Trang 1

HNH THC:
- page set up li.
- c ch: c ni 14, ni 13,
- canh l bn tri cn ln xn.
- lm mc lc d nhn tng qut, cha cn nh trang hoc mc
lc t ng.
- cc mc khng c du :
v d: 1.1.1. phn b sinh thi : -> 1.1.1. Phn b sinh thi
- dng lnh control + h thay shoxlec -> soxhlet.
- sau du ngoc ( vit lin) -> (vit lin).

NI DUNG:
- PHN TNG QUAN:
o b sung cc cng trnh v mp ng, tp trung l nghin
cu ging mnh, ngoi v trong nc.
o b sung cc phng php xc nh hot tnh chng oxi ha.
- PHN KT QU NGHIN CU:
o 1,5 ml thuc th FRAP c trn vi 50 l mu: xem li ci ny:
th nht l khc mnh, th hai l khng thc t v th tch khng
cho mt cuvet.
o Bng 3.1 v 3.2: 3 ln o thu 3 kt qu ( x l s liu) Fe
2+

ri gi tr trung bnh.
o Hai bng trn s liu qu p, v cng l tng ( lp li
qu cao), em xem li hoc gp thy trao i hn.
- PHN BN LUN:b sung nhiu hn v cc cng trnh
nghin cu v so snh.
o Xy dng khong tuyn tnh: hnh 3.1 cho thy phng trnh hi quy
c dng y = 0.017x 0.0363, h s r = 0.9922, cho thy c s tng
quan tuyn tnh cht ch gia nng hot cht vi hp th quang
...
Trang 2

o Xc nh hot tnh chng oxi ha: b sung thm mt s na.
o B sung phn So snh mu ti v mu kh: so snh 2 kt qu
trung bnh ca mu ti v mu sy kh (vi f
tn
>f
lt
), ta c t
tn
>t
lt
.
nh vy, gi tr trung bnh ca mu ti cao hn mu sy kh c
ngha thng k (t t
0.05
).
Kt qu kho st c trn cho thy, hm lng cht chng oxi ha
trong mp ng ti cao hn mp ng sy kh. iu ny cho thy
nhit cng c nh hng n hot tnh chng oxi ha trong mp
ng, nhit cng cao th hm lng cht chng oxi ha gim. - gii
thch ti sao.
b sung thm: mt s nghin cu ngi ta dng 2 nhit khc nhau
v vy, nu mun trch li cc hot cht chng oxi ha trong mp
ng th ta nn chn mp ng dng ti v trong qu trnh trch li
nn hn ch s dng nhit cao v s gy tn tht hot cht sinh hc.
- KT LUN: cha n ni.

PS. sa nhiu trn bi vit tuy nhin save m n khng chu lu.

Trang 3

Chng 1
TNG QUAN
1.1. TNG QUAN V MP NG (Momordica Charantia )[1]
Mp ng cn gi l kh qua, cm l chi, li b o, hng c nng,
lng qua, mp m, chua hao ( Mng- Thanh Ha)
Tn khoa hc: Momordica Charantia L. ( Momordica balsamina Desc.,
Cucumis africanus Lindl )
Thuc h B: Cucurbitaceae .
1.1.1. Phn b sinh thi :
Trn th gii mp ng c mt hu ht cc nc nhit i t Chu Phi
sang Chu v Chu M nh n , mt s nc ng Nam nh M Lai,
Thi Lan, Philipin.. Mp ng cn c trng nhiu cc tnh min Nam Trung
Quc nh: Phc Kin, Qung ng, Qung Ty, Giang T, Trit Giang
Ti Vit Nam mp ng c trng khp ni t min Nam n min Bc,
hu ht cc tnh t ng bng, trung du n min ni. Mp ng thng c
trng xen vi bu, b, [6]
Mp ng c bin sinh thi tng i rng, nhit thch hp cho cy
sinh trng t 20 24
0
C, hoc cao hn. Cy sinh trng nhanh trong ma ma
m, ra hoa v qu sau 7 8 tun gieo trng.
1.1.2. c im thc vt : [1],[2]
Mp ng l mt loi dy leo, thn mu xanh lt c gc cnh, ngn hi
c lng t, cy mp ng c i sng khong mt nm. Kch thc dy mp
ng khong bng ngn tay t, dy b t 5 7 m, dy leo c nh nhiu tua
cun.
Trang 4

L n nhm, mc so le, di 5 10 cm, rng 4 8 cm, phin l chia 5 7
thy hnh trng, mp c rng ca u, mt di l c mu xanh nht hn mt trn,
gn l ni r mt di, phin l c lng ngn.
Hoa mc n c k l, hoa c, hoa ci cng gc, c cung di, cnh
hoa mu vng nht, ng knh ca cnh hoa chng 2 cm, hoa c c ng i
ngn, trng gm 5 cnh mng hnh bu dc, nh (nhy) ri nhau. Hoa ci c i v
trng ging vi hoa c.
Tri hnh thoi di 8 15 cm, gc v u thon nhn, trn mt v qu c
nhiu u sn si ni ln to nh khng ng u. Tri cha chnh c mu xanh, khi
chn c mu vng hng. V th Trung Quc, mp ng (Momordica charantia)
cn c tn gi l hng dng, hng c nng. Khi chn tri tt t u, tch ra lm
ba phn.
Ht mp ng dt, di 13 15 mm, rng 7 8 mm, trng gn ging ht b
ng nhng c kha v c mng bao bc, quanh ht khi chn mng c mu mu
nh mng gc.
1.1.3. Thnh phn dinh dng [3]
Theo ti liu ca Vin i Hc Purdue v cc loi rau qu Chu hi nhp
vo M (Willsetal 1984), thnh phn dinh dng tnh bng gam trong 100g qu
mp ng nh sau (bng 1.1)




