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Đề Tài Trích Ly Collagen Từ Da Cá Basa - Tài Liệu, eBook, Giáo Trình
Đề Tài Trích Ly Collagen Từ Da Cá Basa - Tài Liệu, eBook, Giáo Trình
Lun vn tt nghip
Trang 1
.
1.5 nm
300 nm
Hnh 1.1 Cu trc ca phn t collagen (triple heix)
N bao gm 3 chui polypeptide (gi l chui a) cun li vi nhau. Mi chui a cun thnh ng
xon c theo hng t phi sang tri vi 3 gc trn mt vng xon. Ba chui ny xon li vi nhau theo
hng t' tri sang phi to thnh ng b ba xon c (triple helix).
A
oc
1:
cc
2:
tx 1':
|(RroVaaGly)5| (ProYaaGly) 3
|(RroYaa<3ly)s|I
(ProYaaGly);
Gly
G lytCysOysGly-|(RroVaaGly)5|
(ProYaaGly):
s
s Gly
1 : Vaa = Pro
2: Vaa = Hyp
0,2
Gly -1 - X
0,27
Gly -1 -1
0,09
Procollagen
I
.
COOH
r
C
r
Procollagen
peptidase
N-terminus end cleaved
cleaved
C-terminus end
Tropocollagen 3000
Phn t collagen
Chui polypeptide
T hp thnh
vi si
T hp thnh
si
D-Period
Hnh 1.6 S sp xp ca
phn t collagen trong mt
si
Nh vo cu
vn c ca cc
c
chuyn
collagen,
cung
T hp thnh
b si
trc c th bc, bn
chui xon c
sang cc si
cp cho cc
C vi lin kt bn trong mi chui triple helix v mt s lng bin thin cc lin kt ngang
gia cc chui hnh thnh nn nhng t hp c trt t (nh si collagen). Cc b si ln hn
c to nn nh vo s tr gip ca mt vi loi protein (bao gm nhng loi collagen khc
nhau), glycoprotein v proteolycan hnh thnh cc loi m khc nhau t s sp xp lun phin
ca nhng thnh phn trn. S khng ha tan ca
c nghin cu n cho ti khi ngi ta pht hin tropocollagen c th c trch ly t nhng ng vt cn non do cha
xy ra lin kt ngang hon ton. Tuy nhin, s pht trin ca k thut hin vi (Electron
Microscopy (EM) v Atomic Force Microscopy (AFM)) v s nhiu x tia X cho php cc
nh nghin cu thu c nhng hnh nh chi tit v cu trc ca collagen. Nhng tin b ny
c ngha quan trng trong vic hiu c cch thc m collagen tc ng ln mng li t
bo; s xy dng, pht trin, phc hi, thay i ca m trong qu trnh pht trin.
b
Hnh 1.7 Hnh nh ca b si (a) v si (b) collagen
Tnh n thi im hin ti, c 42 loi chui polypeptide c nhn dng. Chng c m ha
bi 41 loi gene khc nhau, to thnh 27 loi collagen. H collagen c phn loi thnh nhng
phn h khc nhau da vo s t hp ca cc siu phn t [4].
hng loi protein khc nhau th hin tnh a dng v phm vi ng ch , t nhng loi collagen ring bit nh collagen
II, X c trong sn n nhng loi collagen dng si phn b rng khp nh collagen I v V.
Nhng loi collagen khc nhau thc hin nhng chc nng chuyn dng trong cc m v c
phng thc sp xp ring bit ca cc siu phn t. Mt s loi c cu to t mt tam phn
(homotrimer), trong khi nhng loi khc c cu to t nhiu tam phn. Loi collagen phong
ph nht l nhng collagen dng si hnh thnh nhng nn tng cu trc ca da, gn, xng, sn
v nhng loi m khc.
Hin ti, c t nht khong 13 loi collagen c trch chit. Chng khc nhau v
chiu di ca chui xon c cng nh bn cht v kch c ca nhng phn khng xon c
[1].
Bng 1.2 Kt cu chui v s phn b ca cc loi collagen trong
c th
Phn
bngi [ 1 ]
a2(I), trimer (al(I))3
(<xl(II))3
(al(III))3
(al(IV))2a2(IV)
al(V)
a2(V)
a3(V)
(al(VII))3
Cha bit
To ra t t bo mng trong
Sn
(al(X))3
Sn
Sn, a m ct sng, dch thy tinh
(al(XII))3
Gn, dy chng
Cha bit
Da, xng
Hn 90% collagen trong c th l cc collagen loi I, II, III v IV. Nhng bnh tt lin quan
n collagen l do s khuyt tt v gene nh hng n s sinh tng hp, s sp xp cng nh
cc qu trnh khc trong s sn sinh collagen mt cch bnh thng.
1.4 ng dng ca collagen
1.4.1
ng dng trong y hc
Collagen l mt vt liu c kh nng phn hy sinh hc, c tnh tng thch sinh hc cng nh
kh nng cm mu nn c th c ch to thnh nhng dng khc nhau, l mt loi vt liu sinh
hc l tng cho vic sn xut cc sn phm y hc. Mng
collagen c s dng cho hng lot nhng ng dng nh lm cht bt kn sinh hc v ci
thin tnh p ng sinh hc i vi nhng m cy. Collagen c s dng nh mt h thng
phn hy sinh hc cho ra cc loi thuc bao gm thuc trnh thai, khng sinh, insulin, hormone
tng trng, [5]... Collagen c s dng rng ri trong phu thut thm m, cha lnh vt
thng cho cc bnh nhn b phng, ti to xng v nhiu mc ch khc thuc nha khoa, phu
thut, chnh hnh. Collagen cn c dng trong vic xy dng cu trc da nhn to cha tr
cho cc vt bng nghim trng. i khi chng c s dng kt hp vi silicone,
glycosaminoglycan, nguyn bo si, cc tc nhn tng trng v cc hp cht khc. Collagen
cng c bn trn th trng nh mt cht b sung, ci thin tnh linh hot ca khp. Ngoi ra,
collagen cn l phn nn gip tch t calcium trong xng. Neu hm lng collagen gim ng
k, calcium khng th tch t lm cho xng gin v d gy, sn d b hao mn to nn cc cn
au khp gi v hng. Nh vy, xng cng cp hn, ngoi calcium cn b sung mt lng
ln collagen.
Collagen l mt polymer t nhin, n c dng trong
phu thut to hnh nh bm mi, cng da mt,...
Mc d khng th hp th qua da nhng hin nay
collagen ang c dng lm thnh phn chnh trong cc
sn phm m phm.
collagen
1.4.
2.2
Mng
mng
Age 50+
Age 40+
Hnh 1.12 S lo ha da
thng hay trong giai on ti to sau khi iu tr nm, mn trng c, so, rn da v tiu da tha
sau khi gim bo. Sn pham Phials of Collagen and Elastin vi cc thnh phn chnh l collagen,
elastin v vitamin F, trong collagen c chit xut t da heo gip kch thch t bo v sa
cha cc khim khuyt ca m nh phc hi kh nng n hi ca da. Ngoi ra, sn phm cn
c th iu tr cc vt rn do tng cn hay mang thai.
-
n ung.
a phng php trn u l nhng bin php rt an ton khng h nh hng ti da v c th.
Gim thiu so mn
so mn ca collagen
collagen v gelatin dng trong thc phm, dc phm, m phm v cng nghip thuc da.
