 (12):48～ 46
27　B radbu ry RB, W h ite DE. The chem istry of Subterranean clover. Part
Ⅱ:syn thesis and reduction of isoflavones related to gen istein and form-
ononet in. J Chem Soc, 1953:871 ～ 876
28　Geib elM , G eigerH , T reut ter D. Tectochrysin 5-and gen is tein 5-glu-
cosides from the bark of P runus cerasus. Phytochem istry, 1990, 29
(4):1351 ～ 1353
29　Khalid SA, G ellertM , S zend rei K, Duddeck H. Prunetin 5-O-β-D-
glu copyranoside, an isoflavone f rom the pedun cle ofP runus avium and
P. cerasus. Phy tochem istry, 1989, 28 (5):1560 ～ 1561
30　Zhang XR (张晓瑢), L iao X (廖循), D ing LS (丁立生). Chem i-
cal const ituen ts ofCycloca rya pa liuru s. Chin J App lEnvironB iol (应用
与环境生物学报), 2001, 7 (1):90～ 91
31　W atanabe K, K in jo J, N ohara T. The new isof lavonoid glycosides from
Lupinu s luteus and L. polyphyl lus × arboreu s. Chem ＆ Pharm Bu ll,
1993, 41 (2):394～ 396
32　Zhou YS (周燕生), N iMY (倪慕云). C hem ical constituen ts from
fresh leaves ofR ehmann ia g lu tinosa Libosch. Ch ina J Chin Ma tM ed
(中国中药杂志), 1994, 19 (3):162～ 166, 191
33　Li LQ (李良琼 ), LiMR (李美蓉 ), Yang ZH (杨智彪), H u XB
(胡晓斌). S tud ies on th e chem ical con stitu ents of hairy leaf taxi llus
(Taxi llus n igran s). Ch in Trad＆Herba lD rugs (中草药), 1995, 26
(3):118 ～ 121, 116
34　Chen CX (陈昌祥), Ye HY (叶海亚), Shen YL (申永乐), H ao
XJ (郝小江). The chem ical con stitu ents f rom S ch im a wa llich ii. Acta
B otY unnan ica (云南植物研究), 1997, 19 (2):201 ～ 206
35　Actis-G oretta L, O ttavian i JI, K een CL, Fraga CG. Inh ib ition of an-
gioten sin converting enzym e (ACE) activity by flavan-3-ols and p rocy-
an id in s. FEBS Lett, 2003, 555 (3):597 ～ 600
36　T su tsum iY, Sh im ada A, M iyano A, N ish id a T, M itsugnaga T. In vitro
screen ing of angioten sin I-converting enzym e inhibi tors from Japanese
cedar(Cryptom eria japon ica). J Wood Sci, 1998, 44:463 ～ 468
37　M eun ierMT , V illie F , JonadetM , Bastide J, B as tide P. Inh ib ition of
angiotension I converting enzyme by flavano lic compound s:in vitro and
in vivo studies. P lan taM ed ica, 1987, 53 (1):12～ 15
38　Häck l LPN, Cu ttle G , Dovich i SS, L im a-Landm an MT, N icolauM.
Inh ib ition of angiotensin-converting enzym e by qu ercetin alters the vas-
cu lar response to b radyk in in and angioten sin I. Pharm acolog y, 2002,
65 (4):182～ 186
39　M elzig M F, E scher F. Indu ction of neu tral endopep tidase and angio-
ten sin-converting enzyme activity ofSK-N-SH cells in vitro by quercetin
and resveratro.l Pha rmazie, 2002, 57 (8):556～ 558
40　Avallone R, Zanoli P, Pu ia G , K leinschnitzM , S chreier P, B arald i
M. Pharmacological p rofile of ap igen in, a flavonoid isolated f romMa t-
ricaria chamom i lla. B iochem Pharm acol, 2000, 59 (11):1387 ～
1394
41　 Sato Y, Suzak iS, N ish ikaw a T , K iharaM , Sh ib ataH , H igu tiT. Phy-
tochem ical flavones isolated from S cutellaria ba rba ta and an tib acterial
act ivity agains t m eth icillin-resistant S taphylococcus au reus. J E thno-
pha rmaco l, 2000, 72 (3):483～ 488
42　Zhang YH , ParkYS, K im T J, Fang LH , Ahn HY, H ong JT, K im Y,
Lee CK , Yun YP. Endothelium-dependent vasorelaxant and ant ipro lif-
erative effects of ap igen in. Gen Pharmacol:Va scular Syst, 2000, 35
(6):341 ～ 347
43　Gupta S, A faq F , Mukh tar H. Selective grow th-Inhibitory, cell-cycle
deregu latory and apop totic response of ap igenin in norm al versus hum an
prostate carcinom a cells. B iochem ＆B iophysR esComm un, 2001, 287
(4):914 ～ 920
44　Zand RS, Jenk ins DJ, Diam and isEP. S teroid horm on e activity of f la-
vono ids and related com pounds. B reast CancerR es＆Treat, 2000, 62
(1):35 ～ 49
494 　　　　　　　　应 用与 环 境生 物学 报　　Chin J App lEnviron B io l　　　　　　　　　　　　　　 　　　12卷
45　Sugihara N , A rakaw a T, Ohn ish iM , Fu runo K. An ti-and p ro-oxida-
t ive effects of flavonoids on m etal-induced lip id hyd roperoxide-depend-
en t lip id peroxidat ion in cu ltu red hepatocytes loaded w ith α-linolen ic
acid. F reeR adical B io l＆Med, 1999, 27 (11～ 12):1313～ 1321
46　周伯庭 , 李新中. 母菊的化学成分研究. 湖南中医学院学报 ,
2001, 21 (1):27～ 28
47　Ling Y (凌云), H e BZ (何板作 ), Bao YY (鲍燕燕). S tud ies on
the ch em ical con stituents of comm on du cksm eat (Spirodela po lyrrh i-
za). Chin Trad＆H erba lDrug s (中草药), 1999, 30 (2):88 ～ 90
48　张杰. 木犀草素抑制人 HL-60细胞增殖和诱导该细胞凋亡的作
用. 国外医药:植物药分册 , 2003, 18 (5):205
49　Zhao WX (赵稳兴), Liang CL (梁崇礼). Luteo lin inh ib its prolifera-
t ion and collagen syn thesis of hepatic stellate cel ls. Ch in J H epatol (中
华肝脏病杂志), 2002, 10 (3):204～ 206
50　Du SH (都述虎), J in JS (金继曙). B lood coagu lan t effect of th e
main chem ica l con stitu en t of pu rs lane speedw ell(Veron ica peregrina).
