M U
Nghin cu thnh phn ho hc cy m Trn Thanh H - K 50A
M U
Vit Nam c mt thm thc vt rt phong ph vi hn 12.000 loi bao gm 2500
chi v 300 h. Nhiu loi trong c s dng t lu trong cc lnh vc khc nhau
ca cuc sng, c bit l dng lm thuc cha bnh.
Plantago major L. tn thng gi l cy M , thuc h M (Plantaginaceae)
l loi cy mc hoang v c trng ph bin nhiu ni trn khp nc ta v nhiu nc
trn th gii. y l mt cy dc liu c dng ph bin, cc b phn c s dng ring
bit trong iu tr bnh cho cc v thuc khc nhau nh: Xa tin t (Semen
Plantaginis), L m (Folium Plantaginis) hay M tho (Herbal
Plantaginis). N mang nhiu tc dng dc l c gi tr: tc dng li tiu, cha
ho, tc dng khng sinh, cha l cp v mn tnh, tc dng lm hng phn thn
kinh,... ngoi ra li t c vi c th sinh vt. Do , cc v thuc t cy M c
quan tm nghin cu v c tiu chun ho trong s dng, cc tiu chun v kim
nghim dc liu t M c trong t in quc gia.
Nh bit, tc dng cha bnh ca dc liu c quy nh bi hot tnh sinh, dc hc
ca cc hp cht ho hc c trong . Mc d cy M c nghin cu nhiu c v thnh phn
ho hc v hot tnh sinh hc, song cho n nay n vn ang thu ht c s quan tm
c bit ca cc nh nghin cu trn th gii. Vi mong mun gp phn tm hiu cy M
, chng ti thc hin ti: Nghin cu thnh phn ho hc cy M (Plantago major
L., Plantaginaceae).
Theo hng nghin cu ny, lun vn c nhng nhim v sau:1. Tch chit v kho
st s c mt ca cc lp cht c trong cy M
(Plantago major L.).
2. Phn tch cc phn chit v phn lp mt s thnh phn chnh.3. Kho st cu
trc mt s cht phn lp c.
Chng 1: TNG QUAN TI LIU
Plantago major L. c s dng lm thuc t hng th k nay. y l mt cy dc
liu mang nhiu tc dng dc l c gi tr: tc dng li tiu, tc dng cha ho,
khng sinh, cha l cp v mn tnh, tc dng lm hng phn thn kinh, lm lnh vt
thng, chng vim, gim au, chng oxy ho, khng sinh yu, iu bin min dch v
chng lot...ngoi ra li t c vi c th sinh vt. Theo kinh nghim s dng lu
truyn trong dn gian, cc bnh dng m iu tr bao gm: bnh c lin quan ti
da, c quan h hp, c quan tiu ho, di truyn, chuyn ho, chng ung th,
gim au v chng nhim khun. V thnh phn ho hc, trong cy c cha
polysaccarit, lipid, dn xut ca axit caffeic, flavonoid, iridoid
glycosid, alkaloid, terpennoid v cc axit hu c. 1.1. THC VT HC [1,
2, 3 ] 1.1.1. H M (PLANTAGINACEAE).
H M , danh php khoa hc: Plantaginaceae Juss., l mt h thc vt c
hoa trong b Hoa mi (Lamiales). Chi in hnh l Plantago L.
H Plantaginaceae ngha rng (sensu lato) l mt h a dng, ph bin ton
cu, nhng ch yu ti cc khu vc n i. N bao gm ch yu l cc cy thn tho, cy
bi v mt t cy thy sinh c r mc trong t (chng hn chi Callitriche). L
ca chng mc thnh vng xon hoc mc i, cc l c dng l n hay l kp. Cu trc v
hnh dng ca hoa rt a dng. Mt s chi c 4 i hoa v 4 cnh hoa, nhng loi
khc c t 5-8 i/cnh hoa, chng hn chi Sibthorpia. Hoa ca phn ln cc chi
l i xng theo nhiu mt phng. Trng hoa thng thng c 2 mi. Trong mt s
chi, b nh c hnh thnh trc trng hoa. Qu l dng qu nang, thng nt ra ti
cc phn chia gia cc ngn. Trong chi Veronica phn phn chia ny theo
chiu di; trong cc loi ca chi Antirrhinum th khi qu nt n gii phng
phn hoa thng qua cc l u bao phn; hoc n c th xoay chuyn thng qua ng
tun hon ngang xung quanh bao v.
1.1.2. CHI M .H Plantaginaceae (ngha rng) bao gm khong 90 chi v
khong 1.700 loi. Chi M (danh php khoa hc: Plantago) l mt chi cha
khong 200 loi thc vt nh, khng d thy, c gi chung l M . Phn ln cc loi
l cy thn tho, mc d c mt s t loi l dng cy bi nh, cao ti 60 cm. L ca
chng khng c cung, nhng c mt phn hp gn thn cy, l dng gi-cung l. Chng
c 3 hay 5 gn l song song v ta ra cc phn rng hn ca phin l. Cc l hoc
l rng hoc l hp bn, ph thuc vo tng loi. Cc cm hoa sinh ra cc cung
thng thng cao 5-40 cm, v c th l mt nn ngn hay mt cnh hoa di, vi
nhiu hoa nh, c th phn nh gi. Cc loi m b u trng ca mt s loi thuc b
Cnh vy (Lepidoptera) n l.
Cc loi M mc nhiu ni trn th gii, bao gm chu M, chu , Australia,
New Zealand, chu Phi v chu u. Nhiu loi trong chi phn b rng khp th
gii nh l mt dng c di.
1.1.3. M LN (PLANTAGO MAJOR L.). [1,3]M ln, (tn ca loi ny xut
pht t vic n hay mc vt chn nga ko xe; m l nga, l mng chn); cc tn gi
khc nh m tho, nh n, dt (Thi), suma (Th).
Tn khoa hc: Plantago major L., Plantago asiatia L., thuc Gii:
Plantae; Ngnh: Magnoliophyta; Lp Magnoliopsida; B Lamiales; H
Plantaginaceae; Chi Plantago L. M l mt loi c sng lu nm, thn ngn, l
mc thnh cm gc, phin l hnh tha hay hnh trng, c gn dc theo sng l v ng
quy ngn v gc l. Hoa mc thnh bng, c cn di xut pht t k l. Hoa u, lng
tnh, i 4, xp cho, hi nh nhau gc. Trng mu nu gm 4 thu nm xen k gia k
l i. Nh 4, ch nh mnh, di, 2 l non cha nhiu tin non. Qu hp trong cha
nhiu ht nh trn hay bu dc, mu nu en bng. M l cy a sng, a m v c kh
nng chu hn nh. M mc hoang v c trng ti nhiu ni nc ta. Thng trng bng
ht ni t m vo ma xun. Thu hoch M thng vo thng 5-8, thu hi vo lc cy
ang ra hoa, nh c cy v, b gc r v cc l su a, em ra sch, ro nc, sau em
phi hoc sy kh. Thu hoch ht M thng vo thng 7-8, thu hi vo nhng ngy
kh ro, ct ly nhng bng qu gi, em phi hay sy kh, ri v xt, sng sy sch
ri ly ht v tip tc phi kh.