Trang 5


Bng 1.1. Thnh phn dinh dng trong 100 g mp ng [34]
Phn n c 84 g Cht bo 0,1 g
Nc 93,8 g Carbohidrate 0,2 g
Protein 0,9 g Calsium 22 mg
Sinh t A 0,04 mg Potassium 260 mg
Sinh t B1 0,05 g Magnesium 16 mg
Sinh t B2 0,03 g St 0,9 mg
Niacin 0,4 g Zinc 0,2 mg
Sinh t C 50 g

Thnh phn dinh dng trong 100 g mp ng theo bng thnh phn dinh
dng thc phm Vit Nam (2008) (bng 1.2)
Bng 1.2. Thnh phn dinh dng trong 100 g mp ng
T l thi b 20 % St 0,6 mg
Nng lng 16 g Phospho 29 mg
Nc 94,4 g Beta-caroten 40 mcg
Protein 0,9 g Vitamin B1 0.07 mg
Tinh bt 3,0 g Vitamin B2 0.04 mg
Trang 6



1.1.4. Thnh phn ha hc [1], [34]
- Glycosid: momordicin v charantin. Charantin l mt hn hp steroid lm
h ng v mt glycosid khc gc pyrimidin cng c tm thy.
- Allkaloid momordicin, du thc vt.
- Mt peptid ging insulin - polypeptid-P c phn t lng 11.000 trong
qu, ht v cc m trong thn cy
- Vitamin B1, C, Adenin, betain, protein (0,6%).
- Ht cha 32% du vi cc acid bo stearic, limoleic, oleic, cc nucleosid
pyrimidin vicine, Glycoprotein alpha-momorcharin v beta-momoecharin ( tc
dng ph thai) v lectin, cc acid amin t do vi nhm amin bn tri v nhm
axt cacboxylic bn phi, nhm R ty vo tng axt amin c th.
1.1.5. Cng dng v tc dng:
Hu ht cc b phn ca cy mp ng u c cng dng cha bnh [3].
Trong Y Hc C Truyn th ngi ta s dng cc thnh phn ca cy mp
ng cha mt s bnh nh sau:
R
R mp ng dng tr l. Ti n , dch r (cng nh l v qu) mp
ng c dng tr bnh i ng tt, do c tc dng lm gim lng ng
glucose trong mu. R mp ng c th tr bnh gan v ta c th p dng mi
dng bnh [8].
Cht x 1,1 g Vitamin PP 0.3 mg
Tro 0,6 g Vitamin C 22 mg
Calci 18 mg
Trang 7

Thn
Thn cy mp ng c dng tr mt s bnh nh: Ung x lng
(dng cho ph n ung sau khi sinh), bnh gan vng da [9].
L
L c v ng, tnh mt. L non n tr bnh nng bc trong mnh. Gi l, vt
nc, thm cht mui ung tr bnh nng m man. Cha chng n c sng ,
mn nht, au nhc, cha rn cn, gip c th mau bnh phc khi mt mi, ho
kht, hi hp, i ng xa vt v, lao ng qu sc... [9], [7], [12]. C th dng l
mp non nu canh n.
Ngoi ra l cy mp ng cn cha c nht c, sng ty, vt thng
nhim c Dch l mp ng cn c tnh cht hi nhun trng, lm h st,
dit giun [3], [8].
Hoa
Hoa c cng dng cha d dy, cha au mt v cha chng l cp tnh [3],
[6].
Tri
Ngoi cng dng lm thc n, tri mp ng cn c dng tr nhiu
bnh nh: Tr ho, st, kit l, lm lnh da non cc vt thng cc vt lot c tnh
[14].
Tri mp ng c tnh hn, mt khng c. Lc cn xanh n c tnh gii
nhit, lm tiu m, nhun trng, b thn, li tiu, lm bt au khp xng. Khi
chn, tri mp ng c tnh b thn, dng huyt [14].
Trang 8

Trung Quc tri mp ng cn dng tr t qu tim, bnh st, kh
ming, vim hng. n , dch tri mp ng dng tr rn cn. Thi Lan
dch qu dng tr bnh v gan v l lch, c bit l lm h ng mu bnh
nhn tiu ng [8].
Cc nh khoa hc Ai Cp ang c k hoch sn xut thuc vin cha bnh
tiu ng t qu mp ng hay cn gi l kh qua, thay th thuc tim
insulin [15] do Mp ng c kh nng c ch s chuyn ha v hp thu ng,
ngn nga cn bnh tiu ng v n nh ng huyt ngi tiu ng [8].
Hot cht trong nha ca mp ng c mt lp ph c bit chng li s ph hy
ca cc enzym phn hy insulin trong d dy.
Ht
Ht c cht bo, v ng, hi ngt, tnh m, thanh nhit, gii c, gii cm,
li tiu, cha ho vim hng, rn cn, tr ng kinh [9].
Ht mp ng c tnh b dng, trng kh, iu tr cc chng bnh nh ln
cn co git do st cao hoc kinh phong tr em [16].
Ht mp ng dng vi liu 3 gam ht kh, di dng sc ly nc ung, c th
cha ho, lm h st [16].
Ngoi ra, cc hot cht sinh hc trong mp ng cn c tc dng chng th
thai, chng ung th (do s c ch tng hp protein), lm h huyt p, chng
HIV[17].
1.2. VI NT V GC T DO V CHT CHNG OXY HA [35]
Gc T Do ( Free radical ) l bt c phn t ha cht no ch c mt in t
duy nht (electron mang in m) hay mt s l in t.
Trang 9

