Escolar Documentos
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B.TERNAI~~~R!~~ANLEY
CbcmhtryDeparent, hTrobe University.B&oars, Austr& 3083
II. GQGW
InltiM flu cbmie. Unimitat Hobenlleim(LM.). D-7oal Salaprt-70
and
T. J. MABRY
Depament of Botany, Unhmity of Tew at Austin.TX 78712,U.S.A.
glycoside2
~vo&j&7.11.12
The usefuloessof ‘% Nh4R my for llrvonoid Accordingly. C-S has now been assigned
aglycone structure analysis is well e!3tablisbed.Published the Iowa lkld signal (ca 161ppm) amI C-9 the higher
work ioclu&s studks of lIavones,‘4 lIavoDols,‘” gilly- (co. 157ppm). represeat& a reversal of SOme earlier
MvoMids,“” cllalcoW,W eavans,’ is@ lISSigMNOtS.‘J Tlmc clureot aa&nmcots aptxar to be
aavolhes,“” bi&Wntoids’” and auronl!a,Ltog&her with inconsistent with substituho effects observed on
oc4+onal llavoooid glycosides.=” There i!? pnreotly methylatioo of the 54H.’ However, the 54H is strongly
good agreement between d&lent workers 00 tba?assiga- H-bon&dtothe4ketofunctioointhcsecumpoundsand
meat of signals, especially since the as&llmeots for Cb for this reasoo the signal of the adjaceot carbon would
aadC-8,andC-~PadC-9m~hydroxyilavoao~hrvebeeo not ntXessarily obey “normaP substitutioo additivity
ckrilkd as a result of more intlqsive studks.z’.“.‘2 ruks. hked in a comperabk case with 2-hydroxy-
Because flavonoid glyatsides are more important than acetophenonc,itllasbeeoshowntbattlN%crulcsarenot
aglycooes in nature, we initiated a systematic study of applkabk to the C-2 s$nal whicl~ moves upfkld oo
this class of natural products. 10 the present unn- methyl&on of tbc 24H’ rath than downlkld as pre-
munkatioo, the 13C NMR spectra of a wide range of dicted by the substitution ‘hks.
natulauy occWiqt flavowid G-glycoskks and some Tabk2contahsrlistingoftbesignalsassociatedwitb
acylated derivatives are presented and analysed to assess the carbohydrateportion of the flavoooid glycosides. For
the uwfulDess of this tecboique for glycoside structure tbc monoglycosides. assignments were basal 00 those
eluc*n.
. Comparability with previous work in this prc~~,‘Lnade for monosacclmr+ and pLethy1$y-
basbeeomain~tIuuughtbeu!?eof The spectra of the dl- and bl~ycos&s
EzWMimetbylsulpboxide as solveot. were resolved either by “best lh” match& to ap-
propriate monoMccharide spectn, or where possibk,
pglltT8AND- w a comparative study of the simpler mono- or
The glycoside spectra studkd here are preseoted in di-glycosides that coIlstitutc part of the structure. Thus
Tables1~2Tabklcontainsalistineoftbesignals for exampk, assignments in the spectrum of kacmpferol
assochd with the a&one portion of,eacll mokcuk. 7-G-&coside-Ukutiaoriae were deduced by reference
Assi$punentsweremadeprhrilybyonrlopywitbpm- to the spectra of k8empferol7-G-glua~ide, kaempferol
vious studks.‘-’ For reference purpoWs sevarl of these ‘3,7di4+atM and’ Lsempferol 3-Wutinosifk.
aglyconcsand their metbykted derivatives are inch&d Glucose C-l sigh& in the spectra of tbc 3-G.
hthetabk.Oaed&uItyarisingwithtbehvohl3-, ropbaoriaer aBd 74h?ohperidorides were assign4
on the
aud 3,7-G-&o&s is the near equivakoce of the c-2 . . basis of previous work on various
and c-9 sigmls. aod no attempt ha!3 been made to gluahsh .Ian In some of t&e more complex spectra
ditlereotiate between them in this study. accumteass@neotscouldnotbemadeforanumberof
It has recently been &own that the ox& sigoals with similar chemical shifts. This limitation
carbons C-S and C-9 each give signals which bave dis- however, has not affected tbc conclusions reached in this
tilK%VC “C-‘II coupliq# in saydroxyhtal communicatioo.