Cy m ( Plantago major L.) 1.2. NG DNG [1, 2, 10, 23, 26]. 1.2.1.
TC DNG CHA BNH
Cc loi trong chi Plantago thng c s dng lm thuc. Chng c tc dng lm
se, gii c, khng trng, chng vim nhim, cng nh lm du chng vim, long m,
cm mu v li tiu. S dng ngoi, dng thuc p t l l c ch i vi cc vt cn ca
cn trng, chng pht ban do sn c, cc ch l lot nh cng nh nht. S dng
trong, n dng iu tr ho v vim ph qun di dng ch, thuc sc hay xi r. Ht
ca Plantago psyllium l c ch iu tr to bn, rut kt co cng, b sung cht
x dinh dng v bnh ti tha, h thp cholesterol v kim sot bnh i ng. Theo
y hc c truyn, M c v ngt, nht, tnh hn, khng c, vo 3 kinh can, thn v
tiu trng, c tc dng tiu vim, phong nhit, li tiu, cha ho, lm long m,
sng mt, thuc b. M thng c dng cha si niu, nhim trng ng tit niu, vim
thn, cm lnh ho, vim kh qun, vim rut, vim kt mc cp, vim gan. L sc
hoc nu cao ung lm thuc thng tiu, cha ho lu ngy, vim kh qun, t l, au
mt . L ti sc ung cha l cp v mn tnh; Gi nt p mn nht, lm mn nht nhanh
chng v m v mau lnh. Ht M cha ph thng, thp t, mt , ho do ph nhit. C
cy nu cao c bi tr bng hoc ly cy non, ti sc ly nc ung cha cao huyt
p.
Ngoi Trung Quc v Vit Nam, Plantago major L. cng c s dng nhiu nc
khc trn th gii trong iu tr nhiu loi bnh (bng 1) bao gm cc bnh v da,
bnh nhim khun, bnh h tiu ho, h h hp, tun hon, bnh chuyn ho, chng
ung th, gim au v lm lnh vt thng.Bng 1: ng dng P. major trong iu tr
bnh CC LOI BNHPHN S DNG
Cc bnh ngoi da: p xe, mn, chng vim, sng au do cn trng t,cn, thm
tm, bng, vim da, chng nhim khun cho vt thng h, lm mm nhng phn da b
chai cng, ni ban , cm mu vt thng, mn nga, chng to m cho bnh chc l,
lm nhanh lin vt thngh.r, l, ht, ton cy, dch chit nc
Cc bnh h hp: cha ho, hen suyn, vim ph qun, cm lnh, thi tai, tc
dng long m, cc bnh phi, vim hng.l, r, ht, ton cy,dch chit nc
Cc bnh tiu ho: t, to bn, l, vim lot d Dy, vim li, su rng, vim
ming.l, r, ht, ton cy, dch chit nc, dch chit cn
Cc bnh h sinh dc- tit niu: ri lon kinh nguyt, si thn, au bng
quang, nhim khun niu o, vim m o.l, r, ht, ton cy, dch chit nc
Bnh lin quan ti h tun hon v tim: chng tan mu, tng lc mu, dng cho
ngi b tiu ng, tc dng li tiu, ph, tr, cao huyt p.l, r, ht, ton cy,
dch chit nc
Bnh h thn kinh: h st, gim au, cha sc tm l, suy nhc thn kinh, gim
au rng.l, r, ht, ton cy, dch chit nc
Chng ung th: chng u buton cy, l, dch chit nc
Chng k sinh trng, gii c: ty giun, cha st Rt, rn cnton cy, l, dch
chit nc
1.2.2. MT S BI THUC DN GIAN T M
1. Tr vim ng tit niu, i rt i but:
2. Tr tiu chy:
3. Tr au mt sng do can nhit:
4. Tr tiu chy tr em 5. Tr ho. 1.3. THNH PHN HO HC [4-7, 12,
16-19, 22-27] 1.3.1. CACBOHYDRAT.
Trong ht cha mt s monosaccharit: glucoz, fructoz, xyloz v
rhamnoz; disaccharit c sucroz; trisaccharit c planteoz ng vai tr mt
cacbohydrat d tr trong ht. V ngoi ca ht c cha cht polysaccharit c
kh nng to dch nhy c nht cao trong nc. Thnh phn dch chit
polysaccharit ht m c xyloz, arabinoz, axit galacturonic, axit
glucuronic, rhamnoz, galactoz v glucoz. Trong l c cha trisaccharit
raffinoz, tetrasccharit stachyoz, stachyoz ng vai tr l cacbonhydrat
d tr tm thi trong thc vt.
Th nghim trn ng vt thc nghim i vi mt s plantaglucid t M nh
galactoarabinan, galactan cho thy cc cht ny c tc dng cha lot liu
1,5-3 g/ngy. liu 1 mg/kg, cc plantaglucid ny lm gim t l lot d dy
chut gp 20 ln so vi khng iu tr; c tc dng lm tng tit dch v ch; lm
gim t l nhim khun v chng co tht ng rut th; lm gim phn ng vim, ph n
gy bi formalin v dextran.Mt s dng pectin este ho c phn t lng ln
(46-48 kDa) c tc dng hot ho i thc bo ngi (in vitro) lm tng sinh TNF
(tumornecrosis ). 1.3.2. LIPITMt s axit bo phn lp c t P.major c h
thng li bng di y: Bng 2: Axit bo phn lp t ht P.majorAXIT BO HM LNG
(%)
Axit Myristic -
Axit Plamitic -
Axit Stearic -
Axit Oleic 37,4
Axit Linoleic 25,3
Axit Linolenic 0,9
Axit Arachidic -
Axit Behenic -
Axit Lignoceric -
Axit 9-Hydroxy-cis-1octadecenoic 1,5
Bng 3: Axit bo phn lp t l P. major AXIT BO HM LNG (%)
Axit Myristic1,8
Axit Plamitic15,9
Axit Stearic2,1
Axit Arachidic1,3
Axit Behenic1,3
1.3.3. ANKALOIT
T P.major nhn dng v phn lp c indicain v plantagonin
.
