BO MON DUOC LIEU • •

DAI HOC Y DUOC - TP. HO CH MINH

•••

DUftC LiEU CHUA ANTHRANOID

■■

PHAN 1:0?i cuUng vl anthranold

1
 MUC TIEU HOC TAP ■ ■ ■

TRINH BAY DUttC ■

1. Dac diem cau true cua 3 nhom anthranoid

■

(nhom pham nhuom, nhuan tay va dimer).

2. Cac tinh chat can ban cua anthranoid

3. Nguyen tac chiet xuat anthranoid tU dUdc lieu

4. Cac phudng phap djnh tinh, djnh

lUdng anthranoid trong dUdc lieu.
5. Tac dung sinh hoc - Cong dung cua anthranoid

6. 06 dUdc lieu chute anthranoid chinh

■■

(Cassia, Dai hoang, Lo hoi ...)

2
 Noi dung
1- Djnh nghia
2- vj tri phan loai
3- Phan nhom anthanoid (3 nhom)
4- cac loai glycosid
5- Tinh chat cua anthranoid va cac pp djnh
tinh
6- Chiet xuat
7- Phan lap
8- Djnh lu'dng
9- Tac dung sinh hoc
3
 1. OINH NGHIA
■

Anthranoid la nhttng glycosid co phan aglycon la A' cua

9,10 diceton-anthracen.
(= anthracenoid)

4
Trong cau true co 2 nhom chtfc ceton (thUdng la para).
Cac nhOm chtic ceton nay cO the dang oxy hOa hay
dang Khfr

dang oxy
hoa
5
 ✓V /V .

THUAT NGtf
*
anthraglycosid (AG)_>
ANTHRANOID
dang O-/C-glycosid
anthraquinon (AQ) —>
dang aglycon

anthraquinon
anthron, anthranol AQ oxy hoa
dang aglycon —
>
dihydroanthranol AQ khU
v

nhom 1,8 di-OH AQ : 6 C3, C6 thudng co nhom the

CH3, CH3O, CH2OH, CHO, COOH, OH ^ goi chung la
Oxy Methyl Anthraquinon (OMA) 6
1 dang oxy-hoa 3 dang khfr
_-A._
r /— —s

anthraquinon anthror anthranol dihydroanthranol

r AQ

ANTHRANOID + ose

AG

anthraquinon anthron anthranol dihydroanthranol

glycosid glycosid glycosid glycosid

7
 2. VI TRi PHAN LOAI

C6 Benzoquinon

C6 - C1 Phenolic acid

(C6 - Cl)2 Anthraquinon, Stilben

C6 - C1 - C6 Xanthon

C6 - C2 Phenyl acetic acid, acetophenol

n
1 Naphthoquinon
n
-i^

Xanthon Anthraquinon 8
 3. PHAN LOAI ANTHRAQUINON
O

nhom nhuan tay nhom pham nhuom

9
 3. PHAN LOAI ANTHRAQUINON

3.1. nhom pham nhuom 3.2. nhom nhuan tay

OR O OR

O OR

OY 2,OR
O O

sinh tong hop : sinh tong hop :

con duftng acid shikimic con dudng polyacetat

O OH
OH O OH
1 ^ OH

O O

alizarin istizin 10
 3.1. NHOM PHAM NHUOM

O OR

OR

■* 2 nhom -OH (hoac -OR) ke can (Cj va C2)

■-* ^ nhom 1,2 dihydroxy anthraquinon (nhom 1,2)

■-* mau vang tutti, vang cam, do cam den tia.

■* thuftng gap / ho Rubiaceae; trong con trung.

