Resveratrol A

Khoa Ha, i hc Khoa hc T nhin Thnh ph H Ch Minh
RESVERATROL v
Phng php tng hp
GVHD: PGS.TS. Trn L Quan
SV: Nguyn Tn Pht - 0814150
09-02-2012

Nguyn Tn Pht (0814150)
RESVERATROL v phng php tng hp
2

GVHD: PGS.TS. Trn L Quan i hc Khoa hc T nhin Tp.HCM, 09-02-2012

MC LC
PHN I - M U: .................................................................................................. 3
PHN II - KHI QUT V RESVERATROL ....................................................... 3
1. PHT HIN:............................................................................................................... 3
2. TNH CHT: .............................................................................................................. 3
2.1. L tnh: ...................................................................................................... 4
2.2. Ha tnh: .................................................................................................... 4
2.3. Dc tnh: .................................................................................................. 4
3. TN TI TRONG T NHIN: ................................................................................. 5
4. THNG MI HA: ................................................................................................ 6
5. CC HOT CHT LIN QUAN: ............................................................................. 7
PHN III - TNG HP RESVERATROL: .............................................................. 9
1. CC PHNG PHP TNG HP: ......................................................................... 9
2. QUY TRNH CA JING LIU: ................................................................................. 10
2.1. iu ch 3,5-diacetoxybenzoic acid: ............................................................... 10
2.2. iu ch 3,5-diacetoxybenzoyl chloride:.......................................................... 11
2.3. iu ch resveratrol triacetate: ..................................................................... 12
2.4. Thy phn triacetate thu resveratrol: .............................................................. 13
PHN IV - THC NGHIM: ................................................................................... 14
1. IU CH 3,5-DIACETOXYBENZOIC ACID: .................................................... 14
2.1. Tng hp li: ............................................................................................. 14
2.2. Kho st: .................................................................................................. 17
2. IU CH 3,5-DIACETOXYBENZOYL CHLORIDE: ........................................ 20
PHN V - KT LUN V K HOCH: .............................................................. 24
1. KT LUN: .............................................................................................................. 24
2. K HOCH: ............................................................................................................. 24
PHN VI - PH LC: ............................................................................................... 25
PHN VII - TI LIU THAM KHO: .................................................................... 28

3


PHN I - M U:
Cuc sng vnh hng l ch ni tri trong sut qu trnh tn ti v pht
trin vn minh loi ngi. Nhiu cuc tai bin, chin tranh c i cho n hnh ng
chy ua qun s v khoa hc k thut ca cng quc hin i u khng th
khng l h ly ca vn . Con ngi ang u tranh vi nhau v hoc mt loi
thn dc ci t hon ng, mt ci thing ling, thnh tch hay vt bo h mnh
nh Chn Thnh, Cn Phng H Phch, Ngn La Vnh Cu hay Thnh a Ngc
Nhn (Shangri-La), ci Bng Lai ch tn ti trong tr tng tng(!) nhng tt c
dng nh ngy mt kh khn. Ri, cc nh gi kim n thut s, o s v ti cc
nh khoa hc thi i ngh n vic dng thn vo con ng nghin cu iu
ch ra phm thuc thay v u tranh v ngha trn chin trng. Trong s cc thnh
qu thu c c resveratrol, mt hp cht c tin vo kh nng ko di s sng v
chng li bnh tt.

PHN II - KHI QUT V RESVERATROL
1. PHT HIN:
Do vic pht hin resveratrol c bo co trong cc nghin cu ni b nn
cha r ngi u tin c lp c n. Mt nhm nh khoa hc phng Ty tuyn
b pht hin ln u vo nm 1963 trong dch chit c c t cy ct kh (cn gi
l in tht, h trng tn khoa hc: Fallopia japonica, Polygonum
cuspidatum, Reynoutria japonica) tuy c nhng c dng lm thuc trong dn
gian chu (li l mt loi cy xm hi chu u)... khng lu sau b bc bi nghin
cu v cy khuynh dip bn a c (Eucalyptus wandoo) nm 1959.
[8]
Ri s vic
li lp li bi c cc nghin cu trc .
[8]
Theo ghi nhn hin ti, bi bo ca
M. Takaoka nm 1939 l nghin cu sm nht. Takaoka c lp resveratrol t loi
tho dc Veratrum album.
[6]

Tn gi resveratrol c th c ghp t r re es sorcinol-v ve er ra at tr rum-alcoho ol l. Bi
hp phn ca resveratrol l resorcinol, cht ny c tng hp t Veratrum album
v c mang ba nhm OH nh alcohol.
[8]

2. TNH CHT:
Resveratrol (CTPT: C
14
H
12
O
3
; cn gi l 3,4',5-trihydroxystilbene, 3,4',5-
stilbenetriol hay 5-(p-hydroxystyryl)resorcinol) c xp vo nhm stilbenoid,
nhm phenol v nhm phytoalexin, trong thin nhin. C c tnh chng oxy ha,
khng vim v em li nhiu li ch cho h huyt mch. Hp cht c mt trong
nhiu loi thc vt thng xuyn b nm mc, vi khun quy ph nh nho, nhiu
loi tri mng, u phng Gn y ru vang, nc p, scla v b u phng
c cp l cc thc phm ch bin cha di do lng hot cht ny.

