CHNG 1 TNG QUAN V COLLAGEN

1.1 Gii thiu SO lc v Collagen

Collagen l loi protein cu trc chnh yu, chim khong 30% tng lng protein trong
c th cc ng vt c xng sng. Collagen c nhiu trong gn, da, xng, h thng mch
mu ca ng vt v c mt trong cc lp mng lin kt bao quanh cc c. Khong 10%
protein trong c ng vt c v l collagen; cc protein ngoi bo (hn 90% trong gn,
xng v khong 50% trong da) c cha collagen [1]. N c tc dng ging nh mt cht keo
lin kt cc t bo li vi nhau hnh thnh cc m v c quan nn tng trong c th.
Collagen cung cp cho cc m lin kt nhng c tnh ni tri nh' vo s hin din rng
khp v s sp xp mang tnh cu trc ca n. N phn b khp ni trong c th, t ch gn
ni bp chn vi gt chn cho ti gic mc [1]. Trong gn v dy chng, collagen c tc dng
truyn lc t c sang xng v tch tr nng lng n hi. S di chuyn nhp nhng, uyn
chuyn s khng th thc hin c nu thiu nhng tnh cht ny. Collagen cn l cht nn
hu c c trong xng v men rng gip chng chng li s rn nt. N l thnh phn chnh
trong da, mch mu, cc c. Collagen khng ch c chc nng c hc, chang hn nh gic
mc, trt t cu trc ca cc si collagen to nn s trong sut [2].
Collagen c xem nh mt vt liu mang tnh xy dng. S linh hot ca n l nh vo
cu trc cp bc phc tp, to nn s a dng trong tnh cht nhm phc v nhng chc nng
nht nh.
1.2 Cu trc ca collagen
1.2.1
Cu trc phn t ca collagen [1]
Phn t collagen (hay cn gi l tropocollagen) l mt protein hnh tr, di khong 300 nm,
ng knh khong 1,5 nm

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

Hnh 1.2 S hnh thnh chui xon c ni phn t v gia

cc chui polypeptid
e
Mi chui polypeptide c khi lng phn t khong
100 kDa, to nn tng khi lng phn t ca collagen
khong 300 kDa. Chui a c cu to bi khong 1000
amino acid. Cc chui a khc nhau (ai, a 2 v a3) thnh phn
amino acid. S phn b ca cc chui (Xi, a 2 v a3 trong cc

intramolecular 'A' triple-helix formation

phn t collagen khc nhau ty thuc vo s khc nhau v

gene.
Collagen khng c cha tryptophan, giu thnh phn
glycine, proline v hydroxyproline, l mt trong s t nhng
protein c cha hydroxylysine. N cha khong 33%
glycine, 12% proline v 11% hydroxyproline. Cc amino
acid sp xp trong chui xon c theo cc dy vi s phn b
nh sau:
Bng 1.1 S phn b cc amino acid trong chui polypeptide
T l
Triplet
Gly - X - X
0,44
Gly - X -1

0,2

Gly -1 - X

0,27

Gly -1 -1

0,09

Trong , I l imino acid (proline v hydroxyproline), X

l cc amino acid khc. Glycine chim gn 1/3 trong tng s
cc amino acid v n c phn b mt cch u n ti v
tr mi 1/3 xuyn sut trong phn t collagen. Do glycine c
cc nhnh ph nh nht nn s lp li ca n cho php cc
chui polypeptide kt hp cht ch vi nhau hnh thnh nn
mt ng xon c vi khong trng nh phn li.
Ngoi ra, trong phn t collagen c nhng vng gm khong 9
-- 26 amino acid ti cc im u mt amino hay carboxyl ca
chui. Cc vng ny khng sp nhp vi cu trc xon c v
c gi l telopeptide
.
I
H2N ,/ \

Procollagen
I
.
COOH

r
C
r

Procollagen

peptidase
N-terminus end cleaved
cleaved

C-terminus end

Tropocollagen 3000

Hnh 1.3 Cu trc ca procollagen v s chuyn

tprocoagen sang tropocollagen
1.2.2 Cu trc si ca collagen [3]
Phn ln collagen trong mng li ngoi bo c tm
thy dng si, bao gm nhng si mnh, nh. Thng qua qu
trnh to si, cc phn t collagen t hp vi nhau hnh thnh
nn cc vi si (microfibril) bao gm t 4
8 phn t collagen hoc

vi s lng nhiu hn s to thnh cc si (fibril). Nhng si

ny c ng knh t 10 -T- 500 nm ty thuc vo loi m v
giai on pht trin. Cc si collagen s thit lp nn cc si
ln hn (fiber) v cao hn na l cc b si (fiber bundle).

