B MN DC LIU

I HC Y DC TP. H CH MINH

DC LiU CHA ANTHRANOID

MC TIU HC TP
TRNH BY C
1. c im cu trc ca 3 nhm anthranoid
(nhm phm nhum, nhun ty v dimer).
2. Cc tnh cht cn bn ca anthranoid
3. Nguyn tc chit xut anthranoid t dc liu
4. Cc phng php nh tnh, nh lng
anthranoid trong dc liu.
5. Tc dng sinh hc Cng dng ca anthranoid

6. 06 dc liu cha anthranoid chnh

(Cassia, i hong, L hi )
2

Ni dung
1- nh ngha
2- v tr phn loi
3- Phn nhm anthanoid (3 nhm)
4- cc loi glycosid
5- Tnh cht ca anthranoid v cc pp nh tnh
6- Chit xut
7-Phn lp
8-nh lng
9-Tc dng sinh hc
3

1. NH NGHA
Anthranoid l nhng glycosid c
phn aglycon l ca 9,10 diceton-anthracen.
(= anthracenoid)

10

Trong cau truc co 2 nhom chc ceton (thng la para).

Cac nhom chc ceton nay co the dang oxy hoa hay
dang kh
O

OH

OH

OH

dang oxy hoa

dang kh

THUT NG
ANTHRANOID

anthraglycosid (AG)

dng O-/C-glycosid

anthraquinon (AQ)

dng aglycon

anthraquinon

dng aglycon

AQ oxy ha

anthron, anthranol
dihydroanthranol

AQ kh

nhm 1,8 di-OH AQ : C3, C6 thng c nhm th

CH3, CH3O, CH2OH, CHO, COOH, OH
gi chung l Oxy Methyl Anthraquinon (OMA)
6

1 dng oxy-ha
anthraquinon

3 dng kh
anthron

anthranol

dihydroanthranol

AQ
ANTHRANOID

+ ose
AG

anthraquinon

anthron

anthranol

dihydroanthranol

glycosid

glycosid

glycosid

glycosid
7

2. V TR PHN LOI
C6

Benzoquinon

C6 C1

Phenolic acid

(C6 C1)2

Anthraquinon, Stilben

C6 C1 C6

Xanthon

C6 C2

Phenyl acetic acid, acetophenol

C6 C4

Naphthoquinon
O

Xanthon

Anthraquinon

Stilben

3. PHN LOI ANTHRAQUINON

7

6
5

OH

OH

10

OH
1

OH
2

nhm nhun ty

nhm phm nhum

9

3. PHN LOI ANTHRAQUINON

3.1. nhm phm nhum
8

OR

OR
1

3.2. nhm nhun ty

OR

OR
1

sinh tng hp :

sinh tng hp :

con ng acid shikimic

con ng polyacetat

OH
OH

OH

alizarin

istizin

OH

10

3.1. NHM PHM NHUM

8

OR
1

OR

2 nhm OH (hoc OR) k cn (C1 v C2)

nhm 1,2 dihydroxy anthraquinon (nhm 1,2)
mu vng ti, vng cam, cam n ta.

thng gp / h Rubiaceae; trong cn trng.

11

3.1. NHM PHM NHUM

O

OH

O
OH

O glc xyl

purpurin*

alizarin*
O

OH

OH

OH

HOOC

OH

acid ruberythric
OH

OH
OH

Me

glucose
HO

OH

boletol

OH

OH
OH

COOH

acid carminic
12

NHM NHUM MU TNG HP

OH

NH

OH

NH

green dyes

Ph

Ph

yellow dyes

(R = C2H4OH)

13

3.2. NHM NHUN TY

OR

OR
1

2 nhm OH (hoc OR) C1 v C8 ()

nhm 1,8 dihydroxy anthraquinon (nhm 1,8)
mu vng nht vng cam, cam.
gp / h Fabaceae, Polygonaceae, Asphodelaceae
C3, C6: R = CH3, OCH3, CH2OH, CHO, COOH (OMA)

14

3.2. NHM NHUN TY

OH

OH

OH

RO

OH

Me
O

Istizin

Chrysophanol

CH3

Chrysophanol

Aloe emodin

CH2OH

HO

Emodin

Rhein

COOH

MeO

Physcion

15

3.3. NHM DIMER

OH

OH

Glc

OH

OH

Me

OH

COOH

Me

OH

COOH

OH

Glc

OH

diemodin anthron
OH

OH

sennosid A v B

OH

HO

Me
COOH

OH

OH

rheidin A

OH

dng kh ca (emodin + rhein)