Gc t do c tc dng khng tt cho c th lin tc ngay t lc con ngi
mi sinh ra v mi t bo chu s tn cng ca c chc ngn gc t do mi ngy.
Nu khng b kim sot, kim ch, gc t do gy ra cc bnh thoi ha nh ung
th, x cng ng mch, gy suy yu h thng min dch gy d b nhim trng,
lm gim tr tu, teo c quan b phn ngi cao nin.
Gc t do gy tn thng trn cc m, t bo :
- Ph rch mng t bo khin cht dinh dng tht thot, t bo khng tng
trng, cht dn.
- To ra cht lipofuscin tch t di da khin ta c nhng vt i mi trn
mt, trn mu bn tay.
- Tiu hy hoc ngn cn s tng hp cc phn t cht m, ng bt,
m, enzyme trong t bo.
- Gy t bin gene, nhim th, DNA, RNA.
- Gy bin tnh collagen, elastin khin da nhn nheo, c khp cng nhc
Gc t do c th l th phm gy ra ti trn 60 bnh, ng k nht gm c:
bnh va x ng mch, ung th, Alzheimer, Parkinson, c thu tinh th, bnh
tiu ng, cao huyt p khng nguyn nhn, x gan.
Trong c th c rt nhiu loi gc t do, c bit superoxide, ozone,
hydrogen peroxide, lipid peroxy nht l hydroxyl radical, mt gc rt phn ng
v gy ra nhiu tn thng.
Gc t do c to ra bng nhiu cch: c th l sn phm ca nhng stress,
bnh tt, nhim mi trng, thuc l, dc phm, tia phng x mt tri, thc
phm c cht mu tng hp, nc c nhiu chlorine v ngay c oxygen.
trung ha gc t do, ngoi qu trnh t nhin ca c th, ta c th trung
ha gc t do bng cch dng cht chng oxy ha ( antioxidant ) t thc phm,
thuc, m phm.
Trang 10

Hip Hi Tim Mch Hoa K khuyn co: Mt ch dn thn trng v khoa
hc nht v vn ny l dn chng nn n thc phm c nhiu cht chng oxy
ha trong rau, tri cy v cc loi ht, thay v ung thm cht antioxidant. Cc cht
ny c kh nng lm mt hot tnh ca gc t do tch t trong c th, bin chng
thnh nhng phn t v hi, ng thi cng c kh nng duy tr cu trc v chc
nng ca t bo.
Cc cht chng oxy ha gm c -caroten, vitamin C, E, selenium,
bioflavonoids ,lutein, lycopene, coenzyme Q 10, alpha-lipoic acid v ubiquinone
Cc antioxidant c th phn ng ring l hoc cng tc vi nhau loi tr gc
t do. Mi antioxidant c tc dng ring vi tng loi gc t do mi t bo. Do
, cc cht chng oxy ha u c gi tr nh nhau.
1.3. Vai tr ca cc cht chng oxy ha trong mp ng
Theo ng y, mp ng tnh hn, v ng, khng c, nu c dng
thng xuyn s gip gim cc bnh ngoi da, lm cho da d mn mng [3].
Theo y hc hin i, mp ng c tc dng dit vi khun v virus, chng
li cc t bo ung th, h tr c lc cho bnh nhn ung th ang cha bng tia x
[3], [12].
Mp ng cha nhiu thnh phn hu ch, c bit l cc cht chng oxi
ha... nn n c tc dng rt tt trong vic khng khun, nng cao sc khe cho h
min dch [26], lm h ng huyt v cholesterol, kch hot mt lot cc phn
ng hot ha mng t bo lm cho cc t bo ung th v ph n ngng di cn
v tiu dit cc t bo ny [
Mp ng cha mt lng kh ln cis(c)9, trans(t)11 v axit linoic, olenic,
mt lng ln vitamin C, flavonoid[27].
Trang 11

- Nhng cht cis(c)9, trans(t)11 v axit linoic, olenic c tc dng trit tiu cc
mm bnh gy ung th t trng v nhiu chng bnh nan y khc [3], [13] .
- Vitamin C cht c hot tnh chng oxy ha mnh, c kh nng tiu ha gc
t do v ngn khng cho gc ny xm nhp cc phn t cholesterol LDH. Do
, tng cng s bn b ca mao mch, ngn khng cho gc t do xm nhp
qua mng t bo, y mnh tc hi phc vt thng, kch thch sn xut
khng th, acetylcholine, ngn chn tc dng c hi ca oxygen [27].
- Cc flavonoid c tc dng bo v c th, ngn nga x va ng mch, tai
bin, lo ha, thoi ha gan, tn thng do bc x nh kh nng ngn
chn qu trnh oxi ha do cc gc t do, to phc vi cc ion kim loi, c ch
s hot ng ca hyaluronidase. Do , c vai tr nh nhng cht xc tc
ngn cn cc phn ng oxi ha, ci thin tnh trng xuyt huyt di da do
tha enzym hyaluronidase [27].
1.4. NGHIN CU HOT TNH CHNG OXY HA BNG PHNG
PHP FRAP
1.4.1. Nguyn l:
Phng php FRAP c thc hin u tin bi Benzie v Strain o
nng lc kh trong huyt tng [30], [31] nhng cng c s dng
phn tch cc cht chng oxy ha trong thc vt. Phng php ny da
vo s kh phc Fe (III) TPTZ (2,4,6-tripyridyl-s-triazine) thnh phc
Fe (II) TPTZ bn hn bng mt cht kh ( cht chng oxy ha) pH
thp. Fe (II) TPTZ c mu xanh dng m v c th o mt quang
bc sng
max
= 593 hoc 595 nm. Tc ng chng oxy ha c nh gi
bi s tng cng mu ca phc Fe (II) TPTZ (Hnh 1.1.).