Phn t collagen

Hnh 1.4 Cu trc si ca collage

Phn t collagen, di 300 nm, ig knh vng xon 1,5 nm

Hnh 1.5 Qu trnh t hp to si ca cc phn t collagenCc chui collagen sp xp

song song theo chiu dc to thnh cc si vi tnh chu k nht nh. Chng c sp xp so le
nhau mt khong 67 nm v c mt khong trng khong 40 nm gia nhng phn t lin k nhau.

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

m cng, n hi v nhng c tnh c hc ring bit.

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

Si collagen l cc t hp c cu trc bn kt tinh ca cc phn t collagen. Cc vi si kt

hp to nn si ln hn. Cc si collagen c sp xp vi nhng cch thc kt hp v mc
tp trung khc nhau trong cc m khc nhau cung cp cc c tnh khc nhau ca m.
1.3 Phn loi 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.

Hnh 1.8 Mt vi dng sinh hc ca collagen

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

Da, gn, xng, gic mc, men rng, sn,

mch mu

(<xl(II))3
(al(III))3

Mch mu, van tim, h b, rut, nu rng,

thnh t cung
Mng nhy

(al(IV))2a2(IV)
al(V)

a2(V)

a3(V)

(al(V))2a2(V) hoc (al(V))3

al(VI) a2(VI) a3(VI)

hoc Gic mc, xng, mch mu, sn, nu

rng
Da, c tim

(al(VII))3

Da, phi, sn, gic mc, nhau

Cha bit

To ra t t bo mng trong

al(IX) a2(IX) a3(IX)

Sn
(al(X))3

al(XT) a2(XI) a3(X)

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 cng nghip

Neu collagen b thy phn th cc chui polypeptide s tch nhau mt phn hoc hon ton,
hnh thnh nn Gelatin. Gelatin c ng dng trong thc phm, dc phm, m phm v nn
cng nghip phim nh. Cc nh sn xut cc cht b sung da trn nn
l collagencho ch
n kingkhng nh rng sn phm
ca hc th ci
thincht lng ln da,mng tay v sc khe xng khp.
Theo ting Hy Lp, collagen c ngha l ngi
sn xut keo h, ni n qu trnh nu da v gn
ca nga
cng nhng
loi
ng vt khc thu c h.Keo
dn
collagen c ngi Ai Cp s dng cch y
4000 nm v ngi M s dng n cch y
khong 1500 nm.
Hnh 1.9 Keo dn colagen N khng nhng c
dng gi cc dng c trong gia nh m cn rt
hu dng trong vic sn xut cc loi nhc c nh
violin hay guitar; nhng loi ny thng c tho
ra sa cha nn khng thch hp dng cc cht
dnh lu bn, trong khi collagen khi un nng th c th c lm mm mt cch d dng.
1.4.2

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.

Hnh 1.10 Collagen dng trong phu thut thm m

1.4.2.1
Collagen dng si
Collagen dng si c dng trong vic lm lnh cc vt thng, vt rch trong phu
thut. Si collagen c th c x l to cu trc si thng, di dng lm nhng sn phm y
hc cho gn v dy chng. Nhng ng collagen (collagen tube) c s dng thay th cho cc
cu trc nh thc qun, dy thn kinh ngoi bin, niu qun. N cng c s dng trong vic
nui
cy t
bo.

