Professional Documents
Culture Documents
Cay đỏ ngọn
Cay đỏ ngọn
TRNG I HC S PHM
---------------------------------
BI VN BNH
NGHIN CU HO HC V
NHN DNG MT S NHM CHT
C TRONG CY NGN
(CRATOXYLUM PRUNIFOLIUM KURTZ)
LUN V N TH C S HO HC
I HC THI NGUYN
TRNG I HC S PHM
---------------------------------
BI VN BNH
NGHIN CU HO HC V
NHN DNG MT S NHM CHT
C TRONG CY NGN
(CRATOXYLUM PRUNIFOLIUM KURTZ)
Chuyn ngnh: Ho hu c
M s: 60.44.27
LUN V N TH C S HO HC
SKLM
Sc k lp mng
UV
Ultraviolet spectrocopy
MS
Mass Spectroscopy
EI-MS
LC-MS
FT-IR
NMR
H-NMR :
13
C-NMR :
13
DEPT
COSY
Correlated Spectroscopy
HSQC
HMBC
HIV
Unit
http://www.lrc-tnu.edu.vn
M U
Vit Nam nm trong vng nhit i, gi ma, ma thun gi ho nn
h thc vt rt phong ph, a dng. l ngun ti nguyn sinh hc qu gi
thuc loi c th ti to c. Ngay t xa xa cha ng ta bit khai thc
ngun ti nguyn qu bu ny lm n, thc phm, thuc cha bnh, vt
liu xy dng v nhin liu phc v cuc sng.
Trong th gii thc vt y c nhng loi cung cp thc n cho chng ta,
c nhng loi cung cp vt liu, c loi cung cp hng thm, qu ngt, c
nhiu loi c dng lm thuc cha bnh, thuc tr su, cung cp nguyn
liu cho cc ngnh cng nghip thc phm, m phm v dc phm.
Vit nam c mt v tr thun li v thin nhin nh vy nn nn Y hc
dn tc c truyn pht trin t lu i, chim v tr c bit trong vic bo v
sc kho nhn dn trong sut hng ngn nm u tranh dng nc v gi
nc. Tip thu truyn thng qu bu ca cc th h cha ng, ngy nay ng
v Nh nc ta xy dng nhiu chnh sch, nhiu hnh thc ng vin
nhm k tha v pht huy tt ngun ti nguyn qu bu c th ti to c
phc v con ngi c hiu qu nht.
Nhiu cy c c trng dng lm thuc, nhiu loi dng lm
nguyn liu cung cp tinh du cho cng nghip dc phm v m phm nh
bc h, thanh hao hoa vng, hoa hi, c loi c dng lm thc phm
chc nng ung nh cc loi tr, cc loi sm v.v... Trong s c cy
ngn (Cratoxylum prunifolium) thuc loi cy mc hoang di v ph bin
khp cc vng trn lnh th nc ta. c bit cy ngn c nhiu cc
tnh min ni pha Bc, Ty Nguyn v Nam B.
http://www.lrc-tnu.edu.vn
2
Cy ngn v ang c dng lm thuc v lm nc ung
trong phm vi dn gian mt cch kh ph bin cc nc chu , Vit
Nam c bit vng ng bo dn tc min ni pha Bc nc ta. Gn y
ngi ta pht hin dch chit ca l cy ngn c tc dng cha bnh gan,
c tc dng tt vi h thn kinh v tc dng ca n khng thua km g cc
thuc nhp ngoi.
Cy ngn c dng trong dn gian c t lu, nghin cu ho
thc vt cy ngn th mi ch c cc nh khoa hc ch n trong mt
s nm gn y, gp phn lm r thm thnh phn ho thc vt ca cy
ngn to thun li cho vic dng, s dng cy thuc ny lm dc liu v
nguyn liu cho cc mc ch khc, chng ti chn ti: Nghin cu ha
hc v nhn dng mt s nhm cht c trong cy ngn (Cratoxylum
prunifolium). i tng nghin cu l cy ngn mc hoang, thu hi vo
thng 11 nm 2007 ti x Thnh c thnh ph Thi Nguyn .
http://www.lrc-tnu.edu.vn
3
Chng 1
TNG QUAN
1.1. M t thc vt
Theo ti liu v phn loi thc vt, cy ngn cn gi l cy thnh
ngnh, lnh ngnh tu thuc vo tng a phng tn khoa hc l
Cratoxylum prunifolium Kurtz thuc h Ban Hypericaceae [1, 2, 5, 8, 11]
Cy ngn phn b rng ri vng Nhit i, Cn Nhit i, n i,
ch yu vng n i bn cu Bc. Chi Cratoxylum Vit Nam c 5 loi:
Cratoxylum maingayi Kurz c tn l lnh ngnh nh, phn b Bc Trung B
v Lm ng; Cratoxylum prunifolium Kurz lnh ngnh l hp phn b
vng ng Nam B trn lnh th VIt Nam; Cratoxylum prunifolium Kurtz
cy ngn, cn gi l thnh ngnh p mc ph bin vng ng Bc
nc ta; Cratoxylum polyanthum Korth hay Cratoxylum ligustrinum Blume
lnh ngnh; Cratoxylum cochinchinense Blumer cn gi l lnh ngnh Nam
B. Hai loi cui ch yu mc Nam B song cng kh ph bin cc tnh
ng Bc B.
Cy ngn - Cratoxylum prunifolium Kurtz thng thy mc t nhin
hu ht cc tnh min ni v trung du pha Bc Vit Nam c nhiu Xun
Mai - Ba V, H Ty, Sapa, Ph Th, Thi Nguyn, Bc Cn, Lng Sn, Vnh
Ph. Cy ngn cng phn b ph bin cc nc Chu nh: Trung
Quc, Malaysia, Philippin, n , Indonexia, Myanma, Thi Lan.
c im sinh trng v pht trin ca cy ngn:
Cy ngn l loi cy nh, thn g, c gai gc (trong rng lu nm
cy c th cao v to) cnh non c lng t, mu nn gi l ngn [11]. L
hnh mc di 12 - 13 cm, rng 3,5 - 4 cm, mc i xng, cung ngn 3 - 5
mm. Mt gn chnh mu n 1/3 l non, gn l v l c mu n qu
http://www.lrc-tnu.edu.vn
4
na. Hoa mc trn nhng cnh k l mu trng hoc hng c lng mu ta.
Qu nang di 15mm, rng 7 - 8mm. Ht hnh trng di 6mm, rng 3mm
http://www.lrc-tnu.edu.vn
5
Trong nhng nm gn y mt s tc gi nghin cu tc dng cha
bnh ca dch chit ton phn l v thn cy ngn. Nhm nghin cu ca
Hc vin Qun Y - H Ni cho bit cy ngn t c, dnh chit ton phn
ca l v thn cy ngn c tc dng chng xi ho tt, hot tnh t 69%
trong khi tanakan ch t 48% [9] v c tc dng hot huyt, lm lu thng
mu, gim ng nhng trng hp tng ng [12].
B mn Dc hc qun s - Hc Vin Qun Y trn c s nghin cu lm
sng dnh chit ca l ngn Cratoxylum prunifolium trn chut nht trng v
th c so snh vi thuc tanakan do hng Beaufour Ipsen ca Php sn xut
a ra kt lun:
Dch chit ton phn l ngn v dch chit etylaxetat l ngn u
c nh hng ln hot ng ca h thn kinh cc mc khc nhau:
- Dch chit etylaxetat v dch chit ton phn l ngn, Tanakan u
c tc dng hot ho h thn kinh thc vt, th hin s tng r hm lng
Catecholamin trong mu ng vt th nghim sau khi ung cc thuc ny.
