Nghiên C U Bán T NG H P Troxerutin T Rutin

NGUYN TH HONG ANH
B GIO DC V O TO
B Y T
TRNG I HC DC H NI
V BO CH THUC
CHUYN NGNH CNG NGH DC PHM
NGUYN TH HONG ANH
NGHIN CU BN TNG HP
TROXERUTIN T RUTIN
LUN VN THC S DC HC
NM 2013
H NI - 2013
LI CM N
Trc ht cho php ti c by t lng bit n su sc v li cm n
chn thnh nht n PGS.TS. Nguyn nh Luyn - ngui trc tip
hng dn ti, truyn t cho ti nhng kin thc v kinh nghim qu bu
trong sut qu trnh lm thc nghim.
Ti xin chn thnh cm n thy Nguyn Vn Hn cng cc thy gio,
c gio, cc anh ch k thut vin ca b mn Cng nghip dc Trng
i hc Dc H Ni gip v to iu kin thun li cho ti trong qu
trnh hc tp v nghin cu.
Ti xin chn thnh cm n ng u, Ban gim hiu cng ton th cc
thy c gio Trng i hc Dc H Ni lun to iu kin tt nht cho
ti trong sut qu trnh hc tp trng.
Ti xin chn thnh cm n ng u, Ban gim hiu cng cc anh ch
ng nghip trng Cao ng Y t Thi Bnh lun ng vin v to iu
kin gip ti c v vt cht ln tinh thn ti c th hon thnh kho
lun ny.
Cui cng ti xin by t lng bit n ti gia nh, bn b lun l ch
da vng chc, l nhng ngun ng vin to ln i vi ti trong cuc sng,
trong hc tp v trong cng vic.
Xin chn thnh cm n!

Hc vin
Nguyn Th Hong Anh
MC LC
DANH MC CC BNG
DANH MC CC HNH
DANH MC CC S
T VN ............................................................................................................... 1
CHNG 1: TNG QUAN........................................................................................ 3
1.1. Rutin.................................................................................................................... 3
1.1.1. Cng thc, tnh cht ...................................................................................... 3
1.1.2. Ngun nguyn liu v phng php chit..................................................... 5
1.2. Troxerutin........................................................................................................... 7
1.2.1. Cng thc, tnh cht ...................................................................................... 7
1.2.2. Tc dng ca troxerutin (Vitamin P4)........................................................... 8
1.2.3. Mt s sn phm troxerutin trn th trng ................................................ 11
1.2.4. Cc phng php bn tng hp troxerutin t rutin..................................... 13
1.3. Ethylen clorohydrin......................................................................................... 15
1.3.1. Cng thc, tnh cht .................................................................................... 15
1.3.2. nh hng n sc khe con ngi ........................................................... 16
1.3.3. Nhng iu cn ch v an ton lao ng khi lm vic vi EC ................ 16
1.4. Ethylen oxyd..................................................................................................... 17
1.4.1. Cng thc, tnh cht .................................................................................... 17
1.4.2. nh hng n sc khe con ngi ........................................................... 18
1.4.3. Nhng iu cn ch v an ton lao ng khi lm vic vi EO................ 19
CHNG 2: NGUYN VT LIU, TRANG THIT B V PHNG
PHP NGHIN CU................................................................................................ 20
2.1. Nguyn liu ....................................................................................................... 20
2.2. Thit b v dng c .......................................................................................... 21
2.3. Ni dung nghin cu........................................................................................ 22
2.3.1. Xy dng quy trnh bn tng hp troxerutin t rutin.................................. 22
2.3.2. Kim nghim troxerutin .............................................................................. 22
2.4. Phng php nghin cu ................................................................................ 22
2.4.1. Phng php tng hp ha hc................................................................... 22
2.4.2. Xc nh cc thnh phn c trong sn phm............................................... 22

2.4.3. Xc nh cu trc sn phm ........................................................................ 24
2.4.4. nh lng................................................................................................... 24
CHNG 3: KT QU NGHIN CU................................................................. 25
3.1. Nghin cu quy trnh bn tng hp troxerutin ............................................ 25
3.1.1. Tin hnh bn tng hp troxerutin vi tc nhn alkyl ha l ethylen
clorohydrin trn c s phng php ca Stevens ................................................. 25
oxyd trn c s phng php ca F. De carvahho................................................ 32
3.1.3. Xc nh thnh phn ca cc sn phm thu c ....................................... 37
3.1.4. Xc nh cu trc sn phm ........................................................................ 41
3.1.5. xut quy trnh bn tng hp troxerutin .................................................. 44
3.2. nh lng troxerutin trong sn phm ......................................................... 48
3.2.1. Xc nh mt khi lng do lm kh.......................................................... 48
3.2.2. nh lng troxerutin trong sn phm ........................................................ 49
CHNG 4: BN LUN ......................................................................................... 51
4.1. V quy trnh bn tng hp troxerutin ........................................................... 51
4.1.1. V phn ng bn tng hp .......................................................................... 51
4.1.2. V qu trnh tinh ch sn phm sau phn ng............................................. 56
4.2. V xc nh thnh phn hn hp sn phm bng HPLC ............................ 57
4.3. V xc nh cu trc sn phm chnh............................................................ 58
4.3.1. V ph hng ngoi ...................................................................................... 58
4.3.2. V ph khi lng....................................................................................... 59
4.3.3. V ph cng hng t ht nhn .................................................................. 60
4.4. V xc nh hm lng troxerutin..................................................................... 64
KT LUN V XUT ....................................................................................... 65
1. KT LUN.......................................................................................................... 65
2. XUT............................................................................................................. 65
TI LIU THAM KHO
PH LC
DANH MC CC K HIU, CH VIT TT

ADN
ACGHI
CDCl3
CICAD
13
C-NMR
Acid desoxyribonucleic
Hi ngh cc t chc v sinh cng nghip ca M
(American Conference of Governmental Industrial Hygienists)
Cloroform d
Vn bn thm nh ha cht
(Concise Chemical Assessment Document)
Ph cng hng t ht nhn cacbon-13
(Carbon Nuclear Magnetic Resonance Spectroscopy)
COSY
Correlation Spectroscopy
DEPT
Distortionless Enhancement by Polarization Transfer
DMSO
Dimethyl sulfoxid
EC
Ethylen clorohydrin
ESI-MS
Ph khi lng phun m in t

(Electrospray Ionization Mass Spectrometry)
EO
Ethylen oxyd
EtOH
Ethanol
Gi (hour)
HDL
Lipoprotein t trng cao

(High Density Lipoprotein)
HMBC
Heteronuclear Multiple Bonds Correlation
Ph cng hng t ht nhn proton
H-NMR
(Proton Nuclear Magnetic Resonance Spectroscopy)

HPLC
HSQC
Sc k lng hiu nng cao

(High Performance Liquid Chromatography)
Heteronuclear Single Quantum Correlation
IARC
IR
IUPAC
C quan quc t nghin cu ung th

(International Agency for Research in Cancer)
Ph hng ngoi
(Infrared Spectroscopy)
Hip hi Ha hc Quc t
(International Union of Pure and Applied Chemistry)
Lt
LDL
Lipoprotein t trng thp (Low Density Lipoprotein)
MeOH
Methannol
Vin quc gia v an ton sc khe lao ng M
NIOSH- US
(National Institute for Occupational Safety and Health- United

States)
NTP- US
Chng trnh c cht hc quc gia M

(National Toxicology Program - United States)
ppm
n v phn triu (parts per million)
SKLM
Sc k lp mng
SN2
T0nc
US-EPA
US- OSHA
Phn ng th i nhn lng phn t

(Nucleophilic Substitution)
Nhit nng chy
C quan bo v mi trng M
(United States Environmetal Protetion Agency)
Cc qun l sc khe v an ton lao ng M
(United States Occupational Safety and Health Administration)
UV
T ngoi (Ultraviolet)
UV-VIS
T ngoi kh kin (Ultraviolet Visible)
VLDL
Lipoprotein t trng rt thp (Very Low Density Lipoprotein)
DANH MC CC BNG
Bng 1.1. Mt s bit dc c cha troxerutin ca Vit Nam .............................. 11
Bng 1.2. Mt s bit dc c cha troxerutin trn th gii ................................. 12
Bng 1.3. Cc cp tip xc vi ethylen clorohydrin ......................................... 17
Bng 1.4. Gii hn cho php tip xc vi EO ca US-OSHA .............................. 19
Bng 2.1. Cc ha cht, dung mi s dng trong qu trnh thc nghim.............. 20
Bng 2.2. Cc my mc, dng c s dng trong qu trnh thc nghim............... 21
Bng 3.1. Kt qu kho st s nh hng ca nhit n hiu sut phn ng
bn tng hp troxerutin vi tc nhn EC............................................... 26
Bng 3.2. Kt qu kho st s nh hng ca thi gian phn ng n hiu sut
phn ng bn tng hp troxerutin vi tc nhn EC............................... 27
Bng 3.3. Kt qu kho st s nh hng ca t l mol EC/rutin n hiu sut
phn ng bn tng hp troxerutin.......................................................... 29
Bng 3.4. Tnh cht cc sn phm (T1-T4) thu c sau tinh ch ........................ 31
Bng 3.5. Kt qu kho st s nh hng ca nhit n hiu sut phn ng
bn tng hp troxerutin vi tc nhn EO .............................................. 33
Bng 3.6. Kt qu kho st s nh hng ca t l mol EO/rutin n hiu sut
phn ng bn tng hp troxerutin.......................................................... 34
Bng 3.7. Tnh cht cc sn phm (T5-T8) thu c sau tinh ch ........................ 36
Bng 3.8. Kt qu chy HPLC ca T1 ................................................................... 39
Bng 3.9. Kt qu chy HPLC ca T9 ................................................................... 40
Bng 3.10. Kt qu xc nh mt khi lng do lm kh ca T1 v T9 ................. 49
Bng 3.11. Kt qu xc nh hm lng troxerutin ................................................. 50
DANH MC CC HNH
Hnh 3.1.
Sc k troxerutin i chiu (a) .......................................................... 38
Hnh 3.2.
Sc k mu th T650C (EC) ............................................................. 39
Hnh 3.3.
Sc k mu th T1............................................................................. 39
Hnh 3.4.
Sc k mu th T9............................................................................. 40
Hnh 4.1.
Ph ESI-MS negative ca cht T9......................................................... 59
Hnh 4.2.
Ph 13C-NMR v DEPT ca T9 ........................................................... 60
Hnh 4.3.
Ph 1H-NMR gin rng vng t 3,04 4,96 ppm ca T9 .................... 61
Hnh 4.4.
Ph 1H-NMR gin rng vng t 5,11 7,84 ppm ca T9 .................... 62
Hnh 4.5.
Ph COSY gin rng ca T9 ................................................................. 62
Hnh 4.6.
Ph HMBC ca T9 ................................................................................ 63
Hnh 4.7.
Mt s tng tc chnh trn ph COSY ( ) v HMBC () ca T9 ... 63
DANH MC CC S
S 1.
Quy trnh bn tng hp tri(hydroxyethyl) rutin..................................... 45
S 2.
Quy trnh bn tng hp troxerutin ......................................................... 47
T VN
Theo thng bo nm 2010 ca Hi tim mch Vit Nam, hin nay trn ton
th gii bnh tim mch vt xa cc bnh ung th, nhim khun ni chung
v cng l nguyn nhn hng u gy t vong. Mi nm c khong 17,5 triu
ngi t vong do bnh tim mch v cc bin chng ca n [9]. Nu khng c
mt hnh ng tch cc th n nm 2015 s c 20 triu ngi trn ton th
gii cht do bnh tim mch, t l ny tp trung nhiu cc nc ang pht
trin (khong 80%).ng trc nhng thch thc , cc nh khoa hc trn
th gii v ang khng ngng nghin cu tm ra cc thuc c tr cho cc
bnh tim mch.
Rutin l mt flavonoid c tc dng lm bn v lm gim tnh thm ca
mao mch, gim trng lc c v chng co tht. Rutin c dng ch yu
phng nhng bin c ca bnh x va ng mch, iu tr cc trng hp suy
tnh mch [5]. T trc n nay, rutin c bit n nh mt v thuc cha tr
rt tt cc bnh lin quan n mch mu trong ng y cng nh trong Ty y.
Tuy nhin nhc im ca rutin l tan km trong nc nn kh hp thu v
kh bo ch cc dng thuc c kh nng hp thu tt. khc phc nhc
im ny, cc nh khoa hc tm ra mt dn cht flavonoid cng nhm vi
rutin, d hp thu v tc dng dc l mnh hn l troxerutin.
Troxerutin c ngha quan trng trong iu tr suy tnh mch mn tnh
mt bnh l nguy him v ngy cng tr nn ph bin hn nc ta cng nh
trn th gii. Chnh v vy, vic nghin cu bn tng hp cc hot cht c tc
dng iu tr tt nh troxerutin hin ang rt c quan tm.
Troxerutin c bn tng hp t rutin vi tc nhn alkyl ha l ethylenoxyd, ethylen-clohydrin. Vic bn tng hp troxerutin t rutin c thc
hin nhiu quc gia trn th gii nh Anh, M, Trung Quc, Nht
1
BnTuy nhin, hin nay Vit Nam vn cha sn xut c troxerutin,

vic s dng troxerutin cho sn xut dc phm trong nc l t ngun
nguyn liu nhp khu.
Trc nhu cu thc t v tnh hnh sn xut dc phm trong nc, chng
ti tin hnh ti: Nghin cu bn tng hp troxerutin t rutin vi 2
mc tiu sau:
1. Xy dng c quy trnh bn tng hp troxerutin t rutin quy m
phng th nghim.
2. Bn tng hp c troxerutin t tiu chun hm lng theo dc in
Anh 2010.
Vi nhng mc tiu trn, chng ti hi vng kha lun ny s l mt nghin
cu thm d c th ng dng sn xut troxerutin quy m pilot v tin ti
quy m cng nghip ti Vit Nam.
CHNG 1: TNG QUAN
1.1. Rutin
1.1.1. Cng thc, tnh cht
Cng thc cu to
- Cng thc phn t: C17H30O19.
- Khi lng phn t: 610,52 vC.
-Theo s phn loi da vo v tr ca gc aryl th rutin c xp vo nhm
flavonol thuc nhm Euflavonoid [5]. Cng thc cu to nh sau:
OH
O
Flavonol
Rutin
Rutin 3D
Tnh cht
- Bt kt tinh mu vng nht hoc vng hi lc,
ra nh sng c th mu hi sm li. Kt tinh
hnh kim vng nht trong nc. Tinh th ngm 3
phn t nc, tr thnh khan sau khi sy 12h
110oC di p sut gim 10 mm Hg.
- Rutin khan rt d ht m, rutin khan bin mu
nu 125oC, do 195-197oC v phn hy
214-215 oC [3].
- Nng sut quay cc []23D = +13,820.
3
Bt Rutin
- tan: d tan trong pyridin, tan trong methanol v cc dung dch kim
long, hi tan trong glycerin, kh tan trong ethanol ngui v isopropanol, thc
t khng tan trong nc, aceton, ether, cloroform, ether du ha, benzen.
- im chy: 2100C km theo phn hy.
- Ph hp th t ngoi ca rutin trong ethanol c cc i hp th bc sng
259 1nm v 362,5 1; trong methanol l 359 nm.
- Rutin c cu trc l mt glycosid nn rt d b thy phn bi enzym c trong
dc liu l rhamnodistase hoc mi trng acid, nhit cao, dung dch
kim c nhit cao [3].
- Tc dng vi FeCl3 cho mu xanh lc [5].
- Tc dng vi H2SO4 m c: Acid H2SO4 nh ln rutin cho mu vng m
[2].
nh tnh
- o ph IR ca ch phm, so snh vi ph IR ca rutin chun.
- Ha tan 50,0 mg ch phm trong methanol v pha long thnh 250,0 ml vi
cng dung mi, lc nu cn. Pha long 5,0 ml dung dch ny thnh 50,0 ml
bng methanol . o ph hp th t ngoi trong khong t 210 nm n
450nm, dung dch phi cho hai cc i hp th 257nm v 358 nm. hp
th ring bc sng cc i 358 nm phi t t 305 n 330, tnh theo ch
phm khan.
- Phng php sc k lp mng
+ Bn mng: Silica gel G
+ Dung mi khai trin: N-butanol/acid acetic khan/nc/methyl ethyl
ceton/ethyl acetat = 5/10/10/30/50).
+ Dung dch th: Ha tan 25 mg ch phm trong 10,0 ml methanol.
+ Dung dch i chiu: Ha tan 25 mg rutin chun 10,0 ml methanol.
+ Cch tin hnh: Chm ring bit ln bn mng 10 l mi dung dch trn.
Trin khai sc k n khi dung mi i c 10 cm. kh bn mng ngoi
khng kh. Phun ln bn mng hn hp gm 7,5 ml dung dch kali fericyanid
1% v 2,5 ml dung dch FeCl3 10,5%. Quan st bn mng trong vng 10 pht.
Vt chnh trn sc k thu c ca dung dch th tng ng v v tr, mu
sc v kch thc vi vt chnh trn sc k thu c ca dung dch i
chiu [7, 17].
nh lng
- Ha tan 0,200 g ch phm trong 20 ml dimethylformamid. Chun bng
dung dch tetrabutylamonihydroxyd 0,1 M.
- Xc nh im kt thc bng phng php chun in th.
1ml dung dch tetrabutylamonihydroxyd 0,1M tng ng vi 30,53 mg
rutin.
- Yu cu: Ch phm cha 95% - 101,0% rutin tnh theo ch phm khan [7].
Bo qun
ng trong l kn, trnh nh sng [7].
1.1.2. Ngun nguyn liu v phng php chit
1.1.2.1. Ngun nguyn liu
- Rutin c nhiu nht trong cy he (Sophora japonica.L) h u (Fabaceae).
N hoa he kh Nguyn liu
Hoa he ti
chit rutin
5
- Rutin c hu ht trong cc phn xanh ca cy he nhng hm lng cao

