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Ong Sinh Han Hoi Luu PDF
Ong Sinh Han Hoi Luu PDF
MC LC
Trang
Trang ph ba ............................................................................................................i
Li cam oan .......................................................................................................... ii
Li cm n............................................................................................................. iii
Mc lc ....................................................................................................................1
PHN M U......................................................................................................3
CHNG 1: C S L THUYT
1.1. Tm hiu v ngh ...........................................................................................5
1.1.1. M t thc vt ........................................................................................5
1.1.2. Thnh phn ha hc ...............................................................................6
1.1.3. Dc tnh ...............................................................................................7
1.2. Tm hiu v curcumin ....................................................................................9
1.3. Phng php tch bit v tinh ch hp cht hu c......................................11
1.3.1. Phng php chit ................................................................................11
1.3.2. Phng php kt tinh li .......................................................................14
1.4. Tm hiu v HPLC (sc k lng hiu nng cao) ............................................17
1.4.1. Gii thiu phng php HPLC .............................................................17
1.4.2. Tm hiu v h thng HPLC .................................................................17
PHN M U
1. L do chn ti:
Nhiu cng trnh nghin cu th nghim cc nc pht trin trn th gii
khng nh t lu rng curcumin c tc dng hy dit t bo ung th vo loi mnh. Ti
M, i Loan, ngi ta tin hnh th lm sng dng curcumin iu tr ung th v
kt lun: curcumin c th kim hm s pht tc ca t bo ung th da, d dy, rut, vm
hng, curcumin cn l cht b cho d dy, rut, gan, mt, lc mu, lm sch mu,
iu tr vt thng, chng vim khp, d ng, nm, chng vi khun c hiu lc.
Cch y 5000 nm c ngh c bit n nh mt loi gia v, thuc gia truyn
cha c rt nhiu bnh, cha lin so, Tc dng kim hm s pht trin ca t bo
ung th v iu tr nhiu bnh ca ngh l do hot cht trong c ngh gi l curcumin.
Curcumin l thnh phn c bit v l hot cht chnh to nn mu vng c trng cho
c ngh. Trong lng curcumin ch chim khong 0,3 - 1% v khi lng c ngh.
Ch c curcumin t nhin trong c ngh mi c kh nng phng v chng li s pht
trin ca t bo ung th v cc bnh khc cao.
Vit Nam l mt nc nng nghip nhit i nng m vng ng nam Chu
, chnh iu kin trn rt ph hp vi loi cy ly c nh: gng, ngh, ti, hnh,
Trong cy ngh c cy trng rng ri mt s nc ti Chu nh n ,
Trung Quc v Vit Nam cng khng ngoi l. nc ta ni chung v tnh ng Thp
ni ring th cy ngh c trng rt ph bin vi nhiu chng loi a dng v phong
ph. Vi ngun nguyn liu di do v phong ph ni trn cng vi mc ch l vn
dng nhng kin thc hc cng thm nhng thao tc th nghim c hc trong
cc mn thc nghim ha hc. Do ti tin hnh ly trch curcumin t c ngh vng
vi tn ti kha lun tt nghip l: Nghin cu ly trch curcumin t c ngh vng.
2. Mc tiu nghin cu.
Nghin cu ly trch curcumin t c ngh vng ti thnh ph Cao Lnh tnh ng
Thp
3. Nhim v nghin cu.
Tm hiu thnh phn, ng dng ca c ngh v hot cht curcumin.
PHN NI DUNG
CHNG 1: C S L THUYT
1.1. Tm hiu v ngh.[1], [2], [4], [6]
- Ngh cn c tn l ut kim, khng hong, safran des Indes.
- Tn khoa hc Curcuma longa L. (Curcuma domestica Lour).
- Thuc h Gng Zingiberaceae.
- Ta dng thn r gi l khng hong (Rhizoma Curcumae longae) v r c gi l ut
kim (Radix Curcumae longae).
1.1.1. M t thc vt.
Ngh l mt loi c cao 0.60m n 1m. Thn r thnh c hnh tr hi dt, khi b
hoc ct ngang c mu vng cam sm. L hnh tri xoan thon nhn hai u, hai mt
u nhn di ti 45cm, rng ti 18cm. Cun l c b, cm hoa mc t gia cc l ln,
thnh hnh nn tha, l bc hu th khum hnh mng rng, u trn mu xanh lc nht,
l bc bt th hp hn, mu hi tm nht. Trng c phin, cnh hoa ngoi mu xanh lc
vng nht, chia thnh ba thy, thy trn to hn, phin cnh hoa trong cng chia thnh
ba thy, 2 thy hai bn ng v phng, thy di hm thnh mng su. Qu nang 3
ngn, m bng 3 van. Ht c o ht.
c trng khp ni trong nc ta lm gia v v lm thuc ngoi ra cn mc v
c trng cc nc n , Innxia, Campuchia, Lo, Trung Quc, v cc nc
nhit i.