Hnh 1.11 Collagen p dng trong vic iu tr vt bng

vt bng mi, trc khi s
3 ngy sau khi s dng
dng coagen
colagen

collagen

10 ngy sau khi s dng collagen

1.4.
2.2
Mng
mng

Collagen dng mng v lp mng c s dng gi c nh cc vt cht sinh hc chang hn

nh nhn t XI 11 trong mu, dng trong s ti to cc m, ni kt li vng mc, lm mng thm
tch mu, lm vt thay th lp mng cng ca no. N cn c dng cho vic ti to dy thn
kinh, khi phc mng nh, sn v xng, kim sot
s chy mu cc b, khi phc tn thng ca gan, l lp mng chn bo v no, c tc
dng phc hi cc vt thng.
1.4.2.3 Nhng loi collagen c th tim c
Dung dch v huyn ph collagen c dng di dng c th tim c lm thng s tc
nghn ng mch, cha gy xng, ti to ty sng, tr i dm, phc hi chc nng trt ca
gn.
1.4.2.4 S nui cy t bo trn collagen
S pht trin ca t bo vi s h tr ca collagen trong s nui cy t bo va c xem
xt gn y. Vic s dng cht nn collagen cho s pht trin ca t bo da c cng b mt
cch rng ri. Nhng t bo nui cy trn collagen kch thch, lm vt thng mau lnh. Nhng
nghin cn rng ri v s pht trin ca cc t bo da t sinh v khc loi trn nn collagen
chng minh tnh kh thi to ra nhiu chng loi m v cc c quan trong qu trnh nui cy.
S dng ngun nguyn liu t bo nui cy thuc nhiu chng loi khc nhau c tc ng to ln
n vic iu tr cho cc bnh nhn c nhng tn thng v m hoc cc c quan.
1.4.3 ng dng trong m phm
1.4.3.1

Tc dng chng lo ha, ngn nga v ci thin np nhn

Khong 70% cu trc ca da l collagen, phn b ch yu lp h b ca da. Collagen to ra mt

h thng nng , h tr cc c tnh c hc ca da nh sc cng, n hi, duy tr m, lm
cho da c mn mng, ti tn v tr trung. Collagen gip duy tr m ti un cho t bo.
Ngoi ra, collagen cn m bo sc t da, lm sng mu da. S suy gim v cht lng, s lng
collagen s dn n da tr nn kh, mt cng, n hi v thc y qu trnh lo ha ca c th.
Chnh v vy m collagen ng vai tr quan trng gip ci thin cu trc da, kch thch qu trnh
ti to ca ln da, phc hi t bo da b tn thng.
S bin i collagen khi CO' th lo ha
Khi c th lo ha, protein ca t bo kt dnh vi nhau hoc thay i hnh dng lm cho
chng khng cn m nhn c chc nng bnh thng. Collagen b mng i v tnh n hi
gim i r rt. Hu qu l cc np nhn da xut hin. e hn ch s hnh thnh ca np nhn da,
phng php chng lo ha bo v collagen v tng cng tng hp collagen c xem l hu
hiu.
Khi c th bc qua tui 30, tc tng hp collagen b chm li, tc phn hy gia tng.
Trung bnh mi nm, cu trc collagen b mt i khong 1,5% v khi lng; v th cng ln
tui, ln da cng b lo ha nhanh. Trc tui 45, 1/3 trong s tng lng collagen ca c th
c th b mt i. Cc np nhn, np gp xut hin ngy cng nhiu trn trn, quanh ming v
khe mat. Collagen mt i cng nh hng ng k n s nh hnh cc ng nt trn khun
mt, ln da b chng nho, b chy, iu ny c bit thy r khi ph n bc qua tui 40. Ngoi
ra, c rt nhiu nhn t gp phn vo s suy gim ca collagen. Cc cht nhim c cha c to
toxin s kch thch qu trnh oxy ha, to ra cc gc t do. Cc gc t do ny ph hy t bo
bng cch ly i mt electron ca mi t bo. Chng cn l nguyn nhn gy ra cc bnh nghim
trng nh ung th. Ht thuc, ch dinh dng khng cn bng, thiu cc nguvn t vi lng,
cc loi thc phm ng gi cha nhiu cht ha hc c hi v khng tp th dc thng xuyn
l mt s nguyn nhn y mnh s mt mt collagen.

