Scheme S1. Synthesis of Trimer A (1), Trimer B (2) and EC-ECG (5).

(a) Catechin, DMTSF, THF, -35 °C, 1h, and -15 °C, 2h,; (b) epicatechin gallate,

DMTSF, THF, -35 °C, 1h and -15 °C, 2h.

1

Scheme S2. Synthesis of Trimer C (3).

2

NaBH4O

OHOH

HO

OHOH

OH

O

OHOH

HO

OHOH

O

O

OHOH

HO

OHOH

O

OHOH

HO

OHOH

O

OHOH

HO

OHOH

(+)-taxifolin (10) leucocyanidin

add

= (-)-epicatechin (11)

in EtOH

0.1N HCl

= (+)-catechin (12)

= PB4 (9)

= PB3 (8)

Scheme S3. Synthesis of procyanidin B3 (8) and procyanidin B4 (9).

3

0. 0 2. 5 5. 0 7. 5 10.0 12.5 15. 0 17. 5 20.0 22.5 mi n-1. 0

0. 0

1. 0

2. 0

3. 0

4. 0

5. 0

6. 0

mAU(x10)520nm4nm (1. 00)

8.86

19.

987

10.5

7511

.599

13.1

51

Figure S1. HPLC chromatogram of anthocyanins

Upper; Wine polyphenol fraction by HP-2MG, lower; LH-20, 10%MeOH fraction.

HPLC-condition

Column: YMC Polymer-C18, 2 mmφ x 150 mm (YMC CO., LTD.)

Solvent: A; 1%HCOOH/H2O, B; 1%HCOOH/CH3CN

Flow rate: 0.2ml/min

Gradient: B10%(3min), B10%→B40% (15min)、B40%iso（4min）

Column temperature: 40 ºC

Detect: Abs 520 nm

R.T. 6.9 min; delphinidin 3-O-glucoside, 8.9 min; petunidin 3-O-glucoside, 10.0 min;

malvidin 3-O-glucoside,

4

0. 0 2. 5 5. 0 7. 5 10.0 12.5 15.0 17.5 20.0 22.5 mi n

0. 00

0. 25

0. 50

0. 75

1. 00

1. 25

1. 50

mAU(x10)520nm4nm (1. 00)

6.94

1 8.91

810

.017

10.5

6211

.543

12.3

5613

.096

14.9

4215

.537 16

.115

Figure S2. GPC analyses of LH-20 fractions of wine polyphenol mixture and

polyphenol standard.

Samples were applied to connected column of Shodex OH-pak SB806M-HQ and

SB802-5HQ (both column 8mm I.D. x 300 mm, Showa Denko K.K., Japan) and eluted

with 20 mM LiBr in DMF at flow rate of 0.8 ml/min. Column temperature was 40 ºC

and UV detector was at 280 nm.

5

Table S1. 1H and 13C NMR Spectral Data of PB3, 4, and EC-ECG

PB3 (8)a PB4 (9)b EC-ECG (5)c

　ring position δH, (multi; J [Hz]) δC　 δH, (multi; J [Hz]) δC　 δH, (multi; J [Hz]) δC　

A 2 4.54 (ｄ, J= 7) 82.5 4.93 (brs) 79.90 5.22 (brs) 77.22

3 3.78 (ddd, J= 6, 7, 8) 68.0 4.23 (m) 67.42 5.49 (dd, J= 2, 4) 68.99

42.76 (dd, J= 6, 16)

28.02.92 (dd, J= 6, 17)

30.032.88 (dd, J= 2, 17)

26.162.48 (dd, J= 8, 16) 2.82 (dd, J= 3, 17) 3.03 (dd, J= 4, 17)