Hin nay, nhng phn tha t c nh xng, vy v da l nhng ngun thay th cung cp
collagen.
Hnh
1.15 Bt
collagen sn xut t da v vy c
Bt collagen t da v vy c bin (marine fish collagen powder): l loi bt trng, khng
mi, tan hon ton trong nc nhit phng, to thnh dung dch trong sut; c hp th
mt cch d dng, thnh phn protein cao vi hn 90%, hm lng tro v kim loi nng thp.
Collagen
/HI/EIDO ! Collagen
1.7.1
hn minh mn, b sung dinh dng, duy tr cc chc nng v do dai ca khp, gip ngn nga mn v ti sinh
ln da mn mng.
Xut x: Shiseido, Nht B
n
Hnh 1.18 Collagen Update
1.7.4
Xut x: Hn Quc
wL%
collagen
rn FACE SHOP
2 MT S QUY
30 pht
1 ln
20 pht
2 ln
30c
6h X 2 ln (CHBM)
Kettu I pH = 4,6-s-4,7
Ly tm
CH3COOH 0,1M
72h
Ket ta li
Trch ly
I
1 Oh
(isopropyl)
I 20c, 1 tun
4c
Ly tm
Ra sch
Lm kh
v mu
Ra
ru ^\/ 'I nrA,,
I 1 V/,_UV1, Iiga_y
Trch ly
50.000 vng/pht, lh
NaCl 0,8M
L Vi m m m "U
50.000 vng/^ht, lh
Ly tm
Kt ta
Ly tm
Bt collagen
2.4 Hnh 2.3 Quy trnh trch ly collagen t da mcQuy trnh trch ly
collagen t da c tuyt (Baltic cod) [9]
Da c
Dng my nghin () = 3 mm
Khuy
Ct
nh
2h
0 c
0 c
vng/ph
Khuy |^0
24h
4c
2 pht
^ng^nhni^^
0 c
6.000 vng/pht 20
Nguyn liu
Knong tan
Collagen ha tan
Ly tm
pht
1 0 c
10.000 vng/pht
Nhm c hm lng collagen thp: tng lng collagen chim n 5 % so vi tng lng
protein
-
Nhm c hm lng collagen trung bnh: tng lng collagen chim t 5 -- 10 % tng
lng protein
- Nhm c hm lng collagen cao: tng lng collagen trn 10 % tng lng protein
Tht ca nhng loi c khc nhau cha hm lng khc nhau ca collagen ha tan trong
acid (acid soluble collagen) v collagen khng ha tan (insoluble collagen). Hm lng
collagen tng cng dao ng t 0,3 -T- 2,99 % so vi trng lng ti ca cc m, tng ng
tng)
(White pomfret)
mp
(Shark)
C ui
(Ray)
Tng lng collagen trong c th thay i t' 3,26 % ( c thu) cho n 6,97% ( c
chnh Nht Bn). Lng collagen c th ha tan trong acid nm trong khong t
13,1 % (c chnh) n 56,6 % (c bn). Hm lng collagen thu c s thay i ty theo loi
v loi m c s dng trong qu trnh trch ly. Trong tt c cc loi, kh nng ha tan ca
collagen trong tht tng i cao nhng thp i vi nhng loi collagen trong ni tng.
Collagen trong da, vy, xng v vy chim phn ln trong tng lng collagen.
2.7 So snh collagen t c v collagen t cc loi gia sc
Cho n nay, hu ht cc loi m phm trn nn collagen u c cha protein chit xut
t' gia sc. Bi s bc pht ca bnh b in BSE (Bovine Spongiform Encephalopathy - mad
cow disease), collagen t gia sc ang dn rt khi phm vi s dng, thay vo l ngun
collagen c trch t c. N c nhiu c im tt hn cho vic ng dng trong y hc v m
phm. M phm trn nn collagen t gia sc c tc hp th rt chm trn da ngi. i mt
vi vn ny, liu php tim collagen vo da c gii thiu. Tuy nhin, nhng bt li ca
cch thc ny l t tin, au n v i km vi n l nhng ri ro. Gi y, cc nh khoa hc
Phn Lan pht trin mt tin trnh trch ly collagen t' c. Khng ging vi nhng loi
collagen thu c t nhng phng php trc y, collagen t c c hp th hon ton trn
da ngi. C sng trong phm vi rng ln vi cc iu kin v nhit , su v p sut khc
nhau. iu ny c ngha l collagen trch t da c c mt sc chng chu c bit vi cc ph
hy l v ha hc.
Fish-erived
So snh kh nng hp
ol/ |th vo
140 mu gia collagen t c v collagen t ln
120
3*
-e
0.
J3
100
M S
o
>
S >
M
P<j derrvifd
II
a\
80
60
/\\
40
20
0
10
15
20
AV
/ v\ hp th vo mu ca collagen trch ly t c v t ln
Hnh 2.5 th so snh kh nng
Thi sian sau khi s dung
/
2.8 Ngun nguyn liu c cho vic sn xut collagen Vit Nam
Vit Nam l mt nc ang pht trin, c nn nng nghip vi cc sn phm phong ph. Trong
, cc sn phm t c chim mt phn ln trong kim ngch xut khu. Nhng nm gn y, vic
nui c Basa rt pht trin, c bit ng bng sng Cu Long. Trong qu trnh ch bin c,
song song vi nhng chnh phm dng cho vic xut khu (ch yu l fillet) l mt khi lng
ln cc ph phm nh da, xng, vy,... y l nhng ngun giu collagen, c th s dng thay
th cho nhng ngun cung cp collagen c in nh da, xng t ln, b. Vic tn dng nhng
ngun ny s gp phn lm tng gi tr kinh t ca loi c Basa
.
vy hu mn c mu
Trc y, tn loi Pangasus Pangasus c t cho c Basa l khng chnh xc. V thc
t, loi Pangas ius Pangas ins khng phn b lu vc sng Cu Long m ch gii hn n
, Pakistan, Banglades v Myanmar, l loi c th sng vng ca sng v bin ven b trong
mi trng nc l hay nc mn, khi trng thnh ui c mu vng sng. iu ny khng th
B Siluriformes
H Pangasiidae
Ging Pangasius
Loi Pangas ins Pangas ins
c im sinh trig
hanh, sau 60 ngy t c chiu di 8 -- 10,5cm; sau v nui 10 -- 12 thng c th t 1,3 -- 1,5 kg/con. Mt s b
nui c Basa thm 6 -- 9 thng, c c c th t ti 1,8 -* 2,2 kg/con. Nng sut nui hin nay
khong 120kg/m3 b nui v sn lng c thu hoch trung bnh 50 --160 tn/b ty theo c b.
3.2.2
c im sinh dig
C trng thnh ca hu ht cc loi trong h Pangasiidae l n tp v thc n ch yu l t ng vt, ngoi tr 2 loi Pcmgasus sanitwongsei v Pangasus arnaudi n ch yu mn b
hu c. Tng t, c Basa cng c tnh n tp thin v thc n ng vt, nhng t hu n v t
tranh mi n hn c tra. Sau khi ht non hon, giai on ny c n ph du ng vt l chnh. Khi
c giai on ln, c Basa cng d dng thch nghi vi cc loi thc n c ngun gc t ng
vt, thc vt, d kim nh: hn hp tm, cm, rau qu, c vn v cc ph phm nng nghip..., do
rt thun li cho vic nui c trong b.