Trang 12



Hnh 1.1. Phn ng FRAP
1.4.2. Mc ch
Xc nh hot tnh chng oxy ha ca cc cht.
1.4.3. Cc yu t nh hng n kt qu th nghim
- Hot tnh chng oxy ha ca dch chit.
- Phn ng gia thuc th v dch chit.
- Thao tc th nghim.
a) Hot tnh chng oxy ha ca dch chit: dch chit c hay khng hot tnh
khng khun.
b) Phn ng gia thuc th v dch chit:
Phn ng xy ra hon ton hay khng l iu quan trng ca th nghim. Phn ng
xy ra hon ton, lng Fe
2+
c to ra ti a, kt qu o quang tin cy.
c) Thao tc th nghim
Phng php FRAP p dng k thut o quang ph hp th. K thut ny c
nhy cao, ch cn thiu hoc tha mt lng nh phc s to ra mt quang ph
hp th khc. Do cn chnh xc, t m trong tng thao tc.
1.5. CC PHNG PHP NGHIN CU HOT LC DCH CHIT
BNG KHNG SINH
1.5.1 Phng php khuch tn trong mi trng thch a [18]
1.5.1.1 Nguyn l
Trang 13

Nhng l c trn mi trng thch a cy u vi vi khun th nghim
c nng y dch chit, dch chit s khuch tn t l c thnh mt gradient
nng h v vi khun ch mc mt khong cch xc nh, hnh thnh mt vng
c ch. Khong cch ny trong nhng iu kin chun ph thuc vi nhy cm
cu vi khun th nghim. Da vo ng knh vng c ch, ngi ta xc nh
nhy cm ca vi khun i vi dich chit th nghim.
1.5.1.2 Mc ch
K thut c l dch chit khuch tn trong thch thng c p dng
xc nh nhy cm ca vi khun i vi nhng dch chit khc nhau ( nh
tnh) , y dch chit thng c pha long vi hm lng nht nh.
1.5.1.3. Cc yu t nh hng n kt qu th nghim:
- Hot tnh khng khun ca dch chit.
- Mi trng lm khng sinh .
- Nng vi khun.
a) Hot tnh khng khun ca dch chit: dch chit c hay khng hot tnh khng
khun.
b) Mi trng:
Mi trng nui cy dng kim tra hot tnh khng khun ca dch chit
phi l mi trng uc chun ha cao gip cho hu ht vi khun gy bnh
mc tt nh mi trng Mueller-Hinton (MH).
Mi trng MH c chun b theo hng dn ca tng hng sn xut v
vo a peptri v khun vi dy t 3,5- 4,5 mm. Cc a ny phi c t
trn mt mt phng ngang bng m bo cho su ca thch mi v tr trong
a bng nhau.
c) Nng vi khun:
Ch thc hin khng sinh vi vi khun thun chng. Lng vi khun cy
vo mi trng c nh hng ln n ng knh vng c ch nh lng vi khun
nhiu vng c ch nh li, lng vi khun t vng c ch rng ra.
Trang 14

1.5.2. Phng php pha long dch chit trong mi trng [18]
1.5.2.1. Nguyn l
Cy vi khun ln cc a mi trng c ( hoc ng mi trng lng) c
cha cc nng dch chit khc nhau, quan st xem chng vi khun khng pht
trin nng dch chit no.
1.5.2.2 Mc ch
Pha long dch chit vo mi trng thc hoc vo canh thang vi nng
gim dn xc nh nng c ch ti thiu ( MIC) hoc nng dit khun ti
thiu ( MBC) ca dch chit i vi vi khun.
1.6. CC CHNG VI KHUN GY BNH THNG GP
1.6.1. Staphylococcus aureus ( T cu vng)
Staphylococcus aureus c tm thy khp ni, da, tc v hu ca bnh
nhn, nhn vin y t , dng c v cc vt trn bnh vin.
Chng c sc sng lu bn bn ngoi v mi trng, kh nng chu nhit
cao hn cc vi khun khc ( ch sau 60 pht trong 80
0
C) v phi dng ha cht
nng tng i ln mi dit c vi khun ny. V vy S.aureus l tc nhn gy
bnh quan trng, thng gy ra nhim trng vt thng, vt bng v nhim trng
khi thng tnh mch [19].
Mt khc, S.aureus cn v cng thch ng vi p lc khng sinh, l mt
trong cc chng vi khun khng li rt nhiu loi khng sinh thuc h -lactam,
aminosid, macrolid, quinolon nh erythromucin, clindamycin, amynoglycosid
Nhng chng t cu khng methicillin ( Methicillin resistance S.aureus: MSA) l
tc nhn gy nhim khun bnh vin rt ph bin trn th gii [28].
1.6.2. Psedomonas aeruginosa ( Trc khun m xanh)
Psedomonas aeruginosa l trc khun Gram m, c tm thy trong t,
nc, c mt hu ht trong mi trng nhn to, trang thit b y t.
Chng l tc nhn nhim trng c hi, c kh nng ly nhim sang cc m
b h hng, thng ly qua ng phi, ng tit niu, vt bng, vt thng v
c th gy ra cc bnh nhim trng huyt [32]. P. aeruginosa chim khong t
Trang 15

10-11% nguyn nhn gy nhim trng bnh vin. Ch yu l cc chng
P.aeruginosa khng vi -lactam ( ticarcilline, ceftazidime hoc imipenem) hoc
khng aminosid, quinolon [19].
1.6.3. Escherichia coli ( Trc khun Gram m)
Escherichia coli l nhng vi khun l sinh, bnh thng c rut, nhng
ng thi cng l tc nhn gy bnh khi chng xm nhp vo cc c quan khc
nh ng tit niu, dng mu, Mt s chng E. coli c kh nng gy bnh a
chy nh ETEC, EPEC, EIEC, S c mt ca E. coli ngoi cnh v trong thc
n chng t s nhim bn do phn.
Qua nhiu cng trnh nghin cu cho thy mt s ln cc chng E. coli gy
bnh khng cc khng sinh v hin tng 1 chng vi khun E. coli khng vi
nhiu loi khng sinh cng kh ph bin [19].
1.6.4. Streptococci D ( Lin cu rut)
Streptococci to thnh cacbohydrat C c hiu nhm D, pht trin tt
nhit t 10- 45
0
C , nng 6,5% NaCl v thch 40% mui mt. Kh nng
tan mu thay i. Chng c tm thy khun ch bnh thng rut v c th
gy bnh lc xm nhim m, mu, ng tiu hoc mng no, mng trong tim bn
cp, vim tai, vim xoang, vim mng phi [19].









