[1]

DCCC, droplet counter current chromatography (HSCCC , high speed

counter current chromatography)

2-1


1:101:20,

2050
100
100200

2
Vo

V1Vo-V1

 15

2
30

1015

100 C 3

()

10

200g 200ml

()
physical
adsorption,

 Rf

Rf 0.20.3

(7)

resibufogenin(cinobufagin)
(bufalin) 13
2-2
10g 30g
pH 5.3 100200
4.5 60
350ml
2-2.
2-2

1
:5:1 0.373
2-4
:5:1 0.100 1
5-10
:5:1 2.735
11-13 :5:1 0.453
14-15 :5:1 2.636
16-20 :5:1 0.793
21-25 :5:1 0.005 4
26-30 :5:1 0.334
31-32 :5:1 0.307 6
33-35 :5:1 0.296
36-42 :5:1 0.268
43-46 :5:1 0.162
47-48 :5:1 0.084
49-54 :3:1 0.477
55-57
0.388

58-63
0.899 12

9.707

97.07%

O
O

R2

CH3

CH3

CH3
CHO

R3
OH

HO

R1

OH
HO

R1 R2 R3
(hellebrigenin)
H H OCOCH3
bufotlin
H H OH

H H H
bufalin
(telocinobufagin) OH H H
(gamabufotalin) H OH H

).5-1.0
R1 R2
cinobuftalin OH OCOCH3
OH OH

H OCOCH3
(cinobufagin)

OH

H OH

resibufogenin H

kiesilguhr, celite

()

1:1 5
10 510

0.1-0.2

1:0.5-1.0 1 0.5-1

1:1020
1:40

1:1001000

1:10000

0.51.0
3
510
510

2.6cm50cm 0.07ml/min 4.5cm65cm 0.3ml/min

5.5cm90cm0.6ml/min 6.5cm115cm1.2ml/min
4

2-3
,

2-3

,, 0.5

0.05N 0.5N

:() 3:1

: 1:1

1 digoxin

O
R1

CH3
R2

CH3
OH
digitoxose-digitoxose-digitoxose

R1
R2
digitoxin
(gitoxin)
digoxin

H
H
OH

H
OH
H

80100 100 23

1:20
12
0.5 G
80:19:1

2 divaricoside
250g 125ml
1:20
4g 10g
5ml
30ml
2-4
2-4

14-19

20-49

50-70

72-101

102-113

114-153
99:1

154-230
95:5

CH3

HO
CH3

OH
RO

R = L-
divaricoside
sarmentogenin R = H
4

Rf

1,2 1,3

-COOH -C(OH)3 -

1,3

2.5
45:45:4.5:4.5
2-3
-

CH2OH
H

OH

H
CH2OH

(2).

1.

(3).

OH
H

OH
OH

(4).

(5).

(6).

OCH3

OCH3
OCH3

OCH3

OCH3
OCH3

H3 C
O
H

H3CO

CH3

CH3

H3CO
H2C

CH2
C
H

Acorus gramineus Soland.3Kg

50g 530Pa 80C118 C118C
128C128C136C136C138C138C142C 142C 138C
142C 120g
100:2 100:5 100:20
200ml400ml1300ml330ml100ml
100g
20g
110C 4 100:5 1g 20
5:1 5ml ,
20 G 5:11
10 37
42 4856 -6180

azobenzene(p-methoxyazobenzene)(sudan
yellow) III(sudan red III)(p-aminoazobenzene)(phydroxyazobenzene) 20mg 10ml 40ml 60
90C 0.04% 1.5 1015
5 10ml
4:1 20ml 20
30 2-5

26
68 C
55 C 134 C
III 184 C 127 C
153 C
2-5
I
II
III
IV
V

2
3
4
5
6

1
2
3
4
5

1
2
3
4
(1). (2). (3). (4). III (5).
(6).
2-6
I IIa IIb IIIa IIIb IVa IV b IVc V VI
2
3
4
5
6