3.2.3
Ma v sinh sn
Trong t nhin, tui thnh thc ca c Basa t 4 -T- 5 nm. Vo ma pht dc t thng 4 tr
i c c tp tnh bi ngc dng di c tm n cc bi thng ngun sng Mekong, ni c
iu kin sinh thi ph hp cho s pht trien ca tuyn sinh dc v trng. V vy, c khng
t nhin phn sng Mekong ca Vit Nam m bi ca c nm khu vc t a phn tnh
Cratie ca Campuchia tr ln thng ngun, ti y c th bt c c b m nng n 15 kg.
Ma v sinh sn ca c Basa nui bt u t thng 2 ko di n thng 7, c r v tp trung t
thng 3 n thng 5.
3.2.4
Ging c nui
Ging c Basa nui hin nay c t 2 ngun: bt t t nhin v sinh sn nhn to. Trc y
th c Basa ging hon ton c bt ngoi t nhin ti vng bin gii gip Campuchia v Vit
Nam bng cch cu, li hoc cc hnh thc thu bt c giong khc v thng c c ging khong
5 + 6 g/con. Sau khi mua hoc nh bt v, c c chm sc trong b nh trong 3 -- 4 thng cho
n khi t c 80 -i-100 g/con mi a vo b nui c tht. Hin nay c sinh sn nhn to c
c Basa nhng vi sn lng cn rt thp. Theo c tnh, hng nm 2 tnh An Giang v ng
Thp cn 15 triu con ging v ch yu c chuyn trc tip t Campuchia sang.
3.2.5
Mt nui
Ma v nui
Trong qu trnh nui khi thu hoch nn thu hoch 1 ln ht s c. V theo kinh nghim, nu
thu hoch mt phn c th s c cn li d b sc, thng b n dn n hao ht ln.
3.2.7 nh hng ca mt s yu t mi trng i vi c Basa
Ngng oxy ca c Basa rt cao 1,1 0,15 mg/1. C Basa c sc chu ng yu khng th
sng c trong mi trng ngho oxy. Khi hm lng oxy trong mi trng < 1 mg/1 th c ni
u, c th cht trong thi gian ngn.
Kh nng chu mn ca c Basa tng i cao, c c th sng bnh thng trong nc c
mn 1 ,2 % tr xung, khi tng mn th thi gian sng ca c gim xung. Nng mui trong
nc t 3,5% th c ch sng c 1 gi.
Bng 3.2 Sc chu ng ca c Basa trong mi trng nc c mn khc nhau
2,0
1,5
2,5
3,0
3,5
72
1,4
mn (%)
Nhit (C)
pH
nhi. V vy, bn cnh vic khai thc hiu qu ngun tht c, cn tp trung nghin cu s dng
hiu qu ngun m c. C th l nghin cu dng cc phng php tinh luyn ph hp khc
phc cc nhc im vn c ca m c Basa nh c mi tanh v chng i... nhm bin m c
Basa thnh mt ngun cht bo c th s dng rng ri trong thc phm, i sng. y cng
chnh l mc tiu ca cc nh nghin cu l mun th nghim phng php tinh luyn vt l i
vi m c Basa loi tr cc acid bo t do, cc hp cht peroxyde... nhm m bo cho m c
tha mn cc ch tiu v du m thc phm. Bn cnh , vic kh mi tanh ca m c cng l
vn c tp trung nghin cn v mi tanh t nhin ca c gy cn tr rt ln cho qu trnh s
dng trong thc phm. Cc bin php bo qun nhm ko di thi gian tr mi, i ha ca m c
cng c tin hnh nghin cu m bo cho m c c th s dng trong mt thi gian tng
i di.
3.6 Cc sn phm c ch bin t c Basa
Ngun c Basa trong t nhin khng th cung cp cho ch bin cng nghip m ch
yu l ngun c c nui trong lng b. C Basa thng c s dng :
Nu chua, nu lu trong cc nh hng, khch sn.. t c tiu th ph bin trong gia nh v
gi tng i cao.
Ch bin cng nghip: c Basa thng c ch bin dng fillet ng lnh l chnh v ch
yu phc v cho th trng xut khu.
Ngoi sn phm fillet ng lnh, cc sn phm khc ch bin t c Basa c mt trn th
trng trc y cn rt t, c l do quy trnh k thut cha c hon thin to ra cc sn
phm c cht lng cao, p ng nhu cu ca ngi tiu th.
Tuy nhin, vi vic ngy cng ch trng m rng th trng tiu th ni a nn cc sn
phm c ch bin t c Basa c u t nghin cu v pht trin rt nhanh trong giai on
gn nay. Va qua, ti hi ch thu sn Vit Nam nm 2003, cng ty Agifish An Giang (cng ty
xut khu c da trn ln nht ca Vit Nam) gii thiu mt lot cc sn phm mi c ch
bin t c Basa nh :
Dng mn n khai v: c vin Basa, cho Basa, cho mc Basa, ch gi Basa, ch qu Basa,
bnh khoai moan nhn Basa, Basa bnh trn tm bt, nm ng c nhn Basa, Basa ct si tm
bt, bnh ch Basa, honh thnh Basa ...
Dng mn n gia nh ch bin sn: c Basa kho t, Basa fillet st c, Basa ct khoanh mui s
t, c chua dn Basa, kh qua dn Basa, c bu nhi Basa, vy c Basa tm gia v, bong bng
c Basa tm gia v, da c Basa tm gia v ...
Dng mn n t ch bin: Basa ct khoanh, lu Basa, Basa fillet, Basa lt da rt xng, Basa
xin que, bao t Basa, hi sn thp cm, ch tht lt ...
Dng sn phm kh: Basa nguyn con sy kh, kh Basa, kh Basa n lin, ch bng Basa, lp
xng Basa, bnh phng Basa ...
Hin nay tnh hnh th trng xut khu c Basa, c tra Vit Nam vn cha c on nh. V
th cn iu chnh li mc tiu on nh ngnh ngh v pht trin ngh nui loi c ny ca Vit
Nam. Nui c Basa v c tra trc ht dnh cho xut khu, nhng n lc cn quan tm hn
n th trng ni a, ngoi dng s dng c ti, cn nghin cn ch bin mt s sn phm ph
hp vi th trng ny. Nht l nghin cu ch bin tn dng cc thnh phn khc ca c Basa
ngoi thnh phn c gi tr l fillet nh ch bin ngun m c thnh ngun du m s dng c
trong thc phm, ch bin cc ph phm ca c Basa thnh cc sn phm khc ... nhm khai thc
4.1 Mc ch ti
-
H2 02 trong NaOH 0,01 M, thi gian ty mu, t l da/dung dch H2 02 trong NaOH
-
acid v acetic acid), thi gian trch chit v phng php trch chit
-
Ha cht
NaOH
- NaCl
h2 so4
- h2 o2
Citric acid
- Acetic acid
- Na2 HP04
4.2.2
Thit b
Cn hai so
My o m Sartorius MA 35
My ly tm Hermle z 200A
My so mu CR - 300
4.3
4.3.1
4.3.1.1
Phng php chun b mu
Nguyn liu da c Basa c mua v, loi b nhng phn m tha cn st li, ra sch s
b vi nc v tr trong t ng nhit khong -20c s dng cho cc th nghim.