Trang 16

Chng 2
I TNG V PHNG PHP NGHIN CU
2.1. I TNG NGHIN CU:
i tng nghin cu l qu mp ng (Momordica Charantia
Linn.var.abbreviata Ser ). H B Cucurbitaceae.
Mu thu ti huyn akrng tnh Tha Thin Hu vo thng 8 nm 2012 .

Hnh 2.1. Qu mp ng (Momordica Charantia L)
Sau khi thu mu, nguyn liu c ra sch v chia lm hai phn: mt phn
bo qun t lanh, mt phn thi nh, phi, sy kh 50 - 60C, sau xay thnh
bt th v bo qun ni kh thong.
2.2. HA CHT, DNG C:
2.2.1. Ha cht:
- Thuc th TPTZ (2,4,6-tripyridyl-s-triazin ).
- Methanol tuyt i, nc ct.
- Mui FeCl3.6H2O, FeSO4.7H2O, Natriacetat, acid acetic, acic HCl m
c.
- Mi trng Mueller-Hinton c v Muller-Hinton lng.
2.2.2. Dng c:
- T sy Memmert, my xay.
- Cn in t hin s phn tch Shimadzu ( chnh xc 0.0001g).
Trang 17

- B chit Shoxlec, my lc, bp cch thy, my c quayYAmato.
- My o quang ph hp th UV-Vis
- T m nui cy vi sinh vt.
- Phu lc, cc thy tinh, bnh nh mc.
- a peptri, que bng, que cy, pipet pasteur, a pipet.
- Thc o vng v khun
- My hp tit trng.
2.3. PHNG PHP NGHIN CU:
2.3.1. Kho st hot tnh chng oxy ha:
2.3.1.1. Chit xut:
- Mu nguyn liu c ra sach, chia lm hai phn:
+ Thi nh, xay nhuyn bo qun t lnh
+ Phi sy kh, nghin thnh bt mn, bo qun trong iu kin nhit v
m thch hp.
- Chit xut vi methanol tuyt i bng phng php ng su ( The Modified
method of Chang ) [33]
- Dch chit thu c, lc, c quay dung mi di p sut gim n cn.
- Cn thu c c s dng kho st hot tnh khng khun.
2.3.1.2. Nghin cu v hot tnh chng oxy ha:
Kho st hot tnh chng oxy ha ca qu mp ng bng phng php FRAP.
2.3.1.2.1. Nguyn tc:
Xc nh kh nng oxy ha ca cc cht da trn s gim nng lc chng oxy
ha ca ion Fe
2+
theo Benzie Strain(1996) [30], [31].
2.3.1.2.2. Trnh t tin hnh:
+ Chun b dung dich th:
- Dung dch m acetat 300mM (pH = 3,6),
- 10 mM 2,4,6-tripyridyl-s-triazin (TPTZ) trong 40 mM HCl,
Trang 18

- 20 mM FeCl3.6H2O trong nc ct.
T l cc thnh phn trong thuc th FRAP theo th t trn l 10:1:1 theo th
tch.
Dung dch thuc th lun c chun b mi (trong ngy).
+ Tin hnh:
- Chun b thuc th FRAP.
- Trn 1,5 ml thuc th FRAP vi 50 l mu.
- Kho st thi gian phn ng gia thuc th v dch chit: 1,5 ml thuc th
FRAP trn vi 50 l c o quang ti bc sng 595nm sau 30 pht mi ln.
- Xc nh hot tnh chng oxy ha ca mu mp ng ti v kh. Thuc th
FRAP: cn = 1,5ml: 50 l em theo thi gian xc inh, sau o quang vi
my quang ph hp th UV-Vis.
- Cc tnh kt qu:
+ T mt quang D o c ca dung dch.
+ Da vo th ng chun: y=ax+b
+ Tnh hm lng Fe
2+
trong dung dch chnh bng hm lng hot tnh
chng oxy ha c trong mu o : C mol/L
+ Hm lng Fe
2+
hay hm lng hot tnh chng oxy ha trong mu ban
u: C x f ( mol/L) f: h s pha long.
2.3.2. Kho st hot tnh khng khun:
2.3.2.1. Chit xut:
- Mu nguyn liu c phi sy kh, nghin thnh bt mn, bo qun
trong iu kin nhit v m thch hp.
- Chit xut vi methanol tuyt i bng phng php chit Shoxlec.
- Dch chit thu c, lc, c quay di p sut gim n dch chit m
c.
- Cn thu c c s dng nh gi hot tnh khng khun.
Trang 19

2.3.2.2. Nghin cu v tc dng khng khun:
Th nghim xc nh hot tnh khng khun ca dch chit theo phng php
khng sinh trn cc chng vi khun ATCC v vi khun gy bnh. C th
nh sau:
2.3.2.2.1. Nguyn l
ng dng phng php khuch tn trong mi trng thch a.
2.3.2.2.2. Trnh t tin hnh:
- Chun b dung dch th nghim: dch chit mp ng m c.
- iu ch mi trng khng sinh d Muller-Hilton, hp tit khun, vo a
petri v khun ( mi a thch dy 3,5-4,5mm).
- Nui cy cc chng vi khun th nghim.
- Cy tri hn dch vi khun th nghim ( khong 0,1ml) trn mt a thch
Muller-Hiltion, nhit phng khong 30 pht.
- Dng ui popet pasteur tit khun to cc ging trn mt thch.
- Nh y cc ging dung dch chun b.
- rr cc a nhit phng 20-30 pht ( cho dch chit thm thu hoc khuch
tn ra xung quanh), sau chuyn vo t m 37
0
C trong 24 gi.
- Ly a ra v kho st vng v khun (c hau khng, kch thc ng knh
vng v khun).
- Nu dch chit c tnh khng khun mnh vi chng vi khun no th tip tc xc
nh hot tnh khng khun cc nng khc nhau bng cch pha long dch
chit vi chng vi khun .
2.3.3. nh gi mc khng khun ( nu c)
Ch p dng sau khi c kt qu sng lc nh gi nng tc dng.
2.3.3.1. Nguyn l
ng dng phng php pha long dch chit trong mi trng.
2.3.3.2. Trnh t tin hnh
Trang 20