11
 3.1. NHOM PHAM NHUOM

O OH
O-glc—xyl

acid ruberythric

OH O Me

boletol
12
 NHOM NHUOM MAU TONG HOP

OH O NH—R
Ph—S O S— Ph

green dyes (R yellow dyes

= C2H4OH)

13
 3.2. NHOM NHUAN TAY

x 2 nhom -OH (hoac -OR) d va C8 (a)

^ nhom 1,8 dihydroxy anthraquinon (nhom 1,8) x mau

vang nhat ^ vang cam, do cam. x gap / ho Fabaceae,

Polygonaceae, Asphodelaceae
* d C3, C6: R = CH3, OCH3, CH2OH, CHO, COOH (OMA)
 OH 0 OH

III
V^A
0
1

Istizin H

Chrysophanol CH3

Aloe emodin CH2OH

Rhein COOH
 3.2. NHOM NHUAN TAY

OH O OH

H -> < Chrysophanol

HO —) ► Emodin

MeO —i ► Physcion

15
 3.3. NHOM DIMER

OH O OH

diemodin anthron sennosid A va B rheidin A

OH O OH
i
dang khu cua (emodin + rhein)
Me OH
Me OH

- thuftng : 2 ph.tU anthron ^ dianthron,

OH O OH hay ^ dehydro-dianthron
hypericin
- vai truftng hdp la dianthraquinon.
16
OH O OH

O-glycosid

O-glycosid
4. CAC LOAI GLYCOSID

17
 DU'O’NG (OSES)

Phan ose trong AG thi ddn gian.

- Loai ose hay gap : Glc, Rha, Xyl

- it khi gap 2 mach dudng (gan vac

- ft khi gap 1 mach co 3 dudng (gan ) 2 ndi) vao

- Thudng : 1 mach (gom 1 hoac 2 du 1 ndi) dng)

i f

AQ - glc - glc.
AQ - glc - xyl.
18
 / A' "\ r . / /

TINH CHAT VA CAC PHtiQNG PHAP DINH TINH

5.1. Ly tinh chung

5.2. Tinh acid
5.3. Phan ting Borntraeger
5.4. vi thang hoa
5.5. Phan ting vtii Mg acetat / MeOH,
5.6. Phan ting vtii Pyridin / MeOH (1 :
1)
5.7. SKLM (ban silica gel)
5.8. Quang pho IR
19
 5. Tm CHAT - OINH TCNH (OMA)
5.1. Ly tinh chung (chu yeu cua cac OMA)

ca 2 dang : - de tan trong dd. kiem, ROH

- tan dudc / nudc nong, ben nhiet
- kem tan / dd. NaHSO3 (* naphthoquinon)

aglycon : - de tan / dung moi hut cd kem phan cut

- kho tan / acid.
- thang hoa dudc

glycosid : - kho tan / dmhcd kem phan cut.

- khong thang hoa dudc.
20
 5. TiNH CHAT - OINH TCNH (OMA)

5.2. Tinh acid

-OH a (tinh acid yeu hdn)

chi co -OH a : chi tan / kiem manh

co -OH P : tan dutic / kiem yeu

co -COOH : tan dUtic / kiem rat yeu
(bi)carbonat/ ammoniac (Loang)
21
 I AG toan phan
Dai hoang

Dung do H2SO4 25%, A

kiem khac
nhau de djch thuy phan/dm
phan lap huU cd tinh acid
cac chat NaHCO3 5%
(+++)

Na2CO3 5% (++)

► chrysophanol (+)
NaOH 5%

cut 22
 XAC OINH ACID CHRYSOPHANIC

djch AQ / benzen

NH4OH( L) x n lan bo djch NH4OH (AQ acid manh)

v

djch benzen (vang) (AQ acid yeu)

lac vOi NaOH 5%

>t
Ai" ••A

lOp kiem : mau do

Sd bo KL: co acid chrysophanic

(tinh acid yeu, khong tan / NH4OH)

23
 5. Tm CHAT - OINH TCNH (OMA)
5.3. Phan ting Borntraeger ***

NaOH hay KOH

mau thiY < mau do (1,8 di-OH)
1 1 tl I^J xanh tim (1,2 di-OH)
loang
(naphthoquinon cung cho mau do)

f
trong ong nghiem, binh lang
Thut hien tren ban mong, lame
V tren mo thut vat. . .
■■