nht vn l n. Bnh qun trong n he c khong 6 - 30% rutin [10], ring
n he nc ta hm lng rutin lun mc cao 30 - 34% [8] (Hungari
12%, Php 10% [3]).
- Dc in Vit Nam III quy nh trong n he lm nguyn liu sn xut
rutin t nht l c 20% rutin [7].
- nc ta, cy he c trng mt s tnh, nhng nhiu nht l Thi
Bnh [5].
Ngoi cy he cn c mt s dc liu khc cha rutin:
- La mch 3 gc (Fagopyrum esculentum Moench = Polygonum fagopyrum
L., h rau rm Polygonaceae). L kh cha 2-3% rutin. Hin nay to ra
c nhng chng c hm lng 5-8% rutin. T l rutin cao cy trc khi ra
hoa.
- Bch n (Eucalyptus macrorrhyncha F.Muell). Hm lng rutin trong l
trung bnh 10% v c th n 19% nhng l non.
- To ta (Ziziphus mauritiana Lamk) h to ta (Rhamnaceae). L kh cha
1,5% rutin [5].
1.1.2.2. Phng php chit xut
C nhiu phng php c s dng chit xut rutin t hoa he
nh: chit xut bng nc [8], chit xut bng ethanol [8], chit xut bng
dung dch kim, chit xut bng dung dch kim c kt hp siu m [3]. Qua
cc ti liu tham kho cho thy rng chit xut bng dung dch kim c s
dng siu m cho hiu sut chit rutin cao nht.
1.2. Troxerutin
Dc in Anh (2010) xc nh: Troxerutin l hn hp cc dn xut
rutin hydroxyethyl ha c cha ti thiu 80% tri-(hydroxyethyl) rutin, ngoi
ra cn c dn xut mono, bis, tetra-(hydroxyethyl) rutin.
1.2.1.1. Cng thc thnh phn chnh ca troxerutin: tri-(hydroxyethyl) rutin
- Cng thc phn t: C33H42O19.

- Tn khoa hc: 2 [3, 4 bis (2 hydroxyethoxy) phenyl] 5 hydroxyl
7 (2 hydroxyethoxy) 4 oxo 4H chromen 3 yl 6 O (6
deoxy D mannopyranosyl) D glucopyranoside.
1.2.1.2. Tnh cht ca troxerutin
- Dng bt mu vng.
- Tnh tan:
+ Tan trong nc, glycerol, propylen glycol.
+ Khng tan trong ether, benzen, cloroform,
ethanol, methanol lnh.
- T0nc ca hn hp 80% tri-(hydroxyethyl) rutin:
khong 1560C [29].
- T0nc ca tri-(hydroxyethyl) rutin 1810C [17].
Bt Troxerutin
nh tnh
- o ph IR trong vng bc sng t 4000 - 650 cm-1, so snh vi ph IR ca
troxerutin chun.
- Dng sc k: Pic chnh trong sc k ca mu th tng ng v v tr v
kch thc vi pic chnh ca mu chun [17].
Xc nh cc thnh phn hn hp
- Dng sc k lng hiu nng cao (HPLC).
- Tiu chun cho php:
+ T l thnh phn chnh: (tri-(hydroxyethyl)-rutin): 80%.
+ T l cc thnh phn khc: (mono-, bis-, tetra-(hydroxyethyl)-rutin) mi
thnh phn khng qu 5%, c th chp nhn mt thnh phn khng qu 10%
[17].
nh lng
- Tin hnh theo phng php o quang xc nh hm lng mu th.
- Yu cu: Ch phm cha 95105% troxerutin tnh theo ch phm lm
kh [17].
Bo qun:
- ng trong l kn, trnh nh sng [17].
1.2.2. Tc dng ca troxerutin (Vitamin P4)
- Troxerutin c tc dng bo v tnh mch v mao mch rt tt, nh c c
tnh lm gim tnh thm thnh mch, gim ph, ci thin vi tun hon.
- Troxerutin c tc dng kh cc gc oxy v nit, gp phn bo v tnh mch.
Troxerutin c tnh kh mnh, c th v hiu ha cc gc t do. Vi bnh nhn
xut huyt thnh mch, troxerutin c th gip bo v c mng t bo thot
khi s ph hoi bi cc gc t do. Nh , tng c tnh bn ca thnh mao
mch, chng c s xut huyt [8].
- Troxerutin kt hp vi Aescin trong iu tr bnh tai trong. Hiu qu ca

s kt hp ny c th nghim trn 34 bnh nhn. Sau 44 ngy iu tr,
kt qu cho thy 23/34 bnh nhn c thay i ngng hn 10 dB (vic ci
thin ngng nghe hn 10 dB c cho l mt ci thin ng k) [33].
- Theo B.S. Adam v ng nghip, s kt hp coumarin v troxerutin nng
cao hiu qu trong iu tr bnh suy tnh mch so vi vic ch s dng
coumarin n c [14].
- Troxerutin v quercetin c nghin cu trn 4 th nghim th t bin gen.
Kt qu cho thy troxerutin khng gy t bin, quercetin cho phn ng
dng tnh vi cc th nghim t bin gen. S thay th 3 nhm
hydroxyethyl vo quercetin trit tiu kh nng gy t bin gen ca
quercetin [27].
- Nm 2009, Shao-hua Fan v cng s tin hnh nghin cu tc dng ca
troxerutin trn m hnh gy tn thng thn bng D-galactose chut. Kt
qu nghin cu cho thy:
+ Troxerutin c tc dng lm tng hot ng ca cc enzym chng oxy ha
nh cytosolic Cu/Zn superoxid dismutase (SOD-1), catalase (CAT) v
glutathion peroxidase (GPX); ng thi troxerutin cng c tc dng lm gim
nng malondialdehyd (MDA sn phm ca qu trnh peroxy ha lipid).
+ Troxerutin c tc dng lm gim biu hin ca enzym COX-2
(cyclooxygenase -2).
T nhng kt qu ca nghin cu ny cho thy troxerutin c th lm
gim bt tn thng thn gy ra bi D-galactose thng qua c ch chng
vim v chng oxy ha [32].
- Troxerutin c dng iu tr trong bnh tc tnh mch trung tm vng mc.
Troxerutin c tc dng chng kt tp tiu cu, gim qunh ca mu nn
9
c dng iu tr lu di v d phng tai bin mt th hai. Troxerutin ci

thin r rng th lc, ph hong im v tin trin thiu mu ca bnh [12].
- Nm 2010, cc nh khoa hc Trung Quc nghin cu tc dng bo v
thn kinh ca troxerutin. Kt qu cho thy troxerutin c kh nng ci thin
nhn thc v c tc dng bo v thn kinh [23].
- Theo nghin cu ca F. Squadrito cng ng s th s kt hp 150 mg
troxerutin v 1,5 mg carbazochrom cho hiu qu iu tr tt v an ton
nhng bnh nhn mc tr [34].
- Nm 2011, cc nh khoa hc Trung Quc nghin cu v tc dng chng
phng x (tia ) ca troxerutin trn chut v c tnh ca troxerutin trn dng
t bo V79. Kt qu ca nghin cu cho thy:
+ Troxerutin khng c kh nng gy c r rng nng di 20mg/ml.
+ Troxerutin c tc dng chng tia phng x do troxerutin lm tng ng k t
l sng st sau 30 ngy b chiu x (tia gamma) chut nghin cu [41].
- Cng trong nm 2011, Jun Lu tin hnh nghin cu tc dng ca
troxerutin trn m hnh gy i tho ng typ 2 bng ch n giu
cholesterol chut. Trong nghin cu ny, tc gi nh gi hiu qu ca
troxerutin i vi hin tng suy gim nhn thc do s khng insulin no
chut thc nghim. Kt qu thu c t nghin cu cho thy troxerutin c
kh nng phng nga v iu tr bnh suy gim nhn thc do i tho ng
typ 2 v bnh Alzheimer [26].
- Nm 2013, nghin cu ca cc nh khoa hc n cho thy rng troxerutin
c kh nng lm gim ng k lng VLDL, LDL v tng HDL trong mu.
V vy, troxerutin ng vai tr quan trng trong vic iu tit chuyn ha
cholesterol [25].
10
- Trong nghin cu ca Jun Lu cng cng s (2012) cho thy troxerutin c

kh nng c ch biu hin ca enzym cyclin dependent kinase I, tng cng
hot ng ca enzym phosphatase dephosphorylation I dn n c ch s
suy gim tr nh chut thc nghim. V vy, troxerutin c c nh l
mt ng vin tim nng trong phng nga v iu tr bnh thiu ht nhn thc
v mt s ri lon no b khc [24].
1.2.3. Mt s sn phm troxerutin trn th trng
Vit Nam, troxerutin hin ang c s dng vi mt s dng bit dc
v xut x ca cc sn phm ny c trnh by trong bng 1.1.
Bng 1.1. Mt s bit dc c cha troxerutin ca Vit Nam
Stt Tn thuc
Chiona
1
Dopolys-S
2
Ginkotin
3
Thnh phn
Cng ty sn xut
Cao G. biloba 14mg TNHH dc phm

Heptaminol 300mg
Shinpoong Daewoo
Troxerutin 300mg
Vit Nam
Cao G. biloba 14mg
Cng ty c phn
Heptaminol 300mg
xut
Troxerutin 300mg
Domesco
Cao G. biloba 14mg
Cng ty lin doanh
Cao gingseng
BV pharma
nhp
Dng bo ch
Vin nn bao
phim
khu Vin nang
Vin nang
Troxerutin 300mg
4
5
Nebivox
Troxerutin 7g
Neorutin
Troxerutin 7g
CT lin doanh BV
pharma
Cng ty lin doanh
BV pharma
11
Thuc cm
Thuc cm
Trn th gii troxerutin c s dng vi nhiu dng bit dc, cc dng

bit dc ny c trnh by trong bng 1.2.
Bng 1.2. Mt s bit dc c cha troxerutin trn th gii
Stt
1
Tn Thuc
Flebotrat
Ginkor fort
Ronacare
troxerutin
Troxerutin
4
5
6
7
8
9
Troxerutin
injection
Troxerutin
ratiopharm
300 mg
Troxerutin
200mg
Troxeti Eye
drop
Troxevasin
10
Troxivenol
11
Venalot SR
12
Venoruton
Thnh phn
Cng ty sn xut
Coumarin
Troxerutin
Neckerman
Industria
Farmaceutica
Dng bo ch
Vin nn
Cao ginkgo biloba

Heptaminol
HCl Beaufour
Ipsen Vin nang
Troxerutine
(Php)
EMD Chemicals Kem bi da
Inc.
Troxerutin 20mg
Chema Elektromet Gel
Troxerutin
Chengdu PingYuan
Medicine Co.,Ltd
Ratiopharm
Medikamente zum
Schutz der Gefe
Syntenda SP. Z
O.O. P.F.-CH
Hanlim Pharm Co.,
Ltd (Hn Quc)
BalkanpharmaRazgrad AD
Bungari
Rotapharm
Troxerutin 300 mg
Troxerutin 200 mg
Troxerutin
(500 mg/10ml)
Troxerutin 300mg
Troxerutin
Carbazochrom
Coumarin
Troxerutin
Thuc tim
Vin nang
Vin nang
Dung dch nh
mt
Vin nang
Thuc tim
Vin nn
Medica; Vin
gii
& phng ko di
Asta
Schaper
Brummer
Mono-, di-, tri, Novatis Thy S
tetrahydroxyrutosids
12
Gel
Vin nang
1.2.4. Cc phng php bn tng hp troxerutin t rutin

1.2.4.1. Vi tc nhn ethylen clorohydrin
- Phng trnh phn ng:
+ Cl-CH2CH2OH
Xc tc
Phng php Stevens

+ Tin hnh phn ng:
Cho rutin v nc vo bnh phn ng, hn hp c khuy trn bng
my khuy t. Thm NaOH vo hn hp phn ng vi t l mol rutin/NaOH
=1/3. Nng nhit phn ng ln 550C v nh dn EC vo, t l rutin/EC =
1/3. Tip tc nng nhit phn ng ln 750C v duy tr trong 2h. Phn ng
c thc hin di bu kh N2.
+ Kt thc phn ng: lm lnh hn hp phn ng xung di 200C v duy tr
mi trng N2, dng HCl 10 % iu chnh pH = 5. Dung dch phn ng c
gi trong t lnh 24h, rutin cha phn ng s ta li. Lc loi rutin, dch lc
c ct quay ti kh di p sut gim, cn thu c ha tan nng trong
methanol loi b phn mui khng tan. Ct quay dung mi n kh. Cn
thu c ha tan nng trong ethanol. Lc ly dung dch, kt tinh thu c
sn phm [35].
13
Phng php Frantisek Czech

Cho rutin v methanol vo bnh phn ng. Thm acid ascorbic hoc
Na2S2O5 lm cht xc tc. Cho ethylen clorohydrin vo bnh phn ng. Nng
nhit ca khi phn ng ln 85-1000C v duy tr trong 2 gi.
Kt thc phn ng, lm lnh khi phn ng v iu chnh pH = 5 bng
dung dch HCl long. Hn hp c em ct loi dung mi n kh, phn cn
thu c ha tan vi ethanol v lc ly dch lc. Dch lc kt tinh thu sn
phm [21].
Phng php Vladimir Crech
To dng hn dch ca rutin trong nc, sc kh N2 lin tc. Thm
dung dch NaOH v nng nhit ca phn ng ln 75 - 780C. Ethylen
clorohydrin c cho vo, gi nguyn nhit v duy tr phn ng trong
thi gian 2h. Sau khi phn ng c acid ha bng HCl long (a v
pH = 5). lnh, rutin cha phn ng c kt ta. Lc loi rutin v ct dch
lc n kh. Ha tan cn thu c trong ethanol si, lc nng loi mui v
kt tinh dch lc. Lc thu tinh th, sy, thu ly sn phm [40].
1.2.4.1. Vi tc nhn ethylen oxyd
- Phng trnh phn ng
Xc tc
14
Phng php ca Pierre J Courbat

Tin hnh:
To hn dch rutin trong nc v a nhit ca hn hp n 600C.
Thm cht xc tc NaOH vo hn dch vi t l rutin/NaOH= 1/3. Nng nhit
phn ng ln 80 - 850C v sc EO vo trong khong thi gian 6h (30 ml
EO/1 pht). T l mol EO/rutin = 4,5/1.
Sau 6h, hn hp phn ng c pH = 9,5. Hn hp phn ng c lm
lnh v iu chnh pH = 4,5 vi dung dch HCl. lnh, rutin cha phn ng
kt ta. Lc ly dch, ct quay bc hi dung mi. Cn thu c em ha tan
trong ethanol, lc v thu ly dung dch. Lm lnh dch lc kt tinh, lc thu
sn phm kt tinh. Sn phm c kt tinh li bng methanol, sy kh thu
c sn phm troxerutin [18].
Phng php ca F. de carvahho
Tin hnh:
Cho 680 g rutin v 3500ml nc to hn dch rutin trong ni thp
khng g 5l. Thm tip NaOH v un nng n khong 80-850C. EO c
dn vo ni thp. Gi nguyn nhit phn ng v duy tr cho n khi pH
ca phn ng bng 9,5. Kt thc phn ng, lm lnh hn hp phn ng v
dng HCl 10% iu chnh pH = 4,5. Hn hp c lc bi thit b lc nhiu
tng loi tp v rutin cha phn ng, thu dch lc v ct quay loi nc
thu ta. Ra ta bng 5lt ethyl acetat (ra 4 ln). Lc v sy kh thu sn
phm [19].
1.3. Ethylen clorohydrin
- EC c cng thc phn t l C2H5ClO, c cu trc dng mch thng.
15
- Tn khoa hc: 2 cloroethanol.