4.3.1.2
Phng php kho st theo yu t tng phn. Th nghim c b tr ngu nhin, ch thay
i mt nhn t cn cc nhn t khc c nh trong sut qu trnh th nghim. Kt qu ti u ca
th nghim trc c p dng cho th nghim sau.
4.3.2
4.3.2.1
Tin hnh
Cn mt lng mu (G) cho vo chn nung c nung v cn n khi lng khng i
(Gi). a chn nung cha mu vo l nung 500 -- 600
c trong 3 h. Ly
chn nung ra ngui trong bnh ht m v cn. Tip tc nung v cncho n khi khi
lng khng i (G2).
Cng thc
tro c xc nh theo cng thc:
%Tr o = ^ - G ' ) x 1 0 0
G
Trong
G
Gi
G?
4.3.2.1.3
:khi
:khi
:khi
lng mu ban u, g
lng a
lng a + thnh phn tro sau khi sy
Nguyn tc
Dng dung mi k nc trch ly hon ton lipid t nguyn liu c sy kh, nghin
nh. Mt s thnh phn ha tan trong cht bo cng c trch ly theo bao gm sc t, cc
vitamin tan trong cht bo, cc cht mi...tuy nhin hm lng ca chng thp. Do c ln tp
cht, phn trch ly c gi l lipid tng hay du th. Dung mi s c tch khi dch trch bng
cch bc hi v phn cn li chnh l lipid, c em cn xc nh hm lng.
Tin hnh
Cn mt lng mu chnh xc, em sy kh. Dng ethyl ether trch lng mu ny
trong thit b Soxhlet. Dch trch c cho vo mt a bit khi lng lm bay hi dung
mi. Sau , a ny c sy kh, lm ngui trong bnh ht m v c xc nh khi lng.
Cng thc
n / T
_ ( W 7 - W ,) x \00
s
Trong
w I : khi lng a
w2 : khi lng a + thnh phn ca dch trch sau khi sy kh s : khi
lng mu ban u
R-CHNH2-COOH + H2S4
C02
(NH4)2so4
+ h2o
- Ct m
(NH4 )2 S04 + 2NaOH = Na2 SC> 4 + 2 NH3 + 2H20
Phn ng xy ra trong bnh phn ng
NH3 + H7SO4 (NH4)2S04 + H9SO4 d ( bnh hng)
- Chun H2SO4 d bnh hng bng NaOH c nng 0. IN.
Trong
_ ( a - b x F ) x 0,0014x100x1000 O x V
4.3.2.1.5
Dung dch th c chun b vi cng mt cch thc. Pha long dung dch th nng
hydroxyproline nm trong vng t 0,5 -s-1,5 ng/ml, tng ng vi phn tuyn tnh trn ph
quang k. Nng ca hydroxyproline c xc nh da trn ng chun.
Lng collagen c tnh nh sau:
Collagen = hydroxyproline X 14,7
Xc nh bng phng php trch ly s dng lactic acid [13]
Da c sau khi x l s c trch ly bng dung dch lactic acid 0,5 M. Da c c xay nh,
ngm vo lactic acid vi t l 1:6. Tin hnh trch ly trong khong thi gian l 48 h. Phn da c
sau khi trch s c tip tc ngm vo dung dch acid mi trch ly mt cch trit lng
collagen c trong da c. Tng s ln trch ly l 3, thi gian cho mi ln l 48 h. Theo nghin cn
cng b th lactic cho kh nng trch ly cao, hiu sut trch sau 48 h l khong 90 %. Sau 3 ln
trch ly trn cng mt lng mu da, lng collagen trch c l 99,99 %, xem nh l tng
lng collagen c trong da.
4.3.2.1.6
Xc nh sng v mu sc ca da c
c,p/, -
hn mu chun
Nu L* mang du s lch mu theo hng ti hn, mu th ti hn mu chun
Aa* =
khc bit c th nhn thy c gia cc mu. Khng gian mu CELAB c thit lp dng
lp phng. Trc L* chy t nh xung y. Gi tr cc i ca L l 100, tng ng vi mu
trng. Gi tr cc tiu ca L* l 0, tng trng cho mu en. Cc trc a* v b* khng c gii hn
cc gi tr s c th. Gi tr dng ca a* biu th cho mu , gi tr
m
biu
th
mu xanh lc. Gi tr dng
th hin mu vng, gi tr m th hin mu
xanh
lam.
C nhng gi tr delta i km vi h thng CIELAB. Cc gi tr AL*, a*. Ab* cho bit
mu chun v mu th khc nhau nh th no v cc gi tr L*, a*,b*. Nhng gi tr delta ny
thng c s dng cho s qun l cht lng v hiu chnh cng thc, cn thit lp nhng mc
gii hn cho cc gi tr delta ny. Cc gi tr delta nm ngoi gii hn cho bit c qu nhiu khc
bit gia mu chun v mu th.
-
mu chun
-
mu chun
Gii thiu khng gian mu CELCH
Khng gian mu L*c*h* s dng chung gin vi khng gian mu L*a*b* nhng thay v
s dng trc ta vung th n li s dng trc ta hnh tr.
Trong khng gian mu ny, L* biu th sng ging vi L* trong khng gian mu
L*a*b*, c* l cng mu v h* l gc tng mu.
Gi tr cng mu c* bng 0 ngay ti tm v gia tng ty thuc khong cch t ta
mu n tm.
Gi tr gc tng mu c xc nh khi im ti trc a* v c tnh bng n v
0 l +a* ()
90 l +b* (Vng)
180 l -a* (Lc)
270 l -b* (Lam)
Cc gi tr c* v h* c xc nh t a* v b* theo cng thc :
Cng mu c* = yj(a*) +(b*)22
4.3.2.2.1 Xc nh s bin tnh ca collagen theo nhit bng phng php bin i
nht (Dnaturation temperature determined by viscosity method) [14-16]
Dung dch collagen 0,03 % trong acetic acid 0,1 M c np vo a
ca my o
nht vi lng khong 5 ml. Nhit ca a c iu
chnh thng qua biu
nhit. Ci t nhit ca b iu nhit ng vi nhit ban u ca th nghim l 30 c v
khong
7max - ;7min
Trong :
rt :
nht o c ti mi mc nhit
Tmax :
nht ln nht
ng cong bin tnh nhit c xy dng bng cch v th nht tng i theo
nhit .
4.3.2.2.2 Phn tch nhit vi sai (Differential thermal analysis/ Thermogravimetry analysis
(DTA/TG)) [17, 18]
DTA l k thut ghi nhn s khc bit v nhit gia mu th v mt cht i chng theo
thi gian hoc nhit bng mt chng trnh nhit xc nh trong mt mi trng c un
nng hoc lm lnh vi mc c kim sot.
(Thc hin ti Khoa Cng ngh vt liu, trng i hc Bch khoa TP.HCM)
4.3.2.2.3
o ph hng ngoi infrared (IR) spectroscopy [19]
Ph hng ngoi R l mt trong nhng k thut ph c s dng rng ri nht, l phng
php o hp th ca mu th cc bc sng khc nhau ca tia hng ngoi. Mc ch chnh
ca vic phn tch ph R l xc nh cc nhm chc ha hc c trong mu th. Nhng nhm
chc khc nhau s hp th nhng bc sng ring bit. Vi nhiu thit b ph tng, my ph hng
ngoi c th lm vic vi nhiu dng mu t' kh, lng n rn. Phn tch ph IR l mt cng c
ph bin v quan trng lm sng t cu trc v xc nh cc hp cht.