Hnh 1: trans-resveratrol

Hnh 2: cis-resveratrol

2.1.L tnh:
Resveratrol tn ti dng bt trng c ln vng ng: nhit nng chy 259-
261
o
C,
[9]
khi lng phn t 228.24 g/mol. Khng tan trong nc (0.03 g/L), tan
c trong DMSO (16 g/L) v ethanol (50 g/L).
[6]

2.2.Ha tnh:
Resveratrol th hin y tnh cht ca mt phenol, v ca mt alkene.

Resveratrol c hai ng phn: trans-resveratrol v cis-resveratrol. Cu trng
cis thu c t chuyn ha dng trans trong iu kin chiu x UV. Resveratrol bn
vng trong mi trng t nhin, minh chng bi hm lng ca chng sau khi ln
men ru.
[6]

2.3.Dc tnh:
Tc ng ca resveratrol ln c th sinh vt ang gy nhiu tranh ci trong
gii khoa hc. Nhiu bo co cho thy resveratrol mang n mt li l ko di tui
th, ngn nga ung th, u bu v khng vim song, li c bo co phn bc
nhng cng b va nu.

Ko di tui th
-Nm 2003 v khng lu sau , nhm nghin cu khoa hc ca nh sinh hc
David Sinclair v Konrad Howitz ng trn chuyn san Nature pht hin k diu ca
h v kh nng hot cht m rng thi gian sng nm men (Saccharomyces
cerevisiae),
[1]
rui gim (Drosophila melanogaster) v giun trn C. elegans.
[2]

-Mt nhm khc thc nghim li v xc nhn kt qu ca tin s Sinclair trn
C. elegans nm 2007.
[3]
Nhm nghin cu th ba cng b phn bc theo kt qu
vng sng khng c g bin i c C. elegans v D. melanogaster.
[4]

5


-Kt qu li cng gy tranh ci khi gii khoa hc pht hin giun trn C.
elegans mang trong ngi mt gene kim sot thi gian vn lu nay lm khoa hc
gia mt mi tm kim chu k sinh hc loi ny. Giun trn C. elegans gy c
ch bi mi sinh vt u hot ng v ngh ngi theo chu k c nh, gi l nhp
sinh hc. Ring, chng l loi duy nht hot ng khng c nhp sinh hc. Chng
lm vic lin tc ngy v m.
[6][7]

Ngn nga ung th
Cc nh khoa hc c kt qu thc nghim trn ng vt. Cho thy c
kh nng ca hot cht. Cha c nghin cu no c p dng ln ngi tnh n
nay.
[6]

Khc
Nhng li ch khc cng c bo co gm c mi lin quan trong cha tr
i tho ng, khng vim (bnh ph thng), li cho tim mch, tc ng bo v
thn kinh (ca bnh nhn hi chng Alzheimer), khng virus, tng lng
testosterone (gip chut tng tinh dch) y l nhng bo co cha c chng
thc.
[6]

3. TN TI TRONG T NHIN:
Bng di lit k hm lng trans-resveratrol trong cc ging cy trng ph
bin v cc th phm hay chnh phm ca ngnh cng nghip gii kht:
[5]

Ngun Ging / Loi
lng trans-
resveratrol
Ct kh
(Polygonum
cuspidatum)
R. japonica 64
d

R. bohemica 23
d

R. sachalinensis 29
d

HZ, MB 3770
d
, 2960
d

Nc nho Palomino fino 2.4
l

Tri nho Muscadine 5.2-26.4
f

Ht nho
Gamay 3.9
f

Pinot Noir 588
d

V nho
Pinot Noir 118
d

Gamay 6.8f
Palomino fino 15.7
f

B nho
Palomino fino 192
d

Muscadine 22.1-84.2
d

Ru
Muscadine 0.4-2.0
l

Cc loi khc: Cabernet Sauvignon (n = 40),
Tempranillo (n = 12), Cabernet Franc (n = 8), Liatiko
0.2-14.3
l