2
3
4
5
6
1
2
3
4
5

1
2
3
4
5
2-5

III 20mg
30mg

10 Rf
Rf 2-7
III

 2-7 Rf
II
III
IV
V
III
0.000.11
0.120.36
0.370.58
0.58
0.100.26
0.270.58 0.590.78
0.78

0.180.45
0.460.71 0.720.90
0.90

0.610.68
0.690.85 0.860.94
0.94

Rf

550 C 24

9.5 95 III 6000g

45g
6000ml
7 2-8
2-8

ajmaline
99.5:0.5
(reserpine) (canescine)
(yohimbine)(pseuoyohimbine)-corynanthine
99:1
-(-yohimbine)
OH
OH N
N
CH3

H2
C
CH3

R1
R2

N
H

H
H3COOC

R1R2H
R1R2OCH3

CH3
O

N
N
H

O
C
O

H
H3COOC

OCH3
OCH3

OCH3

N
H

R1

OCH3

R2

R3

R -H
R1=-H, R2=-COOCH3,R3=-OH
R -H
R1=-H, R2=-COOCH3,R3=-OH
- R1=-H, R2=-COOCH3,R3=-OH

4 95 III 1000g
10g
1000ml
5 2-9
2-9

112
aricine
2243
99.75:0.25

4448
99.5:0.5
isoreserpiline
4950
99.5:0.5
isoreserpiline
5167
99.5:0.5
reserpiline
6869
99:1
-(- yohimbine)

700800


1 710

pH 56 105
C 8
2 20 510

pH 56
5:10000 1Kg 500mg
60 C 10 pH 67 100
C

6
10
90 C
1:1
pH4
5 100 C
100 C
100 C

()

150 C
45 100 C

 45

1.5 1
9

1:1

2550 1 2ml4ml
100ml 5g10g

25
15715

510203050709515715
3.5

6
(Caloglossa leprieuii (Ment) j. Ag.) 25Kg 70
1.5 6000ml 100
10
pH 56

 300ml
600g 100 C 5
6.7cm 51 cm 6
7ml
5000ml 6000ml 58000ml 10
2000ml 201000ml 506000ml 70
500 ml 15 ml Rf 0.55
513100000
243 C
CH2
H3C

CH2COOH

N
H

COOH

-kainic acid
4

polyamide

6 66 11 1010
6 66
6 66
11 1010

6 66 1600020000 200 C

6 66

1 6 66
10 90 C

pH 4
100 C
80 C
2 3
17 50 4
5 pH 10 pH 7
8

DMF
5:10:30:201:1
90955NaOH 10

9095
34 9095
23 5NaOH 1 23 10

5NaOH

5NaOH

pH 89 2 10

1 6
CONH

2-

H2C

CH2
N

N
CH2

H2C

H2C

CH2
CH2

N
CH2

CH2

H2C

H2C

H2C

C
C

H N

H2C

CH2
CH2

2-
2

HO

OH

OH

>

>

OH

OH

OH

7-
5-
OH

OH

>
OH

>
OH

OH
OH

COOH

>
HO

HO

>

OH

3
4- - -

OH
OH

>

>

OH
HO

OH

OH

OH

>HO

OH

> > > >

> > > >

<0.002cm

100 ml 1.5g2.5g

2030

102030507095 3.5

(DNP-)

5:10:30:20.

1030 5095

Rf

Rf

Rf
Rf

1
90:5:5 4:1
1:11:13:23060
6:4 7:3 1:4:5
65:25:10
2 94:6 96:412:2:1
90:6:4 84:8:860:38:2
70:22:864:28:8
3
10 8:2:1
10:10:5
4
1:1 45:8:4
93:710
5 8:16:2:1:1
6 30:2.5:0.9:0.1--
88:12:0.1
7 8:2 9:5501.5:100
1:1
20:1:10.05mol/L 3:1
5:10:30:208:2
8 4:14:1
9
60:35:5
50:20:25:5

-4-
Sophora japonica L.2Kg,
28g,
60g 5.3cm,
80cm
3050
300ml

 30
-4- genistein-4glucoside,sophoricoside, 257 C 50
genistein, 294296 C

HO
HO

HO

O
OR
OH O

OH

OH

OH

R = H

-4-
R =
()
Psoralea corylifolia L.
5

50
70 50
212 C 70
165 C

1
2
3
4

5
6

300400 50 34

III-V
II-III

50

70100

1
8090 C

II III

10

1:100 1:300

2
5

2-

2
5

 50
100

Schisandra sphenanthera Rehd. Et Wils

III-IV
: 6:1 2-
,,
:
120-200 III-IV 240
2.5 40
1.4 6:1 10

6:1

11.2cm
17.6cm;17.6cm27.2cm;27.2cm33.2cm
17.6cm27.2cm

2-10
0cm5.5cm
5.5cm10.5cm 10.5cm16cm
16cm19.5cm
19.5cm 22.5cm 22.5cm
25cm 25cm27.5cm
27.5cm
32cm
32cm36cm
36cm40cm