- Dch chit etylaxetat l ngn c tc dng tng ng Tanakan
gy hot ho ng b cc t bo no th thc nghim, th hin lm gim
thnh phn sng chm delta, tng thnh phn sng alpha trn in no , cn
dch chit ton phn l ngn khng th hin r tc dng hot ho h thn
kinh trung ng.
- Dch chit etylaxetat l ngn c tc dng lm tt c hai qu trnh
hng phn v c ch c iu kin no b ng vt th nghim (thng qua
hot ng phn x c iu kin), tng ng vi Tanakan v tt hn tc
dng ny ca dch chit ton phn l ngn.
- Mc nh hng lm tng cng chc nng h thn kinh trung
ng ca cc cht theo th t gim dn: dch chit etylaxetat l ngn,
Tanakan, dch chit ton phn l ngn [4].
http://www.lrc-tnu.edu.vn
6
i vi cao nc (cn tng) 30 - 80 mg/kg c hot tnh khng khun
vi cc vi khun staphylococcus auresu, Escherichia coli, Salmonella typhi.
Dch chit tng s t l cy Cratoxylum cochinchinense Thi Lan
c ng-i ta th nghim v cho thy chng c tc dng chng nhim khun,
chng xi ho, nhng hot ng ca t bo [22, 23, 24].
Singapo dch chit cy Cratoxylum cochinchinense c s dng
trong nhn dn c tc dng chng cc gc t do cha cc bnh lin quan n
s cng thng, mt mi [25].
Ti Nht Bn, t r cy ca mt s loi thuc chi Cratoxylum ngi ta
bo ch thnh thuc lm tng tr nh, chng lo ho, mt ng ngi gi.
Trung Quc, l ca cy ngn ngi ta ch bin thnh tr pha
nc ung hng ngy v s dng mt cch rng ri. T phn dch chit
etylaxetat ca l cy ngn tch c cc xanthone c tc dng khng u
trng ca mui gy st da vng hn c cht rotenon, khng nhng th cc
cht ny cn c nhiu trin vng lm thuc chng mi [28].
Phn dch chit etylaxetat ca Cratoxylum cochinchinense c tc dng
km hm peroxit ho lipit mng t bo gan, km hm s xut hin lipit c tnh
gy vim gan v c tc dng bo v gan tt [27].
Theo cc lng y, cy ngn c v ngt, tnh mt b phn dng l
non, v cy, v r. Ngi b au bng n khng tiu, ung nc nu ca l
lnh ngnh gip tiu ho tt thng dng 100g l non nu mt lt nc, thay
nc ung hng ngy. Khi b cm nng, st th dng l non 50g nu vi 1 lt
nc ung.
Trong dn gian Vit Nam, cy ngn mi thy dng trong phm vi nhn
dn lm thuc kch thch tiu ha, phc hi sc kho khi m au, sinh , bo v
thnh mch, chng lo ho, tng tr nh ngi cao tui. Ngi ta thu hi l
ngn vo dp Tt mng 5 thng 5 (Tt oan Ng) lm thc vt nu nc
http://www.lrc-tnu.edu.vn
7
ung hoc c th sau em phi kh mi dng. Gn y c mt vi tc gi
Vit Nam nghin cu v cho thy l ngn c tc dng cha bnh, c tc
dng chng gc t do, bo v thnh mch, bo v t bo gan chng m lo,
chng xi ho v min dch invitro kh r [7].
Ti c s sn xut kinh doanh thuc thnh phm, thc phm chc nng
Y hc c truyn Thi Nguyn, Bc s Hong Sm s dng dch chit ca l
ngn lm thc phm chc nng, thay ch lm nc ung, cha cc bnh
nan y nh: nga, gh l, ronan thn kinh, mt ng, ming ng n khng
ngon, gim tr nh
1.3. Tnh hnh nghin cu ha hc thc vt chi Cratoxylum
1.3.1. Cc hp cht c khung triterpen
Theo ti liu [19] t dch chit n-hexan ca thc vt Cratoxylum
cochinchinense tch c ngoi cc triterpen c 5 vng ngng t nhm
lupeol v -mangostin cn phn lp c cc triterpen c hai vng ngng t.
Sau khi tch bng cc phng php sc k v da vo kt qu phn tch cc
ph ca cc cht c tch ra ngi ta ngh cng thc cu to ca 7 hp
cht triterpen hai vng l:
http://www.lrc-tnu.edu.vn
8
27
28
29
30
25
26
http://www.lrc-tnu.edu.vn
9
COOH
HO
O
HO
O
OH
OH
OH
OH
HO
OH
O
O
O
O
HO
HO
HO
OH
V: Acid dicaffeoylquinic
1.3.3. Cc cht c khung xanthone
Cc xanthone l nhm cht ln nht, ph bin nht c hu ht cc
loi thuc chi Cratoxylum [6, 14, 17, 19, 21, 22, 23, 24, 25, 26, 27, 29, 30].
pht hin ra l cc xanthone v dn xut di dng C-glycoside ca
chng. T loi Cratoxylum cochinchinense [25] Soon Yew Tang cng vi cc
cng s phn lp c C-glicoside ca xanthone v xc nh c cng
thc cu to ca n c dng VI.
http://www.lrc-tnu.edu.vn
10
HO
OH
O
HO
OH
HO
OH
OH
VI:
1,3,6,7-tetrahydroxyxanthone- C2--D-glycoside.
OH
OH
MeO
O
HO
HO
R1
OCH3
VII.a: Dulxis-xanthone
OH
R1
OH
OH
VII.d:
Formoxanthone A; (R 1 =
).
http://www.lrc-tnu.edu.vn
11
R1
OH
MeO
HO
VII.e:
3-isomangostin; (R 1 =
).
).
3,4-dihidro-5,9-dihidroxy-7-(3- hidroxy-3-methybutyl)-8-methoxyOH
).
OH
R1
HO
R2
OH
VII.f:
Isocudraniaxanthone B;
(R1 = H; R2 = OMe).
VII.f1: Gerontoxanthone I;
(R1 =
O
R2
OH
O
OH
VII.g: 10-O-methylmaclur-xanthone;
VII.g1: Macluraxanthone;
; R2 = OH).
O
R1
(R1 =
(R1 =
; R2 = OMe).
; R2 = OH).
http://www.lrc-tnu.edu.vn
12
VII.g2: Xanthone V 1.
(R1 =
VII.g3: 6 - deoxyjacreubin;
(R1 = H; R2 = H).
O
; R2 = OH).
OH
R1
OH
VII.h: Formoxanthone B;
(R1 =
).
OH
HO
OH
http://www.lrc-tnu.edu.vn
13
O
OH
HO
O
R
OMe
8'
9'
OH
2
5 10'
O
4'
8
5
O
8'
OMe
9'
OMe
2
3
10'
4'
OMe
OMe
OMe
R1
O
R2
http://www.lrc-tnu.edu.vn
14
R2
OH
HO
O
R1
OH
OMe
MeO
HO
OH
MeO
O
OMe
X.c: Maclurathone
OMe
HO
OH
OH
HO
O
OH
http://www.lrc-tnu.edu.vn
15
O
OH
OH
HO
HO
OH
OH
OH
2'
O
1
H3CO
8'
B
4''
HO
1'
9'
4'
OCH3
'''
OH
1''
X.i: Fruniflorone;
3''
(R=
5''
2''
) [22]
OH
OR
O
XI.a: 3-geranyloxy-6-methyl-1,8-dihidroxy-anthraquinon;
(R=
XI.a1: madagascin;
)
(R =
http://www.lrc-tnu.edu.vn
16
XI.a2: physcion;
(R = Me)
XI.a3: emodin;
(R = H )
OH
OMe
O
OH
HO
OMe
OH
OH
XI.c: Vismiaquinon
HO
OH
OH
OH
OH
OH
XII.a: ihirokampherol
OH
XII.b: Taxifolin
http://www.lrc-tnu.edu.vn
17
1.4. Nhng nghin cu ho thc vt loi Cratoxylum prunifolium.
T l ca loi Cratoxylum prunifolium, Xue-Li-Cao, Yoichiro Ito
dng phng php sc k kh lng tc cao phn lp c epigallocatechin
gallat (XIII.a) v epicatechin gallat (XIII.b) [27].