6


(n = 4), Xinomauro (n = 10), Muscat Bailey A (n =
5), Zinfandel (n = 5), Agiorgitiko (n = 9), Marzemino
(n = 3), Blaufrnkisch (n = 18), Portugieser (n = 9),
Zweigelt (n = 10), Negroamaro (n = 3), Teroldego (n
= 3), Nero dAvola (n = 5)
Ru trng
Nhiu loi: 21 loi Php, 23 loi M, 18 loi Hungary
v Trung u, 20 loi , 7 loi Canada, 2 loi Nam
M v 8 loi ca Australia
0.1
l

u phng
Jinpoong 3
f

NC-7, om, Gazipaa, Florispan ,
erezlik 5025, erezlik PI-355276
0.03-1.92
d

Cc ging trng khc: Spanette, Pearl, NC-18016,
Early Bunch, Whites Runner, Florunner C1, GA
207-3-4, GA 207-2, NC-9, Florispan C1, NC-17291,
Dixie Runner, PI-337396-FAV70, Florispan C3,
Small White Spanish
0.02-1.79
d

R u phng
Jinpoong 1.19
f

Tainan 9, Tainan 11, Tainan 1215-1330
d

Du ty Allstar 0.09
f
, 0.83
d

Vit qut
(blueberry)
Highbush Michigan 0.03
f
, 0.02
d

Lowbush "wild" Nova Scotia 0.01
f
, 0.02
d

Vit qut en
(bilberry)
Ba Lan 0.02
f
, 0.02
d

Ht d ci
(pistachio)
Ohadi, Uzun, Krmz, Halebi, Siirt 0.09-1.67
d

Scla m mu 2.0
f

Hoa bia
Cc ging trng: Hersbrucker Spat, Spalter, Saphir,
Hallertau Mittlefrher, Smaragd, Hallertau Tradition,
Wye Target, Nugget, Hallertau Magnum, Hallertau
Taurus, Saaz, Sladeck, Premiant, Willamette,
Cascade, Tomahawk, Simcoe, Warrior
0.10-2.28
d

Tettnang (ci to gene) 13
d
, 2.7
f

Rau dip
(lettuce)
?? (ci to gene) 56.4
f

Thuc l W38 (ci to gene) 9.3
f

La m Florida, Combi (ci to gene) 35-190
f

Kiwi Hayward (ci to gene) 182
f

d mg/kg kh f mg/kg ti l mg/L

4. THNG MI HA:
Bt chp tranh ci ca gii khoa hc v cng nng xc thc ca hp cht.
Resveratrol c thng mi ha mnh di dng thc phm chc nng by bn
rng khp th gii. Cc th trng chnh ca n l M, Nht Bn, Hn Quc Hm
7


lng resveratrol trong thc phm chc nng dao ng t 50% n 99%.
[6]
Ngun
sn xut r nht l t cy ct kh (Polygonum cuspidatum).
[8]
Cc cng ty qung b
sn phm nh mt loi tin dc tr c cc bnh nan y (tiu ng, ung th) v
tt cho c nam ln n gii. i vi n gii, thc phm s gip tr ha ln da, duy tr
nt thanh xun. Cn vi nam gii th lm tng ni tit t nam, thch hp cho nhng
ngi tp luyn th hnh.

5. CC HOT CHT LIN QUAN:
Nhng hp cht di y thuc cng nhm stilbenoid v c cu thnh t
cc n phn resveratrol vi nhau hoc resveratrol vi glucose:
[6]

Hnh 3: Piceatannol

Hnh 4: Pallidol

Hnh 5: Hopeaphenol

Hnh 6: Piceid

8


Hnh 7: trans-diptoindonesin B

Hnh 8: -viniferin
9


PHN III - TNG HP RESVERATROL:
1. CC PHNG PHP TNG HP:
[9][10]

Phng php tng hp u tin c Takaoka xut bng cch dng phn
ng Perkin ngng t sodium 3,5-dihydroxyphenylacetate vi 4-
hydroxybenzaldehyde, v vi acetic anhydride. Sn phm thu c l 3,4,5-
triacetoxystilbene--carboxylic acid. Sau , tip tc em kh carboxyl v thy
phn ta thu c resveratrol. Hiu sut cc giai on trong phng php ny khng
c bo co c th.

Hnh 9: Phng php tng hp resveratrol u tin dng ngng t Perkin

Hnh 10: p dng phn ng Wittig
Ri cc phng php tng hp da trn phn ng Wittig v ghp cp Horner-
Emmons (cn gi l phn ng Horner-Wadsworth-Emmons, phn ng Wittig-
Horner) c cng b nhng khng nhn c tn thnh v cho ra sn phm l
hn hp ng phn Z/E v hiu sut t c khng c bit ni tri.

Phn ng Grignard cng c dng lm c s nghin cu tng hp. Nguyn
liu ban u l p-anisaldehyde v 3,5-dimethoxybenzyl magnesium chloride.