Ngh thng c thu hoch vo ma thu.
Tumeron
- Ngoi ra cn tinh bt, canxi oxalat, cht bo. C ngh cha 8 10% nc, 6
8% cht v c, 40 50% tinh bt nha.
1.1.3. Dc tnh.
C ngh c bit n vi nhiu cng dng khc nhau, c bit c n v
nhiu nc, c phng ng ln phng Ty, s dng nh mt loi dc liu tr bch
bnh.
Theo hi ng nghin cu Trung ng, c ngh c th cha c nhiu bnh. C
ngh c th cha bnh hen suyn, ho, tr cm, nghin ru, mn v cc bnh ngoi da,
c ngh c th gim vim nhim, tr to gan v nhim trng bng quang, ri lon kinh
nguyt, tng cng sc khe cho tim. C ngh ngm vi nc v mt ong gip li tiu,
hoc nghin ngh vi b qua lc sch c th cha hiu qu bnh tiu ng.
Ngoi ra c ngh cn gip tr au rng v nga su rng, gim au bao t, gip tiu
ha v to cm gic thm n. Nh nc ngh un si vo mi gip cha au u v
chng mt ng.
C ngh khng ch c cng dng gip lin so nh nhiu ngi bit, m cn
mang li rt nhiu tc dng hu ch, c bit i vi sc khe con ngi.
Gip gim cn, lu thng v lc mu; gip c th chng li cc vi khun sng k
sinh trong rut, c bit tt cho h tiu ha; mi y ngi ta chng minh c rng
c th s dng ngh chng ung th v ngh c kh nng khng vim, gim nguy c
nhim trng; c th dng ngh kh trng v lm mau lnh vt thng.
Khi gp rc ri v tiu ha, nghin cu cho thy, ngh c th kch thch tiu ha v
gii phng cc enzim tiu ha, ph v lin kt cacbonhydrat v cc cht bo. Chnh v
th, trong trng hp b au bng, mt cc tr ngh s gip ch rt nhiu.
Cht curcumin c t nhin trong c ngh tng c cc nh khoa hc chng minh l
mt cht chng oxy ha cc mnh c li cho sc khe, c li cho tim mch, chng
cholesterol v ung th.
Ngh c th lm gim hm lng cholesterol c hi trong mu v c kh nng
chng li chng x va ng mch, curcumin c trong ngh cho php gim nguy c
mc bnh nhi mu c tim, c bit l nhng ngi hay ngy ng. Nu cht curcumin
c ng dng thnh cng i vi con ngi, n s m ra mt hng i mi cho cch
phng v iu tr bnh nhi mu c tim ng thi bo v sc khe tim mch ca chng
ta. Khc vi hu ht cc hp cht t nhin khc vi hiu qu hn ch, cht curcumin c
tc dng trc tip ln nhn t bo bng cch nga nga vic sn sinh qu nhiu protein
bt thng. Tuy nhin, cc chuyn gia cng khuyn nn n ngh mc va phi v
c n qu nhiu cng khng gip tng hiu qu ca tr bnh ca ngh.
Curcumin c tc dng khng ung th, c lp v tiu hy t bo ung th. Curcumin
l mt thnh phn c bit lm nn mu vng c trng ca ngh c kh nng tiu dit
hai loi protein trong cc t bo ung th, cc protein ny chnh l ngun duy tr s tn
ti ca chng. Ngh cng gip ngn chn khng cho t bo ung th lan trn i khp ni
trong c th (chng di cn). Ngoi ra ngh cng rt an ton v khng c phn ng ph.
Cc bc s bnh vin University of Texas MD Anderson Cancer Center, mt bnh
vin chuyn v cha tr cc bnh ung th vo bc ln nht th gii, sau khi b ra
nhiu nm nghin cu v th nghim, u ng thanh xc nhn rng: thuc bo ch
bng c ngh c tc dng trc tip git cht cc t bo ung th, ng thi c ngh cng
l mt loi chng oxi ha cc mnh rt cng hiu chng li s ph hoi ca gc t
do v gip tng cng h thng min nhim cho nn h khuyn bnh nhn ung th
nn dng ngh hng ngy.