24
x ldp kiem do, ldp benzen mat mau AQ

x ldp kiem ^ vang, huynh quang luc mau do

+H2O2

dan chat khd 25

 5. TCNH CHAT - OINH TCNH (OMA)

5.4. Thu’ nghiem vi thang hoa

co the lam true tiep vdi bot dUdc lieu chtfa AQ tu do

tinh the AQ

bot dUOc lieu

-mmm-
R
26
THUC
■ HiEN PHAN UNG BORNTRAGER ■
tren lam kinh (sau khi vi thang hoa)

27
 5. Tm CHAT - OINH TCNH (OMA)
5.5. Phan dng vdi Mg acetat / EtOH, MeOH

r
do cam (1,6 va 1,8 di-OH)
AQ (co OHa) Mg acetat
< tim (1,2 di-OH) do tia (1,4
ROH di-OH)

(deu bathochromic !)
Mg
/\
2
-
■
cd che : tao chelat

P - dng dung : djnh luOng / UV-

vis
\
Mg

28
 5. TiNH CHAT - OINH TCNH (OMA)

5.6. Phan u*ng vdi Pyridin / MeOH (1 : 1)

A' oxy hoa ^ vang cam
A' khU ^ tim violet

^ phan biet dang oxy hoa // dang khU.

(lam thuoc thU hien mau / SKLM).

29
5.7. SKLM (ban silica gel)

De khao sat toan bo cac anthranoid trong duO: lieu:

■ a a

- chiet bang MeOH, EtOH (hay ROH + H2O)

- lay djch chiet ROH cham len ban mong

De khao sat cac aglycon mdi sinh + co sin:

- chiet (+ th.phan + oxy hoa) vdi H2SO4 25% + H2O2
- lac vdi CHCl3, lay djch CHCl3 cham len ban mong

De khao sat cac aglycon tu do (co sin):

- chiet bang MeOH, EtOH (hay ROH + H2O), co
- lac vdi CHCl3, lay djch CHCl3 cham len ban mong
30
 DUNG MOI SKLM ANTHRANOID

doi vdi anthraglycosid : doi vdi anthraquinon :

EtOAc - MeOH - HO Bz - CHCI3 (1:1)

EtOAc - PrOH - HO Bz - EtOAc

CHCI3 - MeOH Bz - EtOAc - AcOH

Bz - EtOAc - HCOOH

Bz - EtOH - AcOH

31
HIEN MAU VET TREN SKLM

a. dutii anh sang thudng

b. dutii UV 254 / 365 nm
c. sau khi xU ly vtii thuoc
thU
- hdi ammoniac
-- KOHPyridin
/ ROH /
MeOH
- Mg acetat /
ROH 32
0 D □ Sic KY DO ANTHRANOID TU* CASSIA TORA

33
Vi tri tudng doi cua cac OMA tren ban Si-gel

Chrysophanol Rf cao 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 thap 1,8,6 tri-OH, 3-COOH

tinh phan cut : COOH > CH2OH > CH3 tri-

OH > di-OH
34
 5.8.Quang pho IR

❖ Do 2 nhom carbonyl ► 1630 cm-1 (chelat)

1670 cm-1 (td do)
bang nay manh, kha dac trdng
nhieu trdOng hop : tao doublet.
QH-Q-HO
❖ Do cac nhom -OH
- OH p ^ 3400 cm-1 (thdOng yeu)
OH

- OH a ^ thuftng rat yeu, kho tha'y

- C-O (carbinol) cho bang ro ret d vung 1100 cm i-1

❖ Do vong thdm : cho bang kha rO 6 vUng 3000, 1570 cm -1

35
 1630 1
T-1-r T-1-r
15DG 5 DO
 V

3400
U
T
v
I DUG 3000 £00D
Emodin
5.9. Pho UV cua anthraquinon

42
 PHO UV-Vis CUA OMA

hRf Hdp chat cac cut dai hap 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 he Bz - EA - AcOH (75 : 24 : 01) 43

 6. CHIET XUAT ANTHRANOID

6.1. Cd sd ly luan

6.2. Chiet cac dang khd

6.3. Chiet dang Oxy-hoa

6.4. Chiet glycosid (AG)

6.5. Chiet aglycon (AQ)

- AQ co sin trong dudc lieu

- AQ mdi sinh tfr glycosid

44
 6.1. CO SO LY LUAN ■

6.1.1- NHAC LAI TCNH CHAT (Ly rtnh chung)

ca 2 dang : - de tan trong dd. kiem, ROH

- tan dUdc / nUdc nong, ben nhiet
- kem tan / dd. NaHSO3 (* naphthoquinon)

aglycon : - de tan / dung moi hud cd kem phan cut

- kho tan / acid.
- thang hoa dudc

glycosid : - kho tan / dmhcd kem phan cut.