Age 50+

Age 40+

Hnh 1.12 S lo ha da

S lo ha t bo c tr hon nh collagen v hyaluronic acid. Cc cht ny lm phang v

lp y cc np nhn ca biu b, xc tin qu trnh ti to t bo da. Nhng np nhn c v
quanh vng mt s gim xung nh vo s ci thin ca biu b.
C 2 l do chnh cc bc s v chuyn gia thm m s dng collagen trong phng
php chng lo ha da hin i.
'S Collagen c hiu qu cao trong qu trnh hi phc v ti to da
-

iu tr collagen phc hi: thng c s dng trong trng hp da b tn

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.
-

iu tr tr ha bang collagen 25% c s dng cho nhng ph n t 35 tui bt

u c du hiu lo ha vi s xut hin ca cc np nhn, da b mt nc, chng nho, chy x

khng cn cng mn nh trc na, mu sc ca da cng tr nn sm li. Sn pham Beautee
Collagen Cell Pure vi hm lng collagen nguyn cht ln ti 25%, c kh nng b sung v duy
tr m cho da, cung cp dinh dng v bo v da chng li mi trng nhim bn ngoi.
Ngoi ra, sn phm cn c kh nng xa b cc np nhn nng, ci thin v lm m cc np nhn
su do tui tc, phc hi li kh nng n hi t nhin cho da, ngn chn s hnh thnh np nhn
trn da, thc y tr ha t bo theo c ch t nhin.
'S iu tr bang collagen c an ton tuyt i
Phn ln cc sn pham collagen nguyn cht u c chit xut trc tip t da ng vt l ln,
b, cu v c da trn; c bit hn, c mt s loi collagen c chit xut trc tip t c th
ngi, chnh v vy chng hon ton khng c cc cht ha hc hay c t, kh nng tng thch
vi c th ngi c th ln ti 100% m
-

khng h c phn ng o thi hay loi b. Tt c mi ngi mi la tui u c

th s dng v s lnh tnh ca chng. Ngoi ra, collagen cn c a vo da bng ba phng

php chnh:

Thoa ln da v nh tc ng ca cc cng ngh xung in hay nh sng dn

chng vo su ti vng da cn iu tr.

Tim trc tip vo da.

n ung.

a phng php trn u l nhng bin php rt an ton khng h nh hng ti da v c th.

(a) Trc khi iu tr (b)

Sau khi iu tr
Hnh 1.13 Tc dng chong lo ha ca collagen
1.4.3.2

Gim thiu so mn

Acne Care (4 week program)

Collagen thc y qu trnh ti to

m ti nhng vng da b so v mn.
Mt trong nhng chc nng ca
collagen l to ra mt mng li gi
cho t bo da c nguycn vn v
gip chng d

(a) Trc khi iu tr

i
Hnh 1.14 Tc dng tr

(b) Sau khi iu tr

so mn ca collagen

cc t bo mi ny di chuyn ra bn ngoi, chng s in y cc l su ca m da, lm gim s xut hin ca so.

Collagen cng gip cng c tnh nguyn vn ca t bo da bng cch kt hp vi cc m lin
kt trong da ci thin cu trc v tnh n hi.

1.5 Cc ngun cung cp collagen

Chng ta c th b sung, lm tng lng collagen cho c th bng cch tim collagen hoc
ung cc loi thuc c b sung collagen. Tuy nhin, cch tt nht gia tng lng collagen
cho da l ly collagen t cc ngun thin nhin nh nhng phn trch t' ng v thc vt.
trong nhng loi thc phm no? Di y l mt s ngun m bn c th tm thy collagen duy tr v b sung cho
c th
- Tri cy thuc h cam qut: cc loi tri cy v rau c c cha vitamin c chnh l cc
ngun t nhin sn xut ra collagen. Bn nn thm vo ch n hng ngy cc loi tri cy
nh cam, chanh v du. Bng ci xanh cng l mt trong nhng ngun ng tin cy tham gia
vo vic tng hp collagen cho da.
-

Ngun collagen t nhng sinh vt bin: collagen t cc loi c mp l mt ngun cung

cp collagen cho nhu cu ca bn. Vi nhng ngi Trung Hoa thch n vi c th y l mt

cch hiu qu b sung collagen.
-

Bn cnh , da v xng ca cc loi gia sc nh ln l nhng ngun chnh cung cp

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.

1.6 Cc dng collagen nguyn liu thu c t da c 1.6.1.

Collagen dng bt

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 dng gel c sn

xut qua mt quy trnh phc tp,
trch ly t da ca loi c chp bc
(silver carp). Loi c ny ch n to
bin nn n rt thch hp lm
nguyn liu th trong sn xut cc
sn phm m phm.
Hnh 1.16 Collagen dng
gelCollagen EX Cng dung:
cap t c v hyaluronic acid, cng
vi nhng thnh phn lm p khc
nh nhn sm, mm sen, qu l gai
n , tt c ha trn li to thnh
mt hn hp mang n cho bn ln
da ti tr v sn chc.

ng cha collagen cao

J]r\E\DO

Collagen

/HI/EIDO ! Collagen

Xut x: Shiseido, Nht Bn.