Plantagonin Indicain 1.3.4. CAFFEIC V DN XUTT dch chit methanol
P.major phn lp c axit caffeic v mt s dn xut ca n:
axit caffeic axit chlorogenic caffeinCaffein (3,7-
Dihydro-1,3,7-trimethyl-1H-purine-2,6-dione) c tc dng kch thch hot
ng ca thn kinh trung ng chn lc trn v no, lm tng kh nng nhn thc, tng
kh nng lm vic tr c, gim cm gic mt mi v bun ng. axit acteoside axit
plantagoside
Plantagoside c tc dng c ch sinh arachidonic axit trong qu trnh
ph n tai chut, chng vim, c ch 7-lipoxygenaza, 15-lipoxygenaza, cAMP
phosphodiesteraza v tc dng chng oxy ho ng thi c tc dng khng sinh vi
sinh vt.
Acteoside c tc dng dit gc t do gy bnh, chng oxy ho v c ch qu
trnh peoxy ho lipit; c ch 15-lipoxygenaza km hn so vi plantagoside;
c ch protein kinaza C; c ch andoz reductaza v s to thnh 5-HETE; tc
dng khng khun, gim au, iu bin min dch. Tc dng chng tng huyt p, liu
10 mg/kg trn chut, acteoside c tc dng gim p sut trn c thi k tm thu,
tm trng v trn ng mch ch. 1.3.5. FLAVONOIT
Mt s flavonoit phn lp t P.major bao gm: lutein 7-glucosit,
hispidulin 7-glucuronit, luteolin 7-diglucosit, apigenin
7-glucosit, nepetin 7-glucosit, luteolin 6-hydroxy 4-methoxy
7-galactosit, plantaginin, homoplantaginin.
Tc dng sinh hc ni bt ca nhm cht flavonoit l kh nng chng oxy ho,
dit tr gc t do hot ng gy bnh, c ch qu trnh peoxy ho lipit, ngoi ra
cn nhiu tc dng quan trng khc bao gm c tc dng chng virut, khng khun,
chng ung th,...c bit, mt s flavonoit t P.major nh planaginin,
homoplantaginin v luteolin 7-glucosit c tc dng c ch HIV- reverse
transcriptaza (in vitro). 1.3.6. IRIDOIT GLYSOSIT
Mt s Iridoit glysosit phn lp c t P.major c tng kt li di y:
asperuloside aucubin
gardoside axit geniposidic
Aucubin c tc dng chng vim, chng co tht, gii c i vi trng hp ng c
nm, tc dng bo v gan gy bi -amanitin v CCl4, c ch virus vim gan B.
1.3.7. TERPENOIT.
Mt s triterpenoit phn lp c t l P.major bao gm: axit oleanolic,
axit ursolic, 18 - glycyrrhetinic axit v sitosterol. Ursolic axit c
tc dng c ch cyclooxygenase-2 v cyclooxygenase-1 l cc enzym xc tc
sinh tng hp prostaglandin (in vitro). Axit ursolic v axit oleanolic
c tc dng bo v gan, chng ung th, h m mu.
axit oleanolic axit ursolic
18-glycyrrhentinic -sitosterolAxit Ursolic (axit
3-Hidroxy-12-ursen-28-oic) c tc dng sinh hc: li tiu, chng ung th,
chng vim lot v x ho, c ch hot ng ca virus HIV, c ch protein kinase
C, km hm HIV-1 protease.Axit oleanolic c tc dng sinh hc: chng vim
khp, chng c, chng ri lon gan v tng cng h min dch ca c th. Gn y, hot
tnh khng HIV c pht hin do kh nng ngn cn qu trnh sao m ca virus
HIV-1 t bo H9. 1.3.8. VITAMINBng 4: Mt s vitamin c trong
P.major
VITAMIN NGUN HM LNG
-caroten ht 6 mg/100g
Vitamin C ht 19 mg/100g
Vitamin C l non 25 mg/100g
Dehydroascobic l non 31 mg/100g
Vitamin K1 l-
1.3.9. AXIT HU C.
T P.major phn lp c mt s axit hu c: axit fumaric, axit syringic,
axit vanillic, axit p-hydroxy benzoic, axit ferulic, axit
p-coumaric, axit gentisic, axit salicylic, axit benzoic v axit
cinamic.
axit fumaric axit syringic axit vanillic
EMBED ACD.ChemSketch.20
axit ferulic axit p-coumaric axit gentisic axit salicylic 1.4.
TC DNG DC L [13, 19-22, 25-30]
Trn th gii, P.major c s dng cha nhiu bnh khc nhau, cc tc dng ny
c quan tm tm hiu bi nhiu nh nghin cu trn khp th gii. Nhiu nghin cu
tc dng sinh hc c tin hnh nhm gii ngha tc dng cha bnh ny. Di y l kt
qu cng b m chng ti thu thp c: 1.4.1. Tc dng chng lot.
Nghin cu nh gi kh nng chng lot ca P.major thc hin trn m hnh gy
lot d dy chut bng cch nhng nc gy stress, sau iu tr bng dch chit t
P.major. Kt qu cho thy nhm chut iu tr bng cn chit (metanol-nc) vi
liu lng 1.2g/kg gim 40% hin tng lot so vi nhm chut i chng khng iu
tr. i vi cn chit t dch chit nc, tc dng ny l 37%, cn t dch chit
metanol l 29%. 1.4.2. Tc dng chng ung th.
Cc nghin cu trn ng vt thc nghim cho thy P.major c tc dng phng
chng ung th, chng di cn ung th, phng ung th v, khng c tc dng trn
bnh bch cu cp. Nghin cu trn chut gy ung th t cung bng cch ly nhim
virus herpes, sau 60 tun 93,3% chut khng iu tr xut hin khi u, trong
khi ch c 18,2% chut iu tr bng dch chit t P.major xut hin khi u. Tc
dng trn ch yu l do s tng cng h min dch hn l tc dng trc tip ln
virus, bi v dch chit t P.major c tc dng tt iu tr nhim virut herpes
trn lm sng song li khng c tc dng ln virus herpes trong nghin cu in
vitro. Tng t i vi vi khun gy bnh, trn lm sng dch chit t P.major c
tc dng tt iu tr cc trng hp nhim khun song khng c tc dng c ch cc vi
khun gy bnh trn in vitro. 1.4.3. Tc dng iu bin min dch.
Theo nghin cu ca Basaran v cng s, dch chit t l P.major c tc dng
iu bin min dch. 1.4.4. Nghin cu chng nhim khun (in vitro).