OH
OH
HO
OH
OH
O
OH
C
O
OH
OH
XIII.a:
Epigallocatechin-3-O- gallate
HO
OH
O
OH
OH
OH
C
O
OH
OH
XIII.b:
Epicatechin-3-O-gallate
http://www.lrc-tnu.edu.vn
18
[6] v cho bit chng u c tc dng dit mui gy st rt v c tc dng tt
trong vic dit mi.
T v v r cy Cratoxylum prunifolium trng tnh Nng Khai min
Nam Thi Lan, Nawong Boonnak v cc cng s chit bng iclometan,
phn lp cc cht bng sc k tch c 10 xanthone vi s khc nhau ch
l cc mch nhnh (R 1 v R2) [22] v tc gi gi tn chng l cc
prunifloron c nh s t 1 n 10. Cng thc ca chng c cu trc XIV.
R1
OH
RO
R2
HO
OR
OH
OH
O
O
XV: Anthraquinon
http://www.lrc-tnu.edu.vn
19
Chng 2
http://www.lrc-tnu.edu.vn
20
2.1.3. Phng php kho st v xc nh cu trc ho hc cc hp cht
Cc cht kt tinh phn lp ra c xc nh nhng tnh cht vt l c
trng: mu sc, mi v, h s Rf, im nng chy, ghi cc loi ph nh: ph t
ngoi (UV), ph hng ngoi (FT-IR), ph cng hng t ht nhn 1H- NMR,
ph 13C- NMR, tu theo tng loi cht. Cc s liu ph thc nghim ca cc
cht sch c dng nhn dng cu trc ho hc ca chng.
2.2. Dng c, ho cht v thit b nghin cu
2.2.1. Dng c v ho cht
Cc loi dung mi dng ngm, chit mu l cc loi tinh khit
(pure), cn cc loi dung mi dng sc k ct, sc k lp mng hay dng
trong phn tch l loi tinh khit phn tch (PA).
Sc k lp mng dng tm mng nhm DC - Alufolien Kiesegel 60
F254 Art.5554 trng sn, dy 0,2mm c s dng xc nh s b s
thnh phn c trong cc dich chit, cc phn on chy ct v kim tra s b
sch ca sn phm thu c.
Cc h dung mi khai trin SKLM:
1. n-Hexan - etylaxetat
90:10
H A
2. n-Hexan - etylaxetat
95:10
H B
3. Clorofom - metanol
90:10
H C
4. Clorofom - metanol
50:10
H D
5. Clorofom - metanol
20:10
H E
Cc tm SKLM sau khi sy kh c soi di n t ngoi (UVBIOBLOCK) bc sng = 254nm v 365nm. Thuc th hin mu l
vanilin 1% trong dung dch metanol-H2SO4 , sau sy nhit trn 1000C
http://www.lrc-tnu.edu.vn
21
Cc gi tr Rf trong h dung mi trin khai c biu thc:
Rf =
http://www.lrc-tnu.edu.vn
22
Nh vy t l cy ngn thu nhn c 04 phn on cn l: cn
trong clorofom (C), cn trong etylaxetat (E), cn trong butanol (B) v
cn trong metanol (M). S 2.1
MU KH
1. Etanol
2. Cn trong nc
CHCl3
EtOAc
Cn CHCl3
(C)
Cn EtOAc
(E)
BuOH
MeOH
Cn BuOH
(B)
Cn MeOH
(M)
E1
C1
C2
C3
Cn chit
(gam)
Clorofom
Etylaxetat
n-Butanol
Metanol
787,4
52,264
34,654
40,411
26,604
100%
6,63%
4,40%
5,13%
3,38%
http://www.lrc-tnu.edu.vn
23
2.3.2. Kho st nh tnh cc dch chit
Dng cc thuc th c hiu pht hin cc nhm hp cht thin
nhin c trong hot tnh sinh l cao trong thc vt [3] chng ti thu c kt
qu th nh tnh vi cc nhm cht, kt qu c ch ra bng 2.2.
Bng 2.2. Pht hin cc nhm cht trong cy ngn
Stt Nhm cht
Thuc th c hiu
Hin tng
Kt qu
Saponin
To bt
Bt bn trong axit ++
Glicozttim
Kelle-Kiliani
Vng nu
Cumarin
Axit v kim
Kt ta bng
++
Poliphenol
Flavonoit
Xianidin
T hng n
++
Steroit
Liberman-Bourchard
Mu xanh vng
++
Ancaloit
Dragendorf
Mu vng da cam -
ng kh Felinh
Cho kt ta mu
gch
+++
++
http://www.lrc-tnu.edu.vn
24
http://www.lrc-tnu.edu.vn
25
Chng ti cn tin hnh th cc ng dng lm thc phm chc nng
ti c s sn xut kinh doanh thuc thnh phm, thc phm chc nng Y hc
c truyn Thi Nguyn. Kt qu th hot tnh cc dch chit th c trnh
by trong bng 2.3.
Bng 2.3. Kt qu th hot tnh sinh hc ca dch chit th
t l cy ngn (Cratoxylum prunifolium)
ng knh vng c ch xung quanh ging thch(mm)
Dch chit
Cn tng (H2O)
Cn EtOAc
Hot tnh
+
+
ng
Hot
knh
tnh
(mm)
26
+
17
Salmonella typhi
ng
knh
(mm)
27
18
Hot
tnh
+
+
http://www.lrc-tnu.edu.vn
26
Cc cht cn cn li khng tan trong cc dung mi trn cho tan trong
MeOH thu oc 32,346g cht (k hiu l cc cht M).
2.4.1. Dch chit clorofom
T 11g cn t dch clorofom ca l ngn k hiu (C) c phn
chia trn sc k ct silicagel vi cc h dung mi n-hexan - clorofom vi cc
t l tng dn clorofom t 0% n 100% kim tra cc phn on trn sc k
lp mng, cc phn on ging nhau em gp li v ct loi dung mi thu
c ba cht C-1, C-2, C-3.
2.4.1.1. Taraxeron (Friendoolean - 14- en-3-on) (C1)
Ra gii vi cc h dung mi n- hexan-clorofom (50:1), sau khi ct loi
dung mi, cn thu c kim tra trn SKLM trong h dung mi B v pht
hin bng vanilin 1% trong dung dch CH3OH- H2SO4, kt tinh li trong dung
mi trn thu c 21mg cht rn, tinh th hnh kim, mu trng c nhit
nng chy 278-2800C v cho cc c trng ca mt tritecpen.
Ph FT-IR (KBr); max (cm-1): 1720 (C=O); 1500 (C=C); 3050 (CH3);
2940 (CH).
Ph LC- MS, m/z (%) 424 [M+]; 408 [M-O]+ .