Hnh 1.17 Collagen EX

1.7.1

Collagen update (nc ung b dung) Cng dung:

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

1.7.2 Ediva collagen, hp 30 gi X 2,2g (bt)

Cng dung:
Ediva collagen bo sung collagen, gip lm chm
qu trnh lo ha ca da, ci thin v lm m np nhn
trn da c bit l vng quanh mt, ngn chn s hnh
thnh np nhn trn da, tng tnh n hi gip da sn
chc, duy tr s tr trung, rng r cho ln da. Ediva
collagen gip da duy tr m v chng li cc tc
nhn gy nhim ca mi trng nh tia uv, khi,
bi,... Ediva collagen ci thin s linh hot ca khp,
ci thin s Hnh 1.19 Ediva collagen dung np
calcium, gip xng cng cp, gn v sn khe mnh,
do dai.
Xut x: CTCP Dc Hu Giang, cn Th, Vit Nam.
1.7.3 Viospring collagen active essence
Cng dung:
Viospring c lm t collagen nguyn cht t c s mang
n cho bn mt ln da p, ti tr. Nhng phn t
collagen nh trch ly t vy c bng cng ngh cao cng
nhng chit xut t cy c v hng thm, c tc dng xa
i np nhn, phc hi nt thanh xun, s sn chc cho da.
Thnh phn chnh: collagen t' c, elastin, silk peptide,
chit xut t cy l hi, cy chamomile,...
Xut x: Trung Quc Hnh
1.20 Viospring collagen
active essence

1.7.4

Collagen essential mask sheet

Cng dung:
Nng collagen trong tm collagen cho hiu qu hp th cao, em n ln da
sn cc, mn mng.

Xut x: Hn Quc

wL%

collagen

rn FACE SHOP

Hnh . 21 Collagen essential mask sheetCHNG

2 MT S QUY

TRNH TRCH CHIT COLLAGEN c CNG

B
2.1 Quy trnh trch ly collagen t da heo [6]

30 pht

1 ln

20 pht

2 ln

30c
6h X 2 ln (CHBM)

0^5% CHBM khn^on

isopropyl
-m -m 4JNP
-m .
+ 1,9% monomethylamine (v/v)
HC15M

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

10.000 vng/ pht 30

pht

4c

Ly tm

Hnh 2.1 Quy trnh trch ly collagen t da heo

Hnh 2.2 Quy trnh trch ly coagen t gn chn g

Ra sch

Lm kh

NaOH 0,1M, 3 ngy

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

2.3 Quy trnh trch ly collagen t da mc

[8]lnh
Sy

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

COOH, 0,1 -h0,5M Ti l 1:6 H-1:40

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

Hnh 2.4 Quy trnh trch ly collagen t da c tuy

pht

1 0 c
10.000 vng/pht

2.5 tTnh hnh sn xut collagen hin nay

Trong nhng nm trc y, p ng nhu cu trong cng nghip, collagen c trch ly
ch yu t da, xng ca cc loi gia sc v ln. Khong thi gian gn y, s bng pht ca cc
loi bnh truyn nhim nh bnh b in (Bovine Spongiform Encephalopathy - BSE,
Tranmissible Spongiform Encephalopathy - TSE) v bnh l mom long mng (Food and Mouth
Disease - FMD) ln v gia sc hn ch phm vi s dng ca collagen c ngun gc t chng
bi v c kh nng ly truyn nhng bnh ny sang con ngi thng qua cc m ca ng vt.
Thm vo , collagen trch ly t' ln khng c s dng v ro cn tn gio.
Vi
nhng nguyn
ng tp trung vo cc nghin cu ca
h tm ra nhng
ng, vy, vy ca c nhng loi c nc ngt v c bin, da g, da ch, da mc,.. .c th c s dng nh nhng
ngun thay th [10]. Trong s nhng ngun thay th, c cung cp mt ngun nguyn liu th tt
nht v:
- D tm, sn c s dng
-

Khng c s ly truyn bnh

Khng gp phi tr ngi v mt tn gio

C kh nng thu c collagen vi hiu sut cao

Khong 70% tng trng lng c th ca c b b i di dng cc ph phm nh da, xng,

vy, u, vy, rut,...trong sut qu trnh ch bin. Vic tn dng nhng cht thi ny c th nng
gi tr kinh t ca cc loi c ln.
2.6 Collagen c ngun gc t c
Da vo thnh phn % ca tng lng collagen tnh trn tng lng protein, cc loi c c
chia thnh 3 nhm:
-

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

vi phm vi t' 1,58 -- 13,39 % so vi tng lng protein.

