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O

HO O

OH

OOH

HO O

OH

O

HO O

OH

H3CO

O

O

OH

O

O

HO

OHHO

OH

O

OH

O

O

O

OH

O

O

HO

OHHO

OC

O

CH3

O

O

OH

O

O

HO

OHHO

OpKa ~ 5.7

pKa ~ 11

0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00

2

34

56

7 8 9

1

2

3 45 6

7 8 91

∼

12% B

30% B

22% B

12% B

30% B

22% B

200 220 240 260 280 300 320 340 360 380

λ

259.4

259.4255.9

248.8

259.4

248.8

259.4

255.9

248.8

5.00

10.00

15.00

20.00

25.00

30.00

3 4 5 6 7 8pH

k' R

eten

tion

Daidzin

Glycitin

Genistin

AcetylGenistin

AcetylDaidzin

AcetylGlycitin

Genistein

GlyciteinDaidzein

IsoflavoneGlucosides

IsoflavoneAcetylGlucosides

AgluconeIsoFlavones

pH of DI < 7, at ~ 5.5

pH 3.4 with 0.1% Acetic Acid pH 6.8 with 10 mM Na Acetate

Daidzin

Glycitin

Genistin

AcetylGenistin

AcetylDaidzin

AcetylGlycitin

Genistein

Glycitein

Daidzein

IsoflavoneGlucosides

IsoflavoneAcetylGlucosides

AgluconeIsoFlavones

0.00

5.00

10.00

15.00

20.00

25.00

30.00

3 4 5 6 7 8 9 10

pH of DI <7, ~5.5

pH 9.1

0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00

∼

6

7

1

9

2

3

4

58

12% B

30% B

22% B

259.4

258.2

248.8

259.4

258.2

268.8

248.8

255.9

253.5

λ

0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00

∼

6

7

1

9

2

3

4

5

8

11% B

22% B

30% B

2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00

1

2 3

4

5

6 7

8

9

2 µg Each in 50% MeOH

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0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00

1 2

3

4

56

7

8

9

∼

Minutes0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00

1500

mA

U

3 5

6

7

8

1 29

6

2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00

2

3

4 5

6

78 9

2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00

440

1 2

3

5

6

78

9

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