PH 1H- NMR (500MHz, CDCl3): (ppm) 0,762ppm (3H, s, CH3);
0,84ppm (3H, s, CH3); 0,885ppm (3H, s, CH3); 0,996ppm (3H, s, CH3);
1,011ppm (3H, s, CH3); 1,02ppm (3H, s, CH3); 1,07ppm (3H, s, CH3)
5,48ppm (1H, dd, 3,21 v 8,17Hz).
Ph
13
217,3 (C3); 47,56 (C4); 55,77 (C5); 19,91 (C6); 33,57 (C7); 38,87 (C8); 48,80
(C9); 33,57 (C10); 17,44 (C11); 36,67 (C12); 37,74 (C13); 157,59 (C14); 117,19
(C15); 34,12 (C16); 35,57 (C17); 48,70 (C18); 40,64 (C19); 28,79 (C20); 35,11
(C21); 37,69 (C22); 29,56 (C23); 21,57 (C24); 14,79 (C25); 25,56 (C26); 29,92
(C27); 26,12 (C28); 33,39 (C29); 21,47 (C30).
http://www.lrc-tnu.edu.vn
27
2.4.1.2. Stigmast- 5,22- ien-24R-3-ol
Ra gii ct vi h dung mi n-hexan- clorofom (50:5), sau khi ct loi
dung mi cn thu c kim tra trn SKLM trong h dung mi A, pht hin
bng vanilin 1% trong dung dch CH3OH- H2SO4, kt tinh li trong h dung
mi trn thu c 29mg tinh th hnh kim, khng mu, khng mi, c nhit
nng chy 152-1550C, Rf 100 =61.
Ph FT-IR (KBr); max (cm-1): 3429,2 (OH); 2864,7 (C-H); 1651 (C=C).
Ph EI- MS; m/z (%): 412 [M+] (7); 300 (7); 255 (11); 231 (4); 213
(8); 173 (7); 145 (20); 133 (20); 83 (49,3).
Ph
2H, H6 ); 515 (1H, dd, j22,23 = 15, Hz, j22,20= 5Hz, H-22); 5,01 (1H, dd, j23,22 =
15Hz, j23,24 = 5Hz, H-23).
Ph 13C- NMR (500MHz, CDCl3); (ppm): 36,4 (t, C1); 29,67 (t, C2);
71,8 (d, C3); 42,3 (t, C1); 139,2 (s, C5); 121,4 (d, C6); 37,3 (t, C7); 31,85 (d,
C8); 51,15 (d, C9); 36,12 (s, C10); 24,35 (t, C11); 42,18 (t, C12); 31,4 (s, C13);
56,83 (d, C14); 25,38 (t, C15); 31,6 (t, C16); 55,9 (d, C17); 12,01 (q, C18); 18,95
(q, C19); 40,47 (d, C20); 21,03 (q, C21); 138,4 (d, C22); 129,2 (d, C23); 50,01 (d,
C24); 33,9 (t, C25); 21,19 (q, C26); 19,79 (d, C27); 28,89 (q, C28); 12,22 (q, C29).
2.4.1.3. - Sitosterol
Tip tc ra gii trn ct vi h dung mi n-hexan- clorofom (40:10),
sau khi ct loi dung mi, cn thu c kim tra trn sc k lp mng trong h
dung mi B, pht hin n bng vanillin 1% trong dung dch CH3OH- H2SO4,
kt tinh li thu c 27mg cht rn, tinh th hnh kim, nng chy 140- 1410C,
Rf .100 = 71.
Ph FT-IR (KBr); max (cm-1): 3450 vn rng (H32, C3); 3010- 1650
(lin kt i).
http://www.lrc-tnu.edu.vn
28
Ph EI- MS; m/z (%): 414 [M+] (20); 413 [M-H]+ (41); 398 (28); 397
(100); 395 (32); 383 (11); 361 (11); 257 (3); 255 (6,3); 151 (5,6); 139 (11).
Ph 1H-NMR (500MHz, CDCl3 ); (ppm): 0,68 (3H, s, CH3-18); 1,01
(3H, s, CH3-19); 0,81 (3H, d, j, 7Hz, CH3-26); 0,88 (3H, d, j, 7Hz, CH3-27);
0,83 (3H, t, 7,32Hz, CH3 -29); 0,92 (3H, d, j, 10Hz, CH3-21); 3,52 (1H, m, H3); 5,42 (1H, d, j, 5Hz, H-6).
Ph
13
C6); 71,8 (d, C3); 56,8 (d, C14); 56,1 (d, C17); 50,2 (d, C9); 45,7 (d, C24); 42,5
(s, C13); 42,4 (t, C4); 39,8 (t, C12); 37,3 (t, C1); 36,5 (s, C10); 36,2 (d, C20);
33,9 (d, C8); 31,9 (t, C7); 31,7 (t, C2); 29,2 (d, C25); 28,3 (t, C16); 26,2 (t, C23);
24,2 (t, C15); 21,1 (t, C11); 19,8 (q, C26); 19,4 (q, C19); 19,1 (q, C27); 18,8 (q,
C21); 11,9 (q, C29); 11,9 (q, C18); 23,2 (t, C28).
2.4.2. Dch chit trong etylaxetat (E)
T 15g cn chit etylaxetat ca l cy ngn (k hiu E) c tin
hnh tch cc cht trn sc k ct silicagel, ra gii ct sc k bng h dung
mi clorofom- metanol c t l theo tng dn ca dung mi phn cc,
metanol 0% -100%, kim tra cc phn on trn sc k lp mng, thuc th
pht hin (F eCl3 + K3[Fe(CN)6]) 1% sau gp cc phn on ging nhau,
ui ht dung mi v kt tinh li thu c cht (E1).
* Axit gallic (E1)
Ra gii trn ct bng h dung mi clorofom- metanol (90:10), sau khi
kim tra bng SKLM vi h dung mi C, ct loi dung mi, thu c cn th,
kt tinh li trong h dung mi trn thu c 70mg cht rn mu vng, tinh th
hnh kim, c tn/c khong 248- 2580C.
Ph FT-IR (KBr); max (cm-1): 1700 (C=O); 3100 (OH) rt t.
Ph 1H-NMR (500MHz, MeOD); (ppm): 7,092 (2H, C2 v C6).
PH 13C-NMR (500MHz, MeOD); (ppm): 121,960 (C1); 139,559 (C4);
110,349 (C2, C6); 146,345 (C3, C5); 170,364 (C7).
http://www.lrc-tnu.edu.vn
29
Chng 3
http://www.lrc-tnu.edu.vn
30
3.2. Phn tch nh tnh v pht hin cc nhm cht
Dng cc thuc th c hiu pht hin cc nhm hp cht thin
nhin c hot tnh sinh l cao trong thc vt [3] cho thy trong dch chit
bng clorofom ca l cy ngn c cha cc axit hu c, streroit,
triterpenoit. dch chit bng etylaxetat ca l cy ngn c cha cc hp
cht polyphenol, cc hp cht flavonoit, cng phn dch chit ny tch
c axit gallic v dn xut ca n. Trong dch chit n-butanol c cha cc
tanin, cc ng kh. Khng pht hin thy c ancaloit, cc xianua trong cc
dch chit ca l ngn. Kt qu ca cc phn ng nh tnh xc nh cc
nhm hp cht thin nhin c hot tnh sinh l cao c ch ra bng 2.2.