Bng 2.1 thnh phn collagen trong tht c

ha tan trong acid Collagen
(so vi trng
khng
lng
hati)
tan (so vi trng lng ti)

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.

ng c cha collagen, collagen t c cng c chng minh l c tc hp th vo mu nhanh hn gp 1,5 ln so vi

collagen t' ln.
im mpl.sma)

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
.

CHNG 3 TNG QUAN V C BASA

3.1 Gii thiu v c Basa

Hnh 3.1 C Basa

auvage 1880) thuc h Pangasiidae, l loi c nc ngt c gi tr kinh t cao, c phn b trong mt vng a l hp
lu vc sng Mekong v sng Chao Phraya (Campuchia, Lo, Thi Lan, Vit Nam). Vit
Nam chng c nui nhiu v tp trung ti Chu c (An Giang) v Hng Ng (ng Thp)
vi nng sut cao.
Trc y, c mt s tc gi nh danh tn khoa hc ca c Basa l Pan^asius
pangasus (Hamiton) v vic phn chia ny da vo ti liu ca Smith (1945), Taki (1974) hay
Panasius nasutus (Bleeker).. . Trong cng trnh nghin cu xut bn nm 1991, Tyson
R.Roberts v Chanvalit Vidthayanon nh danh li mt s loi c trong h Pangasiade. Theo
Nguyn Vn Trng, Nguyn Thanh Tng (1996) cng cho rng c s sai st trong vic phn loi.
Cc tc gi trn thng nht khng nh rng c Basa nui ng bng sng Cu Long c
tn khoa hc l Pangasius bocourti. v ngoi hnh, c Basa rt d phn bit vi cc loi c khc
trong h c tra, c thn ngan, hnh thoi, hi dp bn, ln trn, c bit bng to tch ly nhiu
m, chiu di tiu chun bng 2,5 ln chiu di thn.
ng, ming hp. C Basa c 2 i ru, ru hm trn bng Vi chiu di u, ru hm di bng 1/3 chiu di u. C 40
-- 46 lc mang trn cung mang th nht, vy hu mn c 31--36 tia vy. Chiu cao ca cung
ui hn 7% chiu di chun. Mt lng mu
c
lng v vy ngc c mu xm,

vy hu mn c mu

xm xanh v nht dn xungbng, bng

mu trng
trng trong, mng da gia

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

xy ra i vi c Basa hin ang nui khu vc ng bng sng Cu Long.

Trong h thng phn loi, v tr c Basa c xc nh nh sau:

B Siluriformes
H Pangasiidae
Ging Pangasius
Loi Pangas ins Pangas ins

3.2 c im sinh l, sinh thi ca c Basa

3.2.1

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

S c th nui cho mt b dao ng t 20.000 -T- 50.000 con c ging. Mt th nui

trung bnh 90 -- 150 con/m3 vi c c ging 80 -- 100 g/con.
3.2.6

Ma v nui

Vi iu kin kh hu thun li, m p quanh nm khu vc ng bng sng Cu Long, nn c

th th ging c nui vo bt k thi gian no trong nm.
Bng 3.1 Hai v chnh th c ging vo b

Thng thu hoch

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

Ngng nhit c Basa chu ng c thp nht l - 18 c v cao nht l 39 c. Nu vt

qu gii hn ny th c Basa c th cht.
pH: c Basa c th sng trong mi trng c pH t 5,5 H- 11 ,0.
Bng 3.3 Nhng iu kin mi trng c Basa sng v pht trin