Hnh 11: Tng hp da trn phn ng Grignard
Ni bt ti thi im hin ti l phng php tng hp bng phn ng ghp
cp Heck, nh khoa hc c trao gii Nobel Ha hc 2010 vi tm quan trng c
bit trong pht hin, pht trin v nghin cu s dng Paladium lm tc nhn xc
phn ng ghp cp to ni i cu hnh trans. Tng t phn ng Wittig, tc cht
gc ca phn ng Heck c gi thnh cao (3,5-dihydroxybenzaldehyde). Aldehyde
10


ny c bin i qua nhiu bc thnh alkene (styrene) v sau cho ghp cp vi
4-acetoxyiodobenzene. Sn phm qua thy phn cng to ra resveratrol.

2. QUY TRNH CA JING LIU:
Nghin cu ny tp trung kho st tng hp resveratrol theo phn ng ghp
cp Heck c s kh carbonyl c bo co bi gio s Jing Liu. Quy trnh tng hp
gm bn bc:
[9]

2.1.iu ch 3,5-diacetoxybenzoic acid:
bc u, bng phn ng ester ha Steglich 3,5-dihydroxybenzoic acid
c ester ha bo v hai nhm OH v tr 3 v 5. y l phn ng ca cc tc
cht phenol ( nu), acetic anhydride trong dung mi ethyl acetate. Pyridine c
thm vo hn hp ng vai tr nh mt xc tc Steglich (tng t DMAP: 4-
dimethylaminopyridine). C ch xc tc phn ng c trnh by theo gin :

OH
OH
HOOC
H
3
C O CH
3
O O
O
OH
HOOC
O
O
H
3
C
H
3
C
O
O
OH
HOOC
CH
3
O
H
3
C
O
O
O
O
HOOC
CH
3
O
O
CH
3

Hnh 12: C ch phn ng ester ha phenol
11


H
3
C O CH
3
O O
N
N
CH
3
O
O
O
CH
3
N O
O
H
3
C
N H
O
H
O CH
3
O
H
O
O
CH
3

Hnh 13: C ch c chp nhn ca xc tc pyridine
[11]

u tin, pyridine (hoc DMAP) to vi acetic anhydride mt cp ion acetate
v acetylpyridinium bt n nh. Ion phenolate tc kch vo nhm acetyl sinh ester
trong giai on hai, dn dn lm lin kt pyridinium v acetyl yu i cho ti khi t
la. Trc , i ion acetate v/hoc pyridine de-proton ha phenol cho ra acetic
acid v ion phenolate. Acetic acid mi sinh s tip tc proton ha pyridine theo mt
cn bng. Nh vo tr base (triethylamine), pyridinium c de-proton bc vo
vng xc tc mi.

2.2.iu ch 3,5-diacetoxybenzoyl chloride:
Sau khi bo v hai nhm OH phenol, gio s tip tc tin hnh phn ng
to dn xut acid chloride t 3,5-diacetoxybenzic acid. Cc ha cht cn cho phn
ng bao gm 3,5-diacetoxybenzoic acid, v SOCl
2
(thionyl chloride) vi s c mt
ca DMF trong vai tr tc nhn Vilsmeier. Sn phm sinh ra l mt acid chloride rt
nhy nc v km bn trong iu kin khng kh m ca Vit Nam.
12


Cl
S
Cl
O
R
OH
O
S O
Cl
O
Cl
R
O
S
O
Cl
O
O
R
SO
2
Cl
O
R
Cl

Hnh 14: C ch phn ng to acyl chloride
2.3.iu ch resveratrol triacetate:
Phn ng c tin hnh k sau bc trn do s nhy nc ca tc cht va
tng hp. Phn ng iu ch resveratrol l s ghp cp MizorokiHeck. Bao qut,
phn ng ghp cp Mirozoki-Heck s dng tc cht l mt halide bt bo ha v
alkene, Palladium xc tc kh hydride ca alkene ng thi to ni n carbon-
carbon gia nhnh hydrocarbon ca halide vi phn cn li ca alkene, cho ra
alkene mi cu hnh trans. Mt bin th ca phn ng Heck l phn ng Heck c s
kh trc tip nhm carbonyl, tc cht u ca phn ng l acid halide v alkene.
Nh to phc bn vi Palladium, carbon monoxide thot ra sau phn ng c
ngun gc t nhm carbonyl ca acid halide bn cnh trans-alkene mi sinh. S
hnh thnh phc cng gia tng hiu sut phn ng i cht. Xc tc palladium
thng gp dng mui acetate, hn l chloride.
Chu trnh xc tc c m hnh ha trong mt vng khp kn:
13


Hnh 15: Chu trnh xc tc ca Palladium(II)
Giai on A: Palladium xen vo gia phn t acid chloride, to ni vi aryl v vi
chlorocarbonyl. Alkene c b sung, tip tc to hp cht phi tr (phc ) vi tm
Palladium.
Giai on B: Xy ra qu trnh cng syn ca alkene, trong alkene chen vo ni
PdC hnh thnh ni CC
Alkene
C
Alkene
Pd mi. Do sc cng Pitzer, ni CC
Alkene
C
Alkene
Pd s t xoy sang cu trng trans.
Giai on C: tch/loi -hydride sinh phc mi ca alkene v Palladium
Giai on D: phc hi xc tc Pd, kt thc chu trnh cho ra sn phm transalkene,
CO v HCl

2.4.Thy phn triacetate thu resveratrol:
Bc cui cng ch n gin thy phn resveratrol triacetate trong mi
trng acid mnh thu c resveratrol.
14


PHN IV - THC NGHIM:
Chng ti thc hin tng hp resveratrol theo quy trnh ca Jing Liu. Trong bo co
ny trnh by ch hai giai on u ca quy trnh.