- iu ch mi trng Muller-Hilton lng, ch vo cc ng nghim, mi ng 1ml.
Hp tit khun. ngui.
- Pha long cc dch chit theo nng gim dn t 1/2, 1/4, 1/8, 1/16, 1/32, 1/64,
1/128, 1/256, 1/512, 1/1024 trong cc ng mi trng trn.
- Pha hn dch vi khun 10
6
t bo/1ml.
- Cho vo mi ng trn 0,1ml hn dch vi khun 10
6
t bo/1ml . Trn u.
- m 37
0
C trong 24 gi.
- Dng que cy ly hn dch t cc nng cy ln mt a thch.
- m 37
0
C trong 24 gi.
- Ly a ra v quan st vi khun mc nh gi nng cn kh nng c ch s
pht trin ca vi khun.
2.4. KHNG GIAN V THI GIAN NGHIN CU
- Khng gian nghin cu: Khoa Dc, B mn Vi Sinh- Trng i Hc Y Dc
Hu.
- Thi gian nghin cu : t thng 8/2012 n thng 4/2013.
2.5. CC K THUT THC HIN
- K thut chit ng su ( The Modified method of Chang ) [33]
- K thut chit Shoxlec.
- K thut ct loi dung mi di p sut gim trn my c quay chn khng.
- K thut ly mu bng Micropipet.
- K thut cn, ha tan.
- K thut o quang ph hp th phn t bng my o quang ph hp th UV-VIS.
- K thut mi trng.
- K thut cy tri vi khun.
- K thut c l v nh dch chit.
Trang 21

- K thut o ng knh vng v khun.
2.6. X L S LIU
- S liu c x l bng phn mm Microsoft Excel 2007.























Trang 22

y = 0.017x - 0.0363
R = 0.9922
0
0.2
0.4
0.6
0.8
1
1.2
1.4
0 10 20 30 40 50 60 70 80
A
M
Chng 3
KT QU NGHIN CU

3.1. NGHIN CU V HOT TNH CHNG OXY HA CA QU
MP NG (Momordica Charantia)
3.1.1. Chit xut:
- Cn 1000 g mp ng ti, xay nhuyn em -20
0
C trong 2 ngy th
thu mu v tip tc -42
0
C trong 4 ngy ( -133x10
-3
psi ) . Sau , lc vi
dung dch Methanol bng my lc 37
0
C trong vng 24 gi. Lc dung dch, b
b thu c dch chit. Dich chit ny ct thu hi dung mi di p sut
gim c cn mp ng ti
- Cn 1000g mp ng ti, sy kh, xay v tin hnh tng t vi mu
ti ta c cn mp ng kh
- Cn thu c bo qun t lnh, chun b cho nhng bc tip theo.
3.1.2. Kho st hot tnh chng oxy ha
3.1.2.1. Xy dng ng chun
- Dung dch FeSO4.7H2O c pha thnh cc nng 1000, 750, 500, 250,
100 M phn ng vi thuc th FRAP o ti bc sng 595nm.








Hnh 3.1: th ng chun
Trang 23

0
0.2
0.4
0.6
0.8
1
1.2
1.4
0 1 2 3 4
A
h
3.1.2.3. Xc nh thi gian phn ng gia dch chit v thuc th
- 1,5 ml thuc th FRAP c trn vi 50 l mu.
Tin hnh o bc sng 595 nm sau cc khong thi gian 0,5h, 1h, 2h,,3h ta
c kt qu ( bng 3.1).
Bng 3.1: Tm tt s thay i hp th theo thi gian



Hnh 3.2: th biu din s thay i hp th theo thi gian
Sau 2h hp th hng nh, do chn thi gian phn ng gia dch
chit v thuc th xy ra hon ton: 2h.
3.1.2.4. Kt qu kho st hot tnh chng oxy ha
3.1.2.4.1. Mu ti
1,5 ml thuc th FRAP c trn vi 50 l mu, sau 2h em o quang ph
hp th ti bc sng 595nm. Tin hnh trn 3 mu khc nhau, lp li th nghim 3
ln mi mu .


t (h)
hp
th
0.5 0.6199
1,0 0.9213
1.5 1.0368
2,0 1.2625
2.5 1.2617
3,0 1.2628
3.5 1.2596
Trang 24

Hnh ny l v dung dch khng ng nht, xem li

Hnh 3.3. Thuc th FRAP

Hnh 3.4: Thuc th sau khi trn vi dch chit.




Trang 25



Bng 3.1 Tm tt kt qu kho st kho st hot tnh chng oxy ha ca mu
mp ng ti.

S ln o hp th
Trung bnh
FRAP
(mol Fe/L)
Frap ban u
(mol/L Fe)
Ln o Mu
1 0.7857
0.7857
0.7824 44.77 1119.25
2 0.7859
3 0.7855
1 0.7853
0.7855 2 0.7854
3 0.7857
1 0.7759
0.7759 2 0.7761
3 0.7757

H s pha long: f =25 ln.

3.1.2.4.2. Mu kh
Tin hnh tng t mu ti. Tin hnh trn 3 mu khc nhau, lp li th nghim 3
ln mi mu ta c:

Bng 3.2 Tm tt kt qu kho st kho st hot tnh chng oxy ha ca mu
mp ng kh.