3.3. Phn lp v nhn dng cc hp cht c trong cc dch chit khc nhau
ca cy ngn
Cc dch chit t cy ngn (Cratoxylum prunifolium) u l nhng
hn hp phc tp cha cc hp cht khc nhau. phn lp tng cht ra khi
hn hp s dng cc phng php sc k ct, cht hp ph dng l
silicagel, cc h dung mi ra gii thch hp v thng phi lp li nhiu ln.
Vic tinh ch cc cht thng dng phung php kt tinh li trong
dung mi hoc h dung mi thch hp. Nh cch lm thu c cc n
cht c tinh khit cao, p ng cc yu cu kho st tnh cht ho l v
xc nh quang ph ca chng.
Khi phn lp cc thnh phn ho hc t l cy ngn c thc hin
nh trong s 2.1. Bng phng php phn lp trn, t dch chit bng
clorofom ca l cy ngn chng ti thu c 3 hp cht sch l: mt
tritecpen v hai steroit; cn dch chit etylaxetat phn lp c axit gallic v
hn hp cc xanthone vi flavonoit.
3.3.1. Taraxeron ( hay Frendoolean-14-en-3-on) (C-1)
Cht C-1 l cht rn tinh th hnh kim, mu trng c nhit nng
chy 278-2800C n c tch t phn cn chit clorofom. Bng sc k ct
http://www.lrc-tnu.edu.vn
31
silicagen vi h dung mi n-hexan:clorofom thu c cht C-1, cht ny
cho cc phn ng nh tnh ca terpenoit.
Khi lng phn t ca cht C-1 c xc nh bng ph khi LCMS cho bit khi lng phn t ca [M+] m/z bng 424,2 tng ng vi khi
lng phn t bng 424 u v tng ng vi cng thc phn t C30H48O.
Trong ph LC-MS cn xut hin pic m/z 408,2 iu ny tng ng vi
mnh [M-O]+ vi m/z bng 408
Ph FT-IR: cho thy cc vn hp th ca nhm xeton (C=O) max =
1720cm-1, lin kt i (C=C) max = 1500cm-1, cc nhm CH3 max =
3050cm-1 v cc nhm CH max = 2940cm-1, khng pht hin thy cc vn
hp th ca nhm OH. iu ny c th khng nh c hp cht C-1
khng c chc ru m l c chc xeton.
Ph 1H-NMR v ph DEPT: cho bit hp cht C-1 c tn hiu ca 8
nhm CH3 u dng singlet (xem hnh 3.2 v 3.4) c cc chuyn dch
ho hc nh sau: chuyn dch = 0,762ppm (3H, s, CH3); = 0,841ppm
(3H, s, CH3); = 0,847ppm (3H, s, CH3); = 0,885ppm (3H, s, CH3); =
0,996ppm (3H, s, CH3);
13
http://www.lrc-tnu.edu.vn
32
Nh vy, ph NMR hon ton ph hp vi ph FT-IR khng thy c
vn c trng ca nhm OH m li thy xut hin vn c trng ca C=O.
Cc nguyn t H i vi nhm C=O thy ch c 2H vi = 2,50ppm (2H,
dd, J = 7,1 v 11Hz), cn c 2 proton v tr ca nhm =CH (ti C16) c
chuyn dch ho hc = 2,27ppm (2H, dd, J = 3,3 v 4,08 v 6,39).
Trn c s phn tch cc tnh cht ho hc, nht l phn tch cc thng
tin c c t ph LC- MS, FT-IR, ph cng hng t ht nhn 1 H-NMR,
13
29
23
30
19 20
18
12
11
1
8 14
22
17
28
16
15
13
27
9
10
2
3
24
21
6
25
26
http://www.lrc-tnu.edu.vn
33
Bng 3.1. Ph 13C-NMR v 1H-NMR ca taraxeron (C-1) t l ngn
V tr
cacbon
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
13
C-NMR
(-ppm)
38,35
33,08
217,3
47,56
55,77
19,91
33,57
38,87
48,8
33,57
17,44
36,67
37,74
157,59
117,19
34,12
35,57
48,7
40,64
28,79
35,11
37,69
29,92
14,79
29,86
21,34
25,56
26,12
33,36
21,47
1
ACD/13CH-NMR
NMR (-ppm)
(-ppm)
38,4
Vng (1,2 - 1,6)
34,1
2,5 (2H; dd; 7,1 v 11,0)
217,3
47,6
55,8
(1,99)
20,0
Vng (1,2 - 1,6)
35,2
Vng (1,2 - 1,6)
38,9
48,7
(1,78 - 1,87)
37,6
17,5
Vng (1,2 - 1,6)
35,8
Vng (1,2 - 1,6)
37,7
157,6
117,2
5,48 (1H; d; 3,21; 8,17)
36,7
2,27 (2H; dd; 3,3; 4;1; 6,4)
37,7
48,8
(1,78 - 1,87)
40,7
Vng (1,2 - 1,6)
28,8
33,6
Vng (1,2 - 1,6)
33,1
Vng (1,2 - 1,6)
29,9
14,8
29,9
21,5
0,76 - 1,07
(24H ca 8 nhm CH3)
25,6
26,2
33,4
21,4
http://www.lrc-tnu.edu.vn
34
Hnh 3.1
http://www.lrc-tnu.edu.vn
35
Hnh 3.2
http://www.lrc-tnu.edu.vn
36
Hnh 3.3
http://www.lrc-tnu.edu.vn
37
Hnh 3.4
http://www.lrc-tnu.edu.vn
38
3.3.2. Stingmast -5,22-dien-24R-3-ol (C-2)
Cht C-2 l cht rn kt tinh, c nhit nng chy 1520-1550C,
tch c t dch chit clorofom, phn lp bng sc k ct silicagen, ra gii
bng h dung mi n-hexan-clorofom (90:10).
Ph EI-MS cho pic ion phn t [M+] 412 m/z. Cc ph FT-IR, 1HNMR v 13C-NMR u khng nh s c mt ca nhm hydroxyl, ph FT- IR
c vn 3410cm-1 rng, cn ph NMR cho H-3 = 3,47ppm v C-3 =
71,6ppm). Phn t c hai ni i khng lin hp (IR cho max 1622cm-1).
Ph 1H-NMR ca cc proton thuc nhm metyl (CH) cng khng
nh iu y, H-6 = 5,33ppm (1H, dd, j =15Hz v 2Hz); H-22 = 5,15ppm (1H,
dd, j =15Hz v 5Hz); H-23 = 5,01ppm (1H, dd, j =15Hz v 5Hz). Ph 13CNMR cho thy C-5 = 139,2ppm; C-6 = 121,4ppm; CC-22 = 138,4ppm v C-23
= 129,2ppm. So snh cc s liu ph 1H-NMR v 13C-NMR ca cht C-2
vi s liu ph NMR ca stingmasterol [16] hon ton tng t nhau v c
ch ra bng 3.2.
Da trn phn tch cc s liu v ph EI-MS, FT-IR v cc ph NMR ca
hp cht C-2 hon ton ph hp vi cu trc ca cht Stingmast -5,22-dien24R-3-ol.