Me

OH

Me

OH

OH

OH

hypericin

- thng : 2 ph.t anthron dianthron,

hay dehydro-dianthron
- vi trng hp l dianthraquinon.
16

4. CC LOI GLYCOSID
OH

OH
3

O
Rha

OH

OH

CH2OH

Me

glucose

C-glycosid

O-glycosid
O

OH
O

Glc Xyl

OH

OH
3

CH2O

O-glycosid

Glc

Rha

glycosid hn tp
17

NG (OSES)
Phn ose trong AG th n gin.
- Loi ose hay gp : Glc, Rha, Xyl
- t khi gp 2 mch ng

(gn vo 2 ni)

- t khi gp 1 mch c 3 ng (gn vo 1 ni)

- Thng : 1 mch (gm 1 hoc 2 ng)

AQ glc glc.

AQ glc xyl.
18

TNH CHT V CC PHNG PHP NH TNH

5.1.
5.2.
5.3.
5.4.
5.5.
5.6.
5.7.
5.8.
5.9.

L tnh chung
Tnh acid
Phn ng Borntreger
vi thng hoa
Phn ng vi Mg acetat / MeOH,
Phn ng vi Pyridin / MeOH (1 : 1)
SKLM (bn silica gel)
Quang ph IR
Ph UV ca anthraquinon
19

5. TNH CHT NH TNH (OMA)

5.1. L tnh chung (ch yu ca cc OMA)
c 2 dng : - d tan trong dd. kim, ROH
- tan c / nc nng, bn nhit
- km tan / dd. NaHSO3 ( naphthoquinon)
aglycon :

- d tan / dung mi hu c km phn cc

- kh tan / acid.
- thng hoa c

glycosid : - kh tan / dmhc km phn cc.

- khng thng hoa c.

20

5. TNH CHT NH TNH (OMA)

5.2. Tnh acid

-OH (tnh acid yu hn)

OH

HO

HO

OH
OH

-OH
(tnh acid
mnh hn)

ch c OH

: ch tan / kim mnh

c OH

: tan c / kim yu

c COOH

: tan c / kim rt yu
(bi)carbonat, ammoniac (Long)
21

i hong

Dng
kim khc
nhau
phn lp
cc cht

AG ton phn
H2SO4 25%,
dch thy phn/dm
hu c

tnh acid

NaHCO3 5%

Rhein

Na2CO3 5%

Emodin

NaOH 5%
tp phn cc

chrysophanol

(+++)
(++)

(+)

22

XC NH ACID CHRYSOPHANIC
dch AQ / benzen
NH4OH( L) x n ln

b dch NH4OH (AQ acid mnh)

dch benzen (vng)

(AQ acid yu)

lc vi NaOH 5%

lp kim : mu

OH

O
1

S b KL: c acid chrysophanic

(tnh acid yu, khng tan / NH4OH)

OH

Me

23

5. TNH CHT NH TNH (OMA)

5.3. Phn ng Borntreger ***

mu th

NaOH hay KOH

mu (1,8 di-OH)

long

xanh tm (1,2 di-OH)

(naphthoquinon cng cho mu )

- trong ng nghim, bnh lng
Thc hin

- trn bn mng, lame

- trn m thc vt. . .

24

Trong
ng
nghim

Bt DL + acid + thy phn

ngui,

lc vi benzen

lp benzen
lc vi kim long

lp kim

lp benzen

lp kim , lp benzen mt mu

AQ

lp kim vng, hunh quang lc

mu

dn cht kh

+H2O2

25

5. TNH CHT NH TNH (OMA)

5.4. Th nghim vi thng hoa
c th lm trc tip vi bt dc liu cha AQ t do

tinh th AQ
bt dc liu
R
26

THC HiN PHN NG BORNTRAGER

trn lam knh (sau khi vi thng hoa)

bng kim

AQ

AQ

bng kim chuyn sang mu : 1,8 di-OH AQ

27

5. TNH CHT NH TNH (OMA)

5.5. Phn ng vi Mg acetat / EtOH, MeOH

AQ (c OH)

Mg acetat

ROH

cam (1,6 v 1,8 di-OH)

tm

(1,2 di-OH)

ta

(1,4 di-OH)
(u bathochromic !)

Mg
R

- c ch

: to chelat

- ng dng : nh lng / UV-vis

Mg

28

5. TNH CHT NH TNH (OMA)

5.6. Phn ng vi Pyridin / MeOH (1 : 1)

oxy ha

vng cam

kh

tm violet

orange

phn bit dng oxy ha // dng kh.

(lm thuc th hin mu / SKLM).