(Thc hin ti Phng th nghim trng im quc gia vt liu Polymer v Composite, trng
i hc Bch khoa TP.HCM)
4.3.2.2.4
t tiu chun o lng cht lng 3
4.3.2.2.5
t tiu chun o lng cht lng 3
4.3.2.2.
t tiu chun o lng cht lng 3
4.3.2.2.7
S hnh thnh si ca collagen (Fibril formation experiment - Turbidity)
[20]
Collagen c cho vo dung dch m (0,1M Na2HP04, 0,05 Citric acid, pH = 7,2) vi s c
mt ca NaCl (nng cui cng l 0,15M) 4 c. Nng ca mu collagen c iu chnh
bng 0,5 mg/ml trong dung dch m. Mu c lm nng n 35 c kch thch qu trnh hnh
thnh si. Tin hnh ghi nhn hp thu ca mu theo thi gian, 2 pht/ln, trong vng 30 pht.
hp thu c o bc sng 313 nm.
Hnh thi b mt ca collagen (Scanning Electron Microscopy - SEM
)
4.3.2.2.8
Hnh thi b mt ca sn phm collagen thu c v mu gelatin chun c chp theo
phng php SEM quan st c s khc bit v cu trc ca chng.
(Thc hin ti Phng th nghim chuyn su, Trng i hc cn Th)
4.3.3
4.3.3.1
Da c Basa
X3
ln
X3
ln
X3
ln
Ra
Ngm
0,5% LasNa Dd
Ngm
Ngm
Ra
Citric 0.7%
H?0
Ra
Da c loai bo
Hnh 4.1 Quy trnh loi b
Ngm
mu th
c sc
hn mu chun
-
hn mu chun
Ab* = b]ample - b],M: s khc nhau v ta trn trc vng - lam
Hnh 4.2
trnh
mu
DaQuy
sau khi
loity
lipid
c ngm trong dung dch NaCl 10 % trong vng 24 h, t l da : dung
dch l 1:10 (g/ml), nhit phng, chun b cho qu trnh ty mu. Vic ty mu c tin hnh
di tc dng ca dung dch H2O2 trong NaOH 0,0IM.
4.3.3.3 Quy trnh trch colagen
Xay nh
acetic
0,5M
Trch ly
Lc
Ly tm
NaCl
1:6 (g/ml)
4
(C)
Collagen khns tan
5.000 vng/pht 30
pht
Kt tinh
Ly tm
5.000 vng/pht 1
h
Ket tinh li
Ly tm
5000 vng/pht 1
pht
- Phng php tng phn (B): da c c tin hnh trch qua nhiu ln, mi ln 24
h, sau lng collagen thu c cc ln trch c gp li tnh hiu sut.
Da c sau khi x l c ngm trong cc dung dch acid vi t l da/dung dch l 1:6
(g/ml). Dch trch collagen thu c t qu trnh trch ly c tinh ch theo phng php ca
Piez et al. (1963). NaCl rn c cho vo dch trch collagen vi nng 5% (w/v) kt tinh
collagen. Collagen kt tinh (collagen ha tan trong acid) c tch ra bng phng php ly
tm vi vn tc 5.000 vng/pht trong 90 pht, s dng my ly tm Hermle z 200A. e thu
c collagen tinh khit hn, phn collagen kt tinh c ha tan li trong acetic acid 0,5 M
v kt tinh li theo cc bc miu t trn. Qu trnh kt tinh li c lp li 2 ln.
4.3.4
ncHA ) =
f(r)
f(l; 2; 3; 4; 5; 6; 24)
1:20)
Loi acid
Citric v Acetic acid
p ng
Seafood Joint Stock Company), L 2.17, Khu Cng Nghip Tr Nc II, TP. cn Th.
-
Thnh phn
Protein
31,1
Lipid
6,5
58,7
tro
0,1
3,6
Cm quan
100 = 2,54 %
15.75
_ on I o
75.35
X100 = 48,00 %
Nhn xt
v mt cm quan, da c sau khi x l u c mu trng, sng hn so vi da c ban u
nhng da ch qua x l vi NaOH 0,1M s cho ta cm gic nhp nhp khi s' vo, ng thi
mi hi vn cn trong khi da qua quy trnh x l c acid rt sch v khng c mi.
Quy trnh loi bo c s dng acid c hiu qu hn, hiu sut tng i cao (t
79,24 %), cao gp 31,2 ln so vi hiu sut ca quy trnh ch s dng NaOH 0,1 M. Tuy
nhin, mt im bt li c th thy ca qu trnh loi lipid c s tham gia ca cc
a cc protein, lng protein mt i chim 48 %. Trong khi , quy trnh khng s dng acid ch lm mt i 14,53 %
protein. Mt phn collagen c th mt i trong giai on ny.
=> So snh gia hai quy trnh, ta
g i
iu kin c nh
F (g/ml) = 1:10
tAC) = 30 Kho st
AL* = f(CA),f(r)
AC* = f(C22),f(r)
Ah* = f(C22),f(r)
Kt qu kho st (Ph lc 2
)
5.3.1.2
vhiu sut
bc
25
20
15
Hnh
5.1
th
biu
th nh
10
20
30
hng
gian
0.5%
0.75%
1%
1.25%
1.5%
10
20
Thi gian (h)
30
ln qu trnh tay mu
5.3.1.3 Nhn xt
Trong 4h u tin ca qu trnh ty mu, kh nng ty mu ng bin theo thi gian. S chnh
lch v sng gia da c ty mu v da ban u, hay ni cch khc l sng ca da t
gi tr cc i ti thi im 4h. Sau thi im ny, chnh lc
h
sng c xu hng gim xung v sau gn nh duy tr cho n thi im 24h. Xu
hng trn c th hin r c 5 mc nng . Tuy nhin, mc nng 1 % cho thy chnh
lch v sng l cao nht.
T kt qu trn, ta bit c 2 gi tr ti u ca 2 thng s trong qu trnh ty mu:
- Nng H202: 1%
- Thi gian tay mu: 4h
Mt im cn lu l qu trnh ty mu nu c kt hp vi s khuy trn s cho kt
qu kh quan hn.
5.3.2
T l da/dung dch
tAC)
=4
30
Kho st
AL* =f(F)
AC* =f(F)
Ah* =f(F)
Nhn xt
T th ta thy chnh lch sng tng mnh khi t l da/dung dch tng t 1:10 n 1:15.
Sau , chnh lch ny gim nh khi ta tip tc tng t l n mc 1:20. Da c c ty mu
n mc sng nht tng ng vi t l da/dung dch l 1:15. trng ca da mc t l 1:17,5
khng lch nhiu so vi mc 1:15. Trong khi , bo ha mu ti t l 1:17,5 l thp nht =>
Lun vn tt nghip
Trang 61
chn mc t l 1:17,5
5.4 Tng lng collagen trong da c
5.4.1
Da ban u
Da sau kh
x l
Da c sau khi x l
-
5.4.2
5.4.2.1
iu kin kho st
iu kin c nh
mt(g) = 30 C(0O = 4 F (g/ml) =
1:6
^Lactic acid
0,5
{h) - 48
5.4.2.2
Kt quLn
trch ly
Lun vn tt nghip
(g) Thu c
Trang 61
Qua so snh, ta thy phng php trch ly bang lactic acid khng chnh lch nhiu so vi
phng php nh lng hydroxyproline. Phng php nh lng collagen thng qua s nh
lng hydroxyproline l phng php chun, c tnh chnh xc cao, ng tin cy. Tuy nhin,
phng php ny tng i phc tp v gi thnh rt t. Do , xc nh hiu sut ca qu
trnh trch ly ta khng s dng phng php ny m xc nh lng collagen bng qu trnh
tinh ch.