mn (%)
Nhit (C)
pH

3.3 H vi sinh vt trong c

Khi cn sng, ngoi da c c mt lp nht v l mi trng sinh sng tt cho vi sinh vt. S
lng vi sinh vt trn da t 10 2 -T- 10 5 t bo/lcm2 da c. y tn ti cc loi trc khun sinh v
khng sinh nha bo nh Pseudomonas fhiorescens, Proteus vulgaris, Micrococus roseus, E.Coli
v mt so nm mc, nam men sng trong nc.
Trong mang c c rt nhiu vi sinh vt, y a s l nhm vi sinh vt hiu kh thng gp
l Pseudomonas fluorescens. Trong rut c cng c nhiu vi sinh vt ca nc, ca t v t thc
n mang vo, thng thy Clostridium sporogenes, Clostridium welchii, Vibrio setique v nhm
E.Coi. s lng vi sinh vt trong rut c khong 10 3 -- 10 8 t bo/lg cht cha trong rut.

Lng vi sinh vt trong t chc m c ca c tng i t, thng thy: Proteus vulgaris,

Chromobacterium, E.Co, Bac. Subtius... s lng v thnh phn h vi sinh vt ca c ph
thuc vo iu kin sng, s lng vi sinh vt nhim vo c trong bo qun v ch bin c vai tr
rt quan trng trong qu trnh thi ra.
3.4 Cc yu t nh hng n qu trnh sn xut, cht lng sn phm c Basa
3.4.1
nh hig ca thc n
Thnh phn v cht lng ca thc n tc ng trc tip n thnh phn v cht lng
fillet c Basa. Thc n cho c phi m bo y cht dinh dng ph hp cho tng giai on
pht trin ca c.
3.4.2
nh hng ca mi trig
Cc yu t nhit , hm lng oxy ha tan trong nc, pH, mt nui... u c nh
hng n s pht trin v cht lng c.
3.5 Gi tr thc phm ca c Basa
C Basa khng ch cung cp ngun thc phm cho con ngi, m n cn c th c xem
l mt loi c c sn ca vng sng nc ng bng sng Cu Long v c c tht mm mi,
c bit c hng v ring c th trng trong v ngoi nc rt a chung, nht l th trng
M. C Basa l loi c c gi tr kinh t cao hin nay ca Vit Nam v c kh nng cnh tranh
c vi loi c nheo ca M.
Tuy l loi c quen thuc, c gi tr... nhng nhng nghin cu c v loi c ny ch yu
tp trung cc c im sinh l, sinh thi phc v cho vic nui trng v nh bt, cn nhng
nghin cu c tnh h thng v c tnh k thut v gi tr dinh dng cn rt t, nht l cc
nghin cu s dng cc thnh phn khc ca c Basa nh m c, du c... GS.TSKH Nguyn Vn
Thoa v cc cng tc vin ca Vin nghin cn nui trng thy sn thc hin nghin cu v
cc c tnh k thut ca c Basa nh fillet, da, m l...; gi tr thc phm ca c Basa nh
protein, lipid, m, khong... nhng l s liu trung bnh chung cho c Basa.
Cc nghin cu v vic ch bin cc thnh phn c gi tr khc ca c Basa cha c quan
tm nhiu, nht l vic tn dng thnh phn m c. Vin Thy sn II v Phn vin Cng ngh
Thc phm ti Tp.H Ch Minh nghin cn v thnh phn cc acid bo trong m c Basa, tm
cch ch bin m c Basa thnh cht bo tiu chun dng trong thc phm. Tuy nhin, qu
trnh tinh ch thc hin ch yu bng phng php ha hc theo quy trnh cng ngh tinh luyn
c bn.
Tt c nghin cu u mang tnh thm d, th nghim... nhm tm ra cc gii php ti un
cho vic s dng cc thnh phn khc ca c Basa ngoi thnh phn rt c gi tr l fillet c.
Trong thi gian gn y, cc cng trnh nghin cu ca PGS.TS Hong c Nh cho
thy trong m c Basa c s hin din ca DHA (<decosa hexanenoic acid), mt acid bo khng
no gm 22 nguyn t cacbon, 6 ni i, chui cacbon mch di, thng... rt cn thit cho s pht
trin ca h thn kinh trung ng, pht trin thng minh, pht trin no b v vng mc ca thai

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

ti a ngun nguyn liu ny.