29
28
21
19
12
18
11
14
17
22
23
27
24
25
26
16
15
HO
13
20
10
5
4
Stingmast -5,22-dien-24R-3-ol
http://www.lrc-tnu.edu.vn
39
Bng 3.2. Ph 13C-NMR ca cc cht C-2 v stingmasterol [16]
Ph 13C-NMR ca Stigmasterol
(C-2)
V tr
13
C-NMR(-ppm)
cacbon
1
36
2
29
3
71
4
42
5
140
6
122
7
37
8
31
9
51
10
36
11
24
2
42
13
40
14
57
15
25
16
30
17
56
18
12
19
19
20
40
21
19
22
138
23
129
24
50
25
32
26
21
27
21
28
19
29
12
Ph 13C-NMR ca Stigmasterol
[16]
V tr
13
C-NMR(-ppm)
cacbon
1
37,28
2
31,66
3
71,78
4
42,31
5
140,76
6
121,69
7
31,91
8
31,91
9
50,18
10
36,52
11
21,09
2
39,70
13
42,22
14
56,88
15
24,38
16
28,92
17
55,97
18
19,40
19
12,06
20
40,50
21
21,23
22
138,31
23
129,29
24
51,25
25
31,91
26
21,09
27
19,00
28
25,41
29
12,25
http://www.lrc-tnu.edu.vn
40
Hnh 3.5
http://www.lrc-tnu.edu.vn
41
Hnh 3.6
http://www.lrc-tnu.edu.vn
42
Hnh 3.7
http://www.lrc-tnu.edu.vn
43
Hnh 3.8
http://www.lrc-tnu.edu.vn
44
3.3.3. -sitosterol (C-3)
Cht C-3 l cht rn tinh th hnh kim, khng mu, nng chy 1401410C, n c tch t dch chit clorofom, trong h dung mi n-hexanclorofom (40:10).
Ph EI- MS, m/z (%) cho [M+] = 414 (23), [M-1]+ = 413 (41).
T ph EI-MS cho pic [M+] = 414 ng vi cng thc phn t C29H50O.
Ph hng ngoi, ph 1H-NMR v ph 13C-NMR cho thy trong phn t
c nhm OH ti v tr C-3 (IR c vn 3450 cm-1 rng, H-3 = 3,57ppm v C-3
= 72,1ppm). Mt lin kt i ti C 6 (ph IR c max= 3010cm-1 v 1650cm-1
ng vi dao ng ho tr ca lin kt i; cn trn ph 1H-NMR cho H-6 =
5,42ppm (1H, d, J, 5,2Hz), ph
13
14
17
22
23
27
24
25
26
16
15
HO
13
11
20
10
5
4
-sitosterol
http://www.lrc-tnu.edu.vn
45
Bng 3.3. Ph 13C-NMR ca cc cht C-3 v -sitosterol [16]
Ph 13C-NMR ca -sitosterol
(C-3)
V tr
13
C-NMR(-ppm)
cacbon
1
37
2
31
3
71
4
42
5
140
6
121
7
32
8
32
9
50
10
36
11
21
12
40
13
42
14
57
15
24
16
28
17
56
18
12
19
19
20
36
21
19
22
34
23
26
24
46
25
29
26
19
27
19
28
23
29
12
Ph 13C-NMR ca -sitosterol
[16]
V tr cacbon
1
2
3
4
5
6
7
8
9
10
11
2
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
13
C-NMR(-ppm)
37,27
32,54
71,77
42,29
140,75
121,69
31,92
31,92
50,15
36,51
21,10
39,80
42,23
56,78
24,31
28,25
56,08
19,39
11,89
36,16
18,80
33,96
26,11
45,85
29,18
18,82
19,05
23,08
11,99
http://www.lrc-tnu.edu.vn
46
Hnh 3.9
Hnh 3.9
http://www.lrc-tnu.edu.vn
47
Hnh 3.10
http://www.lrc-tnu.edu.vn
48
Hnh 3.11
http://www.lrc-tnu.edu.vn
49
Hnh 3.12
http://www.lrc-tnu.edu.vn
50
3.3.4. Axit gallic (E-1)
Cc cht thu c t dch chit etylaxetat (E) c phn chia trn sc
k ct vi cht hp ph silicagel, ra gii bng hn hp dung mi clorofommetanol vi t l metanol tng dn t 0% - 100% thu c nhiu phn on
khc nhau trong c phn on CHCl3:MeOH (9:1) cha mt cht sch
c k hiu l cht E-1.
Cht E-1 l cht rn, tinh th hnh kim c nhit nng chy
2480 C-2500 C.
Ph IR cho vn hp th mnh max= 1700cm-1 c trng cho nhm
cacboxyl c max = 1700cm-1, vi vn rt t c s sng max = 3100cm-1 c
trng cho nhm OH ca phenol.
Ph 1H-NMR cho tn hiu 7,092ppm ng vi hai proton hai v tr C2
v C6.
Ph 13C-NMR cho thy cc tn hiu: C-1 = 121,960ppm; C-4 =139,559
ppm; C-2 = C-6 =110,349ppm; C-3 = C-5 =146,345ppm; C-7 =170,364ppm.
Nhng s liu v ph NMR ca cht E-1 hon ton ph hp vi phn
mm ACD/NMR ca axit gallic (xem bng 3.4).
Bng 3.4. S liu ph NMR ca E-1 v s liu ph NMR trong
phn mm ACD/NMR ca axit gallic
V tr
cacbon
1
2
3
4
5
6
7
Ph ca cht E-1
13
C-NMR
(-ppm)
121,96
110,35
146,35
139,56
146,35
110,35
170,36
H-NMR
(-ppm)
7,092
7,092
-
ACD/ 1H-NMR
(-ppm)
7,11
7,11
-
http://www.lrc-tnu.edu.vn
51
Trn c s phn tch cc s liu v ph FT-IR, NMR v phn mm
ACD/NMR. Cu trc ca axit gallic l:
7COOH
1
6
HO
OH
4
OH
Axit gallic
T dch chit etylaxetat, ngoi axit gallic cn thu c mt s phn
on khng sch khc c cha cc cht cho phn ng mu vi xianidin; cho
phn ng to mu xanh en vi FeCl3. Cc mu ny c cha hn hp cc
flavonoit, cc polyphenol c h s R f rt gn nhau v vic tch chng bng
ct silicagen cha t c s phn chia tt.
Khi ra gii bng CH3OH 100% thu c phn on cho cc phn ng
mu ging dch chit tng s bng n-butanol. Khi thc hin phn ng thu
phn cc mu ny trong axit HCl 2N thu c glucoz v axit gallic. T kt
qu thu phn ny c th cho rng cc phn on chit bng etylaxetat v
bng n-butanol u cha cc tannin v l galotanin. Tuy nhin cc tannin
y c to ra bi lin kt C-glycoside hay O-glycoside cu trc v th t
lin kt ca chng nh th no, cn cha c lm r v cha tch chng
thnh cc on sch.
http://www.lrc-tnu.edu.vn
52
Hnh 3.13
http://www.lrc-tnu.edu.vn
53
Hnh 3.14
http://www.lrc-tnu.edu.vn
54
http://www.lrc-tnu.edu.vn
55
http://www.lrc-tnu.edu.vn
56
3.4. Th hot tnh sinh hc
Cc cht tng s c chit, tch bng dung mi c phn cc khc
nhau sau khi lm kh kit, loi b hon ton dung mi c gi ti phng th
nghim vi sinh trng i hc Y khoa Thi Nguyn th tc dng sinh hc
ca chng. Kt qu c nu bng 2.3 v hnh 2.1 cho thy tt c cc cht
thu c t cc dch chit ca l cy ngn u c tc dng khng khun
i vi khun Staphylococcus, E.coli, Salmonella typhi.