5.5 Kho st cc yu t nh hng n qu trnh trch collagen Loi
acid, phng php trch v thi gian trch
5.5.1 iu kin kho st
iu kin c nh
m
t (g)
= 30
4*(
4
F (g/ml)
= 1:6
CACCI
= 0.5
f(Loi acid)
f(r)
Kt qu kho st (Ph lc 7)
90
80
0 70
C 60
50
\ 40
20 10
0
20
30
50
70
Thi gi an (h)
A
ce
ti
c,
to
40
60
80
n
ph
n
A
ce
ti
c,
tn
g
ph
n
Cit
ric,
to
n
ph
n
Cit
ric,
t
ng
ph
n
Hnh 5.4 th biu th nh hng ca loi
acid, phng php trch v thi gian
ln hiu sut trch collage
5.5.3 Nhn xt
phn.
- Loi acid: t th ta thy Acetic acid
thch hp cho vic trch collagen hn v n cho
hiu sut cao hn rt nhiu so vi Citric acid.
5.6
a nht tng phn (fractional viscosity) ca collagen t da c Basa theo s gia tng ca nhit . i vi c 3 loi
collagen thu c t phng php trch ly bng 3 loi acid (acetic, citric v lactic acid), th
cho thy nht ca dung dch collagen c khuynh hng gim khi un nng. iu ny c th
c gii thch do s bin tnh ca collagen. Nhit bin tnh, Td, c xc nh l nhit
m ti s thay i ca nht xy ra c mt na. Nhit bin tnh ca collagen t
da c Basa l khong 37,5 c, nhit ny thay i khng ng k gia collagen trch bng
cc loi acid khc nhau. Nhit ghi nhn c trong th nghim ny l tng i cao so vi
nhit bin tnh ca collagen thu c t nhng loi c khc c bo co trong cc
nghin cn trc y. Nguyn nhn l gii choiu ny l do iu kin mi trng sng ca c
c nh hng n cu trc da v qua nh hng ln tnh cht ca collagen. Collagen thu
c t cc loi c khc nhau s c kh nng bn nhit khc nhau.
nht (cP)
Nhit
30
32,5
35
37,5
40
42,5
45
47,5
50
136
128,2
125,6
115,1
99,4
78,5
70,6
54,9
1.1
1.0
Nhit (C)
0.9
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0.0
47,1
Nhit
(C)
30
32,5
35
37,5
40
42,5
45
47,5
50
Nhit (C)
5.6.1.2
DTA /(uVAng)
DDTA J[uVAngAnin
)
1T
G
%
10
DDTA/(uVAng/min)
0.0
0-
-0.10
-0.20
90
-0.30
80
-0.40
-0.10
70
-0.50
60
-0.60
50
40
30
20
20
40
60
80
100
120
Temperature rc
140
160
180
200
-0.20
-0.3
M
u
coll
age
n
trc
h
bn
g
citri
c
acid
c
ln
tp
cht
,
kh
ng
tinh
khi
t
bn
g
coll
age
n
trc
h t
2
loi
acid
cn
li.
Ng
uy
n
nh
n
c
th
l
do
cc
tinh
th
citri
c
ch
a
c
th
m
tch
a
Thi gian (pht)
Collagen
ha tan
0,1357
0,1363
0,1377
0,1422
0,1446
0,1463
0,1529
0,1565
0,1603
0,1623
0,1637
0,1645
0,1660
0,1660
0,1660
0,1660
Collagen
ha tan
0,0558
0,0558
0,0570
0,0604
0,0658
0,0686
0,0765
0,0794
0,0803
0,0819
0,0821
0,0823
0,0828
0,0837
0,0837
0,0837
Gelatin
-----------------------------------------------------Gelatin chun
-----------------------------------------------------Collagen (trch ly
bang acetic acid)
Collagen (trch bang citric
acid)
-----------------------------------------------------Collagen (trch ly
bang lactic acid)
H
nh
5.7
th
bi
u
di
n
c
c
a
col
lag
en
tr
ch
ly
ba
ng
ac
eti
c
aci
d
(a)
,
cit
ric
aci
d
(b)
v
lac
tic
aci
d
(c)
the
o
th
i
gia
n
Nh
n
xt
- D
a
v
o
th
tr
n,
ta
th
y
c
n
g
p
th
c
a
du
ng
d
ch
co
lla
ge
n
ng
bi
n
v
i
th
i
gi
an
.
h
p
th
A
t
ng
nh
an
h
tr
on
g
kh
o
ng
10
ph
t
u
ti
n,
sa
u
t
c
t
ng
ch
m
li
v
c
kh
uy
nh
h
n
g
du
y
tr
on
nh
sa
u
m
t
kh
o
ng
th
i
gi
an
nh
t
nh
.
T
he
o
th
i
gi
an
du
ng
d
ch
co
lla
ge
n
tr
n
n
c
ch
n
g
t
c
s
h
nh
th
n
h
s
i
x
y
ra
tr
on
g
du
ng
d
ch
.
Tr
on
g
kh
i
,
c
n
g
qu
an
g
c
a
du
ng
d
ch
ge
lat
in
kh
n
g
th
ay
i
th
eo
th
i
gi
an
v
kh
n
g
c
s
t
o
s
i
di
n
ra.
olla
gen
thu
c t
qu
trn
h
trc
h ly
s
dn
g
ace
tic
aci
d
c
hp
th
qua
ng
hc
ln
nh
t so
vi
coll
age
n
trc
h ly
ban
g
citr
ic
v
lact
ic
aci
d
=>
s
hn
h
th
nh
si
di
n ra
m
nh
m
nh
t
i
vi
coll
age
n
trc
h ly
ban
g
ace
tic
aci
d.
5.6.
3
C
u
tr
c
b
m
t
ca
coll
age
n
5.6.
3.1
Kt
qu
ph
n
tc
h
S
kh
c
bit
v
hn
h
thi
b
mt
ca
coll
age
n
so
vi
m
u
gel
atin
chu
n
c
qua
n
st
th
ng
qua
hn
h
ch
p
bn
g
ph
ng
ph
p
SE
M.
D
i
y
l
hn
h
nh
SE
M
ca
coll
age
n
thu
c t
da
c
Bas
a
v
ca
m
u
gel
atin
chu
n
H
nh
5.8
H
nh
th
i b
mt
ca
A.
coll
age
n
trc
h
bn
g
ace
tic,
B.
coll
age
n
trc
h
bn
g
citr
ic,
c.
coll
age
n
trc
h
ban
g
lact
ic
v
D.
gel
ati
n
chu
n
5.6.
3.2 N
h
t
C
s
kh
c
bit
rt
r
rn
g
gi
a
cu
trc
ca
coll
age
n
v
gel
atin
.