1389
1390 K. R. MAKKUAIu al.
TLAVWlOID*
'r APiFb”
lb. A-7-4(. 164.1 103.0 181.7 161.2. 97.8 165.1 92~6
Ic A - 7-Ogld2-lhpiow 164.4 '103.3 181.9 161.3' 98.8 462.8 95.0
k (6-l)rha-
K-3-0-&111 156.6' '133.5 177.5 161.3 98.8 164.2 93.8
24. K-7-O-glu(2-l)rha 147.9 135.9 176.1 160.4 98.8 162.4 94.4
h. g-7-o-glu'3-O-61. 156.1. 133.8 177.7 160.9 99.6 163.0 94.8
21. K-7-O-gin3-0~glo!O-ro> 156.0' 133.6 177.5 160.8 99.5 762.9 94.7
a. m.8
K-7-O-gluM-glci~2-l~glu~O-sin~poy 733.4 in.4 s0.9 99.4 -62.0 94.5
p. K-7-O-glu3-0-g1U(2-1)gh 155.9 733.3 1n.s 1641.9 99.4 162.8 94.6
a. K-7-O-rha3-O-&h 796.0' 133.A ~7.6 160.9 99.4 16'1.8 94.5 .
.
a. K-7-O-rlu3-O-glu(OhI v6.0~ 133.6 777.5 160..9 99.5 161.8 94.6
a. K-7-O-rh 3-O-glo(6-1lrha v&d 133.7 ln.6 160.9 99.4 161.7 94.6
a. K-7-O-glo 147.9 136.0 176.1 160.5 99.2 162.9 94.8
h K-7-O-glu3-O-glu(6-1)~~ '156.0' 133.7 177.6 W.9 99.7 162.9 94.9
da. e-J-o-ga (6'-0-gdlo~l) ~6.3~ 133.7 177.4 161.2 98.8 164.1 93.6
46. e-3-O-uab 156.4' 133.5 '.ln.B 161.2 98.7 164.1 93.5
40. Q-j-o-uab 156.3 134.0 1n.6 161.2 98.7 164.1 93.5
Y. e-3-0-rhm 156.4' 134.4 177.7 161.2' 98.6 164.0 93.5
Most of the glycosides studied are natural products special case.As a result of glycosylationthe S-hydroxyl-
(Table 1) and although tbe points of attachment to the 4-keto H-bond is broken and this has a profound effect
aglycone and tbe sequence of sugars are firmly on the electron density distributionin the molecule.Thus
established in the majority of cases, the ring size of each althoughthe shift observed in the C-S signalis similarto
sugar, the anonwic form of each sugar and the inter- that expected from a study of glycosylation effects at
glycosidic linkages between sugars are generally un- other sites, as also are the shifts of the orfho (Cd, IO)_
known or poorly defined. For example the p-anomeric and porn (C%Wated ca&ns, a marked effect is also
form of the 7-O-gh~sides was recognizedby its lability noticed in the signals of the C-ring carbons due to the
to emulsinand conversly, sophorosideswere detected by release (in Iptaolin)of the 6keto function from H-bond-
their stability to this enzyme. Tbe contriition that “C ing. Also, the C-2 and C4 signals move sharply upfield
NMR spectroscopy can make in these and other problem and that of C-3 downfield, the reverse effect to that
areas will now be considered. observed when H-bondingis introduced. for example by
1. 7% &xts of glycosylation on the aglyconr hydroxylationof flavone at C-5.’
spectrum. The effects of glycosylationon the ‘% NMR In the viciity of the OH group, the effect of methyl-
spectra of llavonoid aglycones are summa&d in Table ation of that OH group appears to be tbe reverse of
3. These data represent averaged values of shifts obser- glycosylation (Table 3). Thus, whereas glycosylation
ved in all relevant models studied and are compared in produces an upfIeldshift in the signalof the oxygenated
the table with similar shifts observed on methylationof carbon, methylation produces an approximately
the same OH function. equivalent downfieldshift. L&ewise the shifts observed
The effects of glycosylation of the 7-OH on the C-7 in the signals of the ortho-related carbons tend to be
signal is a 1.4ppm upfield shift which is in agreement up&ld rather than downfieldas on glycosylation. This
with our earlier observations.’ This is accompanied by reversal is not observed at the pam-position however.