- Ethylen clorohydrin nhit thng tn ti th lng, khng mu.
- tan trong nc: 1 x 106 mg/l.
- p sut hi: 4,9 mm Hg 200C.
- Nhit nng chy: - 670C.
- Nhit si: 128 -1300C.
- Khi lng phn t: 80,52 vC [39].
1.3.2. nh hng n sc khe con ngi
EC c kh nng gy c hi cho c th khi tip xc qua ng h hp
hoc hp thu qua da.
- Trn h h hp: EC c th gy kch ng mi v hng gy ho v kh th. EC
gy kch ng phi, tip xc vi lng cao hn c th gy ph phi cp.
- Trn h tiu ha: EC c th gy nn, bun nn v au bng.
- EC c th gy tn thng gan, thn.
- Trn h thn kinh: c th nh hng n s tp trung v tr nh, gy au
u, chng mt, chong, l ln, co git. Tip xc nng cao c th ph
hy h thn kinh trung ng, gy t vong [28].
1.3.3. Nhng iu cn ch v an ton lao ng khi lm vic vi EC
- Gii hn tip xc vi ethylen clorohydrin ni lm vic nh sau:
+ Theo OSHA: Gii hn php l cho php tip xc vi ethylen clorohydrin
mt ca lm vic 8 gi l 5ppm [30, 39].
+ Theo NIOSH v ACGIH: gii hn tip xc vi khng kh c ngh l 1
ppm, gii hn ny khng nn vt qu bt k lc no [30].
- Theo c quan bo v mi trng M US-EPA thng bo cc cp tip xc
vi ethylen clorohydrin nh sau [37]:
16
Bng 1.3. Cc cp tip xc vi ethylen clorohydrin

Cc cp tip xc vi ethylen clohydrin (ppm [mg/m3])
Phn loi
Cp 1
Khng gy bt k tn thng no.
10 pht 30 pht 1 gi
4 gi
8 gi
Cha c s d liu a ra kt qu
Cp 2
7,0
5,0
4,0
1,6
0,77
Gy tn thng cho c th.
[23]
[16]
[13]
[5,3]
[2,5]
21
15
12
4,7
2,3
[69]
[49]
[39]
[15]
[7,6]
Cp 3
C th gy t vong.
1.4. Ethylen oxyd
- Nm 1859, Wurtz tng hp c EO u tin bng cch s dng KOH

loi b HCl t clorohydrin ethylen. Qu trnh oxy ha trc tip ethylen to
ra EO c Lefort tm ra vo nm 1931 dn thay th qu trnh clorohydrin
trong vic sn xut EO.
- Hin nay EO c sn xut bi qu trnh oxy ha trc tip ethylen vi
khng kh hoc oxy [15].
- EO c cng thc ca phn t l C2H4O, cu trc dng vng 3 cnh.
- Tn IUPAC: oxirane.
- Ethylen oxyd th lng hoc kh khng mu, c v ngt.
- T0nc: -112,690C.
- Nhit si: 9,450C.
- Tnh tan: tan tt trong nc.
17
- Polyme ha: EO d b polymer ha vi s xc tc ca nhit , thi gian

bo qun, v s c mt ca ion kim loi.
- Khc phc hin tng polymer ha: bo qun trong cc thng thp khng g,
thit b cha phi bm kh N2 [15].
1.4.2. nh hng n sc khe con ngi
Theo CICAD Thuc chng trnh quc t ca t chc Y t th gii
v an ton ha cht cnh bo mt s nguy c c th xy ra khi tip xc,
lm vic vi EO.
-Tip xc qua ng h hp cp tnh:
EO c mt ngng mi cao (>250 ppm), vi ngng mi ny th kh
nng c hi khi c ngi thy mi. Thm ch sau , mi ca ethylen
oxyd l ngt, thm, v c th d dng b nhm ln vi mi ca diethyl ether.
Ngng mi qu cao ny gy nn s kh khn trong vic cnh bo mc
c hi nhng ngi tip xc, lm vic vi EO.
Nhng nh hng ca vic tip xc qua ng h hp ca EO ph
thuc vo nng EO v thi gian tip xc.
+ nng vi trm ppm, c th c th dung np c mt vi pht m
khng c tc dng ng k cho sc khe. Tuy nhin, nu tip xc lu hn c
th gy ra nhng tn thng nh kch ng mt, da, mi, c hng, phi, nh
hng n thn kinh trung ng, dn n cc triu chng nh bun ng, bun
nn, mt phng hng, co git cc chi. Nhng triu chng ny c th s mt
i trong vi ngy sau khi ngng tip xc cp tnh.
- Tip xc thng xuyn vi EO nng cao c th gy hen suyn, c
thy tinh th, hn m v t lit tr nh.
18
- IARC v NTP xp EO vo nhm tc nhn gy ung th ngi do EO c th

phn ng vi ADN trong c th to thnh cc bin th ADN khc nhau
[15].
- NIOSH xc nh nng EO nguy him ngay lp tc n tnh mng l
800 ppm [31].
1.4.3. Nhng iu cn ch v an ton lao ng khi lm vic vi EO
- US- OSHA cho php gii hn tip xc ca nhn vin lm vic vi EO c
th hin trong bng 1.4 [24].
Bng 1.4. Gii hn cho php tip xc vi EO ca US-OSHA
Hnh thc tip xc
Thi gian tip xc
Nng EO trong khng kh

(ppm)
Tip xc ngn hn
Tip xc di hn
Tip xc di hn
hn 30 ngy/ nm
15 pht
8 gi
8 gi/ngy
0,5
- H thng x l nc thi, kh thi: Sc vo b nc lnh.
19
CHNG 2
NGUYN VT LIU, TRANG THIT B V
PHNG PHP NGHIN CU
2.1. Nguyn liu
Cc ha cht v dung mi s dng trong qu trnh thc nghim c
thng k trong bng 2.1.
Bng 2.1. Cc ha cht, dung mi s dng trong qu trnh thc nghim
Stt
Nguyn liu
Xut x
Acetonitril (HPLC)
Bn mng Silica gel 60 F254
Merck c
Ethylacetat
Trung Quc
Ethylen clohydrin
Merck c
Ethanol
Vit Nam
Ethylenoxyd
Trung Quc
H3PO4 (HPLC)
Isopropanol
Trung Quc
Methanol (HPLC)
10 NaH2PO4
Trung Quc
11 NaOH
Trung Quc
13 Rutin
Cng ti C phn thng mi Dc - Vt

t Y t Khi H Thi Bnh Vit
Nam
14 Troxerutin i chiu
Trung Quc
20
2.2. Thit b v dng c

Cc my mc, dng c s dng trong qu trnh thc nghim c
thng k trong bng 2.2.
Bng 2.2. Cc my mc, dng c s dng trong qu trnh thc nghim
Stt
Tn thit b/ dng c
Xut x
B lc ht chn khng Buchner
Thy S
Cn k thut XB 320
Thy S
Cn phn tch Mettler AB 240
n soi UV sc k CN6
My ct quay chn khng Buchi
Thy S
My o pH Eutech Instrusments pH 510
My o ph cng hng t ht nhn Bruker AV-500
My o ph hng ngoi FTIR Impact
My o ph khi lng Agilent 6310 Ion Trap MS
10 My o quang ph UV-VIS 2450 Shinmadzu
Nht Bn
11 My HPLC Shimadzu SPD-M10AVP
Nht Bn
12 My khuy t gia nhit Wisd Wise stir MSH 20A
Hn Quc
13 My to kh N2 Nitrox UHP N0751
Anh
14 Nhit k thy ngn
Trung Quc
15 T hotte Unilab
Trung quc
16 T lnh
Hn Quc
17 T sy Wisd Wiseven
Hn Quc
18 Bnh cu, bnh sc k, bnh 2 c, bnh 3 c, bnh nh c

mc 25 ml, bnh nh mc 100ml, pipet, cc c m, ng
nghim, a thy tinh.
21
2.3. Ni dung nghin cu

2.3.1. Xy dng quy trnh bn tng hp troxerutin t rutin
- Thc hin phn ng bn tng hp troxerutin t rutin vi 2 tc nhn l
ethylen oxyd v ethylen clorohydrin.
- Kho st cc yu t nh hng n phn ng bn tng hp troxerutin:
+ Kho st nh hng ca nhit n hiu sut phn ng.
+ Kho st nh hng ca thi gian n hiu sut phn ng.
+ Kho st nh hng ca t l mol rutin/EO (EC) n hiu sut phn ng.
- a ra quy trnh bn tng hp troxerutin.
2.3.2. Kim nghim troxerutin
Tin hnh kim nghim hm lng troxerutin theo chuyn lun
troxerutin ca Dc in Anh 2010.
2.4. Phng php nghin cu
2.4.1. Phng php tng hp ha hc
- S dng phng php tng hp ha hc tng hp ra troxerutin da trn
nguyn tc ca phn ng O-alkyl ha.
- S dng cc phng php vt l v ha hc chit tch v tinh ch sn
phm.
- nh ga s b tinh khit ca sn phm bng SKLM v nhit nng
chy.
2.4.2. Xc nh cc thnh phn c trong sn phm
Thc hin theo chuyn lun troxerutin ca Dc in Anh nm 2010.
S dng sc k lng hiu nng cao (HPLC).
- Chun b:
22
+ Dung dch th: Ha tan 10mg cht th trong 10ml dung mi pha ng (c
th ha tan bng siu m).
+ Dung dch i chiu a: Ha tan 10mg troxerutin chun trong 10 ml dung
mi pha ng (c th ha tan bng siu m).
+ Dung dch i chiu b: Pha long 1ml dung dch i chiu a trong 10 ml
dung mi pha ng c dung dch a1, pha long tip 1 ml dung dch a1
trong 100ml dung mi pha ng c dung dch i chiu b.
+ iu kin sc k:
Ct C18 (octadecylsilyl silica gel): Chiu di ct 0,25 m, ng knh
ct 4,6 mm, ng knh ht nhi 5m.
Dung mi pha ng: Hn hp gm 20 th tch acetonitril v 80 th tch
NaH2PO4 1,56% c iu chnh pH = 4,4 bng dung dch acid
phosphoric hoc NaOH.
Tc dng 0,5 ml/ pht.
Detecter UV 350 nm.
Th tch tim 10 l.
- Tin hnh chy sc k:
Thi gian lu tng i so vi tri-(hydroxyethyl) rutin ca tetra(hydroxyethyl) rutin khong 0,5; mono-(hydroxyethyl) rutin khong 0,8; bis(hydroxyethyl) rutin khong 1,1.
trB/ trA 0,5
Vi trA: thi gian lu ca tri-(hydroxyethyl) rutin (trA = 25
trC/ trA 0,8
pht).
trD/ trA 1,1
trB: thi gian lu ca tetra-(hydroxyethyl) rutin.

trC: thi gian lu ca mono-(hydroxyethyl) rutin.
trA: thi gian lu ca bis-(hydroxyethyl) rutin.
- Tiu chun cho php

+ T l thnh phn chnh: (tri-(hydroxyethyl)-rutin): 80%.
23
+ T l cc thnh phn khc: (mono-, bis-, tetra-(hydroxyethyl)-rutin) mi

thnh phn khng qu 5%, c th chp nhn mt thnh phn khng qu 10%.
2.4.3. Xc nh cu trc sn phm
Cc cht tng hp c xc nh cu trc bng cc loi ph: Ph hng
ngoi (IR), ph khi (MS), ph cng hng t ht nhn 1 v 2 chiu (1HNMR, 13C-NMR, DEPT, COSY, HSQC, HMBC).
- Ph hng ngoi (IR) c ghi trn my o hng ngoi bin i Fourier hiu
FTIR Impact 410 bng phng php nn KBr, Vin Ha hc Vin Hn lm
Khoa hc v Cng ngh Vit Nam.
- Ph khi lng (MS) c o bng phng php ESI trn my Agilent 6310
Ion Trap MS ti Vin Ha hc - Vin Hn lm Khoa hc & Cng ngh Vit
Nam.
- Ph cng hng t ht nhn 1 chiu v 2 chiu c o trn my Bruker AM
500 FT-NMR Spectrometer, Vin Ha hc Vin Hn lm Khoa hc v Cng
ngh Vit Nam vi TMS l cht chun ni.
2.4.4. nh lng
S dng phng php o quang, tin hnh theo chuyn lun troxerutin
ca Dc in Anh 2010.
- Tin hnh:
+ Ha tan 0,200 gam mu th nh lng trong 100ml nc ct.
+ Pha long 10 ml dung dch trn vi nc ct thnh 100 ml. Tip tc pha
long 10 ml ca dung dch mi thu c vi nc ct thnh 100 ml.
+ Tin hnh o ph hp th t ngoi bc sng 350 nm.
+ So snh vi gi tr A11 = 250. Xc nh c hm lng mu th.
- Yu cu: Ch phm cha 95105 % troxerutin tnh theo ch phm lm
kh [17].
24
CHNG 3
KT QU NGHIN CU
3.1. Nghin cu quy trnh bn tng hp troxerutin

clorohydrin trn c s phng php ca Stevens
- Phng trnh phn ng
+HO-CH2-CH2 -Cl
NaOH

3.1.1.1. Kho st nhit phn ng
- Tin hnh phn ng:
Cn 6,10 gam (10 mmol) rutin cho vo bnh cu 3 c, thm vo 20 ml
nc, khuy to huyn ph v sc kh N2 lin tc. Sau cho vo hn hp
phn ng 1,2 gam (30 mmol) NaOH v nng nhit ca khi phn ng ln
550C. Thm vo khi phn ng 4,10 ml (60 mmol) EC. Sau khi cho ht tc
nhn alkyl ha, nng nhit ca phn ng ln nhit kho st v duy tr
nhit trong thi gian 2 gi.
- Kt thc phn ng: Lm lnh khi phn ng bng nc , acid ha dung
dch thu c bng HCl 10% n pH = 5. lnh 24 gi, lc hn hp phn
ng loi rutin cha phn ng thu dch lc. Ct quay chn khng dch lc
25
cho ti kit. Cn troxerutin th thu c c ha tan trong khong 80 ml