1. IU CH 3,5-DI ACETOXYBENZOI C ACI D:

Hnh 16: Phn ng iu ch

2.1.Tng hp li:
Thit b:
Bnh cu 3 c 20 mL, c nt silicon
Bp khuy t
C t
Bong bng cha kh nitrogen

Ha cht:
-Resorcylic acid IUPAC: 3,5-dihydroxybenzoic acid
CTPT: C
7
H
6
O
4

PTK (g/mol): 154.12, rn
MP (
o
C): ---
d (g/mL): ---
pKa: ---
Ngun gc: Aldrich-Sigma
Acetic anhydride IUPAC: Acetic anhydride / Ethanoic
anhydride
CTPT: C
4
H
6
O
3

PTK (g/mol): 102.09, lng
BP (
o
C): 139.8
d (g/mL): 1.082
pKa:
Ngun gc: Merck
Ethyl acetate IUPAC: Ethyl acetate / Ethyl ethanoate
CTPT: C
4
H
8
O
2

15


BP (
o
C): 77.1
d (g/mL): 0.897
pKa: ---
Ngun gc: Trung Quc
Pyridine IUPAC: Pyridine / Azabenzene
CTPT: C
5
H
5
N
BP (
o
C): 115.2
d (g/mL): 0.9819
pKa: 5.25 (acid lin hp)
Ngun gc: Merck
Formic acid IUPAC: Formic acid / Methanoic acid
CTPT: CH
2
O
2

BP (
o
C): 100.8
d (g/mL): 1.22
pKa: 3.77
Ngun gc: Trung Quc
n-Hexane IUPAC: Hexane
CTPT: C
6
H
14

BP (
o
C): 68-69
d (g/mL): 0.6548
pKa:
Ngun gc: Merck
NaCl (khan) IUPAC: Sodium Chloride
CTPT: NaCl
PTK (g/mol): 58.44, rn
MP (
o
C): 801
d (g/mL): 2.165
pKa: 6.7-7.3
Ngun gc: Trung Quc
Na
2
SO
4
(khan) IUPAC: Sodium sulfate
CTPT: Na
2
SO
4

PTK (g/mol): 142.04, rn
MP (
o
C): 884
d (g/mL): 2.664
pKa: ---
Ngun gc: Trung Quc

Tin hnh phn ng:
16


Cho 3,5-dihydroxybenzoic acid (~0.5 g, 3.2 mmol) vo bnh cu, sau ln lt
thm EtOAc (7.1 mL), anhydride acetic (1.5 mL, 16 mmol) v pyridine (0.5 mL, 6.4
mmol). ui ht khng kh bng dng kh nitrogen lin tc. Sau khuy nhit
thng trong 4 gi. Sau thi gian , thm formic acid (0.3 mL, 3.8 mmol) vo, tip
tc khuy trong 1 gi na. hn hp phn ng vo nc ri ra v lng pha hu
c bng nc ct 3 ln, nc mui (NaCl ~10%) 3 ln loi tr mui piridinium,
pyridine tha, v formic acid. Dng Na
2
SO
4
hoc MgSO
4
lm khan, em i c quay
v sau kt tinh li trong h dung mi n-hexane/EtOAc (1:1) thu c tinh th.
[9]

Hnh 17: S h thng thc nghim

Nhn xt sn phm: Tinh th sn phm c mu trng hoc i khi ng mu vng
khng c dng hnh hc nht nh, c mi ln ca pyridine v formic acid. C
th tip tc lm sch bng cch dng phu lc v bnh Buchner, cho tinh th
ti kt tinh ln mt phu, xi ra bng dung mi EtOAc lnh. Sau qu trnh ny,
mi m s mt i.
[10]
Ngoi ra trong trng hp tinh th mu ng, kt qu thu
c s l tinh th mu trng tinh khit. Hiu sut phn ng khng c nh,
thng bin thin trong khong 75-90%.