　 4a 102.5 99.40 98.30

　 5 155.0 155.28 155.05

　 6 6.06 (s) 95.5 5.94 (s) 97.65  5.92 (s) 96.28

　 7 155.7 155.90 155.67

　 8 107.7 108.58 107.38

　 8a 157.0 156.47 153.79

　 1' 131.8 131.65 130.63

　 2' 6.57 (d, J= 2) 115.0 7.08 (d, J= 2) 115.20  7.15 (d, J= 2) 114.18

　 3' 145.8 145.09 145.06

　 4' 145.6 145.74 144.91

　 5' 6.66 (d, J= 8) 115.8 6.76 (d, J= 8) 115.90  6.69 (d, J= 8) 114.95d

　 6' 6.24 (dd, J= 2, 8) 119.5 6.85 (dd, J= 2, 8) 118.92  6.97 (dd, J= 2, 8) 118.20

GA 1” 120.41

2”,6” 7.10 (s) 109.52

3”,5” 145.40

4” 138.96

C=O 166.45

B 2 4.24 (d, J= 10) 84.0 4.41 (d, J= 10) 83.79  5.20 (brs) 76.48

upper 3 4.50 (dd, J= 8, 10) 71.0 4.58 (dd, J= 8, 10) 73.77  3.86 (brs) 72.71

6

　 4 4.74 (d, J= 8) 39.0 4.62 (d, J= 8) 38.80  4.83 (brs) 35.91

　 4a 107.0 107.12 101.20

　 5 154.7 157.34 157.59

　 6 5.88 (d, J= 2) 97.0 5.78 (d, J= 2) 97.65  5.91 (d, J= 2) 94.74

　 7 157.2 154.80 157.22

　 8 5.78 (d, J= 2) 96.0 5.83 (d, J= 2) 96.11  5.97 (d, J= 2) 95.26

　 8a 158.2 158.55 157.33

　 1' 132.2 132.27 131.87

　 2' 6.72 (d, J= 2) 115.5 6.97 (d, J= 2) 116.40  6.95 (d, J= 2) 114.71

　 3' 146.3 146.70 145.06

　 4' 146.0 146.33 144.77

　 5' 6.66 (d, J= 8) 115.8 6.78 (d, J= 8) 115.98  6.70 (d, J= 8) 115.21d

　 6' 6.46 (dd, J= 2, 8) 120.5 6.84 (dd, J= 2, 8) 121.00  6.59 (dd, J= 2, 8) 118.47

aPB3 measured in CD3OH at room temperature on a Bruker AVANCE DMX 750 spectrometer (Bruker BioSpin AG,

Switzerland).

bPB4 measured in CD3OH at room temperature on a Bruker AVANCE III HD 800 spectrometer (Bruker BioSpin AG,

Switzerland).

cEC-ECG measured in Acetone-d6/D2O =9:1 at -30 ºC on a Bruker AVANCE III HD 400 spectrometer (Bruker BioSpin

AG, Switzerland).

dAssignments with the same letter are interchangeable.

7

Figure S3. Comparison of synthetic and natural Trimer A (1) by 1H NMR spectrum in

CD3OD at -20 ºC.

Chemical shifts were reported in parts per million (ppm) relative to residual methanol

peaks (H 3.31 for CHD2OD). Standard Bruker pulse sequences were applied. Water

suppression by presaturation was used. Data analyses were carried out with Bruker

TopSpin 3.2 software (Bruker BioSpin AG, Switzerland).

8

Figure S4. Comparison of synthetic and natural Trimer B (2) by 1H NMR spectrum in

CD3OD at -20 ºC.

Method of measurement of 1H NMR is the same condition as that of Trimer A.

9

Figure S5. Comparison of synthetic and natural Trimer C (3) by 1H NMR spectrum in

CD3OD at -20 ºC.

Spectral analysis of 1H NMR is the same condition as that of Trimer A.

10

Figure S6. Comparison of synthetic and natural EC-ECG (5) by 1H NMR spectrum in

Acetone-d6/D2O =9:1 at -30 ºC.

Chemical shifts were reported in parts per million (ppm) relative to residual acetone

peaks (H 2.05 for CHD2COCD3). Spectral analysis of 1H NMR is the same as that of

Trimer A.

11