sign&ant downfield shifts of the signals of the ortho- Methylationof the S-OH is the exception; here, as with
and pam-related carbons, Cb, 8 and 10.the effect being glycosylation methylation produces an upfield shift of
more noticeable with C-10.Glycosylationwith rhamnose the C-5 signal. Shifts in the C-ring too mirror those
appears to have a more marked effect on the C-7 signal observed on glycosylation, thus c&Inning that rekase
than does glycosylation with other sugars, the average of the CO from H-bondingis the primary cause of these
upfield shift being of the order of 2.35ppm. This shifts. In the A-ring, shifts observed are also all in tbe
diiference is of diagnosticvalue. same direction as those for glycosylation. except for Cd
The same general pattern of signal shifts is also 2. Znre~ycosi& linkages. It has been established that
0pserVedon glycosylationat C-3.3’ or 4’. Glycoaylation glycosylation of sugar hydroxyls produces a sizeable
at C-3 however does produce a larger than expected downtield shift in tbe resonance of tbe hydroxykd
‘“0Meffect on the C-2 signalthus confirmingprevious a&on.” Likewise, this has been observed in a preli-
observations.2Rhamnosylationof the 3-OH, as with the minary studf of flavonoid Odiglycosides. For example
7-OH, appears to produce a Merent shift at C-3 than do in !lavonoid rutinosides (rhamnosyl (l-6) glucosides) the
other sugars.t Thus the C-3 signalis seen to shift upfield glucose C-6 signal was found to appear 4.~5.2ppm
by about l.OSppm on rhamnosylation of the 3-OH downlield from that of glucose itself, and in a flavanone
compared with 2.1ppm for glycosylation with other neohesperitioside (rhamnusyl (I-2) glucoside), the C-2
sugars. Sii only one example each of glycosylationat signal appearedabout 2.6 ppm downfield from that of C-2
the 4’- and Y-OH’swas available for study, the figures in glucose. The present study includes five rntinosides
presented must be treated as prelimimuy.However, the (2&a, 41J) and generally confirms the above, although
general trend of shifts observed is the same a3 that the average downfield shift is 5.8ppm. It is also noted
discussed for the 7-OH. the signal of tbe pam-related thattheghlcosec-5signalundergoesanuptXdshiftof
carbon again showingthe greatest shift. Indeed, this shift about 1.4ppm. The singk neohesperidoside studied (?a)
together with those of the ortho-related carbons may is also consistent with previous work, exhibiting a
well be a more reliable guide to the site of glycosylation 3.9ppm downfield shift of the glucose C-2 signal and a
than the shift of the oxygenated carbon itself (e.g. see 2.1 ppm upfield shift of the glucose C-l signal due to
shifts quoted for 3’Gglycosylation. Table 3). rhamnosylation.Apiose appears to have much the same
Glycosylation of the S-OH group clearly represents a effect on tbe glucose spectrum as does rhamnose(cf. lc).
When gh~se is the glycosylating sugar as in
tThe prcvifmslyexpressedopinion’that 3-O-rbamacwylatbnsophorosides (e.g. zb and a) however, a much larger
of quercetiamay cause a large shift in the C-l’ signal is not downfield shift, of the order of 8 ppm, is evident in the
supportedhowever.The 130.3ppmSW formerlyattributedto C-2 signal. Shifts of this size appear to be typical of
C-l’ wLI in fact tbc C-3’,5’ signalof a small kaempferol3-G &glucosylation generally, in disaccharides’~‘9 and oli-
glycosidcimpurity.A protoncoupkdspectrumhasreveakdthe gosac~harides.‘~In 2b and 2l1ghbcosylationat C-2 is also
C-l’ signalat I21ppm. hidden beneath tbc c0mbinr.d C-Y. 5 detectabk in the glucose C-l signal which shifts up6eld
SigMlS. by about 29ppm. The shift diikences observed above
TETVdXNa9-N
1396 K.RMAUIiMrtoL
Twboi and I. Nishiokx. Tmolrsdnm Lmer~ No. 14. 1231 T. A. Gcixxman). p. 317. Pcrgamon Prexx. Oxford (l%Z).