Dch chit methanol c tc dng c ch Salmonnella typhimurinum, S.
aureus v M.phlei, c ch nm F. tricuictum v M. gypseum, C. albicans v
S. cerevisiae. Dch chit 50% methanol c tc dng c ch S. aureus,
B.subtilis, S.dysenteriae v E. coli. Dch chit 70% methanol c tc dng
c ch S. flexneri, S. aureus, S. sonnei, E. coli, Esherichia crim v
M. smegmatis.
Cha l amip do c tc dng chng giardia
Tc dng chng st rt : c ch Plasmodium falciparum, c s dng Tanzania
cha st rt.
Chng virus: khng chng herpes v polio virus, chng bovine corona
virus, bovine herpes virus type I, bovine parainfluenza virus type
III, bovine rotavirus, bovine respiratory syncytial virus, vaccinia
virus v vesicular stomatitis virus. 1.4.5. Chng vim v gim au
Dch chit nc t bt l P. major c tc dng chng vim, cc nghin cu cng
ch ra rng tc dng ny c lin quan ti tc dng c ch sinh tng hp
prostaglandin, tc dng c ch sinh tng hp cyclooxygenase. Cc nghin cu
cng cho thy dch chit t P.major c tc dng gim au, an thn. 1.4.6. Tc
dng chng oxy ho v loi gc t do gy bnh.
Cc nghin cu cho thy phn chit giu flavonoid t P.major c tc dng
chng oxy ho, c ch peoxy ho lipid mng t bo gan, bo v gan. 1.4.7 Tc
dng li tiu.
Kt qu nghin cu ca Casceres v cng s, cho thy nc tiu chut tng
10844% sau 6h, nghin cu s dng bt l m vi liu lng 1g/kg trng lng.
Chng 2: THC NGHIM 2.1. PHNG PHP NGHIN CU
phn tch v phn tch cc phn chit ca cy s dng cc phng php:
Sc k lp mng (TLC): Thc hin trn bn mng trng sn silica gel Merck
60 dy 0,25 mm trn nn nhm. Thuc th hin mu l dung dch vanilin/H2SO4
1% v n t ngoi bc sng 366 nm.
Sc k ct (CC): Sc k ct oc thc hin di trng lc ca dung mi. Cht hp
ph cho sc k ct l silica gel (c ht 63-100 m), c nhi theo phng php
nhi t.
Kt tinh phn on c s dng trong vic tinh ch cc cht.
Cu trc ca cc hp cht c kho st nh s kt hp cc phng php ph: Ph hng
ngoi (IR) c ghi trn my Impac 410-Nicolet FT- IR.Ph khi lng (EI-MS,
70eV) ghi trn my Hewlett Packard HP 5890, Serie II.Ph cng hng t ht
nhn 1H-NMR, 13C-NMR c ghi trn my Brucker Avance-500 MHz, chun ni
TMS (tetrametyl silan), chuyn dch ho hc c biu th bng ppm, hng s J
tnh theo Hz. 2.2. IU CH CC PHN CHIT T CY M 2.2.1. Mu thc vt
Mu nghin cu l phn trn mt t ca cy m (P. major L.), c thu hi vo
thng 7 ti Qunh Ph, Thi Bnh. Mu sau khi ly v c ra sch, phi trong bng
rm ri em sy kh nhit 40-50C, sau em v tht nh. Phn nguyn liu thu c em
ngm chit vi etanol 95%, sau c phn b chn lc vi cc dung mi thch hp.
Cc dung mi c dng l ete du ho 30-600, etyl axetat, n-butanol. 2.2.2.
iu ch cc cn chit
800 g bt kh nguyn liu c ngm vi etanol 95% nhit phng trong 3 ngy.
Sau lc ly dch chit, cn b li c ngm tip vi etanol 95%. Qu trnh ny c
lp li 2 ln. Cc dch chit c gp li, v ct loi etanol 60C di p sut thp n
khi cn 200 ml. Pha long dch chit etanol vi 200 ml ncct ri chit ln
lt vi cc dung mi c phn cc tng dn: ete du ho 30-600, etyl axetat,
n-butanol. Cc dch chit ete du ho, etyl axetat, n-butanol c lm khan
bng Na2SO4. Lc b Na2SO4 v ct loi ht dung mi ta thu c cc phn cn chit
tng ng.Bng 5: Hiu sut cc phn cn chit t cy m TT Phn chit Khi lng (g)
Hiu sut*(%)
1 Ete du ho344.250
2 Etyl axetat151.875
3 n-Butanol253.125
* Tnh theo khi lng mu kh ban u. 2.3 PHN TCH CC PHN CHIT T CY M .
2.3.1. Phn chit ete du hoPhn tch phn chit ete du ho (PMD) bng
TLC
Dng capila a cht ln bn mng trng sn DC Aluflolien 60F254 Merck.
Sau tin hnh kho st sc k lp mng trong cc h dung mi sau: diclomethan/
etyl axetat; n-hexan/ etyl axetat; diclomethan/ axeton.
S phn tch t kt qu tt khi trin khai trong h dung mi: n-hexan/
etyl axetat: 7,5/2,5 (v/v).Bng 6: Kt qu kho st phn chit ete du ho
bng TLCSTTRfAS thngAS UVVanilin/H2SO4Vanilin/ H2SO4,t
10.875 -- ----Tm
20.8 Xanh XanhXanh
30.75 -- ----Tm
40.7 Xanh XanhXanh
50.625 Vng --VngVng
60.575 -- ----Tm
70.475 Vng --VngVng
80.3 Vng HngVngVng
T kt qu phn tch thnh phn cn chit ete du ho c th a ra cc nhn xt
sau:1) Phn cn chit ete du ho gm nhiu cht phc tp.
2) Mt s vt mu hng tm khi phun vi vanilin- axit sunfuric; h nng c
hm lng tng i ph hp cho vic phn tch.
Phn tch cn chit ete du ho bng phng php sc k ct.Chun b mu: cho
15g silica gel vo mt ci s. Ho tan 10g cn ete du ho trong axeton ri
dng cng t ht nh t t ln silica gel. i cho dung mi bay hi bt, ta dng
chy s trn u cho n khi silica gel ti ra. Tip tc tm cht ln silica
gel, trn u. Qu trnh c lp li cho n khi tm xong cht v silica gel kh
ti c mu xanh en.Chun b ct sc k
Cn 50g silica gel (c ht 63-100 m), cho vo cc, dung mi n-hexan
vo, khuy cho ht bt kh ri nhi vo ct, dng qa bp cao su g nh cho n khi
trong ct khng cn bt kh, dng a thu tinh dn u mt ct, sau a hn hp bt
tm mu cht ln ct, cho ln trn mt t bng thu tinh trnh b khuch tn ngc.