29

5.7. SKLM (bn silica gel)

kho st ton b cc anthranoid trong dc liu:

- chit bng MeOH, EtOH (hay ROH + H2O)

- ly dch chit ROH chm ln bn mng

kho st cc aglycon mi sinh + c sn:

- chit (+ th.phn + oxy ha) vi H2SO4 25% + H2O2
- lc vi CHCl3, ly dch CHCl3 chm ln bn mng
kho st cc aglycon t do (c sn):

- chit bng MeOH, EtOH (hay ROH + H2O), c

- lc vi CHCl3, ly dch CHCl3 chm ln bn mng

30

DUNG MI SKLM ANTHRANOID

i vi anthraglycosid :

i vi anthraquinon :

EtOAc MeOH H2O

Bz CHCl3 (1:1)

EtOAc PrOH H2O

Bz EtOAc

CHCl3 MeOH

Bz EtOAc AcOH

Bz EtOAc HCOOH
Bz EtOH AcOH

31

HIN MU VT TRN SKLM

a. di nh sng thng
b. di UV 254 / 365 nm
c. sau khi x l vi thuc th

- hi ammoniac
- KOH / ROH

- Pyridin / MeOH
- Mg acetat / ROH
32

SC K ANTHRANOID T CASSIA TORA

33

V tr tng i ca cc OMA trn bn Si-gel

Chrysophanol

1,8

di-OH, 3-Me

Physcion

1,8

di-OH, 3-Me, 6-OMe

Emodin

1,8,6 tri-OH, 3-Me

Aloe emodin

1,8

di-OH, 3-CH2OH

Rhein

1,8

di-OH, 3-COOH

Acid emodic

Rf cao

Rf thp

tnh phn cc :

1,8,6 tri-OH, 3-COOH

COOH > CH2OH > CH3

tri-OH > di-OH
34

5.8.Quang ph IR
Do 2 nhm carbonyl

bng ny mnh, kh c trng

1630 cm1 (chelat)

1670 cm1 (t do)

nhiu trng hp : to doublet.

Do cc nhm -OH

OH ....O....HO

- OH 3400 cm1 (thng yu)

- OH thng rt yu, kh thy

OH ....O ....HO

OH

- CO (carbinol) cho bng r rt vng 1100 cm1

Do vng thm : cho bng kh r vng 3000, 1570 cm1

35

anthraquinon

3400

1670
36

OH

OH

Istizin

3400

1630
37

OH
OH

alizarin

3400
38

OH
OH

OH

Purpurin

3400
39

OH

OH

HO

Me
O

Emodin

3400
40

Aloin

OH

OH

glucose

3400

CH2OH

41

5.9. Ph UV ca anthraquinon
220 - 260

270 - 290

430 - 440

200

300

400

500

600 nm

42

PH UV-Vis CA OMA
hRf Hp cht

cc cc i hp thu (EtOH, nm)

76 Chrysophanol

225

258

279

288

432

75 Physcion

226

255

267

288

440

52 Emodin

223

254

267

290

440

36 Aloe emodin

225

258

279

287

430

24 Rhein

230

260

432

18 acid emodic

227

252

274

290

444

hRf trong h

Bz EA AcOH

(75 : 24 : 01)

43

6. CHIT XUT ANTHRANOID

6.1. C s l lun
6.2. Chit cc dng kh
6.3. Chit dng Oxy-ha
6.4. Chit glycosid (AG)
6.5. Chit aglycon (AQ)
- AQ c sn trong dc liu
- AQ mi sinh t glycosid
44

6.1. C S L LUN
6.1.1- NHC LI TNH CHT (L tnh chung)
c 2 dng : - d tan trong dd. kim, ROH
- tan c / nc nng, bn nhit
- km tan / dd. NaHSO3 ( naphthoquinon)
aglycon :

- d tan / dung mi hu c km phn cc

- kh tan / acid.
- thng hoa c

glycosid : - kh tan / dmhc km phn cc.

- khng thng hoa c.

45

6.1.2 C S L LUN - ng dng thc tin

Dc liu mi thu
hi
Dc liu x l
(Thu hi , bo qun)
i tng ch yu

cha nhiu dng kh

kch ng tiu ha
cha nhiu dng oxy ha

glycosid

aglycon

xung rut gi
AQ

b rut non
hp thu

xut hin
tc dng

khng cn
tc dng
46

6.2. CHIT XUT DNG KH

Mang ngha khoa hc (sinh nguyn, sinh )
t mang tnh ng dng thc t
K thut : kh (v sn phm km bn)
C th chit xut, theo iu kin:
- trnh nh sng (nng !)
- thc hin trong kh quyn N2, CO2 lng
- trnh dng dung mi nhit cao
- trnh mu tip xc lu vi silica gel
47

6.3. CHIT XUT DNG OXY HA

C ngha thc t hn

Dng Oxy ha d chit hn dng kh.