D
a
vo
hn
h
trn
ta
th
y
coll
age
n
c
cu
trc
m
ng
l
i
si
vi
b
mt
nh
m,
x
x.
Tr
i
li,
m
u
gel
atin
vi
b
mt
tr
n
pha
ng
hn
do
n
kh
ng
c
cu
trc
m
ng
l
i
nh
coll
age
n.
-
C
u
trc
b
mt
ca
coll
age
n l
gi
ng
nha
u
i
vi
c
3
loi
aci
d
dn
g
tro
ng
qu
trn
h
trc
h
ly.
5.6.
4
P
h
n
g
n
g
o
I
R
5.6.
4.1 K
q
u
p
h
c
h
(
P
h
1
1
)
Ph hng ngoi ca collagen thu c t da c Basa
bn
g
qu
trn
h
trc
h ly
ca
3
loi
aci
d
c
th
hi
n
qua
nh
ng
bi
u
d
i
y
.
Ph IR ca collagen trch ly t da c Basa (s dng citric acid)
Ph
IR
ca
coll
Wavcn
umber
age
cm-1
n
trc
h ly
t
da
c
Bas
a
(s
d
ng
ace
tic
aci
d)
Waven
umber
cm-1
V\v
enurr
fcerc
m-1
H
nh
5.1
0
Ph
FT
IR
ca
coll
age
n
thu
t
c
r
no
n
v
tr
n
g
th
nh
(yo
un
g
and
adu
lt
Nil
e
per
ch)
Wavenumber (cm1)
Acetic
Citric
Lactic
C
non
3434
2924
1650
1542
1235
5.
.
4.
3
N
h
n
x
t
N
h
n
g
v
n
g
a
m
i
d
e
(
b
a
o
g
A
,
B
,
I,
I
I
)
c
a
c
o
ll
a
g
e
n
t
d
a
c
B
a
s
a
l
n
g
n
g
n
h
a
u
k
h
i
s
o
s
n
h
g
i
a
3
l
o
i
a
c
i
d
c
s
g
c
h
o
q
u
tr
n
h
tr
c
h
l
y
.
C
o
ll
a
g
e
n
t
h
u
c
t
d
a
c
B
a
s
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Basa c tn sut thp hn so vi c r v c ng. Tn sut ca collagen trch bang acetic acid nh hn so vi collagen
trch bang citric v lactic acid. Vng amide I c mi lin h n mc tng tc gia cc phn
tcollagen. N cn l thnh phn ca cc chui ngu nhin (random coil). T ta c th a ra
kt lun:
-
Acetic l dung mi trch ly hu hiu nht trong 3 loi acetic, citric v lactic acid.
hu chnh nh sau:
- Khu 1: X l da, da ti c mua t Cng ty TNHH Thu Sn Bnh An, Khu cng
nghip Tr Nc, TP. cn Th, tri qua cc giai on x l bao gm loi bo, ty mu. Sau khi
Lun vn tt nghip
Trang 1
Lun vn tt nghip
Trang 1
H2O2
30 g
4h
1:17,5 g/ml 30
c
mt
c
CH,COOH 0,5M
24 h X 3 ln 1:6 g/ml
4
c
T
i
n
h
c
h
6.2.3.1
So snh tc cht trch ly
Trong quy trnh trch ly c s dng 2 loi acid l acetic v citric acid. Hiu sut ca qu trnh
trch ly s dng acetic acid cao hn gp nhiu ln so vi s dng citric acid. Dch collagen thu
c khi trch ly bng acetic acid gn nh khng mu, trong khi nu trch bang citric acid th
dch trch c nh mu hi vng. Mt c im ni tri na ca acetic l ta c th thu c
collagen tinh khit t dung dch collagen trong acetic bng phng php sy lnh. i vi trng
hp collagen trong dung dch citric acid, sn phm ca qu trnh sy lnh khng nhng ch c
collagen m cn ln cc ht tinh th ca citric. Do , thm tch loi tinh the citric l mt vic
lm cn thit collagen trch ly c c the dng cho nhng ng dng xa hn na. Theo cc
nghin cu cng bo, lactic acid cho hiu sut trch ly cao, khong 90%. Trong dung dch lactic
acid, collagen c ho tan hon ton sau khi trch ly 24 h. Tuy nhin, gi thnh ca lactic acid
tng i cao so vi acetic acid. T c th kt lun, acetic acid l dung mi l tng nht cho
qu trnh trch ly collagen t' da c Basa.
Qu trnh trch ly collagen t da c Basa ch yu s dng cc acid hu c nh acetic, citric
hay lactic. Cc acid v c l nhng acid mnh, chang hn nh HC1, c pH thp lm gim kh
nng ho tan ca collagen vo trong dung dch do hiu sut trch ly khng cao. iu ny c
l gii l do nhng nhm amine mang in tch dng ca protein to lin kt vi anion cr, lm
cu trc si ca collagen cht ch hn, kh nng to lin kt vi nc b hn ch lm cho s ho
tan ca collagen gim. pH nh hn 2 cc protein b bin tnh. Si collagen co li lm cho s
hydrat ho khng th xy ra do gia cc i phn t c t khng gian cho phn t nc.
6.3.1
Qu trnh ty mu
n thch hp rt ra cho qu trnh ty mu l:
- Nng H202
1%
- Thi gian ty mu
4h
- T l da : dung dch
1:17,5 g/ml
- Nhit
30 c
Thi gian l thng s quan trng nht nh hng n qu trnh ty mu. Trong khong thi
gian 4 gi u tin, kh nng tay mu ca dung dch H 202 ng bin theo thi gian. Thi gian tng
th trng, sng ca da c tng theo v t gi tr cc i ti 4
hng gim xung v duy tr cho n 24 h. Hai thng s quan trng tip theo l nng H 2O2 v
t l da : dung dch. Gia cc mc gi tr ca 2 thng s va nu, trng ca da c chnh lch
nhau khng qu nhiu. Trong s th nng H2O2 v t l 1:17,5 cho kt qu kh quan nht.
6.3.2
Qu trnh trch ly
Loi acid s dng c nh hng r rt ln hiu sut trch ly. Hiu sut trch ly bng acetic cao
hn so vi citric. Khi thi gian trch ly tng, lng collagen ho tan tng nn hiu sut cao. Ta ch
kho st 3 mc thi gian 24, 48 v 72 h m khng kho st trong khong thi gian lu hn na v
collagen c th b ct mch mnh xung thnh gelatin. Cn nu thi gian qu ngn th khng
cho acid li ko collagen. Phng php trch ly tng phn vi 3 ln trch ly trn cng mt
lng da, mi ln 24 h cho kt qu kh quan hn phng php trch ly ton phn thc hin 1 ln
trong 72 h. Nguyn nhn c th l do nu ch s dng cng mt lng dung dch acid trong mt
khong thi gian di th dung dch c the b bo ho khng ho tan c thm collagen na. Neu
thay i dung dch nhiu ln th c th trch ly trit hn lng collagen c trong da c. Tuy
nhin, s chnh lch gia 2 phng php ny khng nhiu lm. iu kin ti u cho qu trnh
trch ly thng qua kho st c c l:
Loi acid
Acetic acid
72 h
Tng phn
Nng acid
0,5M
Nhit trch ly
4 c
T l da : acid
1:6 g/ml
c bn ca n. s lng lin kt trong collagen gia tng theo tui. Tuy nhin, phn ln cc m
lin kt trong c c ti to hng nm v cc protein c cu trc vi nhiu lin kt khng c
tm thy trong c. Collagen t nhng ng vt cn non d c trch ly hn nhng cho cu trc
c bn thp. Ngc li, collagen t ng vt trng thnh kh ho tan hn v cu trc ca n
c bn cao.