Gii hp ph gradien bng h dung mi n-hexan: etyl axetat, sau khi kho
st bng sc k lp mng thu c 7 nhm phn on gm nhng cht c gi tr Rf gn
nhau k hiu t I n VII.Bng 7: Kt qu sc k ct phn chit ete du hoSTTT l
n-H/ES ml Phn on
1100% n- H1001-5
29.5: 0.51006-10
39.0: 1.010011-15
48.5: 1.510016-20
58.0: 2.020021-29
67.5: 2.530030-44
77.0: 3.020045-55
86.5: 3.520056-65
96.0: 4.010066-70
Kt hp x l loi mu v kt tinh li, t nhm phn on I, II, III, VII, ta
thu c cc cht, k hiu l PMD-1, PMD-2, PMD-3, PMD-4 kt tinh mu trng.
2.3.2 Phn chit etyl axetatPhn tch phn chit etyl axetat( PME) bng
TLC
Ho tan phn chit etyl axetat trong methanol. Dng capila a cht ln
bn mng trng sn DC Alufoluen 60F254 Merck. Trin khai chy sc k lp mng
vi cc h dung mi toluen/ axeton; toluen/ etyl axetat,
diclomethan/axeton cho thy h dung mi toluen/ etyl axetat (8,0:2,0)
c kh nng tch tt nht. Bng 8: Kt qu kho st phn chit etyl axetat bng
TLC
STTRfAS thngAS UVVanilin/ H2SO4Vanilin/H2SO4,t
10.8NuHngXanhXanh
20.725------Tm
30.65NuHngXanhXanh
40.6Vng nht--VngVng
50.375------Tm
Phn tch phn chit etyl axetat bng sc k ct
Cc bc chun b ct sc k v trin khai sc k c thc hin tng t nh trnh by
mc 3.4.2. Phn chit etyl axetat (10g) c ho tan trong methanol, tm mu
v a ln ct, ra gii vi gradien hn hp dung mi toluen/ etyl axetat chng
ti thu c 60 phn on, mi phn on 25 ml.
Sau khi kho st sc k lp mng v gom cc phn on ging nhau thu c 6 nhm
phn on, k hiu t I n VI
Kt hp x l kt tinh v loi mu, t cc nhm phn on III v V thu c cht v
nh hnh kt tinh mu trng k hiu l PME-1, PME-2.
Bng 9: Kt qu sc k ct phn chit etyl axetatSTTT l To/ES mlPhn
on
1100% To1001-4
29.0: 1.01005-9
38.5:1.520010-19
48.0: 2.040020-39
57.5: 2.520039-49
67.0: 3.010050-55
76.0: 4.010055-60
2.3.3 Phn chit n- butanolPhn tch phn chit n-butanol (PMB) bng
TLC
Ho tan phn chit n-butanol trong methanol. Dng capila a cht ln bn
mng trng sn Alufloluen 60F254 Merck. Trin khai chy sc k lp mng vi
cc h dung mi diclomethan/ methanol; diclomethan/ methanol/ nc;
clorofoc/ methanol/ nc nhn thy h dung mi diclomethan/ methanol
(7/3) c kh nng tch tt nht.Bng 10: Kt qu kho st phn chit n-butanol
bng TLCSTTRfAS thngAS UVVanilin/H2SO4Vanilin/H2SO4,t
10.875Vng--VngNu
20.725vngNu VngVng
30.625------ Nu
40.375------Tm
Phn tch phn chit n-butanol bng sc k ct
10 g mu c ho tan trong methanol v c tm vi 15g silica gel v a ln
ct sc k chun b trc.Ra gii bng gradien h diclomethan/ methanol thu c
125 phn on, mi phn on 20 ml.
Tin hnh kho st sc k lp mng cc phn on thu c, gom cc phn on ging
nhau li ta thu c 8 nhm phn on, k hiu t I n VIII.
Kt hp x l loi mu v kt tinh li, t cc nhm phn on III, IV, VII ta
thu c 3 cht PMB-1 mu trng, PMB-2 mu trng, PMB-3 mu vng.
Bng 11: Kt qu sc k ct phn chit n-butanolSTTT l D/MS mlPhn on
1100% D1001-5
29.5 :1.51006-10
39.0 :1.010011-15
48.5 : 1.510016-20
58.0 : 2.020021-30
67.5 : 2.530031-45
77.0 : 3.040046-65
86.5 : 3.510066-70
96.0 : 4.010071-75
105.0 : 5.010076-80
2.4. C TRNG VT L V D KIN PH CA CC HP CHT PHN LP .Cht PMD-1
Cht PMD-1 c im nng chy 60-61C, Rf = 0.9 (n-hexan/ etyl axetat,
8/2, v/v ), cho vt mu hng sng vi dung dch vanillin/ H2SO4.
Ph khi lng (MS) ca PMD-1c pic ion phn t m/z =436.Cht PMD-2
Cht PMD-2 c im nng chy 61C, R= 0.75 (n-hexan/etyl axetat, 8/2,
v/v), cho vt mu hng vi dung dch vanillin/ H2SO4 .
Ph 1H-NMR ca n c 2 nhm metyl -CH3 (3H, t) 0.88 v 0.89 ppm; cc
nhm -CH2 khong 1.2 v 1.3 ppm; nhm OH khong 1.5 v 1.6 ppm.
Cht PMD-3
Tinh th hnh kim mu trng, kt tinh li trong h dung mi n-Hexan:
EtOAC (8:2,v/v), nhit nng chy 133-135C. Hin mu tm m vi thuc hin
vanillin/ H2SO4 c. Cht PME-1
Tinh th mu trng, kt tinh li trong h dung mi CH2Cl2: MeOH (1:1,
v/v), nhit nng chy 290-291C. Rf = 0.725 trong h To/EtOAC (8/2,
v/v), khng hin t ngoi, hin mu tm m vi thuc hin vanilin/ H2SO4
c.1H-NMR (MeOD, 500 MHz) ( (ppm) 5,24 (1H, t, J=3,5 Hz, H-12); 3,20
(1H, dd, J=9,0; 7,1 Hz, H-3); 2,20 (1H, dd, J=13,5; 4,3 Hz, H-18);
1,98 (1H, dt, J=13,5; 4,2 Hz, H-16); 1,10 (3H, s, H-27); 0,98 (3H,
s, H-26); 0,93 (3H, s, H-25); 0,91 (3H, s, H-30); 0,82 (3H, s,
H-29); 0,78 (3H, s, H-23); 0,77 (3H, s, H-24).13C-NMR (MeOD, 125
MHz) ( (ppm) 180,94 (s, C-28); 138,32 (s, C-13); 125,66 (d, C-12);
79,01 (d, C-3); 52,96 (d, C-5); 47,71 (d, C-9); 42,20 (s, C-17);
39,23 (d, C-19); 42,20 (s, C-14); 55,40 (d, C-18); 39,62 (s, C-8);
39,62 (s, C-4); 37,08 (t, C-1); 36,96 (s, C-10); 26,99 (t, C-21);
21,25 (q, C-29); 30,82 (t, C-7); 28,16 (t, C-22); 39,04 (d, C-20);
23,41 (q, C-23); 30,82(t, C-15); 23,64 (t, C-2); 24,34 (q, C-27);
23,64 (q, C-30); 23,41 (t, C-11); 23,64 (t, C-16); 18,45 (t, C-6);
15,53 (q, C-26); 17,02 (q, C-24); 17,10 (q, C-25).