 2-10
cm Rf

1
05.5
1++
2 5.510.5
2+
3 10.516
0.28
11.2
2+++
4 1619.5
0.44
17.6
3+++++2+
5 19.522.5
3+++++
6 22.525
3++++
7 2527.5
0.68
27.2
4++++3+
8 27.532
5+4+3+
9 3236
0.83
33.2
5+++4+
10 3640
6++
1.2.3. 4.
5.6. 156

OCH3

H3CO

O CO

OH
CH3

H3CO
H3CO

CH3

CH3
CH3

OR
OH

H3CO
H3CO

H3C
H

H3C
H

CO
CH3

CH3
CO

Zipax
254nm

150200
200 200 C 23
60 C
2.9cm 215
50 1
8 5
1.5

 14

14 14 1113 7

Sophora japonica L.2Kg,

3:1

60120 110 3.3

50 5
10 3:1

17cm21cm I 27cm34cm II
I
genistein II

kaempferol
HO

O
OH
OH

OH

HO

OH

OH

1938

1956 Stahl
Merck

Rf Rp18 Rp8
Rp2 Rp

1
2
3
10
100 4
500C 5
6
7
8

10

GType 60 15 G
gypsum Type 60 606 x 10-5m
H
G
HF254 H 254nm

254nm
HF254366Type60 HF254 254nm
366nm

254nm 366nm

60HR

140180

GType60EType 60 606
-5
x 10 m
HType60E E
E T E
HF254Type60E HF254
140

4cm x 20cm
20cm x 20cm 10cm x 10cm

7.5cm x 2.5cm
2 cm3 cm

1 3mm5mm

, 0.4mm
1mm
2-

0.4mm
2-

200

2 6ml8ml
105C

1:563:22:3,
5 45

5 3
105C 2
80200
2

3
2

1 G GF25430
60ml90ml 12

105C 30
128C

1214
G 300 85 15
G 1 G 57 H
H 34 2

 120C 1

10 1
9:1

> > II
2 G 25 5
50ml 12

200C220C II 150C160C

IIII-IV
CMC 0.3%
0.5% 30 50 100 0.5%

28.5 1.5 75

G
30
20 0.02

2-11
II
III
IV
V

0.59
0.74
0.85
0.95
0.16
0.49
0.69
0.89

0.01
0.25
0.57
0.78

0.00
0.10
0.33
0.56

0.00
0.03
0.08
0.19

0.04% 0.5%

5
10.01%

 0.5
1cm1.5cm
2mm
3mm

0.5cm1cm

Neher Von Arx

50:50< 60:40< 20:80< <
95:5< 70:30< 30:70< 99:1<
10:90< < 99:1< 99:1< <
99:1< 50:50< 90:10< < <
< 90:10

10:110:2 1:1

10:110:2 1:1

2
9cm x 28cm
x 5cm,

15
0.5
34

0 v/v

2-12

2-12

-TKedde

()Rf

Rf

Rf

1mg2mg

1 5102
0.5mm1mm 20cm x 20cm

3 20cm x 20cm
10mg50mg

150 20cm x 20cm 15 4

0.3mm5

10:10:1

strychninebrucine
150 200
II
2:1

vinblasteine
(leurosidine)(vincristine)