- khong thang hoa dUdc.
45
 6.1.2 CQ SQ LY LUAN - Ung dung thut tien

DUdc lieu mdi thu chUa nhieu dang khfr

hai
kich Ung tieu hoa
DUdc lieu da xfr ly » chUa nhieu dang oxy hoa
(Thu hai , bao quan) i
L l
doi tudng chu yeu <
glycosid aglycon
~r~
i
bj ruOt
/V' A, . \

xuong ruot gia

^ AQ non hap
thu
i i
khong con
/V', ■ . /V

xuat hien ■
tac dung
tac dung 46
 A' A' ?,

6.2. CHIET XUAT DANG KH^

Mang y nghTa khoa hoc (sinh nguyen, sinh E)

It mang tinh ting dung thut te Ky thuat : kho

(vi san pham kem ben)

Co the chiet xuat, theo dieu kien:

- tranh anh sang (nang !)

- thut hien trong khi quyen N2, CO2 long

- tranh dung dung moi d nhiet do cao

■ Ag /V /V ■ ■ A I /V jT.m ■ I ■ I

- tranh de mau tiep xuc lau vdi silica gel

47
 6.3. CHIET XUAT DANG OXY HOA

Co y nghia thut te hdn

Dang Oxy hoa de chiet hdn dang khu\

Dang AQ de chiet hdn dang AG.

nhtfng tac dung nhuan tay cua AQ << AG.

Tuy muc dfch nghien cuU ma chon muc tieu cu the

(AG hay AQ; dang Oxy hoa hay dang khfr)

48
 6.4. CHIET XUAT GLYCOSID

- Chiet bang 1 dung moi phan cut (ROH + H2O)

- Loai cac tap chat lien quan (dac biet la Tannin)

- Co thu hoi dung moi

- Thu dUdc AG tho toan phan + ft tap

SKC
▼
AG tinh khiet

49
 6.4. CHIET XUAT GLYCOSID

bot ■
dudc
■ ■ lieu

(con + H2O)
/V . I I '/V. '/V

co thu hoi con

cao nudc

aceton
f

AG toan phan tho ket tua

SKC
V
AG tinh khiet
50
 Dl co chfra sac to
+chat beo

ether dich
■ EP
►
petrol
dich
■ Bz
►
benzen
¥ dich
■ MeOH

methanol

ba dutfc
■ ■lieu
6.4. CHIEF XUAT GLYCOSID

Zac sac to , chat beo

+ AQ kern ph.ci/c

cac AG phan cut

51
 6.5. CHIET XUAT AGLYCON

a. Dang aglycon co sin / dudc lieu -chiet anthranoid vdi

con,(hoac dung moi kem phan cut)

/V . I I '/V . '/V

- co thu hoi con.

- them nudc, ^ tua AQ tho

-de lang, loc, thu tua AQ tho

(Neu dung dm kem phan cut thi co den can- Tinh che qua cot)

- tinh che (SKC silica gel) de loai tap phan cut ^ hon
52
hop cac AQ kha sach ^ SKC ^ cac AQ pure.
 6.5. CHIET XUAT AGLYCON

b. Dang AQ mdi sinh tfr AG

- Chiet AG bang 1 dung moi phan cut

- Loai cac tap lien quan (dac biet la tannin)

- Thuy phan bang acid (sulfuric 20%, acetic) nong

- Tach rieng phan AQ bang dm hcd kem phan cut

- Tiep tuc tinh che (acid - base; cot silica gel ...)

- Co dung moi, thu san pham (AQ toan phan).