EI-MS (70 eV): m/z 456 [M]+., 438, 284, 219, 203, 189, 161, 133,
119, 95, 69, 55.Cht PME-2
IR (CHCl3) cm-1: 3477 (OH), 2939 (C-H), 1690 (C=O), 1623
(>C=CH-), 753 (=C-H).EI-MS, m/z (%): 456 (M+, 4), 248 (100), 203
(79), 133 (45), 119 (20), 69 (24), 55 (28). 1H-NMR (CDCl3), : 2,83
(1H, dd, J= 14,0 v 4,2 Hz, H-18); 3,21 (1H, m, H-3); 5,28 (1H, t,
J=3,6 Hz, H-12); 0,78 (6H, s,s, CH3-23,-26); 0,90 (3H, s, CH3-30);
0,91(3H, s, CH3-25); 0,94 (3H, s, CH3-29); 1,01 (3H, s, CH3-23);
1,15 (3H, s, CH3-27) . 13C-NMR (CDCl3), : 38,86 (C-1); 27,88 (C-2);
79,13 C-3); 39,71 (C-4); 55,47 (C-5); 18,49 (C-6); 32,66(C-7);
38,66 (C-8); 47,84 (C-9); 37,16 (C-10); 23,24 (C-11); 122,58
(C-12); 143,94 (C-13); 41,92 (C-14); 25,96 (C-15); 23,55 (C-16);
46,58 (C-17); 41,42 (C-18); 46,17 (C-19); 30,77 (C-20); 34,05
(C-21); 32,93 (C-22); 28,20 (C-23); 15,53 (C-24); 15,39 (C-25);
17,06 (C-26); 24,39 (C-29); 25,96 (C-27); 33,13 (C-30).Cht
PMB-3
Tinh th mu vng, kt tinh li trong h dung mi CH2Cl2: MeOH (1;1,
v/v), nhit nng chy 255-256C. Rf=0.725 trong h CH2Cl2: MeOH (7/3,
v/v).EI-MS (70 eV): m/z (%): 286 (100)[M-C6H10O5]+, 258 (19)
[M-C6H10O5-CO]+, 153 (27,6), 137 (13), 134 (15).Ph 1H-NMR (CDCl3,
500 MHz), (, ppm: 6,60 (1H, s, H-3); 6,50 (1H, d, J=2,2 Hz, H-6);
6,80 (1H, d, J=2,2 Hz, H-8); 7,42 (1H, d, J=8,3 Hz, H-2); 6,90 (1H,
d, J=8,3 Hz, H-5); 7,42 (1H, d, J=8,3 Hz, H-6); 5,05 (1H, d, J=7,5
Hz, H-1); 3,51 (1H, m, H-2); 3,44 (1H, m, H-3); 3,50 (1H, m, H-4);
3,52-3,54 (1H, m, H-5); 3,93 (1H, dd, J=12,1 & 2,2 Hz, H-6a);
3,73 (1H, dd, J= 12,1 & 6,3 Hz, H-6b).Ph 13C-NMR (CDCl3), (,
ppm: 166,3 (C-2); 107,0 (C-3); 184,0 (C-4); 163,2 (C-5); 102,3
(C-6); 164,3 (C-7); 98,0 (C-8); 158,5 (C-9); 104,5 (C-10); 123,6
(C-1); 115,0 (C-2); 147,0 (C-3); 151,6 (C-4); 116,5 (C-5); 120,4
(C-6); 102,0 (C-1); 74 (C-2); 78,3 (C-3); 71,6 (C-4); 78,0 (C-5);
62,1 (C-6).
Chng 3: KT QU V THO LUN
3.1. IU CH CC PHN CHIT
Cy M (Plantago major L.) c chit phn lp cht theo phn cc tng dn ca
dung mi cho cc cn chit vi hiu sut so vi nguyn liu th l: ete du ho
(4.250%), etyl axetat (1.875%), n-butanol (3.125%). Quy trnh chit c
tm tt trong s di y:
S 1: Quy trnh chit cy M 3.2. PHN TCH CC PHN CHIT 3.2.1. Phn chit
ete du ho.
Phn chit ete du ho c kho st bng SKLM nh hng cho vic phn tch cc
thnh phn ca n. S phn tch c thc hin trn bn mng silica gel vi cc h
dung mi khc nhau v nhn thy phn chit ny kh phc tp. S phn tch SKLM
trong h n-hexan-etyl axetat (7,5:2,5) cho kt qu tch tt nht (xem bng
6, phn Thc nghim).
Phn chit ete du ho c phn tch bng SKC thng trn cht hp ph silica
gel, v ra gii vi hn hp n-hexan/etyl axetat. Da trn SKLM, cc phn on
ra gii ging nhau c gom li thnh cc nhm phn on, k hiu t I n VII.
Nhm phn on I c ra vi dung mi axeton sau cho kt tinh li trong h
dung mi n-hexan-axeton cho mt cht kt tinh dng v nh hnh mu trng (50
mg), k hiu l PMD-1.
Nhm phn on II c ra vi dung mi axeton sau cho kt tinh li trong h
dung mi methanol-axeton cho mt cht kt tinh dng v nh hnh mu trng
(100 mg), k hiu PMD-2.
Nhm phn on III c ra vi dung mi n-hexan v kt tinh li trong axeton
cho mt cht kt tinh dng tinh th hnh kim, mu trng (200 mg), k hiu l
PMD-3.
Nhm phn on VI c ra vi dung mi axeton v kt tinh li trong h
methanol-axeton cho mt cht kt tinh dng v nh hnh mu trng (55 mg), k
hiu l PMD-4.
