Pure & Appl. Chem., Vol. 61, No. 3, pp. 357-360, 1989. Printed i n Great Britain.

@ 1989 IUPAC

Isolation and structure elucidation of tannins

G e n - i c h i r o Nonaka F a c u l t y o f Pharmaceutical Sciences, Kyushu U n i v e r s i t y , 3-1-1 Maidashi , H i g a s h i - k u , Fukuoka 812, Japan

A b s t r a c t - Chemical examinations show t h a t a new c l a s s o f t a n n i n s d e s i g n a t e d as 'complex t a n n i n ' occurs w i d e l y i n t h e p l a n t s c o n t a i n i n g b o t h condensed and h y d r o l y z a b l e t a n n i n s . The complex t a n n i n s a r e shown t o c o n t a i n i n t h e molecules a h y d r o l y z a b l e t a n n i n m o i e t y connected through a carbon-carbon l i n k a g e t o flavan-3-01 ( f l a v a n o - e l l a g i t a n n i n ) , p r o c y a n i d i n ( p r o c y a n i d i n o e l l a g i t a n n i n ) and f l a v o n o i d g l u c o s i d e ( f l a v o n o - e l l a g i t a n n i n ) m o i e t i e s . Some o f t h e s i g n i f i c a n t f i n d i n g s o b t a i n e d i n t h e f i e l d s o f h y d r o l y z a b l e and condensed t a n n i n s a r e a l s o b r i e f l y d e s c r i b e d h e r e i n .
INTRODUCTION

D i f f i c u l t i e s i n i s o l a t i n g v e g e t a b l e t a n n i n s f r o m an e x t r e m e l y complex m i x t u r e have been overcome i n r e c e n t y e a r s b y development o f new s e p a r a t i o n procedures based m a i n l y on r e v e r s e phase chromatography, and t h e r e b y enormous p r o w e s s has been made i n o u r u n d e r s t a n d i n g o f t h e c h e m i s t r y o f v e g e t a b l e t a n n i n s . I n t h e c l a s s i c a l work, t a n n i n s were i n p r i n c i p l e d i v i d e d on t h e b a s i s o f t h e i r chemical p r o p e r t i e s i n t o two groups; t h e h y d r o l y z a b l e and t h e nonh y d r o l y z a b l e o f condensed. However, r e c e n t accumulated d a t a o f t h e i r s t r u c t u r a l s t u d i e s have shown t h a t t h e y may be c l a s s i f i e d i n t o t h r e e m a j o r groups based m a i n l y on t h e s t r u c t u r a l f e a t u r e s r a t h e r t h a n on t h e chemical p r o p e r t i e s . The t h i r d group i s now d e s i g n a t e d h e r e as 'complex t a n n i n ' , and i t s i s o l a t i o n and s t r u c t u r e a r e t h e m a j o r s u b j e c t s o f t h i s paper. I n a d d i t i o n , some o f t h e s i g n i f i c a n t f i n d i n g s o b t a i n e d i n t h e f i e l d s o f condensed and h y d r o l y z able tannins are b r i e f l y discussed herein.
COMPLEX T A N N I N

The complex t a n n i n i s a s e r i e s o f compounds f i r s t i s o l a t e d f r o m a Fagaceous p l a n t ( r e f . 1) and now found t o o c c u r w i d e l y i n t h e p l a n t s c o n t a i n i n g b o t h h y d r o l y z a b l e and condensed t a n n i n s ( r e f . 2-6). The i s o l a t i o n o f these t a n n i n s i s a l m o s t i n v a r i a b l y achieved b y a combination o f Sephadex LH-20 d e x t r a n g e l ( w i t h a m i x t u r e o f w a t e r and methanol) and reverse-phase (such as h i g h - p o r o s i t y p o l y s t y r e n e , ODs, e t c . w i t h a w a t e r and methanol m i x t u r e ) chromatographies. The f o r m e r chromatography a l l o w s s e p a r a t i o n f r o m more m o b i l e l o w e r - m o l e c u l a r - w e i g h t p o l y p h e n o l i c s , w h i l e t h e l a t t e r i s e f f e c t i v e f o r t h e s e p a r a t i o n o f each s t r u c t u r a l isomer, The s t r u c t u r e e l u c i d a t i o n i s based on chemical means h y d r o l y s i s , d e g r a d a t i o n , s y n t h e s i s , e t c . ) i n c o n j u n c t i o n w i t h h i g h - r e s o l u t i o n 1H and 3C NVR spectroscopy.