Tuy nhin, khng nn xem y l thn dc, v n ch c tc dng khi ung u n
v va phi trong mt thi gian di.
1.2. Tm hiu v curcumin. [8], [9], [10], [11]
- Tn IUPAC: (1E, 6E) -1,7-bis (4-hydroxy-3-metoxyphenyl) -1,6-heptadien3,5-dion.
- Cng thc phn t: C21H20O6
- Phn t khi: 368,38 g / mol.
- Nhit nng chy: 183C (361 K).
- Curcumin l tinh th mu nu , l hot cht c chit ra t ca ngh vng
thuc h gng. Hin ti ngi ta tm thy curcumin tn ti 4 dng hp cht:
+ Curcumin l hp cht chnh chim 60%:
Curcumin
+ Demetoxy-curcumin chim 24% c cng thc cu to sau:
10
Demetoxy curcumin
+ Bis-demetoxy-curcumin chim 14%:
Bis-demetoxy curcumin
+ V mt hp cht mi pht hin l xiclocurcumin chim khong 1%:
Xiclocurcumin
- Curcumin l mt polyphenol v l sc t to nn mu vng c trng ca c
ngh.
- Curcumin c th phn ng c vi axit boric to nn hp cht c mu cam
nn c ng dng dng nhn bit mui ca nguyn t bo.
- Chnh v curcumin l sc t to nn mu vng sng nn curcumin c dng
lm cht ph gia thc phm. Trong cht ph gia thc phm curcumin c k hiu di
m s E100.
- Dc tnh:
+ Curcumin l cht hy dit ung th vo loi mnh nht theo c ch hy
dit tng bc cc t bo c tnh. Chng lm v hiu ha t bo ung th v ngn chn
khng cho hnh thnh cc t bo ung th mi. Trong khi , cc t bo lnh tnh khng
b nh hng. Curcumin c coi l cht tiu biu nht cho th h mi cc cht chng
11
12
K=
S1 C1
=
S 2 C2
S1
cng ln th vic ly cht rn ra rt kh khn, phi dng dung
S2
kV + S
13
14
15
v li phi gii phng khi lc cng nh khi ra. Vic la chn dung mi hay h dung
mi phi da vo mi quan h cu to phn t ca cht kt tinh v dung mi, thng
thng cht phn cc th ha tan tan vo dung mi phn cc v ngc li. Khi cht kt
tinh cha r cu to th phi th ha tan trong dung mi t khng phn cc n dung
mi phn cc. Cch lm nh sau: ly mt vi tinh th vo ng nghim, nh mt t dung
mi vo ri un si, quan st tan cho n khi tinh th tan ht, nu m bo yu cu
trn th cht s kt tinh li khi lnh. Dung mi c xem l tt nu c 0,1g cht kt tinh
tan trong 1ml dung mi nng.
Khi khng chn c dung mi th bt buc phi chn h dung mi. Nguyn tc:
Ly mt dung mi ha tan cht kt tinh ngay nhit thng, sau chn mt dung
mi khng ha tan hay km ha tan cht tinh ch nhng phi tan trong dung mi trn.
Cch lm nh sau: Ly mt vi tinh th vo ng nghim, thm dung mi cho n khi
tan ht, ghi ly th tch dung mi. Nh t t dung mi khng tan cht tinh ch vo hn
hp trn cho n khi vn c, ghi ly th tch dung mi, un hn hp cho tan ht, lc
nng ngui, cht rn s kt tinh li. T l hai th tch o c coi l t l th tch
dung mi chn. Thng chn h dung mi etanol nc, etanol benzen, axeton
ete du ha, axit axetic nc,
Nu c nhiu cht c tan khc nhau nhit khc nhau vo mt dung dch,
ngi ta dng phng php kt tinh phn on. mt nhit xc nh, mt cht no
tan qu bo ha s kt tinh li khi lnh, cn cht kia cha bo ha s li trong
dung dch.
Nu dung mi ha tan cht kt tinh, cn cht bn khng tan, ngi ta s lc nng
loi b cht bn, cn cht kt tinh li trong dung dch s kt tinh khi lnh.
1.3.2.3. Cc thao tc khi kt tinh.
- Chun b dung dch kt tinh hay dung dch nc ci.
Cho mt lng cht cn kt tinh vo bnh cu hai c c lp ng sinh hn hi lu
v phu chit ng dung mi hoc vo bnh tam gic c sinh hn hi lu, thm bt
ri cho dung mi vo t hn mt lng t theo lng tnh ton, un si. Nu si m
16
cha tan ht th thm mt t dung mi vo cho n khi cht tan hon ton, cht bn s
khng tan.