methanol si v lc nng loi mui v c. Dch lc thu c em ct quay
chn khng bc hi dung mi n khi hn hp thu c cn khong 40 ml,
kt tinh 24h. Lc thu ly tinh th, sy kh 700C.
- Kt qu kho st phn ng bn tng hp troxerutin cc nhit khc nhau
c trnh by trong bng 3.1.
Bng 3.1. Kt qu kho st s nh hng ca nhit n hiu sut phn
ng bn tng hp troxerutin vi tc nhn EC
Nhit
Khi lng
T0nc
phn ng (0C)
sn phm (gam)
ca sn phm (0C)
65
2,37
132 -134
70
3,86
150 - 152
75
5,64
153 - 154
80
5,62
153 - 154
90
5,56
152 - 153
Stt
Nhn xt:
+ nhit thp (650C), hiu sut ca phn ng thu c thp nht
(31,94%), nhit nng chy thu c cng khng ph hp vi nhit
nng chy ca cht d kin. Khi tng nhit ca phn ng ln 700C th hiu
sut ca phn ng cao hn khi thc hin 650C.
+ nhit 750C, hiu sut ca phn ng thu c cao nht l 76,01%.
+ Tip tc tng nhit ca phn ng ln 800C, 900C th hiu sut ca phn
ng gim i so vi khi thc hin phn ng 750C.
T cc d liu thu c v kt hp vi phng php sc k lp mng
vi h dung mi n-butanol/methanol/nc (10/1/3) cho thy nhit thc
26
hin phn ng ti u nht l 750C. Chnh v vy chng ti la chn nhit

thc hin phn ng l: 750C.
3.1.1.2. Kho st thi gian phn ng
Sau khi chn c nhit ti u ca phn ng l 750C, chng ti tin
hnh kho st thi gian phn ng bn tng hp troxerutin.
- Th nghim c tin hnh nh sau:
550C. Thm vo khi phn ng 4,10 ml (60 mmol) EC. Sau khi cho ht tc
nhn, nng nhit ca phn ng ln 750C v duy tr nhit trong cc
khong thi gian kho st.
- Kt thc phn ng: khi phn ng c x l nh trong mc 3.1.1.1.
- Kt qu kho st phn ng bn tng hp troxerutin cc khong thi gian
khc nhau thu c nh sau:
Bng 3.2. Kt qu kho st s nh hng ca thi gian phn ng n hiu
sut phn ng bn tng hp troxerutin vi tc nhn EC
Stt
Thi gian
phn ng (gi)
Khi lng
sn phm (gam)
T0nc
ca sn phm (0C)
2,74
132 -134
5,64
153 - 154
5,60
153 - 154
4,75
153 - 154
4,01
152 - 153
Nhn xt:
27
+ Hiu sut ca phn ng tng ln (~2 ln) sau khi tng thi gian thc hin
phn ng t 1h - 2h.
+ Khi tng thi gian phn ng ln 3h, 5h, 7h th thy rng hiu sut ca phn
ng gim dn (bng 3.2).
- T cc kt qu kho st trn, kt hp kim tra sn phm bng phng php
sc k lp mng vi h dung mi n-butanol/methanol/nc (10/1/3), chng ti
nhn thy rng thi gian thc hin phn ng ti u nht l 2h.
3.1.1.3. Kho st t l mol cc cht tham gia phn ng
Sau khi chn c nhit ti u ca phn ng l 750C, thi gian phn
ng l 2 gi, chng ti tip tc tin hnh kho st nh hng ca t l mol
EC/rutin n phn ng bn tng hp troxerutin.
- Th nghim kho st c tin hnh nh sau:
550C. Thm vo hn hp phn ng EC vi cc lng khc nhau kho st
s nh hng ca t l mol cc cht tham gia phn ng. Sau khi cho ht tc
nhn phn ng, nng nhit ca phn ng ln 750C v duy tr nhit
trong thi gian 2 gi.
- Kt thc phn ng: Hn hp phn ng c tin hnh x l nh trong mc
3.1.1.1.
- Kt qu kho st phn ng bn tng hp troxerutin vi cc t l mol cc cht
tham gia phn ng khc nhau c trnh by trong bng 3.3.
28
Bng 3.3. Kt qu kho st s nh hng ca t l mol EC/rutin n hiu sut

phn ng bn tng hp troxerutin
T0nc
Stt
Rutin
(gam)
EC
(ml)
T l
EC/rutin
Khi lng
sn phm (g)
sn phm (0C)
6,1
2,05
3/1
4,61
154 -155
6,1
2,73
4/1
5,34
156 - 158
6,1
3,42
5/1
5,41
153 - 155
6,1
4,10
6/1
5,42
158 - 159
6,1
6,15
9/1
5,49
154 - 156
Nhn xt:
+ Nhit nng chy ca cc sn phm thu c t cc th nghim kho st
cc t l mol khc nhau khng c s chnh lch ng k.
+ Vi t l EC/rutin l 3/1, hiu sut ca phn ng thu c l thp nht
(62,13%). Khi tng t l EC/rutin ln 4/1, hiu sut phn ng tng thm
khong 10% so vi t l 3/1. Tip tc tng t ln mol EC/rutin ln ln lt l
5/1; 6/1; 9/1 th hiu sut ca phn ng thu c tng ln khng ng k.
T cc kt qu kho st trn, kt hp kim tra sn phm bng phng
php sc k lp mng vi h dung mi n-butanol/methanol/nc (10/1/3)
chng ti la chn t l mol EC/rutin l 4/1 thc hin phn ng bn tng
hp troxerutin.
3.1.1.4. Kho st mt s phng php tinh ch sn phm
Sau khi la chn c cc iu kin ti u cho phn ng bn tng
hp troxerutin, chng ti tin hnh kho st mt s phng php tinh ch sn
phm th thu c, xut phng php tinh ch ti u cho quy trnh bn
tng hp troxerutin vi tc nhn alkyl ha l EC.
29
- Th nghim kho st cc phng php tinh ch c tin hnh nh sau:

550C. Nh t t 2,73 ml EC (40 mmol) vo hn hp phn ng. Sau khi cho
ht tc nhn phn ng, nng nhit ca phn ng ln 750C v duy tr nhit
trong thi gian 2h.
dch bng HCl 10% n pH = 5. lnh 24 gi, lc hn hp loi rutin thu
dch lc. Ct quay chn khng dch lc di p sut gim thu c cn
troxerutin th. Ha tan cn troxerutin thu c trong khong 80 ml methanol
si v lc nng loi mui v c thu dch lc T.
+ Phng php 1: Dch lc T thu c em ct quay chn khng loi bt
dung mi cho n khi hn hp thu c cn khong 40 ml, kt tinh trong
ngn mt t lnh. Sau 24h, lc thu ly tinh th ri sy kh trong t sy 700C
thu c 5,34 gam sn phm T1.
+ Phng php 2: Dch lc T thu c em ct quay chn khng loi bt
dung mi cho n khi hn hp thu c cn khong 40 ml, thm khong 5-7
ml isopropanol, lc nhanh thu ly dung dch, ri kt tinh trong ngn mt t
lnh. Sau 24h, lc thu ly tinh th ri sy kh trong t sy 700C thu c
4,82 gam sn phm T2.
+ Phng php 3: Dch lc T thu c em ct quay chn khng bc hi
dung mi n kit thu c cn troxerutin th. Cn troxerutin th c ha
tan trong khong 80 ml ethanol nng. Lc thu ly dung dch ct quay chn
khng loi bt dung mi cho n khi hn hp thu c cn khong 40 ml,
kt tinh ngn mt t lnh trong 24h. Lc thu ly tinh th, sy kh 700C,
thu c 5,12 gam sn phm T3.
30
+ Phng php 4: Dch lc T thu c em ct quay chn khng bc hi

dung mi n kit thu c cn troxerutin th. Cn troxerutin th c ra
trong ethyl acetat 4 ln (mi ln 20 ml). Sau lc thu ly sn phm, sy kh
700C thu 6,79 gam sn phm T4.
- Kt qu: Cc sn phm T1-T4 thu c u tan tt trong nc. Tnh cht
cc sn phm thu c t cc phng php tinh ch trn c trnh by trong
bng 3.4.
Bng 3.4. Tnh cht cc sn phm (T1-T4) thu c sau tinh ch
Stt
Cht
Cm quan
Khi
Nhit
lng
nng chy
(g)
(t0C)
T1
Bt mn, mu vng
5,34
156 - 158
T2
Bt mn, mu vng m
4,82
149 - 150
T3
Bt mn, mu vng nu
5,12
150 - 152
T4
Bt min, mu vng nu
6,79
140 - 142
Ghi ch
D ht m
D ht m
Nhn xt: Sn phm T4 c khi lng ln nht nhng khong nhit

nng chy thp, khng ph hp vi nhit nng chy ca cht d kin. T1
c khi lng ln nht trong 3 cht cn li v khong nhit nng chy ph
hp vi cht d kin. Kt hp vi vic kim tra bng phng php sc k lp
mng vi h dung mi n-butanol/methanol/nc (10/1/3), chng ti nhn thy
phng php 1 l phng php tinh ch n gin v cho sn phm c t l
cht tri(hydroxyethyl) rutin cao nht, v vy phng php tinh ch chng ti
la chn l phng php 1.
- Sn phm T1 c tin hnh chy HPLC xc nh thnh phn c th
(mc 3.1.3).
31

oxyd trn c s phng php ca F. De carvahho
- Phng trnh phn ng
NaOH
3.1.2.1. Kho st nhit phn ng

- Tin hnh phn ng:
nc, khuy to huyn ph v sc kh N2 ui ht khng kh trong bnh.
Sau cho vo hn hp phn ng 0,12 gam (3 mmol) NaOH v nng nhit
ca khi phn ng n nhit kho st. Sc t t 2,44 lt kh EO (100 mmol
EO) vo hn hp phn ng nhit kho st. Thi gian sc kh EO trong
vng 6h.
- Kt thc phn ng: Sau khi kt thc thi gian sc kh (6h), lm lnh khi
phn ng bng nc ri acid ha hn hp phn ng bng HCl 10% n
pH= 5. Sau , hn hp phn ng c ngn mt t lnh trong 24 gi ri
lc loi rutin cha phn ng. Ct quay chn khng dch lc di p sut
gim thu c cn troxerutin th. Ha tan cn troxerutin th thu c trong
khong 80 ml methanol si v lc nng loi mui v c. Dch lc thu c
em ct quay chn khng cho n khi hn hp thu c cn khong 40 ml,
32
lc ly dung dch, kt tinh 24h trong ngn mt t lnh. Sau tin hnh lc
thu ly tinh th, sy kh 700C thu c sn phm troxerutin.
- Kt qu kho st phn ng bn tng hp troxerutin vi tc nhn alkyl ha l
EO cc nhit khc nhau c trnh by trong bng 3.5.
Bng 3.5. Kt qu kho st s nh hng ca nhit n hiu sut
phn ng bn tng hp troxerutin vi tc nhn EO
Nhit
Khi lng
T0nc
phn ng (0C)
sn phm (gam)
ca sn phm (0C)
65
2,74
147 149
70
3,92
162 164
75
5,80
169 170
80
5,82
168 169
90
5,79
168 170
Stt
Nhn xt:
+ khong nhit phn ng t 65 - 700C, hiu sut phn ng thu c thp
hn so vi khong nhit phn ng t 75 - 900C.
+ Trong khong nhit phn ng t 75 - 900C, hiu sut phn ng thay i
khng ng k, nhit nng chy ca cc sn phm thu c trong cc th
nghim kho st khong nhit ny cng c s tng ng.
+ T cc d liu thu c v kt hp vi kim tra sn phm bng phng
php sc k lp mng vi h dung mi n-butanol/methanol/nc (10/1/3) cho
thy khong nhit thc hin phn ng ti u nht l 75 - 800C.
Da vo nhng kt qu nu trn, chng ti la chn khong nhit ti
u cho phn ng bn tng hp troxerutin vi tc nhn EO l 75 - 800C.
33
3.1.2.2. Kho st t l mol phn ng

Sau khi chn c nhit ti u ca phn ng l khong 75 - 800C,
chng ti tip tc tin hnh kho st nh hng ca t l mol EO/rutin n
phn ng bn tng hp troxerutin.
ca khi phn ng n 75-800C. Sc dn kh EO vo khi phn ng theo t l
mol cn tin hnh kho st, duy tr nhit phn ng 75 - 800C trong 6 gi.
- Kt thc phn ng: tin hnh x l khi phn ng nh trong mc 3.1.2.1.
- Kt qu kho st s nh hng ca t l mol EO/rutin n hiu sut to sn
phm c trnh by bng 3.6.
Bng 3.6. Kt qu kho st s nh hng ca t l mol EO/rutin n
hiu sut phn ng bn tng hp troxerutin
Rutin
EO (l)
T l
Khi lng sn
T0nc
EO/rutin
phm (g)
sn phm (0C)
Stt
(gam)
6,1
0,73
3/1
3,91
133 135
6,1
1,23
5/1
5,34
151 152
6,1
1,47
6/1
5,41
169 170
6,1
1,71
7/1
5,42
170 171
6,1
1,95
8/1
5,49
170 171
Nhn xt:
34
+ Nhit nng chy ca cc sn phm thu c t cc th nghim kho st

cc t l mol khc nhau khng c s chnh lch ng k.
+ Vi t l mol EO/rutin l 3/1, hiu sut ca phn ng thu c l thp nht
(52,69%), mt khc khong nhit nng chy ca sn phm thu c cng
ph hp vi khong nhit d kin.
+ t l mol EO/rutin tng t 5/1 n 8/1, hiu sut phn ng thu c tng
ln so vi t l 3/1, mt khc khong nhit nng chy ca sn phm thu
c cng ph hp vi khong nhit d kin.
T cc kt qu kho st trn, kt hp kim tra sn phm bng phng
php sc k lp mng vi h dung mi n-butanol/methanol/nc (10/1/3),
chng ti la chn t l mol EO/rutin l 7/1 thc hin phn ng bn tng
hp troxerutin.
3.1.2.3. Kho st mt s phng php tinh ch sn phm
Sau khi la chn c cc iu kin ti u cho phn ng bn tng
hp troxerutin, chng ti tin hnh kho st mt s phng php tinh ch sn
phm th thu c xut phng php tinh ch ti u cho quy trnh bn
tng hp troxerutin vi tc nhn alkyl ha l EO.
- Th nghim kho st cc phng php tinh ch c tin hnh nh sau:
ca khi phn ng 75-800C. Sc dn vo khi phn ng 1,71 lt (70 mmol)
kh EO. Thi gian sc kh EO trong vng 6h.
dch thu c bng HCl 10% n pH = 5. Sau lnh 24 gi, lc hn hp
35
loi rutin cha phn ng thu dch lc, ct quay chn khng cho ti kit. Cn
troxerutin th thu c c ha tan trong khong 80 ml methanol si v lc
nng loi mui v c thu dch T.
+ Dch lc T c tin hnh tinh ch theo phng php 1, 2, 3, 4 (nh trong
mc 3.1.1.4) ln lt thu c cc sn phm T5, T6, T7, T8.
- Cc sn phm T5 T8 u tan tt trong nc. Tnh cht cc sn phm thu
c t cc phng php tinh ch trn c th hin bng 3.7.
Bng 3.7. Tnh cht cc sn phm (T5-T8) thu c sau tinh ch
Stt
Cht
Cm quan
Khi
Nhit
lng
nng chy
(g)
(0C)
T5
Bt mn, mu vng
5,42
170-171
T6
Bt mn, mu vng m
4,93
154-156
T7
Bt mn, mu vng nu
5,22
154-156
T8
Bt min, mu vng nu
6,84
150-152
Ghi ch
D ht m
D ht m
Nhn xt:
+ Sn phm T8 c khi lng ln nht nhng khong nhit nng chy
thp, khng ph hp vi nhit nng chy ca cht d kin. T5 c khi
lng ln nht trong 3 cht cn li v khong nhit nng chy ph hp vi
cht d kin.
Kt hp vi vic kim tra bng phng php sc k lp mng vi h
dung mi n-butanol/methanol/nc (10/1/3), chng ti nhn thy phng
php 1 l phng php tinh ch n gin v cho sn phm c t l cht
tri(hydroxyethyl) rutin cao nht, v vy phng php tinh ch chng ti la
chn l phng php 1.
36
- Chn T5 l sn phm c cht lng tt nht (c v mt cm quan, khong

nhit nng chy v kim tra bng phng php sc k lp mng vi h
dung mi n-butanol/methanol/nc (10/1/3)), chng ti tin hnh kt tinh li
sn phm T5 trong methanol 1 ln na thu c sn phm T9. Sn phm T9
3,49 gam) thu c c dng bt mu vng sng v c nhit nng chy 180
- 1810C.
- Da vo cc kt qu nu trn, phng php tinh ch ti u chng ti la
chn cho quy trnh bn tng hp troxerutin vi tc nhn EO l phng php 1
(mc 3.1.1.4).
- Sn phm T9 c nhit nng chy ging vi tri-(hydroxyethyl) rutin c
tin hnh chy HPLC xc nh thnh phn c th (mc 3.1.3).
3.1.3. Xc nh thnh phn ca cc sn phm thu c
S dng sc k lng hiu nng cao (HPLC) xc nh s c mt v t
l cc thnh phn mono, bis, tri, tetra(hydroxyethyl)- rutin c trong sn phm
tng hp [17].
Chng ti tin hnh xc nh t l thnh phn cc dn cht trn mu th
T650C (EC), T1 v T9.
Tin hnh:
- iu kin mi trng:
+ Nhit : 280C
+ m:
70%
- iu kin sc k:
+ Pha ng: Acetonitril/ dung dch natri dihydrophosphat 15,6 g/l, iu chnh
pH v 4,4 bng acid phosphoric long (15/85).
+ Ct: C18 (150 x 4,6 mm, 5m).
+ Detector: UV 350 nm.
+Tc dng: 0,5 ml/pht.
+ Th tch tim: 5 l.
37
- Dung dch i chiu:

+ Dung dch i chiu (a): Cn 25,0 mg troxerutin chun, pha thnh 25 ml,
dung mi l pha ng, lc qua mng 0,45 m.
+ Dung dch i chiu (b): Pha long 1 ml dung dch i chiu (a) thnh 10
ml vi pha ng, sau ly 1 ml dung dch to thnh pha long trong 100 ml
pha ng.
- Dung dch th:
Ha tan chnh xc khong 25 mg ch phm trong pha ng, pha long
bng pha ng va 25 ml, lc qua mng 0,45 m.
- Chy sc k:
+ Tim dung dch i chiu (a) vo h thng.
+ Thi gian chy: hai ln thi gian lu ca thnh phn chnh ca troxerutin
(tris(hydroxyethyl)rutin). Thi gian lu tng i vi tris(hydroxyethyl)rutin
(thi gian lu l 32,7 pht): tetra(hydroxyethyl) rutin khong 0,46;
mono(hydroxyethyl) rutin khong 0,73; bis(hydroxyethyl) rutin khong 1,06.
- Kt qu thu c nh sau:
Sc k troxerutin i chiu (a):
Hnh 3.1. Sc k troxerutin i chiu (a)

Tin hnh chy dung dch th
38
Mu th T650C (EC)
Hnh 3.2. Sc k mu th T650C (EC)

Nhn xt: Mu th T650C (EC) c pic chnh l mono, chim 80%.
Ghi ch: T650C (EC): mu th bn tng hp vi tc nhn EC nhit phn ng l 650C (bng 3.1).
Mu T1
Hnh 3.3. Sc k mu th T1
Kt qu c th qu trnh chy sc k lng hiu nng cao ca mu T1
Bng 3.8. Kt qu chy HPLC ca T1

39
Stt
Tn pic
Tetrakis (hydroxyethyl)
rutin
Mono (hydroxyethyl)
rutin
Tris (hydroxyethyl)
rutin
Bis (hydroxyethyl) rutin
2
3
4
Thi gian
lu
Din tch
pic
T l %
15,003
1497550
9,38
23,793
111757
0,69
32,708
13885071
86,97
34,563
470978
2,95
Tng
15965356
Nhn xt: T kt qu chy sc k mu T1, so snh vi sc k

troxerutin i chiu hnh 3.1 c th kt lun mu T1 t tiu chun v t l
cc thnh phn troxerutin theo Dc in Anh 2010.
Mu T9
Hnh 3.4. Sc k mu th T9
Kt qu c th qu trnh chy sc k lng hiu nng cao ca mu T9
Bng 3.9. Kt qu chy HPLC ca T9
40
Stt
1
2
3
4
Tn pic
Thi gian Din tch
Tetrakis
(hydroxyethyl) rutin
T l %
lu
pic
15,148
222459
1,11
32,908
19347338
97,25
34,670
323817
1,64
Mono (hydroxyethyl)
rutin
Tris (hydroxyethyl)
rutin
Bis (hydroxyethyl)
rutin
Tng
19893614
Nhn xt:
+ T kt qu chy sc k mu T9, so snh vi sc k ca troxerutin i
chiu hnh 3.1 c th kt lun mu T9 t tiu chun v t l cc thnh phn
troxerutin theo Dc in Anh 2010.
+ Thng qua sc k khi chy HPLC v nhit nng chy ca T9 c th
khng nh mu T9 iu kin o ph khng nh cu trc sn phm.
3.1.4. Xc nh cu trc sn phm
xc nh cu trc ca cht tng hp c chng ti tin hnh ghi v
phn tch d liu ph hng ngoi (IR), ph khi lng (MS), ph cng hng
t ht nhn 1 v 2 chiu (1H-NMR,
13
C-NMR, DEPT, COSY, HSQC,
HMBC). Kt qu ph ca cc cht c trnh by c th nh sau:
41
A'
5'
6'
HOH 2 CH 2CO
1'
10
OH
A''
3'
B''
OCH 2CH 2OH
2'
6
B'
OCH 2CH 2OH
4'
2''
O HO
3''
OH 4''
OH
5''
1''
6'''
5'''
1'''
HO
4'''
6''
HO
3'''
2'''
OH
3.1.4.1. Ph hng ngoi (IR)

FT-IR (KBr) max (cm -1): 3474, 2920, 1651, 1693, 1498, 1457, 1319, 1212,
1041, 810, 524.
Ghi ch: l dao ng ha tr.
Nhn xt: cc ph IR khng c nhiu tn hiu khng nh cu trc

mt cch chc chn. Tuy nhin, da trn ph trong phn ph lc cng c
th nhn dng s b s hin din ca mt s nhm th nh OH, CO, cc
nhm th gn trn vng benzen, thng qua cc di hp th c trng.
3.1.4.2. Ph khi lng (MS)
S dng phng php o ph khi lng ch ESI-MS chng ti
thu c kt qu nh sau:
Cht
T9
CTPT
C33H42O19
KLPT
742,67
m/z (ESI-MS)
741,10[M - H]743,14[M + H]+
Nhn xt: C th nhn thy cht kho st (T9) u c gi tr pic ion

phn t ng bng s khi ca phn t d kin.
3.1.4.3. Ph cng hng t ht nhn
khng nh chc chn cu trc ca T9, chng ti tin hnh ghi ph
cng hng t ht nhn 1 chiu v 2 chiu. Ph c o trn my Bruker AM
42
500 FT-NMR Spectrometer, Vin Ha hc Vin Hn lm Khoa hc v Cng

ngh Vit Nam vi TMS l cht chun ni. Sau y l kt qu phn tch ph :
1
H-NMR (500 MHz, DMSO): (ppm) 12,50 (1H, s, OH-5); 7,84 (1H,
br s, H-2); 7,69 (1H, dd, J=1,5; 8,5 Hz, H-6); 7,12 (1H, d, J=8,5 Hz, H-5);
6,72 (1H, d, J=2,0 Hz, H-8); 6,37 (1H, d, J=2,0 Hz, H-6); 5,42 (1H, d, J=7,5
Hz, H-1); 5,37 (1H, d, J=4,5 Hz, OH); 5,16 (1H, d, J=4,5 Hz, OH); 5,12
(1H, d, J= 5,5 Hz, OH); 4,96 (1H, t, J= 5,5 Hz, OH); 4,92 (2H, q, J= 5,5 Hz, 2
x OH); 4,52 (1H, d, J= 5,0 Hz,OH); 4,43 (1H, d, J= 4,0 Hz, OH); 4,40 (1H, br
s, H-1); 4,39 (1H, d, J= 5,0 Hz, OH); 4,11 (4H, t, J= 5,0 Hz, CH2-A +
CH2-A); 4,05 (2H, m, CH2-A); 3,75 (7H, m, CH2-B + CH2-B+CH2-B +
H-6a); 3,32 (2H, m, H-3 + H-6b); 3,23 (4H, m, H-2 + H-3 + H-5 +
H-5); 3,06 (2H, m, H-4 + H-4), 0,96 (3H, d, J=6,5 Hz, CH3-6).
Ghi ch: dch chuyn ha hc (ppm), s: singlet vch n, d: doublet vch i, m:
multiplet a vch, dd: double of doublet vch i - i; br s: broad singlet vch n rng. J:
hng s tng tc (Hz).
13
C-NMR (125 MHz, CDCl3): (ppm) 177,5 (C-4); 164,7 (C-7); 160,9
(C-5); 156,6 (C-9); 156,5 (C-2); 150,9 (C-4); 147,6 (C-3); 133,7 (C-3);
122,6 (C-1); 122,4 (C-6); 114,5 (C-2); 112,8 (C-5); 105,1 (C-10); 101,3
(C-1); 100,9 (C-1); 98,4 (C-6); 92,9 (C-8); 76,4 (C-3); 76,0 (C-5);
74,2 (C-2); 71,8 (C-4); 70,6 (C-3); 70,6 (C-2); 70,5 (C-4), 70,4
(CH2-A + CH2-A), 70,2 (CH2-A); 68,3 (C-5); 67,2 (C-6); 59,6 (CH2B), 59,6 (CH2-B), 59,3 (CH2-B); 17,7 (CH3-6).
Ghi ch: l dch chuyn ha hc.
Nhn xt: Cn c vo ph phn ph lc v kt qu phn tch ph 1

chiu v 2 chiu trn chng ti nhn thy: ging nh tin cht rutin, T9 c
tn hiu ca 2 ng trong c 1 ng l glucose v 1 ng l rhamnose,
tn hiu ca 5 nhm metin vng thm, 1 nhm OH c trng cho lin kt cu
hydro ni phn t v tr C-5, 1 nhm cacbonyl v 9 cacbon bc 4. Khc vi
43
tin cht rutin, ph NMR ca T9 cn xut hin thm tn hiu ca 6 nhm

metylen gn vi oxy. iu ny ph hp vi kt qu d kin l gn c 3
nhm hydroxyethyl vo hp cht rutin.
Nh vy, t kt qu o nhit nng chy, sc k khi chy HPLC, kt
qu ph hng ngoi, ph khi, ph cng hng t ht nhn mt chiu v 2
chiu ca T9 kt hp vi mt s ti liu tham kho [42, 22] c th khng nh
chc chn cu trc ca T9 chnh l tri(hydroxyethyl) rutin. Trn c s ,
chng ti xut a ra quy trnh bn tng hp troxerutin.
3.1.5. xut quy trnh bn tng hp troxerutin
3.1.5.1. Quy trnh bn tng hp tri-hydroxyethyl-rutin vi tc nhn EO
44
Rutin 6,10 g
un nng 75 - 800C
6 gi
iu chnh pH =5
Kt tinh lnh 24h
Rutin cha phn ng
20 ml H2O
0,12 g NaOH
Kh N2
Ethylen oxyd
HCl 10%
Lc
Ct quay n kh thu
ta
Nc
Ha tan ta trong
MeOH nng
Mui v c
MeOH
Lc
Kt tinh lnh 24h
Nc ci
Lc
Kt tinh li trong
MeOH
MeOH
Lc thu ta
Sy kh 700C
Sn phm
S 1. Quy trnh bn tng hp tri(hydroxyethyl) rutin
45
Cch tin hnh

- Tin hnh
nc, khuy to huyn ph v sc kh N2 ui ht khng kh. Sau cho vo
hn hp phn ng 0,12 gam (3 mmol) NaOH v nng nhit ca khi phn
ng n 75 - 800C. Thm vo dn vo khi phn ng 1,57 lt (70 mmol) EO
v duy tr nhit 75 - 800C (thi gian sc kh EO ht khong 6 gi).
- Kt thc phn ng: Sau khi kt thc thi gian sc kh (6h), lm lnh khi
phn ng bng nc ri acid ha hn hp phn ng bng HCl 10% n pH
= 5. Sau hn hp phn ng c ngn mt t lnh trong 24 gi ri lc
loi rutin cha phn ng, ra ta bng nc lnh (2 ln x 5 ml). Dch lc
cng dch ra ta c ct quay chn khng (60 - 700C) di p sut gim ti
kit thu c cn troxerutin th. Ha tan cn troxerutin th thu c trong 80
ml methanol si v lc nng loi mui v c. Dch lc thu c em ct
quay chn khng cho n khi hn hp thu c cn khong 40 ml, lc ly
dung dch, kt tinh 24h trong ngn mt t lnh. Lc thu ta, tip tc kt
tinh li ta trong 30 ml methanol (24h) ln 2, lc thu ta, sy kh 700C thu
sn phm tri(hydroxyethyl) rutin.
Kt qu
Sn phm thu c c cc tnh cht c bn nh sau:
- Dng bt mu vng, mn, kh ti.
- Khi lng thu c: 3,49 gam.
- Nhit nng chy: 180 - 1810C.
- Tan trong nc, tan trong ethanol nng, tan trong glycerol.
46
3.1.5.2. Quy trnh bn tng hp troxerutin vi tc nhn EC

Rutin 6,10 g
Rutin cha phn ng

Nc
un nng 750C
2 gi
Ethylen
clorohydrin
(2,7 ml)
iu chnh pH =5
Kt tinh lnh 24h
HCl 10%
Lc
Ct quay n kh
thu ta
Ha tan ta trong
MeOH nng
Mui v c
20 ml H2O
0,12 g NaOH
Kh N2
MeOH
Lc
Kt tinh lnh 24h
Nc ci
Lc
Sy kh 700C
Sn phm
S 2. Quy trnh bn tng hp troxerutin
47
Cch tin hnh

nc, khuy v sc kh N2 lin tc. Sau cho vo hn hp phn ng 1,2
gam (30 mmol) NaOH v nng nhit ca khi phn ng ln 550C. Thm
vo khi phn ng 2,7 ml (40 mmol) ethylen clorohydrin. Sau khi cho ht tc
nhn phn ng, nng nhit ca phn ng ln 750C v duy tr nhit
trong thi gian 2 gi.
Kt thc phn ng: Lm lnh khi phn ng bng nc ri acid ha
hn hp phn ng bng HCl 10% n pH = 5. Sau hn hp phn ng c
ngn mt t lnh trong 24 gi ri lc hn hp phn ng loi rutin, ra
ta bng nc lnh (2 ln x 5 ml). Dch lc cng dch ra ta c ct quay
chn khng (60 - 700C) di p sut gim ti kit thu c cn troxerutin th.
Ha tan cn troxerutin th thu c trong 80 ml methanol si v lc nng
loi mui v c. Dch lc thu c em quay ct chn khng cho n khi hn
hp thu c cn khong 40 ml, lc ly dung dch, kt tinh 24h trong ngn
mt t lnh. Lc thu ta, sy kh 700C thu sn phm troxerutin.
Kt qu
Sn phm thu c c cc tnh cht c bn nh sau:
- Dng bt mu vng, mn, kh ti.
- Khi lng thu c: 5,34 gam.
- Nhit nng chy: 156 - 1580C.
- Tan trong nc, ethanol nng, glycerol.
3.2. nh lng troxerutin trong sn phm
3.2.1. Xc nh mt khi lng do lm kh
Tin hnh [17]:
- Chun b b: b c sy kh n khi lng khng i.
- Cn khong 1,0000 gam bt cho vo b, sy nhit 1050C trong 4 gi.
- Kt qu xc nh mt khi lng do lm kh ca hai mu th T1 v T9
48
Bng 3.10. Kt qu xc nh mt khi lng do lm kh ca T1 v T9