More t h a n t h i r t y t a n n i n s o f t h i s c l a s s have so f a r been i s o l a t e d and s t r u c t u r a l l y e l u c i d a t e d , and f r o m t h e i r s t r u c t u r a l f e a t u r e s , t h e y a r e f u r t h e r d i v i d e d i n t o t h r e e groups. The most commonly o c c u r r i n g i s ' f l a v a n o - e l l a g i t a n n i n ' , w h i c h possesses i n t h e molecule a flavan-3-01, t h e component u n i t o f condensed t a n n i n s , connected t o a h y d r o l y z a b l e t a n n i n m o i e t through a carbon-carbon l i n k a g e . As t y p i c a l examples, t h e s t r u c t u r e s o f s t e n o p h y l l a n i n A ( l f ( r e f . 11, . ) and stenophynin A ( 4 ) ( r e f . 2) a r e shown a c u t i s s i m i n B ( 2 ) ( r e f . 3), m o n g o l i c a i n A ( 3 ) ( r e f . 4 i n the t e x t .
HO

OH
HO HO

HO

OH

1 357

OH

OH

2 -

358

G. NONAKA

OH

HO

HO HO

OH

3 -

4 -

The second i s ' p r o c y a n i d i n o - e l l a g i t a n n i n ' A1 though o n l y one compound, m o n g o l i c a n i n ( 5 ) , has so f a r been known f o r t h i s t y p e t a n n i n ( r e f . 6 ) , i t i s c o n s i d e r e d t o e x i s t r a t h e r w i d e l y i n t h e p l a n t s c o n t a i n i n g b o t h p r o a n t h o c y a n i d i n s and e l l a g i t a n n i n s . The s t r u c t u r e o f 5 p r i n c i p a l l y f o l l o w s f r o m t h i o l y t i c d e g r a d a t i o n and subsequent d e s u l f u r i z a t i o n t o y i e l d a c u t i s s i m i n A ( 6 ) and a l s o f r o m t h e p r e p a r a t i o n o f 5 by condensation o f p r o c y a n i d i n 6-3 ( 7 ) and v e s c a l a g i n ( 8 ) .

HO

OH

0 The 'flavono-ellagitannins', m o n o o l i c i n s A ( 9 ) and B ( l o ) , i s o l a t e d f r o m t h e p l a n t o f t h e genus Quercus ( F a g a c e a e ) ( r e f . 5 ) have n o v e l s t r u c t u r e s i n w h i c h a f l a v o n o i d g l u c o s i d e i s l i n k e d t o an e l l a g i t a n n i n m o i e t y through a carbon-carbon bond. The s t r u c t u r e s have been e s t a b l i s h e d on t h e b a s i s o f a n a l y s i s o f t h e 1H-13C long-range COSY s p e c t r a and p r e p a r a t i o n b y condensation o f t h e c o r r e s p o n d i n g f l a v o n o i d and e l l a q i t a n n i n . The co-occurrence w i t h ( + ) t a x i f o l i n 3 - 0 - g l u c o s i d e and 8 (and c a s t a l a g i n , t h e C-1 epimer o f 8) i n t h e same p l a n t suggests t h a t 9 and 10 a r e b i o s y n t h e t i c a l l y d e r i v e d by condensation o f these compounds.

5 -

OH OH

10 9 -

lsolation and structure elucidation of tannins

359

HYDROLYZABLE TANNIN

D u r i n g t h e p a s t decade, more t h a n two-hundred h y d r o l y z a b l e t a n n i n s and r e l a t e d compounds have been i s o l a t e d f r o m about f i f t y p l a n t m a t e r i a l s i n o u r l a b o r a t o r y . From t h e i r s t r u c t u r a l work, i t i s shown t h a t g a l l i c a c i d i s metabolized, i n t h e f o r m o f t h e e s t e r a t t a c h e d t o t h e p o l y a l c o h o l ( m o s t l y D-glucopyranose) m o i e t y , by i ) o x i d a t i v e c o u p l i n g l e a d i n g t o o l i g o m e r i z a t i o n and i i ) o x i d a t i o n o f t h e a r o m a t i c r i n q as shown i n C h a r t 1 .

i) O x i d a t i v e coup1 ing
HO
HO

COOH

Flavogallonyl

HO

OH

Ho
HO HO

Hexahydroxydiphenoyl Sanguisorboyl
HO
HO

Gallagyl

galloyl

Valoneayl
HO

Dehydrodigalloyl

HO
HO

HO

HO OH

Terchebuloyl
OH

H O G c o
HO

Tergalloyl
I 1

i i ) Oxidation o f aromatic r i n g occo

H O* , ,
HO HO OH

I 1

*
OH

Chebuloyl

Hcxahydroxydiphenoyl

OC OH

Dehydrohexahydroxydiphenoyl
OH

O
HOOC

S OH

HO * : '

HO

HO OH

OH

Flavogallonyl

Trilloyl
I

&

OH

OH

oc co
H O m O & O

I 1

HO

HO H O

HO OH

$ :

bd0

OH

HO OH

OH HO

oc
I
I

co
.