Di y l s phn tch phn chit ete du ho.
S 2 : S phn tch phn chit ete du ho 3.2.2. Phn chit etyl
axetat.
Phn chit etyl axetat c ho tan trong dung mi methanol v cho kho
st SKLM nhn thy trong h dung mi toluen/ etyl axetat (8,0/2,0) cho
kt qu tch tt nht.
10 g phn chit etyl axetat (PME) c ho vi methanol va cho tan ht.
Trn dung dch nhn c vi silica gel, lm bay hi dung mi n kh, thu c hn
hp bt ti c mu vng nu.
Hn hp ny c em chy SKC thng trn cht hp ph l silica gel, ra gii vi
gradien h dung mi toluen/etyl axetat. Da trn SKLM cc phn on ra gii
ging nhau c gom li thnh cc nhm phn on, k hiu t I n VI.
Nhm phn on III c ra vi dung mi axeton v kt tinh li trong
diclomethan/ methanol thu c mt cht dng v nh hnh mu trng (0,5 g), k
hiu PME-1
Nhm phn on V c ra vi dung mi diclomethan v kt tinh li trong h
dung mi methanol/ axeton thu c mt cht kt tinh mu trng xanh (1,5 g),
k hiu PME-2.
Di y l s phn tch cn chit etyl axetat.
S 3: S phn tch phn chit etyl axetat
3.2.3. Phn tch phn chit n-butanol.
Phn chit n-butanol (PMB) c ho tan trong methanol v chy SKLM cho
thy h dung mi diclomethan/ methanol (7:3) c kh nng phn tch tt hn
c.
10 g phn chit PMB c ho tan trong methanol v cho trn u vi silica
gel n khi nhn c hn hp bt ti mu trng t.
Hn hp ny c a ln SKC thng vi cht hp ph l silica gel, ra gii vi
gradien h dung mi diclomethan/ methanol. Da trn SKLM cc phn on ra
gii ging nhau c gom li thnh cc nhm phn on, k hiu t I n VIII.
Nhm phn on III c ra v kt tinh li trong h dung mi n-hexan/ axeton
thu c cht kt tinh mu trng (150 mg), k hiu PMB-1.
Nhm phn on IV c ra v kt tinh li trong h dung mi diclomethan/
axeton thu c cht kt tinh mu trng (125 mg), k hiu PMB-2.
Nhm phn on VII c ra bng diclomethan v kt tinh li trong h dung mi
methanol/ axeton thu c mt cht kt tinh mu vng (0,5 g), k hiu
PMB-3.
Di y l s phn tch phn chit n-butanol.
S 4: S phn tch phn chit n- butanol 3.3. CU TRC MT S CHT PHN LP
3.3.1. Cht PMD-1Cht PMD-1 c im nng chy 60-61C, Rf = 0.9 (n-hexan/
etyl axetat, 8/2, v/v ), cho vt mu hng sng vi dung dch vanillin/
H2SO4.Ph khi lng ca PMD-1c pic ion phn t m/z =436. So snh vi th vin
ph cht ny trng vi cht c xc ng l Untriacontane C31H64 vi trng lp ti
98%. 3.3.2. Cht PMD-2
Cht PMD-2 c im nng chy 61C, R= 0.75 (n-hexan/etyl axetat, 8/2,
v/v), cho vt mu hng vi dung dch vanillin/ H2SO4 . Ph 1H-NMR ca n c
2 nhm metyl -CH3 (3H, t) 0.88 v 0.89 ppm; cc nhm -CH2 khong 1.2 v
1.3 ppm; nhm OH khong 1.5 v 1.6 ppm. Ta c th thy PMD-2 l mt ancol
C40H82O2 mch di.
Cht PMD-1 v PMD-2 l hp cht no thng khng c nhiu hot tnh sinh hc ,
do chng ti khng kho st tip. 3.3.3. Cht PMD-3Hp cht PMD-3 c dng tinh
th hnh kim mu trng, kt tinh li trong h dung mi n-Hexan: EtOAC
(8:2,v/v) , nhit nng chy 133-135C. Hin mu tm m vi thuc hin
vanillin/ H2SO4 c.
T nhng d kin vt l trn chng ti gi thit PMD-3 l mt steroid c tn l
-sitosterol. khng nh chc chn iu ny chng ti cho PMD-3 chy sc k lp
mng vi cht chun l -sitosterol th cho cng mt gi tr Rf . T chng ti rt
ra kt lun hp cht PMD-3 l -sitosterol hay tn gi khc l 3- Stigmast-5
en-3 ol. 3.3.4. Cht PME-1Hp cht PME-1: kt tinh trong h dung mi
Diclomethan/ Methanol (1:1; v/v), tinh th mu trng, im nng chy l
290-291C.
Ph khi lng (EI-MS) ch ra pic phn t m/z 456 M+ (C30H48O3) v cc
mnh phn r 248, 203, 189 tng ng vi s phn r retro Diels-Alder t
tritecpen urs-12-en vi nhm OH vng A (xem s trang 34)Trn ph 1H c tn
hiu ca proton ca nhm hidroxymetin [ 3,20 ppm (1H, dd, J=9,0; 7,1
Hz, H-3], 79,01 ppm (d, C-3) trn ph 13C-NMR v nhm cacbonyl 180,94
ppm (s, C-28). S c mt ca nhm metin olefinic th hin qua tn hiu 5,24
ppm (1H,t, J=3,5 Hz, H-12) gn vo nguyn t C-12 ( 125,66 ppm) trn ph
13C-NMR. Phn tch cc ph 13C, DEPT v HMQC cho thy PME-1 c 30 nguyn t
cacbon vi 7 nhm CH (trong c mt nhm lin kt vi nhm OH); 7 nhm CH3; 9
nhm CH2 v 7 cacbon bc bn. T nhng d kin ph trn suy ra PME-1 l mt
tritecpen nm vng. Phn tch ph 1D v 2D-NMR cng nh so snh vi cc s liu
ph cng b cho php kt lun PME-1 l axit ursolic.
Cu trc phn t ca PME-1 ph hp vi s phn mnh ca n trong khi ph mt s
mnh chnh nh sau:
S 5 : S phn mnh ca PME-1 (axit ursolic) 3.3.5. Cht PME-2Hp cht
PME-2 kt tinh trong h dung mi Diclomethan/ Methanol (1:1; v/v),
tinh th mu trng xanh, im nng chy l 307-308C.