Xc minh sn phm:
*o nhit nng chy: Tin hnh ba ln trong phng th nghim trn tng 2, dy A
ca trng i hc Khoa hc T nhin. Tinh th c nhit nng chy 159
160
o
C.
*HRMS: Tin hnh phng th nghim B16, dy B ca trng i hc Khoa hc
T nhin. Pht hin mi M+Na vi m/z = 261.0370 (tnh ton cho C
11
H
10
O
6
l
238.0477). [ PHN VI - ]
17


*
1
H-NMR (CDCl
3
, 500 MHz): 7.72 (d, J = 2 Hz), 7.20 (t, J = 2 Hz), 2.32 (s).
Vi 7.72 v 7.20, l hai mi ca proton nhn hng phng c hng s ghp J =
2 Hz, do nhn hng phng c hai loi proton ghp meta vi nhau. Vi t l tch
phn ti ci mi 2.32 : 7.72 : 7.20 = 1 : 0.33 : 0.16 = 6 : 2 : 1, chng t nhn
thm c 3H v c 6H (ca mt loi proton) nhnh hydrocarbon ngoi nhn thm.
Tng cng c 9H trong nhn hng phng v nhnh hydrocarbon, thiu 1H ca
nhm carboxylic acid. Tn hiu ca proton acid khng pht hin c c l do s
trao i nhanh vi vt nc. [ PHN VI - ]

2.2.Kho st:
Phn ng c kho st li da theo thi gian, theo nng acetic anhydride, v
theo cc cht ph gia nhm nng cao hiu sut phn ng trong iu kin thc t ca
phng th nghim Vit Nam.

Theo thi gian phn ng
V c bn, hn hp phn ng gm cc cht nu theo ng t l thc trn phng
trnh phn ng. Trong s mol acetic anhydride v s mol xc tc pyridine ln
lt gp i nng 3,5-dihydroxybenzoic acid; lng dung mi s dng theo ng
t l gio s Jing Liu xut (tc c 110 mL EtOAc cho phn ng ca 7.71 g 3,5-
dihydroxybenzoic acid).

*Cch tin hnh
Cho a mol 3,5-dihydroxybenzoic acid vo bnh phn ng, cng vi 2a mol acetic
anhydride v 2a mol pyridine. Thm tip lng dung mi theo t l nu, ri dng
a khuy hoc my khuy t lm ng nht ha hn hp phn ng. Thc hin ui
khng kh bng dng nitrogen lin tc, nt kn bnh phn ng yn
ti nhit phng v ch thi gian. Sau thc hin li cc bc
chit, lng, lm khan, v c quay (khng thc hin kt tinh li)
c sn phm. Kt qu, sinh vin thc hin by phn ng theo thi
gian c tm lc bng th:
t (h) H%
4 80.83
6 84.91
13 83.54
14 87.54
15 88.08
19 92.77
25 85.98
18


th kho st
hiu sut theo thi gian (h)
80
82
84
86
88
90
92
94
0 5 10 15 20 25 30
h
H%

*Nhn xt: Theo th phn ng t hiu sut tt vi thi gian khong t 14-20 gi
ng h. Bn phn ng vi bn thi im (4h, 6h, 19h, 15h) c thc hin trong
cng mt iu kin nhit , thi im thc hin, dng c ch khc nhau thi
gian. Ba th nghim cn li (13h, 14h, 15h) c thc hin trong mt hon cnh
khc (ngy khc, nhit phng khc) nn c th mang sai s theo b.

Theo nng acetic anhydride
*Cch tin hnh
Thc hin tng t th nghim v thi gian, trong ch
thay i t l gia acetic anhydride vi 3,5-dihydroxybenzoic
acid (gi tt l phenol) t 2.0 cho n 3.0. Thm tip lng
dung mi theo t l nu, ri dng a khuy hoc my
khuy t lm ng nht ha hn hp phn ng. Thc hin
ui khng kh bng dng nitrogen lin tc, nt kn bnh
phn ng yn ti nhit phng v ch thi gian. Sau
thc hin li cc bc chit, lng, lm khan, v c quay
(khng thc hin kt tinh li) c sn phm. Kt qu,
sinh vin thc hin by phn ng theo thi gian c tm
lc bng th:
anhydride
/
phenol
H%
2.0 80.83
2.2 89.19
2.4 75.35
2.8 96.09
3.0 94.88
19


th kho st
t l acetic anhydride/phenol
70
75
80
85
90
95
100
2 2.2 2.4 2.6 2.8 3
H%

*Nhn xt: Nhm su phn ng c thc hin cng lc: cng nhit , cng b
dng c th nghim vi t l 2.4 t hiu sut thp hn c th do mt mt sn
phm qu trnh chit lng. Khi sinh vin cha hai pha tch tt vi loi b
pha nc v tip tc ra pha hu c vi nc. Theo th, t l tt nht t hiu
sut cao c th l anhydride/phenol=2.6