T1
1
2
3
KL b
25,8495
26,9487
26,4387
Mt khi lng do lm kh
KL bt KL b + bt KL sau sy
1,0039
26,8534
26,8158
0,9982
27,9469
27,9112
1,0006
27,4393
27,4022
T9
1
2
3
27,8374
25,5377
26,3641
1,0024
1,0056
0,9984
28,8398
26,5433
27,3625
28,7998
26,5035
27,3218
KL mt
0,0376
0,0357
0,0371
% mt
3,7454
3,5764
3,7078
3,68
0,0400
0,0398
0,0407
3,9904
3,9578
4,0765
4,01
Ghi ch: KL: khi lng (gam).
Nhn xt: Kt qu xc nh mt khi lng do lm kh cho thy mu

T9 c hm m ln hn mu T1.
3.2.2. nh lng troxerutin trong sn phm
- Tin hnh:
+ Ha tan khong 0,200 gam mu nh lng trong 100ml nc ct trong
bnh nh mc 100ml.
+ Pha long 10ml dung dch trn vi nc ct thnh 100ml. Tip tc pha
long 10ml ca dung dch mi thu c vi nc ct thnh 100 ml.
+ Tin hnh o ph hp th t ngoi bc sng 350 nm.
+ So snh vi gi tr A11 = 250.
Mi mu th lm 3 ln v ly gi tr trung bnh.
- Cng thc tnh:
+ p dng nh lut Lamber-beer
A= A1% 1cm x C% x l
Trong :
A:
hp th quang bc sng 350nm.
A1% 1cm:
hp th ring ca dung dch c nng 1% trong cuvet

c b dy 1cm.
C%:
Nng phn trm (g/100ml).
l:
B dy cuvet = 1cm.
49
+ Cng thc tnh hm lng troxerutin c trong mu th

Y% =
Ath x PL th
A1% 1cm x mth
Trong : Y%:
Hm lng troxerutin c trong mu th.
PLth:
pha long ca mu th (=10000 ln).
mth:
Khi lng cn ca mu th tnh bng gam.
+ Cng thc tnh hm lng troxerutin c trong mu th tnh theo khi lng
Y x 100
lm kh (T%):
T% =
(100 - m)
- Kt qu tin hnh th hm lng troxerutin ton phn ca hai mu th T1
v T9 c trnh by c trnh by c th trong bng 3.11 di y:
Bng 3.11. Kt qu xc nh hm lng troxerutin
T9
T1
AT9 th 1 AT9 th 2 AT9 th 3
AT9
KL cn
(g)
A11
PL
m
T%
T%
AT1 th 1 AT1 th 2 AT1 th 3
0,4857
0,4859
0,4883
0,4969
0,4977
0,4977
0,5128
0,5127
0,5130
0,5086
0,5087
0,5086
0,5020
0,5020
0,5029
0,4869
0,4870
0,4869
0,4866
0,4974
0,5128 AT1
0,5086
0,5023
0,4869
KL cn
0,2199 (g)
A11
PL
m
97,18
0,2136
250
10000
3,68
98,88
0,2106
0,2048
99,05
98,73
0,2102
250
10000
4,01
96,47
0,2104
98,55
97,42
98,89
Ghi ch: A: hp th trung bnh; T%: hm lng phn trm trung bnh; KL: khi lng.
Nhn xt: mu T1 v T9 c hm lng troxerutin trong mu th nm

trong tiu chun cho php (95 - 105%) ca Dc in Anh 2010.
50
CHNG 4
BN LUN
4.1. V quy trnh bn tng hp troxerutin

4.1.1. V phn ng bn tng hp
- Phn ng bn tng hp c thc hin trong mi trng N2 v rutin c cha
4 nhm OH phenol nn d b oxy ha. S c mt ca O2 trong khi phn
ng cng vi kim v nhit cao c th dn n phn ng phn hy, chuyn
ha rutin to ra nhiu tp.
- Phn ng tng hp troxerutin t rutin v tc nhn EC hoc EO l phn ng
th i nhn SN2. Phn ng xy ra qua cc giai on:
+ Giai on 1 to tc nhn nucleophin (phenolat) bng cch cho rutin phn
ng vi kim (NaOH).
+ Giai on 2: tc nhn nucleophin tn cng vo carbon dng in ca EC/
EO theo c ch th lng phn t.
- Khi tham gia phn ng, c cht s c hot ha thnh trung tm

nucleophin bng cht xc tc l mt kim mnh. Bn trung tm c hnh
thnh trn 4 v tr phn ng trn v c sp xp theo th t gim dn tnh
nucleophin, gim dn kh nng th: 7> 4> 3> 5. Do sn phm to thnh
c th l mt dn cht hoc mt hn hp cc dn cht sau [18]:
51
7-mono-O-(-hydroxyethyl) - rutin
7, 4-bis-O-(-hydroxyethyl) - rutin
7, 4, 3-tris-O-(-hydroxyethyl) - rutin
7, 4, 3,5-tetra-O-(-hydroxyethyl) - rutin
- S hnh thnh cc dn cht hydroxyethyl rutin ph thuc vo nhit , thi
gian phn ng, t l mol EO (EC)/rutin v s c mt ca xc tc kim.
- Hn hp vi cc t l cc thnh phn khc nhau c nhit nng chy khc
nhau v khong nhit nng chy ny l: 156 1810C.
4.1.1.1. V vai tr ca xc tc kim
S c mt ca xc tc kim l yu t quan trong trong vic to tc nhn
nucleophin, nhng nu lng NaOH qu nhiu c th dn ti phn ng thy
phn cc dn cht to thnh.
Phn ng to troxerutin tin hnh theo th t ln lt to dn cht
mono, di, tri, tetra. Nh vy, lng NaOH ban u va to phenolat
mt nhm OH vng thm, cht trung gian monophenolat ny phn ng vi
EO/EC to dn cht mono-hydroxyethyl-rutin v hon tr xc tc NaOH ban
u. Xc tc NaOH va c gii phng li tip tc tn cng vo nhm OH
vng thm th 2, v phn ng tip tc to thnh cc dn cht bis-, tri-, tetrahydroxyethyl-rutin.
- Vi tc nhn alkyl ha l EC, dng lng xc tc kim nhiu hn so vi
phn ng dng tc nhn EO do cn c kim trung ha HCl sinh ra sau
phn ng.
4.1.1.1. V nhit tin hnh phn ng
- Theo tc gi Carvalho, phn ng bn tng hp troxerutin s khng din ra
nu nh nhit phn ng di 500C [19].
52
- T d liu bng 3.1 (tc nhn EC) v bng 3.5 (tc nhn EO) nhn thy
rng:
+ khong nhit thp: sn phm thu c t phn ng alkyl ha
vi c hai tc nhn EC (650C thu 2,37 gam) v EO (650C thu 2,74 gam) c
hiu sut thp. Mt khc, khong nhit nng chy ca hn hp tng ng
thu c (132-1340C) cng khng ph hp vi khong nhit nng chy d
kin. Kt qu kim tra trn sc k lp mng vi h dung mi n-butanol/
methanol/ nc (10/1/3) cho thy t l cht tri(hydroxyethyl) rutin thp.
T kt qu ny chng ti nhn thy rng c th khong nhit t
65-700C, sn phm thu c c t l dn cht mono-, bis- chim a s. Kt
qu ny cng c khng nh trong sc k mu th T650C (EC), sc k
cho thy t l mono chim 80% (mc 3.1.3).
+ Khi nng nhit phn ng ln 75 - 800C, nhn thy hiu sut phn
ng v nhit nng chy ca sn phm c s thay i ng k. Bn cnh ,
d liu cng cho thy nhit nng chy cc sn phm thu c nm trong
khong d kin (154-1700C). Kt qu kim tra trn sc k lp mng vi h
dung mi n-butanol/methanol/nc (10/1/3) cng cho thy t l cht
tri(hydroxyethyl) rutin cao hn nhiu.
+ Tip tc nng nhit phn ng ln 90, hiu sut phn ng cng
khng tng thm v t l tri(hydroxyethyl) rutin khng c s khc bit.
iu ny c th kt lun khi duy tr nhit t 750C 800C th iu
kin phn ng l tt nht. iu ny cng ph hp vi cc ti liu tham kho
[18, 19, 35, 21].
- Tuy nhin vi tnh cht l nghin cu thm d tin ti sn xut quy m
cng nghip, nn nhit ti u chng ti chn cho quy trnh bn tng hp
53
troxerutin vi tc nhn EC l 750C v vi tc nhn EO l khong 75 - 800C

c th m bo an ton v tit kim chi ph trong sn xut hn.
4.1.1.3. V thi gian tin hnh phn ng
- Thi gian tin hnh phn ng cng nh hng n kh nng to thnh sn
phm chnh (tri- hydroxyethyl-rutin). Nu thc hin phn ng qu thi gian
ti u c th s c hin tng thy phn cc dn cht hydroxyethyl gii
phng cc phenolat ban u. Nu thc hin phn ng trong thi gian ngn
hn thi gian ti u th hiu sut to thnh sn phm chnh cng khng cao do
a phn cc sn phm to thnh l cc dn cht mono, di- hydroxyethyl-rutin.
- Nhn thy thi gian phn ng vi tc nhn EC l 2h, thi gian phn ng vi
tc nhn EO l khong 6h. L gii cho iu ny, chng ti cho rng c th vi
tc nhn EC, phn ng bn tng hp troxerutin l ng pha (lng) trong khi
vi tc nhn EO, phn ng l d pha (lng kh) nn thi gian phn ng xy
ra lu hn so vi tc nhn EC.
- Vi tc nhn EC, t d liu bng 3.2 cho thy:
Khi thi gian phn ng thay i 2h, 3h, 5h, khng c s thay i ng
k v hiu sut phn ng thu c cng nh nhit nng chy ca sn
phm.
Khi thi gian phn ng l 1h, sn phm thu c c khi lng thp
(2,74 g) v nhit nng chy thp (132-1340C), kt qu ny gn ging vi
mu th T650C (EC). iu ny cho php chng ti s b kt lun vi thi gian
phn ng < 2h, sn phm to thnh c t l dn cht mono l chnh.
Khi ko di thi gian phn ng ln 7h, khi lng sn phm thu c
c xu hng gim c th l do hin tng thy phn hoc oxy ha cc dn
cht hydroxyethyl. V vy, thi gian phn ng chng ti la chn l 2h. iu
ny cng trng vi mt s nghin cu trc y c cng b [35, 21].
54
4.1.1.4. V t l mol tc nhn alkyl ha/rutin

- T cng thc cu to ca EC v EO, nhn thy rng tc nhn EC c kh
nng tham gia phn ng SN2 mnh hn tc nhn EO trong phn ng to dn
cht hydroxyethyl. Vi tc nhn EO, phn ng bn tng hp troxerutin l d
pha nn t l mol tham gia phn ng cng cao hn do c th c hin tng
tht thot tc nhn phn ng. V vy, t l mol EC/rutin thp hn so vi t l
mol EO/ rutin.
- Vi tc nhn EC - t d liu thu c bng 3.3 cho thy:
+ Khi tng t l mol EC/rutin ln 4/1, hiu sut phn ng thu c tng ln
hn so vi t l 3/1. Khi tng t ln ny ln ln lt l 5/1; 6/1; 9/1 th hiu
sut phn ng thu c tng ln khng ng k. Mt khc sn phm thu c
t t l 4/1 c nhit nng chy ng vi nhit nng chy ca troxerutin
(1560C). Kt qu kim tra trn sc k cng cho thy hm lng ca dn
cht chnh tri(hydroxyethyl) rutin l cao nht.
+ Trong cng b ca mt nghin cu trc y [35], t l mol EC/rutin la
chn l 3/1, tuy nhin nghin cu ny cha cng b t l thnh phn cc dn
cht mono, bis, tri, tetra- to thnh.
T cc l do nu trn, t l mol chng ti xut cho phn ng bn tng
hp troxerutin l EC/rutin = 4/1.
- Vi tc nhn EO - t d liu thu c bng 3.6 cho thy:
+ t l mol EO/rutin phn ng tng t 5/1 n 8/1, khi lng sn phm thu
c tng ln so vi t l 3/1, mt khc khong nhit nng chy ca sn
phm thu c khng c s chnh lch qu nhiu v cng ph hp vi
khong nhit d kin. Cn nhc n khi lng sn phm thu c cc t
l mol 5/1; 6/1; 7/1; 8/1 v kt hp vi kim tra sn phm thu c trn sc k
lp mng, chng ti xut la chn t l mol EO/rutin l 7/1.
55
+ Theo nghin cu ca F.De Carvalho v cng s [19] th t l EO/rutin l

8/1, tuy nhin nghin cu ny cng cha cng b t l cc thnh phn dn cht
thu c. Theo nghin cu ca Jian-Dong Xu v cng s [22] th t l ny l
7.5/1; vi tinh khit c xc nh bng HPLC (Shimadzu) l 99,84%.
4.1.2. V qu trnh tinh ch sn phm sau phn ng
- Vi tc nhn alkyl ha l EC, dng methanol kt tinh thu c sn phm
c khong nhit nng chy gn vi nhit nng chy ca troxerutin hn
(~1560C), mt khc sn phm thu c c th cht mn v kh ti hn so vi
cc phng php tinh ch cn li.
- Vi tc nhn alkyl ha l EO, vic dng methanol tinh ch cng em li
nhng kt qu tch cc hn so vi cc phng php tinh ch bng dung mi
khc. Cc hp cht thu c c nhit nng chy (~1710C) cao hn so vi
cc sn phm thu c t phn ng vi tc nhn EC v gn vi nhit nng
chy ca dn cht tri-(hydroxyethyl) rutosid. S khc bit v nhit nng
chy ny c th l do dn cht tri-(hydroxyethyl) rutosid chim t l chnh
trong hn hp.
Nh vy, t cc kt qu trn, kt hp vi vic kim tra sn phm bng
sc k lp mng c th s b khng nh: vi tc nhn phn ng l EO, sn
phm thu c c t l thnh phn chnh tri-(hydroxyethyl) rutosid cao hn so
vi tc nhn EC.
- khng nh iu d on , chng ti tin hnh kt tinh li sn phm T5
(t0nc = 170-1710C) trong methanol thu c sn phm T9 (t0nc = 1801810C). Sn phm T9 v mt s sn phm khc c tin hnh chy HPLC
xc nh thnh phn.
56
4.2. V xc nh thnh phn hn hp sn phm bng HPLC

HPLC l k thut phn tch da trn c s ca s phn tch cc cht trn
mt pha tnh cha trong ct, nh dng di chuyn ca pha ng lng di p
sut cao [1]. Php th xc nh thnh phn troxerutin s dng sc k phn b
pha o (ct dng l C18).
Mu phn tch c ha tan trong pha ng. Pha ny l mt cht lng
c cho qua ct C18 mt cch lin tc. Cc cht tan l thnh phn ca mu
s di chuyn qua ct theo pha ng vi tc khc nhau ty thuc vo h s
phn b gia cht tan vi pha tnh v pha ng [1, 2]. Nh tc di chuyn
khc nhau cc thnh phn ca mu s tch ring bit thnh di, lm c s cho
php phn tch xc nh thnh phn troxerutin .
- Troxerutin i chiu c cha 82,41% tri-hydroxyethyl [ph lc 28].
- Kt qu chy sc k ca mu T9(EO), T1(EC) cho thy c s tng ng v
thi gian lu ca cc pic trong mu th vi thi gian lu ca cc pic trong
mu i chiu. iu ny cho php s b xc nh T9, T1 l troxerutin c t l
cc dn cht mono, bis, tri, tetra(hydroxyethyl)rutin khc nhau; trong sc
k ca T9 khng c s xut hin ca pic mono(hydroxyethyl) rutin.
Mu
Thi gian lu (pht)