Ph khi lng (EI-MS) ch ra pic phn t m/z 456 M+ (C30H48O3) v cc
mnh phn r 248, 203, 189 tng ng vi s phn r retro Diels-Alder t
tritecpen urs-12-en vi nhm OH vng A
Ph 1H- v 13C-NMR cho thy PME-2 c 48 proton v 30 nguyn t cacbon.
Nh vy cng thc phn t ca PME-2 l C30H48O3.
Cu trc ca PME-2 c xc nh da vo cc tn hiu xut hin trn cc ph NMR.
Ph DEPT ca PME-2 cho thy trong 30 nguyn t cacbon, c 7 nguyn t
cacbon bc bn, 5 nhm CH, 10 nhm CH2 v 7 nhm CH3. V iu ny ph hp vi
nhn xt rng cht PME-2 l mt hp cht tritecpen nm vng. Kt hp phn tch cc
tn hiu trn cc ph 1H- v 13C-NMR, v HMQC nhn thy rng proton ca nhm
hidroxymetin xut hin 3,20 ppm (1H, dd, J=9,0 v 7,1 Hz, H(-3) c gn
vo nguyn t >CH c (c 79,1 ppm (d, C-3). S c mt ca nhm metin
olefinic th hin qua tn hiu 5,28 ppm (1H, t, J=3,5 Hz, H-12 c gn vo
nguyn t C-12 c c 125,66 ppm. Ngoi ra, tn hiu ca nhm cacboxyl c pht
hin d dng trong ph 13C-NMR ( 180,94 ppm (s, C-28).
Trn c s nhng phn tch trn cng nh so snh vi cc s liu ph cng b cho
php kt lun cht PME-2 phn lp c l axit oleanolic. 3.3.6. Cht
PMB-3
Cht PMB-3 kt tinh mu vng t hn hp dung mi diclomethan/ methanol,
nc. 256C. Kho st cc ph IR, EI-MS, NMR v kt hp so snh vi cc d liu ph
tng ng trong ti liu nhn dng PMB-3 l luteolin-7-O--D-glucopyranozit
hay c tn gi khc l cynaroside
Cynaroside Di y l s phn mnh ca PMB-3
S 6: Cc mnh phn r ca hp cht PMB-3KT LUNT cc kt qu thu c chng ti
rt ra c mt s kt lun sau
1. iu ch cc phn chit t l cy M (Plantago major L.) theo phn cc ca
dung mi nh, ete du ho (PMD) vi hiu sut l 4,250%; phn chit etyl
axetat (PME) hiu sut l 1,875% v phn chit n-butanol (PMB) vi hiu sut
l 3,125%.2. T phn chit PMD phn lp c ba cht kt tinh, k hiu l PMD-1,
PMD-2, PMD-3. T phn chit PME phn lp c hai cht kt tinh k hiu l PME-1
v PME-2. T phn chit PMB phn lp c 3 cht kt tinh PMB-1, PMB-2 v
PMB-3.3. S dng cc phng php ph ( IR, EI-MS, NMR, DEPT, COSY, HMBC,
HSQC) v sc k lp mng vi mu i chng xc nh c cu trc ca mt s cht phn lp
c nh sau:PMD-1: l mt ankanol mch di, cha xc nh c cu trc.PMD-2: l mt
ankanol mch di, cha xc nh c cu trc.PMD-3: l -sitosterolPME-1: l
axit ursolicPME-2: l axit oleanolicPMB-2: cha xc nh c cu trc.PMB-3:
l luteolin 7-O--D-glucopyranoz.4. S dng phng php kt tinh phn on
trong kho lun ny chng ti thnh cng trong vic phn lp ring r hai hp
cht ng phn cu to l axit ursolic v axit oleanolic. C hai axit ny u c
hot tnh sinh hc mnh trong vic chng ung th gan, chng vim khp, chng
c, c hot tnh khng HIV, tng cng h min dch cho c th. V th chng c s
dng rng ri trong cc sn phm m phm v y t.TI LIU THAM KHO
Ting Vit
1. V Vn Chi (1996), T in cy thuc Vit Nam, NXB Y hc, tr 703.
2. Dc in Vit Nam III (2002), B y t, tr. 39.
3. Tt Li (1986), Nhng cy thuc v v thuc Vit Nam, NXB Khoa hc k
thut H Ni, tr 231-233,. Ting Anh
4. Ahamad, M.S., Ahmad, M.U, Osman, S.M., (1980), A new
hydroxyolefinic acid from Plantago major seed oil, Phytochemistry,
19, 217-239.5. Ahamad, M., Rizwani, G.H.,Aftab, K., Ahmad, UV.,
Gilani, A.H., Ahmad, S.P., (1995), Acteoside: a new
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EMBED ACD.ChemSketch.20
EMBED ACD.ChemSketch.20
plantaginin
EMBED ACD.ChemSketch.20
homoplantaginin
Luteolin 7-O--D-glucopyranozit
EMBED ACD.ChemSketch.20
Nguyn liu
Cn cha kit
Dung dch H2O + etanol
Dch chit ete du ho
Dch nc+etanol
Cn ete du ho
Dch n- butanol
Dch EtOH + H2O
Cn n- butanol
Dch EtOAC
Dch EtOH + H2O
Cn EtOAC
Ngm vi ETOH 95%, 2 ln, mi ln 3 ngy
Gom dch chit, c ui ETOH
1.Chit vi ete du ho
2. Lm khan bng Na2SO4
3. Lc b Na2SO4
C ui ete du ho
1. Chit vi EtOAC
2. Lm khan bng Na2SO4
3. Lc b Na2SO4
C ui EtOAC
1.Chit vi n-butanol
2. Lm khan bng Na2SO4
3. Lc b Na2SO4
C ui n-butanol
PMD (10g)
1-7
8-15
16-27
27-36
36-50
51-70
PMD-1
cht kt tinh mu trng (50mg)
nc.58-60C
PMD-2 cht kt tinh mu trng (100mg)
nc.64-66C
PMD-3
cht kt tinh mu trng (200mg)
nc.133-135C
PMD-4
cht kt tinh mu trng (55mg)
PME (10g)
1-6
7-12
13-18
19-25
26-46
47-60
PME-1
cht kt tinh mu trng (0,5g)
nc.290-291C
PME-2
cht kt tinh mu trng xanh (1,5g)
nc.307-308C
PMB (10g)
1-19
20-21
22-23
24-45
46-53
54-59
PMB-2
cht kt tinh mu trng (125mg)
nc.131-133C
PMB-1
cht kt tinh mu trng (150mg)
nc.148-150C
PMB-3
cht kt tinh mu vng (0,5g)
nc.198-200C
60-69
70-80
Axit ursolic
Axit oleanolic
PAGE 27
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