Trang 26

S ln o hp th
Trung bnh FRAP
(mol/L Fe)
Frap ban u
(mol/L Fe) Ln o Mu
1 0.6683
0.6684
0.6692 39 780
2 0.6683
3 0.6686
1 0.6701
0.6702 2 0.6705
3 0.6701
1 0.6689
0.6688 2 0.6689
3 0.6687

H s pha long f = 20 ln

Hnh 3.4 th biu din hm lng chng oxy ha mu
mp ng ti v kh

y = 0.0196x - 0.0952
R = 0.9988
0
0.2
0.4
0.6
0.8
1
1.2
0 20 40 60 80
Series1
Linear (Series1)
Trang 27

3.2. NGHIN CU HOT TNH KHNG KHUN
3.2.1. Chit xut
Nguyn liu sau khi thu hi, ra sch, ro, sy kh nhit 60
0
C, sau
nghin nh thu c bt th. Bt ny c chit vi methanol tuyt i bng
phng ph p chit Shoxhlec.
Dich chit ct thu hi dung mi p sut gim thu c dch chit m c.
Dch chit c bo qun t lnh khoa Vi Sinh, chun b cho cc bc
nghin cu hot tnh khng khun.
3.2.2. Kt qu kho st hot tnh khng khun
S dng a thch v khun chun b sn. Cy tri mi chng vi khun ln mi
a thch. Mi a thch c 1 l.Tng cng c 4 a thch th nghim trn 4
chng vi khun ATCC. Nh dch chit vo cc l thch.
Kt qu 4 a thch ca 4 chng vi khun ATCC th hin cc hnh 3.5 n 3.8
v bng 3.3.

Hnh 3.5: Vng v khun ca dch chit trn S.aureus ATCC.
Trang 28


Hnh 3.6: Vng v khun ca dch chit trn P. aeruginosa ATCC.




Hnh 3.7: Vng v khun ca dch chit trn E.coli ATCC.


Trang 29



Hnh 3.8: Vng v khun ca dch chit trn Streptococcic ATCC.

Bng 3.3: Kt qu kho st hot tnh khng khun ca dch chit mp ng

Chng vi khun ng knh vng khng khun (mm)
Streptococci ATCC 17
S.aureus ATCC 15,5
P. aeruginosa ATCC 14
E.coli ATCC 14

Nhn xt:
Kt qu cho thy hot tnh khng khun ca mp ng (Momordica Charantia
Linn ) u c tc dng trn c 4 chng vi khun gy bnh thng gp. V ng
knh vng v khun khc nhau ty thuc chng vi khun. Tuy nhin, nhn chung
hot tnh khng khun ca mp ng trn cc chng ny vn cha cao, do ,
Trang 30

vic tip tc nghin cu hot tnh khng khun bng phng php pha long dch
chit ngha thc tin s khng cao.

























Trang 31

Chng 4
BN LUN


Hin nay, tnh trng nhim mi trng, thc phm nhim cht c hi..
ngy cng tr nn trm trng. Do , qu trnh xm nhp to ra cc gc t do trong
c th cng tr nn thun li gy nhiu bnh ngi, lm c th nhanh chng b
lo ha. ng thi, vi s lm dng thuc, s dng khng sinh mt cch khng
hp l khng ch l nguyn nhn lm tng cc ADR nghim trng m cn l
nguyn nhn lm tng ng k chi ph cho ngi bnh [20].
V vy, qu trnh nghin cu cc hp cht chng oxy ha cng nh nghin
cu tm ra cc khng sinh mi an ton hn, hiu qu hn l mt phng php hu
hiu gip con ngi nng cao sc khng, min dch, phng chng lo ha v
chin thng c cc vi khun gy bnh.
Nc ta, mt nc kh hu nhit i gi ma vi ngun dc liu v cng
phong ph. Ngay c nhng rau qu lm thc phm thng ngy cng c tnh nng
chng oxy ha v khng khun: ta t, ti, ch xanh, mp ng..
Do , nghin cu hot tnh chng oxy ha v khng khun ca nhng thc
phm hng ngy gip chng minh bng thc nghim cng dng phng cha bnh
ca nhng thc phm ny, khng nh tm quan trng ca chng ng thi gip
ngi dn la chn cch s dng hiu qu nht.






Trang 32


4.1. NGHIN CU HOT TNH CHNG OXY HA
4.1.2. So snh vi cc kt qu nghin cu khc
4.1.2.1. ng hc phn ng FRAP
Theo Benzie v Strain [28], phn ng Frap kt thc sau 4 pht vi cc cht
kho st v kh nng chng oxy ha l bilirubin, acid uric, vitamin C v vitamin E.
Tuy nhin, vi cc polyphenol v mt s hp cht trong dch chit thc vt phn
ng i khi xy ra rt chm nn phn ng FRAP c th ko di trong vi gi [23].

bit c thi gian hon thnh phn ng vi cc mu th nghim, cc
cao chit c o ng hc phn ng trn my quang ph UV-Vis v kt qu cho
thy thi gian phn ng FRAP ko di 2 gi. Nh vy, cc mu phn ng hon
ton, chn thi gian phn ng l 2 gi cho tt c cc mu. Kt qu khng c s sai
bit ng k no so vi ti liu nghin cu [ 21],[23].
4.1.2.2. Kt qu chng oxy ha theo phng php FRAP
4.1.2.2.1. So snh kt qu nghin cu vi cc kt qu nghin cu khc.
Bng 4.1: So snh vi nghin cu khc
Mu Hot tnh chng oxy ha
FRAP
( mol/L Fe
2+
)
Phng php chit
Ti liu [21] 1079 Chit shoxlec
Sinh vin thc hin 1119.25 ng su (The Modified
method of Chang )