150 200 IV V
10:10:1 1

Pseudolarix kaempferi Gorden

2-13
105C 4:1

2-13

1
2
3
4
5
6

0.92
0.77
0.70
0.47
0.37
0.28

141C142C

20 15
2-14 180 250
III 3:7 3


2-14 15

x 100

70 02

2
-
70 02

3
63 02

4
35 02

5
85 02

6
50 02

7
55 03

8
56 02

9
36 02

10
39 03

11
60 02

12
79 04

13
68 03

14
25 01

15
65 03

1 IIIII

2 IIIII
9:1 9:1 9:1
9:1 10 70
3 100:2 1:1
10 105C 36:9:5 15
105C
4:1:5 5:1:3 30 3 6:2:7 5

4 III 99:1 95:5

70:20:5 2 3,5- 5

5 105C 1:1:1
G105C 1 10:3
20:1 1:1 5 0.5
90C 5
6 IIIII 5:1
1:1 1:1
7 G120C 2 95:5
90:10 85:15 5
5
8 IIIII 99:1 97:3
95:5 90:10 25 140C 5
510


9 G 10:1.6:1.2 8:0.4:1.6
90:5:3 45:8:14 0.05%
0.1 5 0.1% 15 0.1 20
( 100C )
10 G105C 1:1 10:4
3:1:1 75:25 105C
4:1:5 7:3 10:10:5:2
80:20:4 0.1% 80 10
11 105C 65:35
0.5 9 0.5 0.1
G105C 6:3:1 65:20:15
70:15:15 6:3:1 5:3:2

22 6

2
3


1
2
3
4

2-155

6 pH

pH

pH
pH
pH

pH
2-15

1
90
0.17
150120
0.08
2
90
0.16
12091
0.08 1
3
90
0.15
9060
0.08
4
180
0.34
151121
0.08
5
180
0.32
12091
0.08 3
6
180
0.30
9060
0.08
()

20cm
30cm 2-
2-

2- 2-
2:7:1

2-

20cm30cm 2
2 5
0.5
3mm5mm

23 2-
5:1:1:1
15:3:2
12
1cm2cm 510

1
0.050.85
0.05 0.4
0.6 23
4


56

4:1:5

10ml 2ml 8ml

8cm x 30cm
2 2

2 7 20C
0.710.60
0.36

4090ml 1010ml 2020ml

4cm x 30cm
4 1.5mm2mm

2 6 30C

 2-16
2-16

(4:1:2)
9:1:2
(lycorine)
0.60
0.73

0.53
0.67
galanthamine

0.51
0.68
lycoramine

0.72
0.84
(tazettine)

0.69
(pseudolycorine) 0.55

0.58
0.72
1

0.71
0.82
2

0.53
0.71
3

0.77
(homo- lycorine) 0.59

0.72
0.88
(papyramine)

lycorenine
0.72
0.92

CH3
N

CH3
N
O

O
CH2OH

NCH3

O C C
O H

HO

R OH
R OCH3

CH2OH

O C C
O H

4:1:5 2:1:1
4:1:5 120pH48

4:1:5 2:1:1

4:3:3
5

4:1:1:2 35:35:30


1:1 5:5:1 4:3:3
T

4:1:5 2:1:5

4:1:5 4:1:5
6:4:3 2:7:1 2:1:5

4:1:5 4:1:5
14:4:1:1

SO3HCOOH

H
C

C
H2

H
C

H
C

C
H2

SO3H
SO3H
H
C

SO3H
C
H2

C
H

H
C

C
H2

SO3H

SO3H

1 x 77321
Amberlite IR-120Dowex 50Lewatit S100Permutit QWofatit KZeo Karb225

5
0.10.1mol/L

42 Amberlite IR-100Dowex 30Wofatit KSZeo Karb215

OH
C
H2

OH
C
H2

CH2

C
H2
SO3H

SO3H

OH
C
H2

H2
C

H2
C

C
H2
OH

CH2

pH
2

 101 x 128724101
Amberlite IRC-50Wofatit CZerolit226
CH3
H
C

C
H2

C
H2

COOH

CH3
C
H

C
H2

CH3
C
H2

COOH

C
H2

H
C

H
C

C
H2

C
H2

H
C

H
C

C
H2

CH2

COOH

C
H

H2C

C
H2

N (CH3)3Cl HC

C
H2

H2 C

N (CH3)3Cl

1
201 201 x 7
717 Amberlite XE-98Amberlite IRA-400Amberlite IRA-410Dowex 1Dowex
2Lewatit MIINalcite SARPermutit SIPermutit SIIZerolit FF
OH
Cl 40C45C
2
330701 311 x 2704
321301330
Amberlite IR-4BDowex 3Lewatit MPermutit WWofatit MWofatit N