Tetra(hydroxy Mono(hydroxy
ethyl) rutin
ethyl) rutin
Tri(hydroxyl
ethyl) rutin
Bis(hydroxy
ethyl) rutin
Troxerutin
i chiu
15,034
23,898
32,738
34,678
T1
15,003
23,793
32,708
34,563
T9
15,148
32,908
34,670
- V t l cc thnh phn c trong hn hp cng c bo co trong sc k ,

c th nh sau:
57
Mu
T1
T9
Din tch pic (t l %)

tetra(hydroxy mono(hydroxy tri(hydroxyl
bis(hydroxy
ethyl) rutin
ethyl) rutin
ethyl) rutin
ethyl) rutin
1497550
111757
13885071
470978
(9,38%)
(0,69%)
(86,97%)
(2,95%)
222459
19347338
323817
(1,11%)
(97,25%)
(1,64%)
Tng
15965356
19893614
T kt qu chy HPLC cho thy mu T9 thnh phn chnh

tri(hydroxyethyl)rutosid chim 97,25%, mu T1 t l ny l 86,97%. Kt
qu ny cho thy sn phm tng hp c t quy trnh vi tc nhn EC v EO
u t t tiu chun v t l cc thnh phn theo Dc in Anh 2010.
Trong mt s nghin cu trc y th t l ny l 88% [20] v 99,84% [22].
Sn phm T9 cng c tin hnh kim nghim xc nh thnh phn
cc dn cht ti Trung tm kim nghim nghin cu dc Cc Qun Y
Vit Nam. Kt qu cho thy T9 c pic chnh chim 97,58 % (ph lc 25).
T cc kt qu trn chng ti nhn thy rng sn phm T9 (vi t l pic
chnh chim 97,58%, nhit nng chy 180-1810C) iu kin o ph
xc nh cu trc sn phm.
4.3. V xc nh cu trc sn phm chnh
4.3.1. V ph hng ngoi
Ph IR ca T9 cho nh hp th c trng ca cc nhm chc
max3474 cm-1 (OH) v 1651 cm-1 (C=O). Tuy nhin, khng nh chc chn
cu trc ca cc cht th cc d liu ca ph khi lng v ph cng hng t
ht nhn mi thc s c ngha quyt nh.
58
4.3.2. V ph khi lng

Ph khi lng ng vai tr quan trng vic xc nh khi lng phn
t cc cht tng hp v thng l loi ph c la chn o u tin kim
chng xem sn phm phn ng c to thnh c l cht d kin hay khng.
Ty theo ch o, trn ph ca T9 xut hin pic ion c s khi
bng [M+H]+ (ch ESI (+)) hoc [M-H]- (ch ESI (-)). Pic ion phn t
ca cc hp cht hu c ni chung khng phi l vch ring l m l mt cm
pic v cc nguyn t cha trong hp cht u tn ti cc ng v khc nhau.
Bi vy, bn cnh vch chnh ng vi pic ion phn t M+ cn c cc vch
[M+1]+, [M+2]+,... vi cng ph hp quy tc a thc ng v [28].
Cht T9 c cng thc phn t d kin l C33H42O19. tng ng vi s
khi 742,67. Trn ph ESI-MS negative xut hin pic ion [M-H]- cng
ln nht c s khi 741,10 (hnh 4.2), ph ESI-MS positive xut hin pic ion
[M+H]+ cng ln nht c s khi 743,14. Nh vy, s b cho thy cht
T9 c s khi ng nh s khi d kin.
Pic
M-1
Pic M
Pic
M+ 1
Hnh 4.1. Ph ESI-MS negative ca cht T9

Trn ph ESI-MS positive cn cho pic ion m/z 597 [M-rhamnose +
H]+ v pic ion m/z 435 [M-glucose-rhamnose + H]+.
59
4.3.3. V ph cng hng t ht nhn

Ph NMR l phng php hin i trong vic phn tch cu trc cc hp
cht ha hc, da trn nguyn tc cng hng ca cc ht nhn ca cc nguyn
t khi c t trong mt t trng. Trong ph NMR c hai thng s c trng
lin quan n cu trc ha hc ca mt phn t l dch chuyn ha hc v
hng s tng tc spin spin J. T cc d liu phn tch cc ph 1 chiu v 2
chiu NMR cho php xy dng cu trc phn t ca mu phn tch.
D liu ph cng hng t ht nhn ca cc cht c trnh by trong
mc 3.1.3.3 v ph trong phn ph lc. Sau y l mt s bn lun da
trn s liu ph cng hng t ht nhn ca T9.
Ph 13C-NMR v DEPT ca T9 rt ging vi tin cht rutin, ch khc l
thy xut hin thm 6 nhm metylen gn vi oxy nm trong khong C 59,370,4. Ngoi ra trn ph
13
C-NMR cn c tn hiu ca 1 nhm cacbonyl
C177,5(C-4); 5 nhm metin vng thm C122,4 (C-6); 114,5 (C-2); 112,8
(C-5); 98,4 (C-6) v 92,9 (C-8); 1 nhm metylen gn vi oxy C 67,2 (C6); 1 nhm metyl C 17,7 (CH3-6); 10 nhm metin sp3 v 9 cacbon bc 4.
C8
C2
C5
C5
C2
C7
C4
C2
C6
C5
C6
C8
Hnh 4.2. Ph 13C-NMR v DEPT ca T9

60
Ph 1H-NMR ca T9 cng cho thy nhiu hn ph 1H-NMR ca rutin

6 nhm metylen gn vi oxy nm trong khong H 3,75-4,11.
CH B + CH B + CH B + H-6a
2
CH A + CH A
2
CH2 A
Hnh 4.3. Ph 1H-NMR gin rng vng t 3,04 4,96 ppm ca T9

Cng ging nh rutin, trn ph 1H-NMR ca T9 xut hin tn hiu ca
1 nhm OH H 12,50 c trng cho lin kt cu hydro ni phn t v tr C5, 3 proton vng thm tng tc kiu ABX H 7,84 (1H, br s, H-2); 7,69
(1H, dd, J=1,5; 8,5 Hz, H-6); 7,12 (1H, d, J=8,5 Hz, H-5) v 2 proton vng
thm tng tc dng meta H 6,72 (1H, d, J=2,0 Hz, H-8); 6,37 (1H, d,
J=2,0 Hz, H-6). Ngoi ra cn c tn hiu ca 2 proton anomeric thuc 2
ng glucose v rhamnose H 5,42 (1H, d, J=7,5 Hz, H-1) v 4,40 (1H,
br s, H-1), tn hiu ca 1 nhm metyl thuc ng rhamnose H 0,96 (3H,
d, J=6,5 Hz, CH3 - 6) v tn hiu ca 8 nhm metin, 1 nhm metylen ca 2
ng nm trong khong H 3,06 - 4,40. Da vo cc d liu ph trn cho
php xc nh hp cht T9 c gn 3 nhm hydroxyethyl.
61
H8
H6
H2
H5
H1
H6
Hnh 4.4. Ph 1H-NMR gin rng vng t 5,11 7,84 ppm ca T9

Trn ph COSY phn ng cho thy h tng tc ca mt n v
ng glucose H-1/H-2/H3/H-4/H-5/ CH2-6 v mt n v ng
rhamnose H-1/H-2/H3/H-4/H-5/ CH3-6. ng thi tng tc
gia cc nhm metylen CH2-A/CH2-B, CH2-A/CH2-B v CH2-A/CH2-B
cng c quan st thy trn ph COSY.
Hnh 4.5. Ph COSY gin rng ca T9

62
Ging nh tin cht rutin, ng glucose gn vi khung flavonoit v

tr C - 3 v ng rhamnose gn vi glucose v tr C - 6 c xc nh da
vo tng tc xa trn ph HMBC: proton H-1 tng tc vi C- 3, C-2, C3; proton H-1 tng tc vi C-6 v CH2-6 vi C-1.
Hnh 4.6. Ph HMBC ca T9
Hnh 4.7. Mt s tng tc chnh trn ph COSY (
63
) v HMBC () ca T9
Kt hp cc d liu ph IR, MS, 1H-NMR, 13C-NMR, DEPT, COSY,

HSQC, HMBC, v so snh vi ti liu tham kho [42,22] cho php xc nh
cht T9 chnh l 2-[3,4-bis(2-hydroxyethoxy) phenyl]-5-hydroxy-7-(2hydroxyethoxy)-4-oxo-4H-chromen-3-yl 6-O-(6-deoxy--D-mannopyranosyl)
--D-glucopyranoside.
4.4. V xc nh hm lng troxerutin
Hm lng troxerutin c trong mu th T1 v T9 c xc nh theo
phng php o quang ph hp th t ngoi bc sng 350 nm [17].
Kt qu o c tng hp trong bng 3.10 v bng 3.11 cho thy sn
phm T1 v T9 c hm lng troxerutin tnh theo ch phm lm kh nm
trong tiu chun cho php (95 - 105%) ca Dc in Anh 2010.
Mu T1 v T9 cng c kim nghim xc nh hm lng
troxerutin ti Trung tm kim nghim nghin cu dc Cc Qun Y Vit
Nam, kt qu cho thy T1 t 98,8%, T9 t 96,7% hm lng troxerutin
[ph lc 26, 27].
64
KT LUN V XUT
1. KT LUN
V xy dng quy trnh bn tng hp troxerutin
xy dng c quy trnh bn tng hp troxerutin t rutin vi 2 tc
nhn alkyl ha l EO v EC quy m phng th nghim.
- Bn tng hp troxerutin vi tc nhn EO theo quy trnh xy dng: T 6,10g
rutin ban u, qua quy trnh bn tng hp thu c 3,49 gam T9, l dng
bt mn mu vng, nhit nng chy 180-1810C, thnh phn chnh
tris(hydroxyethyl)rutosid chim 97,58%; kt qu bin gii ph cho php xc
nh T9 chnh l tris(hydroxyethyl)rutin.
- Bn tng hp troxerutin vi tc nhn EO theo quy trnh xy dng: T 6,10g
rutin ban u, qua quy trnh bn tng hp thu c 5,34 gam T1, l dng
bt mn mu vng, nhit nng chy 156-1580C. Kt qu chy HPLC cho
php s b khng nh T1 l troxerutin vi t l cc thnh phn t tiu chun
BP 2010(pic chnh tris(hydroxyethyl)rutosid: 86,97 %)
Quy trnh BTH troxerutin vi tc nhn EC d dng thc hin hn so
vi quy trnh BTH t tc nhn EO. Tuy nhin, sn phm thu c t quy trnh
BTH troxerutin vi tc nhn EO c t l tri(hydroxyethyl) rutin cao hn.
V nh lng sn phm troxerutin thu c
Sn phm thu c c hm lng troxerutin trong T1 t 98,8%, T9
t 96,7% - t tiu chun hm lng theo Dc in Anh 2010.
2. XUT
- Nghin cu cc phng php tinh ch sn phm thu c t tiu chun
Dc in Anh 2010.
- Tip tc nghin cu kho st cc yu t nh hng n quy trnh bn tng
hp troxerutin quy m bn pilot v pilot.
65
TI LIU THAM KHO
TING VIT
1. Trn T An (2007), Kim nghim dc phm, Nh xut bn y hc, tr
84-110.
2. Trn T An, Thi Nguyn Hng Thu (2006), Ha phn tch II, Nh
xut bn y hc, tr 123.
3. on Th Bnh (2009), Nghin cu chit xut Rutin t hoa he bng
phng php siu m, Kha lun tt nghip dc s i hc kha 20042009, tr 8.
4. B mn Cng nghip Dc (2009), Thc tp k thut sn xut Dc
phm, Trng i hc Dc H Ni, tr 62.
5. B mn Dc liu (2006), Bi ging dc liu I, Trng i hc Dc
H Ni, tr 290-293, 261.
6. B Y t, Dc in Vit Nam III (3)
7. B Y T, Dc in Vit Nam IV, tr 555-557.
8. Nguyn Th Xun Hiu (2009), Nghin cu tng hp troxerutin, Kha
lun tt nghip dc s i hc kha 2004 -2009, Trng i hc Dc
H Ni.
9. Hi tim mch hc Vit Nam (2010), Khuyn co 2010 v cc bnh l
tim mch & chuyn ha, NXB Y hc chi nhnh TP. H Ch Minh, tr 9.
10. Tt Li (1999), Nhng cy thuc v v thuc Vit Nam, NXB Y
hc, tr 298-299.
11. V Th Thy (2005), La chn mt s thng s trong quy trnh chit
xut rutin t hoa he, Kha lun tt nghip Dc s i hc kha 20002005, Trng i hc Dc H Ni.
12. Trn L Thy Vn (2010), nh gi kt qu iu tr bnh tc tnh

mch trung tm vng mc ti bnh vin mt trung ng trong 5 nm,
Lun vn Thc S Y hc, Trng i Hc Y H Ni.
13. Vin dc liu (2004), Cy thuc v ng vt lm thuc Vit Nam,
NXB Khoa hc k thut, Tp 1, tr 971- 972.
TING ANH
14. Adam B.S. et al (2005), Troxerutin protects the isolate perfused rat
liver from a possible lipid peroxidation by coumarin, Phytomedicine,
12 (1-2), p. 52-61.
15. American Chemistry Council (2007), Product stewardship guldance
manual- Ethylene oxide.
16. Boccalon H. et al (1997), Characteristics of chronic venous
insufficiency in 895 patiens followed in general practice, Int
Angiology, 16(4), p. 226-234.
17. British Pharmacopoeia (2010), vol 2, p. 2169.
18. Courbat P. J et al (1969), Quercertin and quercertin glycoside,
United States Patent Office, 3420815.
19. De carvalho Levy Rebelo F. et al (2005), The production of tri hydroxyethylrutin (troxerutin) in a pilot plant, 2nd Mercosur Congress
on Chemical Engineering, p.1-9.
20. Estanove C. et al (2005), Troxerutin with a high content of
trihydroxyethylrutoside and process for its preparation, US 6,855,697
B1.
21. Frantisek Czech et al (1982), Mixture of O-hydroxyethylated rutin
derivative, CS, 225440.
22. Jian-Dong Xu et al (2013), Synthesis and antioxidant activities of

flavonoids derivative, troxerutin and 3,4,7-triacetoethoxyquercetin,
Chinese Chemical Letters, 24, p. 223-226.
23. Jun Lu et al (2010), NGF-Dependent Activation of TrkA Pathway: A
Mechanism for the Neuroprotective Effect of Troxerutin in DGalactose-Treated Mice, Brain Pathology, 20, p. 952-965.
24. Jun Lu et al (2012), Troxerutin Counteracts Domoic acid- Induced
Memory Deficits in Mice by inhibiting CCAAT/ Enhancer Binding
Protein - Mediated Inflammatory Response and Oxidative Stress, The
Journal of Immunology 1202862.
25. Leelavinothan Pari, Perumal Elangovan (2013), Troxerutin protects
nickel-induced alterations in lipids and plasma lipoprotein in rats,
International Journal of Basic and Life Sciences, 1, p. 39-49.
26. Lu. J et al (2011), Troxerutin protects against high cholesterolinduced cognitive deficits in mice Brain - Oxford Journals, 134(3),
p.783-797.
27. Marzin D. et al (1987), Study of mutagenic activity of troxerutin, a
flavonoid derivative, Toxicol Letter, 32 (2-3), p. 297-305.
28. Mc Lafferty F.W., Turecek F. (1993), Interpretation of mass spectra,
University science books, 4th edition, USA.
29. Merck & Co.Inc. (2006), The Merck index, published by Merck & Co.
Inc., 14th edition.
30. New Jersey Department of Health and Senior Services (1999), Ethylen
chlorohydrin, New Jersey.
31. NIOSH Chemical Listing and Documentation of Revised IDLH Values
(1994), Documentation for Immediately Dangerous to Life or Health
Concentrations (IDLH).
32. Shao-hua Fan et al (2009), Troxerutin protects the mouse kidney from
d-galactose-cause injury through anti-inflammation and anti
oxidation, International imhumopharmacology, 9, p.91-96.
33. Siegers C.P. et al (2008), Aescin and troxerutin as a successful
combination for the treatment of inner ear perfusion disturbances,
Phytomedicine, 15(3), p. 160-163.
34. Squadrito F. et al (2000), Double-blind, randomized clinical trial of
troxerutin-carbazochrome inpatients with hemorrhoids, European
Review for Medical and Pharmacological Science, 4, p.21-24.
35. Stevens A. et al (1958), A tri- (Hydroxyethyl) Ether of rutin and
process for the production thereof, GB, 833174.
36. Universitaire Pers Maastricht (2011), The antioxidant flavonoid 7mono- O- (-hydroxyethyl)-rutoside, from clinic to concept, Maastricht
- Dutch.
37. US Environmental
Protection Agency
(2008),
Acute
exposure
guideline levels Ethylene chlorohydrin (2- chloroethanol), USA.