Phn ng vi s c mt ca ph gia:
Cc ph gia c b sung vo phn ng ln lt l EtONa, Et
3
N v hn hp
EtONa+Et
3
N cng t l vi acetic anhydride. Hiu sut phn ng thu c khng
my ni tri tng cho thm cc ph gia ny xut pht t c ch phn ng c
xc tc pyridine: c giai on chuyn dch cn bng pyridine & acetic acid v
pyridinium & acetate. Thm EtONa v Et
3
N y chuyn dch cn bng theo chiu
sinh pyridine cho vng xc tc mi.
20


2. IU CH 3,5-DI ACETOXYBENZOYL CHLORI DE:

Hnh 18: Phn ng iu ch
Thit b:
21


Bnh cu 3 c 20 mL, c nt silicon
Bp khuy t
C t
ng hon lu
Bong bng cha kh nitrogen
Ni un nc
Pipette

Ha cht:
3,5-diacetoxybenzoic acid IUPAC: 3,5-diacetoxybenzoic acid
CTPT: C
11
H
10
O
6

PTK (g/mol): 238.05, rn
MP (
o
C): 161-162
d (g/mL): ---
pKa: ---
Ngun gc: [t chun b]
DMF IUPAC: Dimethylformamide
CTPT: C
3
H
7
NO
BP (
o
C): 153
d (g/mL): ---
pKa: ---
Ngun gc: Aldrich-Sigma
Thionyl Chloride IUPAC: Sulfurous dichloride
CTPT: SOCl
2

BP (
o
C): 74.6
d (g/mL): 1.638
pKa: ---
Ngun gc: Merck
Toluene IUPAC: Methylbenzene
CTPT: C
7
H
8

BP (
o
C): 110.6
d (g/mL): 0.8669
pKa: ---
Ngun gc: Merck

*Cch tin hnh: Cho 3,5-diacetoxybenzoic acid (0.3664 g, 1.54 mmol), DMF (2
git) vo bnh cu phn ng lp sn ng hon lu, ri thm vo thionyl chloride (0.7
mL, 10 mmol), thc hin ui khng kh bng dng nitrogen lin tc. Dung dch
22


c khuy nhit thng trong 15 pht trong kh nitrogen; sau tin hnh
un hon lu 80
o
C trong 2 gi. Dung dch t trong sut chuyn sang mu vng,
ri m mu dn theo thi gian. Dung dch ng rn sau khi ngng un. Sau ,
thc hin ui thionyl chloride bng rt chn khng. Cho dung dch toluene vo v
khuy ha tan cht rn cn li. Lc ny xut hin cht rn/vt du khng tan
trong toluen mu nu, gn b. ui toluene, ta thu c cht rn mu vng nht
mi hc (0.3583g).
[9]

Hnh 19: S h thng thc nghim

*Nhn xt sn phm: Sn phm c mu vng, tnh n mn mnh.

*Xc minh sn phm:
+ HRMS: Tin hnh phng th nghim B16, dy B ca trng i hc Khoa hc
T nhin. Khng pht hin thy mi cp mi M+/M+2 (do Chlorine c hai ng v
35
Cl v
37
Cl t l 3:1) hay M+Na/M+2+Na vi m/z ln lt l 253/255 v 276/278
(tnh ton cho C
11
H
9
O
5
Cl l 256.0139). Ch nhn thy mi 238.0561 ca cht acid
gc. [ PHN VI - ]
+
1
H-NMR (CDCl
3
, 500 MHz): Khng c p dng do c bn cu trc ca acid v
dn xut acyl chloride ca n c khung sn tng t, ch khc bit nhm OH
23


carboxylic acid v nhm th Cl dng nh khng lm thay i nhiu dch
chuyn ha hc ca cc proton. Trong trng hp tng hp acid bn trn, ph
1
H-
NMR cng khng cho tn hiu proton acid.
+ Trc nghim nhanh: phng php nh tnh nhm chc acyl chloride theo Vogel
[phin bn 5 tr. 1212, nm 1989
[12]
] cho kt qu sau:
-Dung dch AgNO3: cho vo ng nghim dd AgNO3, nh nh cho chy ln thnh
ng nghim vi git dd acid chloride/toluene c hai lp dd trong sut trong
ng nghim. Lc ng nghim, lp tc lp dd nc bn di ha c. [DNG
TNH vi Cl]
-Th vi BaNO
3
: pht hin c hay khng SOCl
2
tha. Cho vo ng nghim dd
BaNO
3
, nh nh cho chy ln thnh ng nghim vi git dd acid chloride/toluene
c hai lp dd trong sut trong ng nghim. Lc u v yn, quan st khng c
s vn c cc lp. [M TNH vi SOCl
2
, do SOCl
2
trong mi trng nc s b
phn hy to HCl v H
2
SO
3
cho phn ng to kt ta trng vi BaNO
3
. Th nghim
c th cha ng do HCl to mi trng acid cn s to ta BaSO
3
. C th kim tra
li bng cch dng Ba(OH)
2
thay vai tr.]
-Th vi alcohol tuyt i: cho vi tinh th acid chloride vo ng nghim tht kh,
khong 1mL alcohol tuyt i c thm vo (Quan st xem c hay khng s xut
hin ca bt kh HCl hay khng). ngay vo hn hp 2mL dd NaCl bo ha trong
nc, thy hnh thnh mt lp hu c trn mt ng thi ngi thy mi thm
ngt/tri cy ca cc ester. [DNG TNH c trng vi acyl chloride, do SOCl
2