Kt qu khng c s chnh lch nhiu so vi kt qu ti liu [21]. S chnh
lch y ch yu l do phng php chit. iu ny chng t, phng php chit
Shoxlec tuy mt phng php chit kit trong qu trnh chit xut nhng di tc
Trang 33

ng ca nhit nn mt phn cht chng oxy ha b phn hy, do kt qu
thp hn so vi xc nh hot tnh chng oxy ha s dng phng php chit ng
su.
Qu trnh chit ng su lm ph v cu trc sinh hc ca nguyn liu, gip
nguyn liu d dng thm dung mi khi lc nhng vn m bo nguyn liu bo
ton tt c cc hot cht.
4.1.2.2.2. So snh hot tnh chng oxy ha ca mp ng vi mt s dc liu



Hnh 4.1: Biu hot tnh chng oxy ha
Mp ng kh: A Mp ng ti: B
C nh ni, c mc: C Ti: D
Xuyn tm lin: E

0
500
1000
1500
2000
2500
A B C D E
Series 1
Series 2
Series 3
Trang 34

Xuyn tm lin c tc dng khng vim, h nhit, khng khun, thng c
dng lm thuc iu tr cc bnh: nhim khun h hp ( vim mi, vim xoang
mi, vim gan, vim rut, thng hn, lao phi, vim tai gia, vim phn ph,
vim tt tnh mch, nhim khun tiu ha (vim rut cp, kit l), vim tit niu (
vim niu o, vim thn b thn), vim da, mn nht v nhng bnh xon trng.
Ti cng l mt loi dc liu c tc dng chng oxy ha cao, trong thnh
phn ti c cha alliin, cht chng oxy v mt s thnh phn nh selenium,
vitamin C, vitamin Ec kh nng ngn chn s pht trin ca cc khi ung bu.
Da vo biu hnh 4.1 ta thy, hot tnh chng oxy ha ca mp ng
chim hn 2/3 hot tnh chng oxy ha ca ti ( 1508 mol/L Fe
2+
) v xp x bng
mt na hot tnh chng oxy ha ca xuyn tm lin ( 2151 mol/L Fe
2+
) [21]
.iu ny c chng t, mp ng ngoi l mt thc phm hng ngy, n cng l
mt dc liu c kh nng chng oxy ha cao.
Hot tnh chng oxy ha ca mp ng sau khi sy kh ( 780 ( mol/L
Fe
2+
) ) chim 2/3 so vi mu ti (1119.25 ( mol/L Fe
2+
))
.
. iu ny c
ngha, pht huy ht tc dng ca mp ng ta nn dng mp ng cng ti
cng tt. Tuy nhin, iu ny cng ng thi chng t rng, mp ng dng
di dng phi kh, sc nc ung vn cn tc dng chng oxy ha tng i
cao.
4.3. NGHIN CU HOT TNH KHNG KHUN:
Kt qu cho thy hot tnh khng khun ca mp ng (Momordica
Charantia Linn ) u c tc dng trn c 4 chng vi khun gy bnh thng gp.
V ng knh vng v khun khc nhau ty thuc chng vi khun. Nhn chung,
chng Streptococcus d nhy cm nht so vi cc chng cn li di tc dng ca
dch chit hay ni cch khc tc dng khng khun ca dch chit c hiu qu cao
nht i vi chng Streptococci. Tip n l chng S.aureus v P. aeruginosa ,
E.coli.
Trang 35

Hot tnh khng khun ca mp ng trn cc chng ny vn cha cao, do
, vic tip tc nghin cu hot tnh khng khun bng phng php pha long
dch chit ngha thc tin s khng cao.
























Trang 36

KT LUN

Qua mt thi gian nghin cu, ti thu c cc kt qu sau:
1. to c cn v dch chit m c ca qu mp ng a vo kho
st hot tnh chng oxy ha v khng khun.
2. Kt qu nghin cu hot tnh chng oxy ha cho thy, qu mp ng c
hot tnh chng oxy ha tng i cao, c ngha thc t.
3. La chn cch dng, cch bo qun m bo s dng mp ng vi hm
lng chng oxy ha tt nht.
4. Kt qu nghin cu hot tnh khng khun cho thy:
Dch chit qu mp ng u c tc dng khng khun trn cc chng vi
khun gy bnh thng gp















Trang 37

KIN NGH

Tip tc nghin cu hot tnh chng oxy ha v khng khun trn cc b phn
khc ca cy mp ng: l, r, ht.. c ci nhn ton din hn v tc dng
chng oxy ha v khng khun ca mp ng.






















Trang 38

TI LIU THAM KHO

TING VIT
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v K Thut, tr. 734.
2. V Vn Chi (1999), T in Cy Thuc Vit Nam, Nxb Y hc.
3. Hi n (2002), Mn n v thuc, Nxb Thun Ha.
4. Lin Hng (2004), u im ca khng sinh thc vt, Tp ch Cy thuc
qu, (25), tr. 17.
5. B mn Dc liu (2007), Bi ging dc liu tp I, trng i hc Dc
H Ni, Nxb Y hc
6. T Duy Chn (1999), Nhng phng thuc hay cha bnh bng hoa,
Nxb Ngh An.
7. Thi Xun Mi (2002), Tho dc cha bnh, Nxb Thanh Ha.
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d tm v cc bi thuc gia truyn, Nxb Thanh Ha.
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Ho hc hu c ton quc ln th t, NXB i hc Quc gia H Ni.
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14. Nguyn Thanh Hng (2007), Cc phng php ph trong ha hc hu
c, Nxb Khoa Hc v K Thut. Phm Vn Thanh, Phm Kim Mn,
15. on Th Nhu, Nguyn Thng Dong, V Kim Thu, Nguyn Kim
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21.
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properties of Momordica charantia L. (bitter melon) fruit extract,
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34. http://www.medinet.hochiminhcity.gov.vn/data/news/2009/7/6700/muopdan
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htm
36.















:




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