Chng ti cng gi mu chit bng etylaxetat v chit bng etanol
sau khi lm sch dung mi n c s sn xut kinh doanh thuc thnh
phm, thc phm chc nng Y hc c truyn Thi Nguyn. Cc mu u c
tc dng tt i vi vic chm sc sc kho con ngi, cc cht c trong cc
dch chit trn u khng gy c i vi con ngi.
http://www.lrc-tnu.edu.vn
57
KT LUN
1. Nghin cu sng lc ho thc vt ca l cy ngn (Cratoxylum
prunifolium) pht hin thy nhiu nhm hp cht thin nhin c hot tnh
sinh l cao; l ng kh, tritecpenoit, steroit, saponin, cumarin, axit hu
c, cc flavonoit, cc cht polyphenol, cc tannin, cc cht xanthone.
2. T l cy ngn ln u tin phn lp v da vo cc c trng
ho l, cc s liu ph EI-MS, NMR, FT-IR nhn dng c cu trc ca 4
hp cht hu c thuc cc nhm polyphenol, steroit, terpenoit.
3. Phn tch cc ph IR, UV, MS v NMR cc cht tinh khit c
tch ra ni trn nhn dng c chng l: taraxeron, stigmasterol , sitosterol v axit gallic.
4. T dch chit tng ca l ngn (Cratoxylum prunifolium) chng
hon ton khng gy c hi vi con ngi, c th ch bin thnh thc ung
hng ngy nh mt thc phm chc nng c tc dng khng cc khun
Staphylococcusauresu (T cu vng), E.coli (Thc khun ng rut) v
Salmonella typhi (Thng hn).
KIN NGH
Cy ngn l mt cy thuc qu c nhiu tc dng trong y hc v vy
ti ngh vi cc cp c thm quyn tip tc cho nghin cu mt cch su
rng hn v cy ngn nhm phc v tt trong i sng nhn dn.
http://www.lrc-tnu.edu.vn
58
http://www.lrc-tnu.edu.vn
59
Ting Anh
13. A.K.Ratty, N.P.Das (1988), Biochemical medicine and metabolic
biologo, 39, 69-79.
14. Anake Kijjoa, Maria Jose, T.G.Gonzalez, Madalena M.M.Ana M.Damas,
Ing-on Mondranondra, Llartur M.S.Silva and Werner Herz (1998),
Xanthones from Cratoxylum Maingayi. Phytochemistry Vol 49, No.7,
2159-2162.
15. G.M.Kitanov, I.Assenov I and Dam The Van (1988), Flavonols and
Xanthones from Cratoxylum prunifloum kurz, Pharnazie 43, 879 .
16. Goat J.L, Akihisa T.Analysis of steroit frested, pp.324. Chapman & Hall.
London, New York - Tokyo.
17. Guat-Lee Sia, Graham J.Bennett, Leslie J.Harrison and Keng-Yeow Sim
(1995), Minor xanthones from the Bark of Cratoxylum cochinchinense.
Phytochemistry, Vol 38, No.6, pp.1521-1528.
18. J. Graham (1993), et al, triterpenoids, tocotrienols and xanthones from the
Bark of cratoxylon cochinchinense, phytochemistry, Vol 32, No 5, 1245.
19. Lien Hoa Dieu Nguyen, Le Li J. Harrison (1998), ritecpenoid and
xanthone constituents fo Cratoxylum conchinchinese, phytochemisty
50, 471-476.
20. Macias, M.Ulloa, M.Ulloa, M. Gamboa, A. and Mata, R. Phytotoxic
comopounds from the new coprophilous fungus Gnanomyces polytlrix.
21. Munekazu Iinuma, Hideki, Ttero Ito, Toshiyuki Tanaka and Domingo
A.Madulid (1996), Two xanthones from roots of Cratoxylum
formosanum. Phytochemistry, Vol, 42, No 4, PP.1195-1198.
22. Nawong Boonnak, Chatchanok Karalai, Suchada Chantrapromma, Channita
Ponglimanont, Hoong-Kun Fun (2006), Bioac tive prenylated xanthones
and
anthraquinones
Tetrahedron 62 ,8850-8859.
http://www.lrc-tnu.edu.vn
60
23. Pitchaon Maisuthisakul, Rungnaphar Pongsawatmanit, Michael H.Gordon
(2007), Charaterization of the phytochemicals and antioxidant properties
of extracts from Teaw (Cratoxylum fromosum Dyer). Food chemistry
100 1620-1629.
24. Sompong Boonsri, Chatchanok Karalai, Chanita Ponglimanont, Akkharawit
Kanjana-Opas, Kan Chantrapromma (2006), Anibac terial and cytotoxic
xanthones from the roots of Cratoxylum fomosum. Phytochemistry 67,723-727.
25. Soon Yew Tang, Matthew Whiteman, Zhao Feng Peng, Andrew Jeniner, Eu
Leong Yong and Barry Halliwell (2004), Charcterition of Antioxidant and
Antiglycation Properties and Isolation of Active Ing redients from
Traditional Chinese medicines. Free Radical Biology & Medicine,
Vol.36, No.12, pp.1575-1587, .
26. Soon Yew Tang, Matthew Whiteman, Andrew Jeniner, Zhao Feng Peng
and Barry Halliwell (2004), Mechanisin of cell death induced bi an
antioxidant extract of Cratoxylum cochinchinense, Free Radical
Biology and Medicine, Vol 36, No.12,pp, 1588-1611.
27. Xue- Li Cao, Yu Tian, Tian-You Zhang, Yoichiro Ito (2000), Supercritical
fluid extraction of catechins Cratoxylum prunifolium Dyer. Joumal of
Chromatography A, 898, 75-81.
28.