- SKC ^ cac AQ tinh khiet

53
 7. PHAN LAP OMA
■

Dang aglycon : kem phan cut hOn dang glycosid.

^ de phan lap hdn dang glycosid

Dang monomer : de phan lap hdn dang dimer

7.1. PhUOng phap sac ky cot Cot hap phu (Silica gel,

Nhom oxyt)

Dung moi khai then :

- dang AQ : dung he dung moi kem phan cut

- dang AG : dung he dung moi phan cut hdn

54
 7.2. Thay doi pH
I AG toan phan
Dai hoang

H2SO4 25%, A

djch thuy phan tinh

acid

NaHCO3 5% * Rhein (+++)

Na2CO3 5% * Emodin (++)

NaOH 5% ► chrysophanol (+)

cut 55
 8. OINH LUONG OMA

8.1. PhUdng phap can (Deals Kroeber)

8.2. PhUdng phap so mau (Auterhoff)

8.3. PhUdng phap the tich (Tschirch - Schmitz)

8.4. PhUdng phap tao phut mau, do quang

8.5. PhUdng phap SKLM

8.6. PhUdng phap HPLC

8.7. PhUdng phap sinh vat

56
 IS

8.1. PhUdng phap can (Deals Kroeber)

Nguyen tac
- Thuy phan AG bang H2SO4 25% nong

- Tach rieng AQ (co sin + mdi sinh) bang CHCl3

- Loai tap (naphthoquinon . . .) vdi NaHSO3 thua

- Loai NaHSO3 thfra bang HCl loang

- Boc hdi dung moi CHCl3 den can, say 60OC ^ 80OC

- Can, tinh ham lUdng %

57
 8 gngra nigNg|

8.2. PhUdng phap so mau (Auterhoff)

Nguyen tac
- Vu’a thuy phan (vdi AcOH bang)
vda chiet AQ (vdi Et2O) trut tiep ttt bot dUdc lieu
- Djch Et2O dUdc lac vdi kiem (NaOH + NH4OH)
- Ldp kiem (do) chda AQ dUdc do Abs (X = 540 nm)
- So sanh vdi dudng cong chuan :
* istizin hay chrysophanol / (NaOH + NH4OH)
* CoCl2. 6H2O / H2O
- Suy ra ham ludng AQ / mau
58
 IS

Chu y trong phudng phap so mau (Auterhoff)

• Cac dan chat khfr ^ vang / moi trudng kiem

(can trd sd do mau) ^ can Oxy hoa (BM 20'),

de nguoi roi mdi do quang.

• Hieu ket qua trudc va sau khi BM ^ [dan chat khu* ].

• muon xac djnh ham lUOng aglycon co sin / dUOc lieu

^ chiet ngay bang dung moi httu cd kem phan cut

(khong can thuy phan bang AcOH bang)

59
 IB

8.3. Phutfng phap the tich (Tschirch - Schmitz)

! ! AQ

KOH 0,1 N thua

<- HCl 0,1 N

Chu y

- ban than AQ la chi thj mau (do / kiem; vang / acid)

- sai so thua do polyphenol (chu yeu do Flavonoid)

 8 gngra
8.4. Phudng phap tao phut mau vdi Mg acetat 0,5% / MeOH,
do quang

- chiet AG bang nudc nong + laHCO3.

- oxy hoa + thuy phan (= FeCl3 + HCl) nong ^ AQ

- chiet AQ td do bang Et2O.

- can Et2O + Mg acetat 0,5% / MeOH ^ mau

- do quang d 515 nm (mau trang = MeOH)

- ^ [anthranoid] / mau

(thudng ap dung / Dai hoang, ket qua quy ve Rhein)

61
 MS IDjil
8.5. PhUOng phap SKLM

Muc dfch : Ban djnh lUOng // chuan

I
Nguyen tac :
0
tren cung 1 ban mong,
cham dong the tfch chuan (S) // thU (T)
So sanh (S) va (T) ve
- dien tich vet, I

- cudng do mau (co / khong th' thU)

Suy ra [C] co trong mau thU. ST
62
 8 gBNH [LlffiNg

8.6. PhUdng phap HPLC Tin cay, thong dung.

Nguyen tac : Trong cung 1 dieu kien,

chuan (X) va (X/mau) ^ peak co cung thdi gian lUu.