Dng AQ

d chit hn dng AG.

nhng tc dng nhun ty ca AQ << AG.

Ty mc ch nghin cu m chn mc tiu c th
(AG hay AQ; dng Oxy ha hay dng kh)

48

6.4. CHIT XUT GLYCOSID

- Chit bng 1 dung mi phn cc (ROH + H2O)
- Loi cc tp cht lin quan (c bit l Tannin)
- C thu hi dung mi
- Thu c AG th ton phn + t tp
SKC
AG tinh khit

49

6.4. CHIT XUT GLYCOSID

bt dc liu
(cn + H2O)

c thu hi cn

cao nc

aceton
AG ton phn th kt ta
SKC
AG tinh khit

50

6.4. CHIT XUT GLYCOSID

Dl c cha sc t
+cht bo

ether
petrol

dch EP

benzen

dch Bz

methanol

dch MeOH

Cc sc t , cht bo +
AQ km ph.cc

cc AG phn cc

b dc liu
51

6.5. CHIT XUT AGLYCON

a. Dng aglycon c sn / dc liu

-chit anthranoid vi cn,(hoc dung mi km phn cc)

- c thu hi cn.
- thm nc, ta AQ th
- lng, lc, thu ta AQ th
(Nu dng dm km phn cc th c n cn- Tinh ch qua ct)
- tinh ch (SKC silica gel) loi tp phn cc
hn hp cc AQ kh sch SKC cc AQ pure.

52

6.5. CHIT XUT AGLYCON

b. Dng AQ mi sinh t AG

- Chit AG bng 1 dung mi phn cc

- Loi cc tp lin quan (c bit l tannin)

- Thy phn bng acid (sulfuric 20%, acetic) nng

- Tch ring phn AQ bng dm hc km phn cc

- Tip tc tinh ch (acid base; ct silica gel )

- C dung mi, thu sn phm (AQ ton phn).
- SKC cc AQ tinh khit
53

7. PHN LP OMA
Dng aglycon

: km phn cc hn dng glycosid.

d phn lp hn dng glycosid

Dng monomer : d phn lp hn dng dimer

7.1. Phng php sc k ct

Ct hp ph (Silica gel, Nhm oxyt)
Dung mi khai trin :
- dng AQ : dng h dung mi km phn cc
- dng AG : dng h dung mi phn cc hn
54

i hong

AG ton phn

7.2. Thay i pH

H2SO4 25%,
dch thy phn

tnh acid

NaHCO3 5%

Rhein

Na2CO3 5%

Emodin

NaOH 5%
tp phn cc

chrysophanol

(+++)
(++)

(+)

55

8. NH LNG OMA
8.1. Phng php cn

(Deals Kroeber)

8.2. Phng php so mu (Auterhoff)

8.3. Phng php th tch (Tschirch - Schmitz)
8.4. Phng php to phc mu, o quang
8.5. Phng php SKLM
8.6. Phng php HPLC

8.7. Phng php sinh vt

56

8. NH LNG OMA
8.1. Phng php cn (Deals Kroeber)
Nguyn tc
- Thy phn AG bng H2SO4 25% nng
- Tch ring AQ (c sn + mi sinh) bng CHCl3
- Loi tp (naphthoquinon . . .) vi NaHSO3 tha

- Loi NaHSO3 tha bng HCl long

- Bc hi dung mi CHCl3 n cn, sy 60OC 80OC

- Cn, tnh hm lng %

57

8. NH LNG OMA
8.2. Phng php so mu (Auterhoff)
Nguyn tc

- Va thy phn (vi AcOH bng)

va chit AQ

(vi Et2O) trc tip t bt dc liu

- Dch Et2O c lc vi kim (NaOH + NH4OH)

- Lp kim () cha AQ c o Abs ( = 540 nm)
- So snh vi ng cong chun :
* istizin hay chrysophanol / (NaOH + NH4OH)
* CoCl2. 6H2O / H2O
- Suy ra hm lng AQ / mu
58

8. NH LNG OMA
Ch trong phng php so mu (Auterhoff)
Cc dn cht kh vng / mi trng kim
(cn tr s o mu) cn Oxy ha (BM 20),
ngui ri mi o quang.
Hiu kt qu trc v sau khi BM [dn cht kh ].
mun xc nh hm lng aglycon c sn / dc liu
chit ngay bng dung mi hu c km phn cc
(khng cn thy phn bng AcOH bng)
59