Institute
of
New
Technology
Application,
Xizhimen
http://www.lrc-tnu.edu.vn
61
PH LC
I. Cht taraxeron
Trang
Ph LC-MS ca taraxeron. 62
Ph FT-IR ca taraxeron................................................................................65
Ph 1H-NMR ca taraxeron66
Ph 13C-NMR ca taraxeron...71
Ph C13CPD & DEPT ca taraxeron.73
Ph C13CPD-COSYGP ca taraxeron...78
Ph C13CPD-HSQC ca taraxeron81
Ph C13CPD-HMBC ca taraxeron...82
II. Cht stigmasterol
Ph EI-MS ca stigmasterol........84
Ph FT-IR ca stigmasterol ...85
Ph 1H-NMR ca stigmasterol86
Ph 13C-NMR ca stigmasterol...87
III. Cht - Sitosterol
Ph EI-MS ca - Sitosterol88
Ph FT-IR ca - Sitosterol.89
Ph 1H-NMR ca - Sitosterol90
Ph 13C-NMR ca - Sitosterol...95
Ph C13CPD & DEPT ca - Sitosterol.97
IV. Cht axit gallic
Ph FT-IR caaxit gallic 98
Ph t ngoi UV ca axit gallic.................................................................. 99
Ph 1H-NMR ca axit gallic..100
Ph 13C-NMR ca axit gallic101
http://www.lrc-tnu.edu.vn
62
http://www.lrc-tnu.edu.vn
63
http://www.lrc-tnu.edu.vn
64
http://www.lrc-tnu.edu.vn
65
Ph FT-IR ca taraxeron
http://www.lrc-tnu.edu.vn
66
Ph 1H-NMR ca taraxeron
http://www.lrc-tnu.edu.vn
67
http://www.lrc-tnu.edu.vn
68
http://www.lrc-tnu.edu.vn
69
http://www.lrc-tnu.edu.vn
70
http://www.lrc-tnu.edu.vn
71
Ph 13C-NMR ca taraxeron
S ha bi Trung tm Hc liu i hc Thi Nguyn
http://www.lrc-tnu.edu.vn
72
http://www.lrc-tnu.edu.vn
73
Ph DEPT ca taraxeron
http://www.lrc-tnu.edu.vn
74
http://www.lrc-tnu.edu.vn
75
http://www.lrc-tnu.edu.vn
76
http://www.lrc-tnu.edu.vn
77
http://www.lrc-tnu.edu.vn
78
Ph COSYGP ca taraxeron
http://www.lrc-tnu.edu.vn
79
http://www.lrc-tnu.edu.vn
80
Hnh
http://www.lrc-tnu.edu.vn
81
Ph HSQC ca taraxeron
S ha bi Trung tm Hc liu i hc Thi Nguyn
http://www.lrc-tnu.edu.vn
82
Hnh
Ph HMBC ca taraxeron
S ha bi Trung tm Hc liu i hc Thi Nguyn
http://www.lrc-tnu.edu.vn
83
http://www.lrc-tnu.edu.vn
84
http://www.lrc-tnu.edu.vn
85
http://www.lrc-tnu.edu.vn
86
http://www.lrc-tnu.edu.vn
87
http://www.lrc-tnu.edu.vn
88
http://www.lrc-tnu.edu.vn
89
http://www.lrc-tnu.edu.vn
90
Ph 1H NMR ca - Sitosterol
S ha bi Trung tm Hc liu i hc Thi Nguyn
http://www.lrc-tnu.edu.vn
91
http://www.lrc-tnu.edu.vn
92
http://www.lrc-tnu.edu.vn
93
http://www.lrc-tnu.edu.vn
94
http://www.lrc-tnu.edu.vn
95
http://www.lrc-tnu.edu.vn
96
http://www.lrc-tnu.edu.vn
97
Ph DEPT ca - Sitosterol
S ha bi Trung tm Hc liu i hc Thi Nguyn
http://www.lrc-tnu.edu.vn
98
Ph IR ca axit gallic
S ha bi Trung tm Hc liu i hc Thi Nguyn
http://www.lrc-tnu.edu.vn
99
Ph UV ca axit gallic
S ha bi Trung tm Hc liu i hc Thi Nguyn
http://www.lrc-tnu.edu.vn
100
http://www.lrc-tnu.edu.vn
101
http://www.lrc-tnu.edu.vn
102
http://www.lrc-tnu.edu.vn
103
http://www.lrc-tnu.edu.vn
104
http://www.lrc-tnu.edu.vn
105
MC LC
M U.................................................................................................... 1
Chng 1. TNG QUAN........................................................................... 3
1.1. M t thc vt .................................................................................. 3
1.2. Mt s cng dng ca chi Cratoxylum............................................... 4
1.3. Tnh hnh nghin cu ha hc thc vt chi Cratoxylum ...................... 7
1.3.1. Cc hp cht c khung triterpen ................................................. 7
1.3.2. Cc cht axit hu c .................................................................. 8
1.3.3. Cc cht c khung xanthone ....................................................... 9
1.3.4. Mt s i din ca khung anthraquinon.................................. 15
1.3.5. Mt s i din ca khung flavonoit......................................... 16
1.4. Nhng nghin cu ho thc vt loi Cratoxylum prunifolium. .......... 17
Chng 2. PHN THC NGHIM ........................................................ 19
2.1. i tng v phng php nghin cu ............................................ 19
2.1.1. Thu mu cy, xc nh tn khoa hc v phng php x
l mu ...................................................................................... 19
2.1.2. Phng php phn lp cc hp cht t cc dch chit ................ 19
2.1.3. Phng php kho st v xc nh cu trc ho hc cc
hp cht................................................................................... 20
2.2. Dng c, ho cht v thit b nghin cu ......................................... 20
2.2.1. Dng c v ho cht ................................................................ 20
2.2.2. Thit b nghin cu .................................................................. 21
2.3. Cc dch chit t cy ngn (Cratoxylum prunifolium Kurtz) ........ 21
2.3.1. Cc dch chit .......................................................................... 21
2.3.2. Kho st nh tnh cc dch chit .............................................. 23
2.3.3. Th hot tnh sinh hc.............................................................. 23
http://www.lrc-tnu.edu.vn
106
2.4. Chit sut, phn lp v tinh ch cc cht t cy ngn................... 25
2.4.1. Dch chit clorofom.................................................................. 26
2.4.1.1. Taraxeron (Friendoolean - 14- en-3-on) (C1) ................... 26
2.4.1.2. Stigmast- 5,22- ien-24R-3-ol .......................................... 27
2.4.1.3. - Sitosterol....................................................................... 27
2.4.2. Dch chit trong etylaxetat (E) ............................................... 28
Chng 3. THO LUN KT QU NGHIN CU.............................. 29
3.1. Nguyn tc chung........................................................................... 29
3.2. Phn tch nh tnh v pht hin cc nhm cht ................................ 30
3.3. Phn lp v nhn dng cc hp cht c trong cc dch chit khc
nhau ca cy ngn ............................................................................ 30
3.3.1. Taraxeron ( hay Frendoolean-14-en-3-on) (C-1) .................... 30
3.3.2. Stingmast -5,22-dien-24R-3-ol (C-2) .................................... 38
3.3.3. -sitosterol (C-3) ................................................................... 44
3.3.4. Axit gallic (E-1) .................................................................... 50
3.4. Th hot tnh sinh hc .................................................................... 56
KT LUN .............................................................................................. 57
KIN NGH ............................................................................................. 57
TI LIU THAM KHO........................................................................ 58
http://www.lrc-tnu.edu.vn
107
DAN H MC C C B N G
Bng 2.1. Khi lng cht tng s c chit tng phn on l cy
ngn (Cratoxylum prunifolium Kurtz) ..................................... 22
Bng 2.2. Pht hin cc nhm cht trong cy ngn ................................. 23
Bng 2.3. Kt qu th hot tnh sinh hc ca dch chit th t l cy
ngn (Cratoxylum prunifolium) ................................................... 25
Bng 3.1. Ph 13C-NMR v 1H-NMR ca taraxeron (C-1) t l ngn...... 33
Bng 3.2. Ph 13C-NMR ca cc cht C-2 v stingmasterol [16] ............... 39
Bng 3.3. Ph 13C-NMR ca cc cht C-3 v -sitosterol [16] .................. 45
Bng 3.4. S liu ph NMR ca E-1 v s liu ph NMR trong phn
mm ACD/NMR ca axit gallic .................................................. 50
http://www.lrc-tnu.edu.vn
108
DAN H MC C C H N H
Hnh 1.1. nh cy ngn (Cratoxylum Prunifolium Kurt, Hypericaceae) ... 4
Hnh 2.1. nh c gy c ch xung quanh ging thch............................ 24
Hnh 3.1 ................................................................................................... 34
Hnh 3.2 ................................................................................................... 35
Hnh 3.3 ................................................................................................... 36
Hnh 3.4 ................................................................................................... 37
Hnh 3.5 ................................................................................................... 40
Hnh 3.6 ................................................................................................... 41
Hnh 3.7 ................................................................................................... 42
Hnh 3.8 ................................................................................................... 43
Hnh 3.9 ................................................................................................... 46
Hnh 3.10 ................................................................................................. 47
Hnh 3.11 ................................................................................................. 48
Hnh 3.12 ................................................................................................. 49
Hnh 3.13 ................................................................................................. 52
Hnh 3.14 ................................................................................................. 53
Hnh 3.15. Ph 1H-NMR ca cht axit gallic ............................................. 54
Hnh 3.16. Ph 13C-NMR ca cht axit gallic ............................................. 55
DAN H MC C C S
S 2.1. Quy trnh ngm chit ................................................................ 22
http://www.lrc-tnu.edu.vn