[X] se ty le thuan vdi S (dien tich peak).

biet nong do chuan, tU S do dUdc d (X) va (X/mau)

^ [X]% trong M

63
 X uenip

inijd u _ 3S
X
Xx 1—CO

(X snip) N m n§w ir

DldH dBi|d 6up^ii|d

 8 ragra
8.7. Phu'dng phap sinh vat

- Dda tren tac dong lam tang nhu dong ruot

■ ■ m m

l
m tang toc do bai xuat phan.

- Thdc hien tren chuot / nhuyen the.

- Lam song song vdi lo chting, lo trang

Ldu y :
❖ Sd dap ting ctia chuot co the rat khac nhau
(Sennosid / Phan ta diep >> Aloin / Lo hoi)
65
 9. TAC DUNG SINH HOC CUA OMA
■■

9.1. Cua nhom 1,2 di-OH AQ

Chu yeu : chat nhuom mau. Cac chat thong dung :

alizarin *, purpurin *,

acid carminic, do carmin

acid kermesic, acid laccaic A, B, C, D.

acid ruberythric

mot so : Khang khoi u (antitumor)

Damnacanthal, nor-damnacanthal trong Morinda.

66
 9. TAC DUNG SINH HOC CUA OMA

■■

9.2. cua nhom 1,8 di-OH AQ

* dang AG (chu yeu la cac p-glucosid)

- khong bj chuyen hoa tai ruot non.

- tai ruot gia : bj thuy phan ^ AQ (dang oxy-hoa)

- tiep tuc bj khd ^ anthron, anthranol (co hoat tinh)

- lam tang nhu dong cd trdn ^ nhuan tay

-■* dang AQ : bj hap thu d ruot non ^ khong nhuan tay.
67
 TAC DUNG DU(X LUt CUA OMA

AQ gay
kich u*ng !

AQ bj
AG ^ AQ ^ >
hap thu ! <■
^ dang khfr

mat tinh
nhuan NHUAN TAY
tay
 9. TAC DUNG CUA OMA
■

Tang nhu dong ruot va co bop cO trdn ( bang quang, td ung . . .)

tac dung cham (uong : sau 6 - 10 h) : nhuan xo tay phu trd

dieu trj soi than (lieu trung binh !)

■ \■ • m A'. | /V . A. , ■ /\? ANA 'W | A ■ A i /V \ \

bai tiet qua phan, nUOc tieu, sda, mo hoi (nhuom mau) tranh

dung lau dai (le thuoc thuoc, giam Kali-huyet) khOng dung cho

phu nd co thai, cho con bu . . .

■ I/V ■V | »'A 41A'iA.« ,\.■ A ? ■ A /V | ■ A \

khong dung lieu cao doi vdi ngUdi co soi (gan mat, nieu) than trong :

ngUdi gia, tre nho, benh tri mot so co tac dung khang nam da (hac

lao), thOng mat mot so co tac dung dieu hOa / kich thich mien djch

(aK) 69
 PHAN BO TRONG TU1 NHIEN

a. Trong con trung, thud vat bac thap : it

b. Trong thuc vat bac cao : chiem dai da so

a a a a

bl. Ldp 1 la mam : Asphodelaceae (chi Aloe)

b2. Ldp 2 la mam : kha hep, chu yeu d :

- Rubiaceae (chi Rubia, Morinda)

- Fabaceae (chi Cassia)

- (chi Rheum, Rumex)
Polygonaceae (chi Rhamnus) . . .
- Rhamnaceae 70
Ho Fabaceae

1. Phan ta dieo

2. Thao quyet

minh

3. Muong trau

Ho Liliaceae
■

4. Lo hoi
CAC Dl/GC LiEU DANG CHU Y

Ho Polygonaceae

5. Dai hoanq

6. Ha thu 6

do

Ho Rubiaceae
■

7. Ba kich

8. Nhau
71