8. NH LNG OMA
8.3. Phng php th tch (Tschirch - Schmitz)
AQ
KOH 0,1 N tha
HCl 0,1 N
Ch
- bn thn AQ l ch th mu ( / kim; vng / acid)
- sai s tha do polyphenol (ch yu do Flavonoid)
60

8. NH LNG OMA
8.4. Phng php to phc mu vi Mg acetat 0,5% /
MeOH, o quang
- chit AG bng nc nng + NaHCO3.
- oxy ha + thy phn (= FeCl3 + HCl) nng AQ
- chit AQ t do bng Et2O.
- cn Et2O + Mg acetat 0,5% / MeOH mu
- o quang 515 nm (mu trng = MeOH)
- [anthranoid] / mu
(thng p dng / i hong, kt qu quy v Rhein)
61

8. NH LNG OMA
8.5. Phng php SKLM

Mc ch

: Bn nh lng // chun

Nguyn tc :
trn cng 1 bn mng,
chm ng th tch chun (S) // th (T)
So snh (S) v (T) v
- din tch vt,
- cng mu (c / khng th th)

Suy ra [C] c trong mu th.

S T
62

8. NH LNG OMA
8.6. Phng php HPLC
Tin cy, thng dng.
Nguyn tc : Trong cng 1 iu kin,
chun (X) v (X/mu) peak c cng thi gian lu.
[X] s t l thun vi S (din tch peak).
bit nng chun, t S o c (X) v (X/mu)
[X]% trong M
63

8. NH LNG OMA
Phng php HPLC
XT
mu th M (cha X)
ST

ST

SC

XT
=
XC

tR
pht

SC

chun X
XC
64

8. NH LNG OMA
8.7. Phng php sinh vt
- Da trn tc ng lm tng nhu ng rut
lm tng tc bi xut phn.
- Thc hin trn chut / nhuyn th.

- Lm song song vi l chng, l trng

Lu :

S p ng ca chut c th rt khc nhau

(Sennosid / Phan t dip >> Aloin / L hi)

Thc t t s dng.

65

9. TC DNG SINH HC CA OMA

9.1. Ca nhm 1,2 di-OH AQ
Ch yu : cht nhum mu. Cc cht thng dng :

alizarin *,

purpurin *,

acid carminic,

carmin

acid kermesic,

acid laccaic A, B, C, D.

acid ruberythric
mt s : Khng khi u (antitumor)
Damnacanthal, nor-damnacanthal trong Morinda.
66

9. TC DNG SINH HC CA OMA

9.2. ca nhm 1,8 di-OH AQ
dng AG (ch yu l cc -glucosid)
- khng b chuyn ha ti rut non.
- ti rut gi : b thy phn AQ (dng oxy-ha)
- tip tc b kh anthron, anthranol (c hot tnh)
- lm tng nhu ng c trn nhun ty

dng AQ : b hp thu rut non khng nhun ty.

67

TC DNG DC LC CA OMA

AQ gy
kch ng !
AQ b
hp thu !

mt tnh
nhun ty

AG AQ
dng kh

NHUN TY

9. TC DNG CA OMA
Tng nhu ng rut v co bp c trn ( bng quang, t
cung . . .)

tc dng chm (ung : sau 6 10 h) : nhun x ty

ph tr iu tr si thn (liu trung bnh !)
bi tit qua phn, nc tiu, sa, m hi (nhum mu)
trnh dng lu di (l thuc thuc, gim Kali-huyt)
khng dng cho ph n c thai, cho con b . . .
khng dng liu cao i vi ngi c si (gan mt, niu)
thn trng : ngi gi, tr nh, bnh tr

mt s c tc dng khng nm da (hc lo), thng mt

mt s c tc dng iu ha / kch thch min dch (aK)

69

PHN B TRONG T NHIN

a. Trong cn trng, thc vt bc thp : t
b. Trong thc vt bc cao : chim i a s

b1. Lp 1 l mm : Asphodelaceae (chi Aloe)

b2. Lp 2 l mm : kh hp, ch yu :
- Rubiaceae

(chi Rubia, Morinda)

- Fabaceae

(chi Cassia)

- Polygonaceae

(chi Rheum, Rumex)

- Rhamnaceae

(chi Rhamnus) . . .
70

CC DC LiU NG CH
H Fabaceae

H Polygonaceae

1. Phan t dip

5. i hong

2. Tho quyt minh

6. H th

3. Mung tru
H Liliaceae

H Rubiaceae

4. L hi

7. Ba kch
8. Nhu
71