NHCH2CH2CH2NHCH2CH2NH
HN

HN

C
H2

N
H
CH2
NH

N
H
CH2

C
H2

N
H

C
H2

HN

H OH
RCOONa H2O
RCOOH NaOH

COOH
H SO3H

1 60 120

116
8 4
2

3
36
1 x 7 4.5
4.5 x 89.09 = 400.95mg = 0.4g 89.09
pH
pH
12
1413
4 50

1 pH
pH

pH 2
pH 6 pH
pH 12 7
2

4 0.1

20 35 0.83mm0.42mm 50

H OH
12
1 20
7101
H 10 4

H H
10 4
H
2 20
4 OH 10 10
4
OH OH 10 4 OH
OH
OH
3 10
4 H 10 4
10
H H
10 4 H
4

10

1cm2cm


pH

pH

Polygonatum 4kg
1000ml Zeo Karb 215 H
3000ml 1200ml 0.5mol/L
1000ml pH 5 Zeo
Karb 215 H 1500 ml 0.5mol/L
2 ml
130 31459 -2-460505
506534 615667
-668750
NH2

NH2
HOOC C C COOH
H2 H

NH2
HOH2C C C COOH
H2 H

HO

C C
H2 H

COOH

O
NH2
H2N C C C COOH
H2 H

H3C

H
C
OH

NH2
C
H

COOH

H2N C C C COOH
H2 H2 H2

NH2
NH

H2N

(CH2)4 C
H

COOH

H2N

NH2
N
H

(CH2)3C
H

COOH

N
H

COOH

-2-

[23]

Amberlite XAD-4XAD-2Daion HP20 ADS-52

COOR3
CONRRSO
4
C5H4NNRR 2-17
2-17

/nm
Amberlite XAD-2 - PS

330
4.0
Amberlite XAD-3 - PS
526
4.4

Amberlite XAD-4 - PS
750
5.0

ADS-5
PS
550
2025

ADS-3
PS
1000
56

Amberlite XAD-6 - -COOR

498
6.3
Amberlite XAD-7 - -COOR
450
8.0
ADS-8
-COOR
140
25.0

Amberlite XAD-9 -
250
8.0

Amberlite XAD-10 -
69
35.2

ADS-16
50

Amberlite XAD-11 -
Amberlite XAD-12 -
ADS-7
-NRR
200

170
25

21.0
130

H103X-5 ASSI-2AB-8

D DA

Amberlite XAD-7

[4]

5nm20nm 60nm500nm

2/3

102030507095

B12

1:1 pH3 1:1

90 3[5]

10

ADS-7 S-038
S-038
70

[6]70

[7]
N

H3C N
H3C

OH O

CH3
N

N
N
CH3

[8]

(molecular sive filtration)

gel filtration(restricted
diffusion chromatography)(exclusion chromatography)(gel permeation
chromatography)
bio-gel P sepharose,bio-gel
ACM-Sephadex
DEAE-SephadexSP-Sephadex
QAE-Sephadex
Sephadex LH20

-1,6 95 1,3 3--1,2-

0.02mol/L
sephadex G-25 0.25mol/L 60C
0.5mol/L ( 0.5moi/L )

110C 40 120C
CH2O
O

CH2

OH
O

O
CH2

OH

OCH2CHOHCH2O

CHOH

OH
OH

CH2

OH

CH2
O

O
OH
OH

O
CH2O

CH2

OH

HOHC

CH2

OH

CH2

O
OH

OH
O

OH

O
OH
CH2

G
10 G-25 2.5
2-18
2-18 Sephadex

ml/g ml/g ml/g g/ml

G-10 2
0.8
1.0
1.24 <700
<700
3 1
G-15 3
1.1
1.5
1.19 <1500
<1500
3 1
G-25 5
2.0
2.5
1.13 1000-5000 100-5000 6 2
G-50 10
4
5
1.07 1500-30000 500-10000 6 2
G-75 13
5
7
1.05 3000-70000 1000-50000 24 3
G-100 17
6
10
1.04 4000-150000 1000-100000 48 5
G-150 24
8
15
1.03 5000-400000 1000-150000 72 5
G-200 30
9
20
1.02 5000-800000 1000-200000 72 5