Ch n ung
mt s loi c, lng collagen v s lng lin kt trong n gia tng cng vi s thiu
thn thc n. Theo nghin cu, nhng con c thiu n s sn sinh ra nhiu collagen, c bit l
collagen c nhiu lin kt, hn l nhng con c n ung y .
Thnh phn imino acid
Kh nng bn nhit ca collagen lin quan n thnh phn imino acid (proline v
hydroxyproline). Thnh phn imino acid cng cao th cu trc chui xon c ca collagen cng n
nh. Thnh phn proteoglycan v glycoprotein cng nh hng n s bn nhit ca collagen.
.3.3.3 S hnh thnh si ca collagen
Collagen c th kt hp li to thnh si di nhng iu kin sinh l hc. Qu trnh ti to
si trong quy m phng th nghim c th chia ra lm 3 bc bao gm giai on khi mo ph
thuc v nhit , giai on pht trin ph thuc nhit v giai on pht trin khng ph thuc
nhit . Nhng loi collagen khc nhau vi nhng cu trc khc nhau ca cc chui xon c nh
hng n thi gian v mc to si. Nhng quy trnh iu ch khc nhau ca cng mt loi
collagen cng cho nhng p ng v iu kin khc nhau trong qu trnh to si. S ti to si
ca collagen chu nh hng ca loi collagen, phng php trch ly v iu kin trong qu trnh
to si (cht m, nhit v nng protein,...)- Gelatin c iu ch iu kin nhit cao,
cc nhm amide b thu phn v cu trc chui xon c collagen b ph hu. cu trc ca gelatin
khng c trt t v khi lng phn t phn b trong mt khong rng l cho s ti to si khng
th xy ra. Thm vo , s t hp ca cc n phn collagen trong qu trnh to si lin quan
n s tng tc tnh in, k nc qua li gia cc chui collagen. Qu trnh iu ch collagen
da vo s gia tng ca cc in tch m, iu ny gp phn lm cho s tng tc gia cc phn
t kh khn do lc y gia cc in tch m. Do , gelatin khng c kh nng hnh thnh si.
.3.3.4 Ph hng ngoi ca collagen
IR l phng php o chiu di sng v cng hp th bc x hng ngoi ca mu.
Nhng thnh phn polypeptide v protein cho ta 9 loi di hp th l amide A, B v I - VII. Trong
s , amide I v II l 2 di ni bt nht trong chui protein. Vng ph nhy nht trong thnh
phn cu trc bc hai ca protein l di amide I (1700 -T-
Sizeimicions)
c. Lactic acid
Kho st
Kho st
nhit trch ly
1.
2.
4.
6.
Shuai Zhao, M.Z., Guoying Li, Preparation of collagen from the pigskin shavings by
alkali extraction.
7.
Rume Sakai, S.I., Toshizo Isemura, Soluble collagen of chicken leg tendon; Its
9.
Maria Sadowska, I.K.O., Celina Niecikowska, Isolation of collagen from the skins of
Baltic cod (Gadus morhua). 2002.
10.
L.s. Senaratne, P.-J.P., Se-Kwon Kim, Isolation and characterization of collagen from
brown
13.
Elzbieta Skierka, M.S., The influence of different acids and pepsin on the
extractability of collagen from the skin of Baltic cod. 2006.
14.
J.H. Muyonga, C.G.B.C., K.G. Duodu, Characterisation of acid soluble collagen from
skins of young and
V. Vijayasundaram, V.R. and P.R. Palaniappan, The study of the changes in the
thermal properties ofLabeo rohita
Yun Shana, Y.Z., Yong Caoa,Qunhua Xua, Huangxian Jub, Zonghan Wuc,
Preparation and infrared emissivity study of
HaiYing Liu, D.L., ShiDong Guo, Studies on collagen from the skin of channel
catfish (Ictalurus punctaus). 2006.
20.
MC LC
CHNG 1 TNG QUAN V COLLAGEN.......................1
1.1
1.2
Cu trc ca collagen....................................................................................................1
1.2.1
1.2.2
1.3
1.4
ng dng ca collagen................................................................................................10
1.4.1
1.4.2
1.4.3
1.5
1.6
1.6.1.
1.6.2.
1.7
1.7.1
Mt s sn phm collagen...................................................................................19
Collagen EX.........................................................................................................19
1.7.2
1.7.3
1.7.4
1.7.5
2.2
2.3
2.4
2.5
2.6
2.7
2.8
3.2
3.2.1
c im sinh trng...........................................................................................31
3.2.2
c im sinh dng............................................................................................31
3.2.3
Ma v sinh sn....................................................................................................32
3.2.4
Ging c nui........................................................................................................32
3.2.5
Mt nui...........................................................................................................32
3.2.6
Ma v nui..........................................................................................................33
3.2.7
3.3
3.4
3.4.1
3.4.2
3.5
3.6
cu...................38
4.1
Mc ch ti..............................................................................................................38
4.2
4.2.1
Ha cht................................................................................................................38
4.2.2
Thit b..................................................................................................................38
4.3
4.3.1
4.3.2
4.3.3
4.3.4
4.3.5
p ng.................................................................................................................52
CHNG 5
5.1
nghim....................................................................39
KT QU TH NGHIM.........................53
nh gi nguyn liu.....................................................................................................53
5.1.1
5.1.2
5.2
liu ................................................................53
5.3
5.3.1
ty mu.............................................................55
5.3.2
T l da/dung dch.................................................................................................57
6.1
6.2
6.2.1
6.2.2
6.2.3
6.3
nh gi kt qu............................................................................................................79
6.3.1
Qu trnh ty mu..................................................................................................79
6.3.2
6.3.3
6.4.1
Kin ngh
84
85
......................................................................................................................................................59
Hnh 5.4 th biu th nh hng ca loi acid, phng php trch v thi gian n
hiu sut trch collagen.............................................................................................................62
Hnh 5.5 ng cong bin tnh theo nhit ca collagen..................................................64
Hnh 5.6 ng phn tch DTA/TG ca collagen..................................................................66
Hnh 5.7 th biu din c ca collagen trch ly bng acetic acid (a), citric
acid (b) v lactic acid (c) theo thi gian..................................................................................68
Hnh 5.8 Hnh thi b mt ca A. collagen trch bang acetic, B. collagen trch bng
citric, c. collagen trch bng lactic v D. gelatin chun........................................................69
Hnh 5.9 Ph R ca collagen...................................................................................................71
Hnh 5.10 Ph FTIR ca collagen thu c t c r non v trng thnh (young and
adult Nile perch).........................................................................................................................72
Hnh 6.1 Quy trnh sn xut collagen t da c.......................................................................76
Hnh 6.2 Ket qu o kch thuc ht ca insoluble collagen x bang acetic acid... 83
Hnh 6.3 Ket qu SEM ca insoluble collagen x l bang 3 loi acid A. Acetic acid,
B. Citric acid, c. Lactic acid.....................................................................................................84