38. USA Occupational Safety and Health Administration (2002),
Ethylenoxyd, OSHA fact sheet.
39. USA Occupational Safety and Health Administration (2013), Ethylen
chlorohydrin- OSHA 07, OSHA fact sheet.
40. Vladimir Czech et al (1982), Isolation of O--hydroxyethyl rutosids,
CS, 199923.
41. Xu Ping et al (2011), Radioprotective Effects of Troxerutin against
Gamma Irradiation in V79 Cells and Mice, Asian Pacific J Cancer
Prev, 12, p. 2593-2596.
42. Yong Mei Xiao et al (2010), Regioselective enzymatic acylation of
troxerutin in nonaqueous medium, Chinese Chemical Letters, 21, p.
59-62.
PH LC
Ph lc 1
Ph IR ca T9
Ph lc 2
Ph ESI-MS negative ca T9
Ph lc 3
Ph ESI-MS positive ca T9
Ph lc 4
Ph 1H-NMR ca T9
Ph lc 5
Ph 1H-NMR gin rng ca T9
Ph lc 6
Ph 1H-NMR gin rng ca T9
Ph lc 7
Ph 13C-NMR ca T9
Ph lc 8
Ph 13C-NMR gin rng ca T9
Ph lc 9
Ph 13C-NMR gin rng ca T9
Ph lc 10
Ph DEPT ca T9
Ph lc 11
Ph DEPT gin rng ca T9
Ph lc 12
Ph COSY ca T9
Ph lc 13
Ph COSY gin rng ca T9
Ph lc 14
Ph lc 15
Ph lc 16
Ph HSQC ca T9
Ph lc 17
Ph HSQC gin rng ca T9
Ph lc 18
Ph HSQC gin rng ca T9
Ph lc 19
Ph HMBC ca T9
Ph lc 20
Ph HMBC gin rng ca T9
Ph lc 21
Ph lc 22
Ph lc 23
Sc k cht T1
Ph lc 24
Sc k cht T9
Ph lc 25
Phiu kim nghim thnh phn troxerutin
Ph lc 26
Phiu kim nghim hm lng troxerutin T1
Ph lc 27
Phiu kim nghim hm lng troxerutin T9
Ph lc 28
Phiu chng nhn phn tch ca troxerutin i chiu
A'
5'
HOH 2 CH 2CO
1'
10
OH
3
4
A''
3'
B''
OCH 2CH 2OH
2'
6
B'
OCH 2CH 2OH
4'
6'
2''
HO
3''
OH
OH
5''
1''
6'''
6''
5'''
1'''
HO
4'''
4''
HO
3'''
2'''
OH
Ph lc 1: Ph IR ca T9
Ph lc 2: Ph ESI MS negative ca T9
Ph lc 3: Ph ESI MS positive ca T9
A'
5'
6'
HOH 2 CH 2CO
1'
10
OH
3
4
A''
3'
B''
OCH 2CH 2OH
2'
6
B'
OCH 2CH 2OH
4'
2''
HO
3''
OH
OH
5''
1''
6'''
6''
5'''
1'''
HO
4'''
4''
HO
3'''
2'''
OH
Ph lc 4: Ph 1H-NMR ca T9
A'
5'
6'
HOH 2 CH 2CO
1'
10
OH
3
4
A''
3'
B''
OCH 2CH 2OH
2'
6
B'
OCH 2CH 2OH
4'
O HO
2''
3''
OH
OH
5''
1''
6'''
6''
5'''
1'''
HO
4'''
4''
HO
3'''
2'''
OH
Ph lc 5: Ph 1H-NMR gin rng ca T9
A'
5'
HOH 2 CH 2CO
1'
10
OH
3
4
A''
3'
B''
OCH 2CH 2OH
2'
6
B'
OCH 2CH 2OH
4'
6'
O HO
2''
3''
OH 4''
OH
5''
1''
6'''
5'''
1'''
HO
4'''
6''
HO
3'''
2'''
OH
Ph lc 6: Ph 1H-NMR gin rng ca T9
A'
5'
6'
HOH 2 CH 2CO
1'
10
OH
3
4
A''
3'
B''
OCH 2CH 2OH
2'
6
B'
OCH 2CH 2OH
4'
O HO
2''
3''
OH 4''
OH
5''
1''
6'''
5'''
1'''
HO
4'''
6''
HO
3'''
2'''
OH
Ph lc 7: Ph 13C-NMR ca T9
A'
5'
4'
6'
HOH 2 CH 2CO
1'
10
OH
3
4
A''
3'
B''
OCH 2CH 2OH
2'
6
B'
OCH 2CH 2OH
O HO
2''
3''
OH
OH
5''
1''
6'''
6''
5'''
1'''
HO
4'''
4''
HO
3'''
2'''
OH
Ph lc 8: Ph 13C-NMR gin rng ca T9
A'
5'
6'
HOH 2 CH 2CO
1'
10
OH
3
4
A''
3'
B''
OCH 2CH 2OH
2'
6
B'
OCH 2CH 2OH
4'
2''
HO
3''
OH
OH
5''
1''
6'''
6''
5'''
1'''
HO
4'''
4''
HO
3'''
2'''
OH
Ph lc 9: Ph 13C-NMR gin rng ca T9
Ph lc 10: Ph DEPT ca T9
Ph lc 11: Ph DEPT gin rng ca T9
A'
5'
HOH 2 CH 2CO
1'
10
OH
3
4
A''
3'
B''
OCH 2CH 2OH
2'
6
B'
OCH 2CH 2OH
4'
6'
2''
HO
3''
OH
5''
1''
6'''
6''
5'''
1'''
HO
4'''
4''
OH
HO
3'''
2'''
OH
Ph lc 12: Ph COSY ca T9
A'
5'
HOH 2 CH 2CO
1'
10
OH
3
4
A''
3'
B''
OCH 2CH 2OH
2'
6
B'
OCH 2CH 2OH
4'
6'
O HO
2''
3''
OH
OH
5''
1''
6'''
6''
5'''
1'''
HO
4'''
4''
HO
3'''
2'''
OH
Ph lc 13: Ph COSY gin rng ca T9
A'
5'
6'
HOH 2 CH 2CO
1'
10
OH
3
4
A''
3'
B''
OCH 2CH 2OH
2'
6
B'
OCH 2CH 2OH
4'
2''
HO
3''
OH
OH
5''
1''
6'''
6''
5'''
1'''
HO
4'''
4''
HO
3'''
2'''
OH
A'
5'
4'
6'
HOH 2 CH 2CO
1'
10
OH
3
4
A''
3'
B''
OCH 2CH 2OH
2'
6
B'
OCH 2CH 2OH
2''
HO
3''
OH
OH
5''
1''
6'''
6''
5'''
1'''
HO
4'''
4''
HO
3'''
2'''
OH
A'
5'
6'
HOH 2 CH 2CO
10
3
4
8
OH
O
9 O
O
6'
1'
A''
3'
B''
OCH 2CH 2OH

A'
2'
5'
HOH 2 CH 2CO
1'
B'
OCH 2CH 2OH
4'
B'
2''
CH 24''
OH
2OH
4' OCH
3''
HO
OH
O
A'' B''
3'
6''
5''
O
1'' OCH CH OH
2
2
2'
6
10
OH
3
4
2''
HO 6'''
HO
1''
5'''
3''
O
4''' HO
6'''
3'''5''
5'''
4''
1'''
OH
6'' 2'''
O
OH
1'''
HO
4'''
OH
O
HO
3'''
2'''
OH
Ph lc 16: Ph HSQC ca T9
A'
5'
6'
HOH 2 CH 2CO
1'
10
OH
3
4
A''
3'
B''
OCH 2CH 2OH
2'
6
B'
OCH 2CH 2OH
4'
O HO
2''
3''
OH
OH
5''
1''
6'''
6''
5'''
1'''
HO
4'''
4''
HO
3'''
2'''
OH
Ph lc 17: Ph HSQC gin rng ca T9
A'
5'
6'
HOH 2 CH 2CO
1'
10
OH
HOH 2 CH 2CO
3
4
O
O
B''
OCH 2CH 2OH

2''
A'3'' OH
B' 4''
OH
OCH
CH
2
2OH
O
6''
5''
O
1'' 3'
A'' B''
OCH5'''2CH 2OH
O HO
5'
4'
6'
2
A''
3'
2'
B'
OCH 2CH 2OH
4'
1'
2'
6'''
6
10
OH
3
4
O HO
HO
2''
4'''
3''
O
HO
OH
O
OH
3'''
2'''
5''
1''
6'''
6''
OHO
5'''
1'''
HO
4'''
1'''
4''
HO
3'''
2'''
OH
A'
5'
6'
HOH 2 CH 2CO
1'
10
OH
3
4
A''
3'
B''
OCH 2CH 2OH
2'
6
B'
OCH 2CH 2OH
4'
O HO
2''
3''
OH 4''
OH
5''
1''
6'''
5'''
1'''
HO
4'''
6''
HO
3'''
2'''
OH
Ph lc 19: Ph HMBC ca T9
Ph lc 19: Ph HMBC ca T9
A'
5'
4'
6'
HOH 2 CH 2CO
1'
10
HOH 2 CH 2CO
OH
3
4
O
O
6'
10
OH
3
4
B''
A'2CH
OCH
B'2OH
OCH
CH
2OH 24''OH
4'
O HO
1'
2''
3'
2' 1''
6
A''
3'
5'
2'
B'
OCH 2CH 2OH
3''
OH
A'' B''
O
6''
OCH 2CH
OH
5'' 2
O
O HO
6'''
2''
HO1''
4'''
3''
5'''
OH 4''
HO
6'''
5''
6''
2'''
OH
5'''
1'''
HO
4'''
3'''
OH
1'''
HO
3'''
2'''
OH
Ph lc 20: Ph HMBC gin rng ca T9
A'
5'
HOH 2 CH 2CO
1'
10
OH
3
4
A''
3'
B''
OCH 2CH 2OH
2'
6
B'
OCH 2CH 2OH
4'
6'
O HO
2''
3''
OH
OH
5''
1''
6'''
6''
5'''
1'''
HO
4'''
4''
HO
3'''
2'''
OH
A'
5'
6'
HOH 2 CH 2CO
1'
10
OH
3
4
A''
3'
B''
OCH 2CH 2OH
2'
6
B'
OCH 2CH 2OH
4'
O HO
2''
3''
OH
OH
5''
1''
6'''
6''
5'''
1'''
HO
4'''
4''
HO
3'''
2'''
OH
Ph lc 23: Sc k cht T1
Ph lc 24: Sc k cht T9
Ph lc 25: Phiu kim nghim thnh phn troxerutin
Ph lc 26: Phiu kim nghim hm lng troxerutin/ T1
Ph lc 27: Phiu kim nghim hm lng troxerutin/ T9
Ph lc 28: Phiu chng nhn phn tch ca troxerutin i chiu

Nghiên C U Bán T NG H P Troxerutin T Rutin

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Nghiên C U Bán T NG H P Troxerutin T Rutin

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NGUYN TH HONG ANH

CHUYN NGNH CNG NGH DC PHM

NGUYN TH HONG ANH

Xin chn thnh cm n!

Nguyn Th Hong Anh

2.4.2. Xc nh cc thnh phn c trong sn phm............................................... 22

DANH MC CC K HIU, CH VIT TT

Distortionless Enhancement by Polarization Transfer

Ph khi lng phun m in t

Lipoprotein t trng cao

Heteronuclear Multiple Bonds Correlation

Ph cng hng t ht nhn proton

(Proton Nuclear Magnetic Resonance Spectroscopy)

Sc k lng hiu nng cao

C quan quc t nghin cu ung th

(International Union of Pure and Applied Chemistry)

Lipoprotein t trng thp (Low Density Lipoprotein)

(National Institute for Occupational Safety and Health- United

Chng trnh c cht hc quc gia M

n v phn triu (parts per million)

Phn ng th i nhn lng phn t

T ngoi kh kin (Ultraviolet Visible)

Lipoprotein t trng rt thp (Very Low Density Lipoprotein)

Sc k troxerutin i chiu (a) .......................................................... 38

Sc k mu th T650C (EC) ............................................................. 39

Ph ESI-MS negative ca cht T9......................................................... 59

Ph 13C-NMR v DEPT ca T9 ........................................................... 60

Ph 1H-NMR gin rng vng t 3,04 4,96 ppm ca T9 .................... 61

Ph 1H-NMR gin rng vng t 5,11 7,84 ppm ca T9 .................... 62

Ph COSY gin rng ca T9 ................................................................. 62

Mt s tng tc chnh trn ph COSY ( ) v HMBC () ca T9 ... 63

Quy trnh bn tng hp tri(hydroxyethyl) rutin..................................... 45

Quy trnh bn tng hp troxerutin ......................................................... 47

BnTuy nhin, hin nay Vit Nam vn cha sn xut c troxerutin,

CHNG 1: TNG QUAN

N hoa he kh Nguyn liu

- Rutin c hu ht trong cc phn xanh ca cy he nhng hm lng cao

- Cng thc phn t: C33H42O19.

- Troxerutin kt hp vi Aescin trong iu tr bnh tai trong. Hiu qu ca

c dng iu tr lu di v d phng tai bin mt th hai. Troxerutin ci

- Trong nghin cu ca Jun Lu cng cng s (2012) cho thy troxerutin c

Cao G. biloba 14mg TNHH dc phm

Cao G. biloba 14mg

Cao G. biloba 14mg

Cng ty lin doanh

khu Vin nang

Trn th gii troxerutin c s dng vi nhiu dng bit dc, cc dng

Cao ginkgo biloba

1.2.4. Cc phng php bn tng hp troxerutin t rutin

Phng php Stevens

Phng php Frantisek Czech

Phng php ca Pierre J Courbat

- Tn khoa hc: 2 cloroethanol.

Bng 1.3. Cc cp tip xc vi ethylen clorohydrin

Gy tn thng cho c th.

- Nm 1859, Wurtz tng hp c EO u tin bng cch s dng KOH

- Polyme ha: EO d b polymer ha vi s xc tc ca nhit , thi gian

- IARC v NTP xp EO vo nhm tc nhn gy ung th ngi do EO c th

Thi gian tip xc

Nng EO trong khng kh

- H thng x l nc thi, kh thi: Sc vo b nc lnh.

Bn mng Silica gel 60 F254

Cng ti C phn thng mi Dc - Vt

2.2. Thit b v dng c

B lc ht chn khng Buchner

Cn phn tch Mettler AB 240