(1977). “E. Bnitanier and W. Valta. Tumbdmn 29).227 (1973).
‘bv. M. Chxri. M. Ilyu. H. Wagwr. A. Ncumelyi. P.C. Chen. “J. B. Stotlurx. Cti-I3 NMR Spectmscopy Vol. 24. of
L-K. Cben. Y.-C. tin unJ Y.-M. tin. fiytocb. 18. 1273 Orsonic Chemistry, A S&u of Monomwbs.D. 46b.Academic
Wm. P&s. NewYotijl972).
“Chinl-i# Chang, 1. Org. Chem. 41.1881 (1976). mH. G&r and B. Krumbein. Z. Nafutiorxc~. 21r. 773 (1973).
‘21.Y. L&mand aml M. rhltcil. &Panic hfmic ResoMnce 9. nH. P. “&. H. Geiger, U. !&uckcr. -H. Waldrum, G: V&r
I79 (l9zq. Veldt and T. J. Mabry. plryfwh. 14. 1613(1975).
“V M. Chari, M. Jordan, H. Wagner and P. W. Tbeis. fiy- 9. Geiger. U. Lxn&*B. Britixch. T. J. Ma@. U. Schwkcr-
&em. 16. Ill0 (1977). Suhr. G. Vandcr Veldt and H. Waldrum, Ibid, in press.
“G. C. Levy and Cl. L. Nelson. In C&on-l3 NMR for Ornoric ‘PRcfs. contained in, 1. B. Harborne. Compamtlw B&hem. of
Chcmisfs b. 81. Wiky-Interxcicnce, New York (l97&. - fhr Flowwids. Academic Press, London il%7).
“hi. Kobavashi. Y. Tcmi. K. Tori and N. Tsuii. e Tetmhdmn =N. A. M. Saleh. W. Maiak and G. H. N. Towers. flvtocihnrr.
_
htfus N-o.8. aI9 (1976). 11. lo95 (1972). -
“P. A. J. Goren and M. Maxurck. Can. 1. Clrem.53, 1212(1975). “S. Beckman and H. Geiger. Ibid. 7.1667 (1968).
“R G. S. Ritchic. N. QT. B. Korsch, H. J. Koch and A. S. =S. Beckman and H. Geiger, Ibid. 2,281(1%3).
P&in. Ibid 53, 1424(1975). “N. Kawano. H. Miura and E. Matsuishi. Chen. fionn. Bull.
‘k. Yamoaka, T. Usui, K. Mawlx, K. Tuximura, H. Sngi- Japan 14.299 (1966).
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“P. C&on, H. J. Jcn&x aod 1. C. P. Smith, L Am. Crkan. 73, I33 (1953).
Sot. 96.8681 (1974)and Refs. therein. “c.f. J. B. Harbornc. Pbyfocbem. 4, I07 (l%5).
‘DM.R. Vignon and Ph. J. A. Vottcro. Tetmhedmrr Lettux No. %H. Geiger and S. Beckman. 2. Natwfoncb 2&. II39 (l%5).
28.2445 (1976). “H. Pacbxco and A. Grwiller. Bul. Sot. Cbim. Fr. 3212(1966).
“H. Geiger. U. Schuckcr, H. Waldrun. G. Vandcr Vcldc and T. “H. Kramer. Diswtation. M&hen (1%6).
J. Mabry. 2. Naturfonch. 3k. 296 (1975). *cf. R. Teouk. G. Grenicr. H. Pa&co lad J. &pin. Buff. Sot.
=B. Steugel and H. Geiger. Ibid 31~.622 (1976). Chim. Fr. 546 (I%I).
=S. Hattori. The Cbaism of Iknmoid Compounds (Edited by