Nu dng hn hp dung mi th cho dung mi tan tt vo trc cho n khi cht
rn tan hon ton ri thm dn dung mi ha tan km vo cho n khi cht rn kt ta
ri tan nhit si. Nu dung dch c mu, phi thm cht ty mu nh than hot
tnh, than xng hay silicagel vi t l 1/20 hay 1/50. Khi cho cht kh mu phi
dung dch lnh, khng c thm cht kh mu khi dung dch ang nng trnh dung
dch b tro ra ngoi. Sau , un si li dung dch trong 2 3 pht.
- Lc nng dung dch.
Lc dung dch ang nng loi b cht bn, cht ph khng tan ra khi dung
dch. Cn lc nng bng phu lc nng, lc nhanh trnh cht kt tinh trn phu.
- Thc hin kt tinh cht.
Dung dch nc ci c y ming khng nt cht, ngui hay lm lnh bng
nc lnh hay nc th tinh th cht s kt tinh.
Ch nu dung dch cha kt tinh, cha t n bo ha th phi c ui bt
dung mi ri mi lm lnh. Kch thc ca tinh th ph thuc vo tc kt tinh v
lm lnh. Nu lm lnh nhanh th thu c tinh th nh s hp th dung mi v cht
bn. Tinh th cng ln khi lm lnh kt tinh chm.
Nu khng thy kt tinh th c th gy mm kt tinh bng cch cho vo dung
dch mt vi tinh th ca cht tinh ch hoc ly a thy tinh c vo thnh bnh.
Nhng cht c nhit nng chy thp thng tch ra dng du nn phi lm
lnh chm v su tinh th kt tinh chm.
- Tch lc tinh th.
Tinh th tinh khit c lc nhanh trn phu Bucsne p sut thp. Khi lc cn
ra li khi kt ta bng dung mi lnh. Mt s trng hp ngi ta s gn, lng. Trong
trng hp cht ho nc v d b oxi ha th dng phng php ring.
- Lm kh tinh th.
17
18
Degasse
Pump
3
Column
5
Tim
mu 4
Detector
19
- Tuy nhin theo kinh nghim th chng ta t khi s dng 4 ng dung mi cng
mt lc m chng ta ch s dng ti a l 3 v 2 ng cho h pha ng lun c
pha trn ng nht hn, h pha ng n gin hn qu trnh ra gii n nh.
- Hin 4 ng dung mi phc v ch yu cho vic ra gii Gradial dung mi
theo thi gian v cng tc xy dng tiu chun.
Lu : + Tt c dung mi dng cho HPLC u phi l dung mi tinh khit v c
ghi r trn nhn l dng cho HPLC hay dung mi tinh khit phn tch.
+ Tt c cc ha cht dng pha mu v pha h m phi c s dng
l ha cht tinh khit phn tch.
S dng ha cht tinh khit nhm mc ch trnh hng ct sc k hay nhiu
ng nn, to nn cc peak tp trong qu trnh phn tch.
2. B kh kh Degasse.
Mc ch ca b kh kh nhm loi tr cc bt nh cn st li trong dung mi
pha ng.
Nu nh trong qu trnh phn tch m dung mi pha ng cn st cc bt kh th
mt s hin tng sau y s xy ra:
+ T l pha ng ca cc ng dung mi khng ng s l cho thi gian
lu ca Peak thay i.
+ Trong trng hp bt qu nhiu b kh kh khng th loi tr ht c
th c th bm cao p s khng ht c dung mi khi p sut khng ln v my sc
k s ngng hot ng.
Trong bt c trng hp no nu trn cng cho kt qu phn tch sai.
3. Bm cao p.
Mc ch bm pha ng vo ct thc hin qu trnh chia tch sc k. Pump
phi to c p sut cao khong 250 at 500 at (1at = 0,98 bar) v pump phi to
dng lin tc. Lu lng bm t 0,1 9,999 ml/pht.
My sc k lng hin nay thng c p sut ti a 412bar (khong 420at). Tc
dng 0,1 9,999 ml/pht.
20
A l tn hiu o c.
C l nng cht phn tch.
21
22
2,7
0,4
23
Bt ngh
Dch chit
24
25
n v tnh
Kt qu
Curcumin
69,5
Phng php
HPLC HPLC Liquid
Chromatography
26
Sn phm kt tinh
Curcumin chun
4,811
4,818
27
28