Ch in Trad＆H erba lD rugs (中草药), 1996, 27 (7):416 ～ 417
51　何丽娜 , 何素冰. 木犀草素体外抗柯萨奇 B3病毒的作用. 中国现
代应用药学 , 2000, 17 (5):362～ 365
52　Li B (李波), Zh eng CC (郑传痴), L in ZW (林中文), Sun HD
(孙汉董). Ch em ical stud ies of Ajuga lupu lina var. M ajor. Na t P rod
R＆D (天然产物研究与开发), 2000, 12 (3):4445 ～ 4447
53　丁利君 , 吴振辉 , 蔡创海 , 陈欢玲. 金银花中黄酮类物质最佳提
取工艺的研究. 食品科学 , 2002, 23 (2):62～ 66
54　Wu L M (吴丽明), Zhang JZ (张锦周), Zhuang ZX (庄志雄 ).
M easu ring the es trogenoid activ ity of stam en nelum b in isw ith uterotro-
ph ic assay. J Mod Clin Med B ioengin (现代临床医学生物工程学杂
志), 2003, 9 (2):83 ～ 87
55　X iao D (肖东), Gu ZL (顾振纶 ). E ffects of qu ercetin on p latelet
and reperfusion-indu ced arrhy thm ias in rats. Acta P harm aco l S in (中
国药理学报), 1993, 14 (6):505～ 508
56　Feng CG (冯长根), W uWX (吴悟贤), L iu X (刘霞), Li SC (李
生才 ). Research advan cem en t of onion′s chem ical ingred ien t and
pharm aceu tical action. Shanghai J Trad Ch in M ed (上海中医药杂
志), 2003, 37 (7):63 ～ 65
57　W u YH (吴燕华), LiuWL (刘文力), Yan H (阎红), Li N (李
楠), C ai TY (蔡同一). Determ ination of the phenolic com pounds in
app le ju ice by h igh perform ance liqu id ch rom atog raphy. Ch in J Ana l
Chem (分析化学), 2002, 30 (7):826～ 828
58　 Sun L (孙莲), M eng L (孟磊), C hen J(陈坚), M a J(马季), H u
R (胡瑞), Jia DZ (贾殿增). Determ ination of rut in and quercetin in
mu lberry leaves by high perform ance cap illary electrophoresis. Chin J
Chroma togr (色谱), 2001, 19 (5):395～ 397
59　Xu XL (徐雄良), Zhang ZR (张志荣), Ke ZH (柯尊洪 ), T ian F
(田锋). Quan titative analysis of quercet in in bud ofSophora japon ica
by RP-HPLC. Ch in Trad＆Herba lD rug s(中草药), 2003, 34 (6):
565 ～ 567
60　Liu L, Yang T M , S im on S A. The protein tyrosine k inase inhib itor,
genistein, decreases excitabi lity of nocicep tive neurons. P ain, 2004,
112 (1～ 2):131 ～ 141
61　Zhu L, Jiang ZL, K rn jevi K , W ang FS, Ye JH. G enistein d irectly
b locks g lycine receptors of rat neu rons fresh ly isolated from the vent ral
tegmental area. Neuropharm acology, 2003, 45 (2):270 ～ 280
62　顾晓华 , 杨爱华 , 宗志敏 , 魏贤勇. 大豆异黄酮. 精细与专用化学
品 , 2001 (6):11～ 12
63　Zeng HY (曾虹燕), Zhou PH (周朴华), H ou TZ (侯团章). Ad-
vances in s tudies on chem ical con stituen ts of Trifolium pretense. Ch in
Trad＆Herba lD rug s(中草药), 2001, 32 (2):189～ 190
495　 4期 MA Chunhu i, et a l. :C hem ical S tudy on th e S tem ofLon icera land eo lata