2-18


-P(Bio-Gel-P)
CH2=CH-CONH2

T 100
C
8 x 25 100 6 2
T 525 T 5
T 25 C 125

pH211
2-19
2-19 bio-gel P

(ml/g) (g/g)
(h)
bio-gel P-2
3.8
1.5
1702600
24
bio-gel P-4
5.8
2.4
6003500
24
bio-gel P-6
8.8
3.7
10005000
24
bio-gel P-10
12.4
4.5
250040000
24
bio-gel P-30
14.8
5.7
300050000
1012
bio-gel P-60
19.0
7.2
500065000
1012
bio-gel P-100
19.0
7.5
5000100000
24
bio-gel P-150
24.0
9.2
5000200000
24
bio-gel P-200
34.0
14.7
40000250000
48
bio-gel P-300
40.0
18.0
50000600000
48
C C
H2 H

C C
H2 H

C C
H2 H

C C
H2 H

CO

CO

CO

CO

NH2

NH

NH2

NH

CH2
C NH
H2
CO
C C
H2 H

CH2

NH

NH2

NH

CO

CO

CO

C C
H2 H

C C
H2 H

C C
H2 H

-D 3,6- -L-
agarose
(agaropectin)

 Sepharose
;Sagavac;Bio-Gel A;Gelarose;Super Ago-Gel 220

2-20

x 103 stokes
Sepharose
Sephadex
cm x 10-8
4B
6B G-200

13.7
19.2
0.86
0.78
0.75
45
27.3
0.72
0.62
0.53

71
36.1
0.68
0.53
0.40

186
0.60
0.42
0.27

670
82.5
0.45
0.27
0.00

-
1000
0.38
0.22
0.00
LH-20
LH-20Sephadex LH-20(Sephadex )
Sephadex G-25 Sephadex LH-20

2-21 LH-20
pH 2
2-21 LH-20

ml/g
ml/g

2.2
4.04.5
2.1
4.04.5

1.9
4.04.5

1.8
3.54.5

1.8
3.54.5
1
1.6
3.03.5

1.6
3.03.5

1.4
3.03.5

1.4
3.03.5

0.8

0.4

0.2

LH-20 1002000
10020000

pH

 2-22
2-22

meq/g(g/g) pH
CM-sephadex G-25
CH2COO
4.50.5
0.4
6-10
CM-sephadex G-50
CH2COO
9
+
DEAE-sephadex A-25 (CH2)NH (C2H5)2 3.50.5 0.5
DEAE-sephadex A-50 (CH2)NH+(C2H5)2
5
QAE-sephadex A-25
3.00.4
0.3
10-2
QAE-sephadex A-50
6
SE-sephadex C-25
(CH2)2SO3
2.30.3
0.2
2-10
SE-sephadex C-50
(CH2)2SO3
3
SP-sephadex C-25
(CH2)2SO3
2.30.3
0.2
2-10

SP-sephadex C-50
(CH2)2SO3
7

CM-sepharose CL-6B
CH2COO
132
10.0
3-10
+
DEAE-sepharose CL-6B (CH2)NH (C2H5)2 122
10.0

9-2

3-10

:(CH2)2N (C2H5)2CH2CHOHCH3

Vt
Vo,ViVg,

Kd,

Ve
Ve = Vo + Kd Vi
Kd = (Ve Vo)Vi
Kd 0 Ve = Vo
Kd 1 Ve = Vo + Vi
0<Kd<1
Kd

C
d
1.5d c

Sephadex G-10G-15
Sephadex G-25
100 150

2 3

100

5ml10ml/min
1cm2cm

blue dextran 2000 C(cytochrome C)

2000 0.2% 0.02moi/L
0.5ml-1ml/cm2 0.02moi/L

Vo

Sephadex G-200 0.3mg0.5mg

0.02 Sephadex G-75
0.2mg 0.01
0.25

3
1mm2mm 1

pH

G-25
1:1:1

280nm260nm 230nm

0.8%
0.5mol/L


50C 2% 0.5mol/L

709095
60C80C

0.02%,
(0.01%0.02%)(0.005%-0.01%)(0.001%-0.01%)

[1]
198373163, 191-201, 215-222
[2] 200293
108
[3] 199713365
[4] 1997338
[5] 1995112145
[6] 199392174
[7] 200016116
[8] 1994112-129