Valoneayl

Chart 1. Y e t a b o l i s m o f q a l l i c a c i d

360

G. NONAKA

Among v a r i o u s h y d r o l y z a b l e t a n n i n s , those w i t h a dehydrohexahydroxydiphenoyl e s t e r group r e p r e s e n t a r e l a t i v e l y r a r e group o f compounds which o c c u r r a t h e r w i d e l y i n t h e p l a n t kingdom, m o s t l y p r e d o m i n a t i n g i n t h e p l a n t e x t r a c t s . The i s o l a t i o n o f t h e n o v e l t a n n i n , e l a e o c a r p u s i n ( 1 1 ) , w h i c h possesses an a s c o r b i c a c i d m o i e t y connected t o t h e dehydrohexahydroxydiphenoyl group, suggests t h a t dehydroascorbic a c i d i s i n v o l v e d i n t h e o x i d a t i v e metabolism o f t h e hexahydroxydiphenoyl qroup t o t h e dehydrohexahydroxydiphenoyl group ( r e f . 7, 8).

CONDENSED T A N N I N

A p p l i c a t i o n o f t h e above-mentioned chromatographic procedures has l e d t o t h e i s o l a t i o n o f s e v e r a l o l i g o m e r i c p r o c y a n i d i n s i n c l u d i n g t r i m e r s , t e t r a m e r s , pentamers and hexamers i n t h e i r p u r e forms. As t y p i c a l examples, t h e p r o c y a n i d i n s i s o l a t e d f r o m Ceylonese c a s s i a b a r k ( r e f . 9) and Areca seed ( r e f . 1 0 ) a r e shown here. I t s h o u l d be n o t e d t h a t t h e t r i m e r i c proanthoc y a n i d i n s , cinnamtannins B-1 ( 1 2 ) and D-1 ( r e f . 11) possess a s t r o n g l y sweet t a s t e , whereas o t h e r compounds t a s t e a s t r i n g e n t , s u g g e s t i n g t h a t t a n n i n s combine w i t h p r o t e i n s s p e c i f i c a l l y .

HO

no

Cinnamtanni ns

Arecatannin A-series

no

Arecatannin B-series
REFERENCES
G.Nonaka, H.Nishimura and I . N i s h i o k a , J.Chem.Soc. P e r k i n Trans. 1 163-172 ( 1 9 8 5 ) . H.Nishimura, G.Nonaka and I . N i s h i o k a , Chern.Pharm.Bul1. 3 , 3223-3227 ( 1 9 8 6 ) . K.Ishimaru, G.Nonaka and I . N i s h i o k a , Chem.Pharm.Bul1. 3 , 602-610 ( 1 9 8 7 ) . G.Nonaka, K . I s h i m a r u , K.Mihashi, Y.Iwase, Y.Ageta and I . N i s h i o k a , Chem.Pharm.Bul1. 2,, 857-869 ( 1 9 8 8 ) . K . I s h i m a r u , M.Ishimatsu, G.r\lonaka, K.Mihashi, Y.Iwase and I . N i s h i o k a , Chem.Pharm.Bul1 ,. " i n p r e s s " . K . I s h i m a r u , M.Ishimatsu, G.Nonaka, K.Mihashi, Y.Iwase and I . N i s h i o k a , Chem.Pharm.Bul1 ,. " i n p r e s s " . T.Tanaka, G.Nonaka and I . M i s h i o k a , J.Chem.Soc. P e r k i n Trans. 1 369-376 ( 1 9 8 6 ) . 941-943 ( 1 9 8 6 ) . G.Nonaka, S.Morimoto and I . N i s h i o k a , Jhem.Pharrn.Bul1. 2, 9. G.Nonaka, S.Morimoto and I . N i s h i o k a , J.Chem.Soc. P e r k i n Trans. 1 2139-2145 ( 1 9 8 3 ) . 10. G.Nonaka, F.-L.Hsu and I . N i s h i o k a , J.Chem.Soc. Chem.Comm. 781-783 ( 1 9 8 1 ) . 4338-4345 ( 1 9 8 5 ) . 11. S.Morimoto, G.Nonaka and I . Y i s h i o k a , Chem.Pharm.Bul1. 2, 1. 2. 3. 4. 5. 6. 7. 8.