d khng th to ester c mi thm.]
24


PHN V - KT LUN V K HOCH:
1. KT LUN:
Phn ng to 3,5-diacetoxybenzoic acid t 3,5-dihydroxybenzoic acid c hiu
sut cao, trong khong t 75%-90%. Phn ng c s dng cc tc cht l acetic
anhydride, 3,5-dihydroxybenzoic acid (phenol), pyridine v dung mi ethyl
acetate. Hiu sut ti u khi t l mol anhydride:phenol = 2.6:1 trong khong
thi gian t 14-20h nu khng dng my khuy t. Sn phm thu c thng
dng tinh th trng hoc ng vng. Nhng tinh th ny c ra li bng ethyl
acetate lnh trong phu lc Buchner s tinh khit hn v mang mu trng.

Phn ng iu ch 3,5-diacetoxybenzoyl chloride phi c tin hnh trong iu
kin khan nc. Cc dng c cn c lm kh sn. Sn phm thu c cn tin
hnh ngay phn ng ghp cp Heck (giai on 3) nhm trnh sn phm b h
hao do hi nc khng kh. bo qun sn phm, phi tr trong chai l kn v
sc kh tr nh Nitrogen hoc tt hn l Argon, t v tr kh thong khng c
nh sng.

2. K HOCH:
Trong iu kin c th, sinh vin s tip tc kho st phn ng iu ch 3,5-
diacetoxybenzoic acid theo nng pyridine v nng dung mi.

Tin hnh kho st chi tit phn ng iu ch 3,5-diacetoxybenzoyl chloride nu
c.

Bc n giai on th ba l ghp cp Heck bng xc tc Palladium.

Sau thc hin qu trnh thy phn resveratrol triacetate c resveratrol
thnh phm.
25


PHN VI - PH LC:

Ph lc 1: Ph
1
H-NMR (CDCl
3
, 500 MHz) ca 3,5-diacetoxybenzoic acid
26


Ph lc 2: Ph HRMS ca 3,5-diacetoxybenzoic acid vi mi M
+
Na l 261.0370
27


Ph lc 3 Chy ph HR-MS cho 3,5-diacetoxybenzoyl chloride
ch thu c mi M+ 238.0561 ca acid gc
28


PHN VII - TI LIU THAM KHO:

1. Howitz, K.T., et al., Small molecule activators of sirtuins extend
Saccharomyces cerevisiae lifespan. Nature, 2003. 425(6954): p. 191-196.
2. Wood, J.G., et al., Sirtuin activators mimic caloric restriction and
delay ageing in metazoans. Nature, 2004. 430(7000): p. 686-689.
3. Gruber, J.A.N., S.Y. Tang, and B. Halliwell, Evidence for a Trade-Off
between Survival and Fitness Caused by Resveratrol Treatment of
Caenorhabditis elegans. Annals of the New York Academy of Sciences, 2007.
1100(1): p. 530-542.
4. Bass, T.M., et al., Effects of resveratrol on lifespan in Drosophila
melanogaster and Caenorhabditis elegans. Mechanisms of Ageing and
Development, 2007. 128(10): p. 546-552.
5. Zhang, A., et al., Occurrence and estimation of trans-resveratrol in
one-year-old canes from seven major Chinese grape producing regions.
Molecules, 2011. 16(4): p. 2846-61.
6. Resveratrol - Wikipedia English
7. "Tm ra gen kim sot thi gian Giun trn C.elegans",
khoahoc.com.vn, ngy: 21-12-2010
8. "Resveratrol", www.biologie.uni-freiburg.de
9. Jing Liu, et al., Synthesis of resveratrol and its analogs, phase-transfer
catalyzed asymmetric glycolate aldol reactions, and total synthesis of 8,9-
methylamido-geldanamycin. Department of Chemistry and Biochemistry,
Brigham Young University, 2007: p.3-5, 101-104.
10. Dustin Sprouse, The synthesis of Resveratrol. Department of Biology
and Chemistry, School of Science and Engineering, Oral Roberts University
2009: p. 33-45.
11. DMAP, Steglich's catalyst - Wikipedia English
12. Vogels Textbook of Practical Organic Chemistry (Fifth Edition),
John Wiley & Sons, Inc., New York, 1989: p. 1212.

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