CHNG 4 PHNG PHP NGHIN cu

4.1 Mc ch ti
-

Kho st cc yu t nh hng n qu trnh loi mu da c Basa: nng dung dch

H2 02 trong NaOH 0,01 M, thi gian ty mu, t l da/dung dch H2 02 trong NaOH
-

Kho st nh hng ca cc yu t ln qu trnh trch chit collagen: loi acid (citric

acid v acetic acid), thi gian trch chit v phng php trch chit
-

nh gi tnh cht ha l ca sn pham collagen thu c

4.2 Ha cht - Thit b

4.2.1

Ha cht

NaOH

- NaCl

h2 so4

- h2 o2

Citric acid

- Acetic acid

Sodium Lauryl Sulphate (LasNa)

- Na2 HP04

4.2.2

Thit b

Cn hai so

My xay Philips HR 1361

My o m Sartorius MA 35

My ly tm Hermle z 200A

My so mu CR - 300

My o nht Brookfield HDBV - II

My quang ph Varian Carry

4.3

Phng php nghin cu

4.3.1

Phng php tin hnh th nghim

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

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

Phng php phn tch

4.3.2.1

Phng php phn tch nguyn liu

4.3.2.1.1 Xc nh hm lng m trong nguyn liu (Moisture)

Cn mt lng mu c khi lng khong 1 gram. Cho mu vo a nhm v t vo my
o m Sartorius MA 35. c gi tr m trn ng h hin th ca my. Tin hnh o 3 ln
ly gi tr trung bnh.
4.3.2.1.2 Hm lng tro tng (Total ash content) [11]
Nguyn tc
Khi nung 550
thnh phn tro.

600 c, cc cht hu c chy han ton, cht v c cn li gi l

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

Hm Iig bo (Fat content) [11]

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

% Lipid tng (du th) = -----------------

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

4.3.2.1.4 Hm Iig m (Protein content) [11]

nh lng protein bng phng php Kjeldahl trn c s xc nh hm lng nitrogen
protein (thng chim khong 16 % trong cc protein khc nhau). Nhn gi tr nitrogen xc nh
c vi h s 6,25 (100:16) s ra hm lng protein tng s. e nh lng nitrogen protein
ngi ta thng xc nh nitrogen tng s v nitrogen phi protein, sau ly hiu s gia hai
dng ny.
Xc nh nitrogen tng
Nguyn tc: di tc dng ca H2 SO4 c nhit cao, cc hp cht hu c c cha
nitrogen b phn hy v b oxy ha n C0 2 v H2 0, cn nitrogen chuyn thnh amoniac v tip
tc kt hp vi H2SO4 to thnh mui amoni sulphate.

Quy trnh c tin hnh theo cc bc sau:

- V c ha nguyn liu

C 11SO 4: K 2SO 4 (1:3)

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.

Hm lng Nitrogen trong mu c tnh bng cng thc

v

Trong

_ ( a - b x F ) x 0,0014x100x1000 O x V

X : hm lng Nitrogen tnh bng g/1

A
: s ml H S04em hp th NH3
B
: s ml NaOH 0,1N tiu tn khi chun
H2 SO4 tha
V
: s ml mu em v c ha
0,0014 : s gram nitrogen tng ng vi
lml H2 SO4 0,1N
F : h s hiu chnh nng dung dch NaOH 0,1 N (F l t s gia nng thc t v nng
l thuyt ca NaOH)
2

Xc nh nitrogen phi protein

Dng cc dung mi thch hp chit rt tt c cc dng nitrogen phi protein, tuy nhin
trong dch chit vn cn ln mt vi loi protein. V vy, cn dng cht kt ta tch phn
protein ha tan trong qu trnh chit rt.
> nh lng protein trong nguyn liu
Protein(mg) (Ntng s - Nphi protein) x 6,25

4.3.2.1.5

Xc nh tng lng collagen trong da c

Xc nh bng phng php nh lng hydroxyproline [12]

> Nguyn tc
Trong tng lng amino acid c trong collagen, hydroxyproline chim khong 14 %. Loi
amino acid ny gn nh ch c tm thy trong collagen m khng c trong bt k mt loi
protein no khc ca c th, ngoi tr' mt lng nh trong elastin. Do , c th nh lng
collagen thng qua nh lng hydroxyproline.
Cu trc ca hydroxyproline c cha vng pyrrolidine, n c th b oxy ha hnh thnh
vng pyrrole. Vng pyrrole ny tc dng vi tc cht Ehrlich, 4-(N,N
dimethylamino)benzaldehyde to thnh hp cht quinoid c mu (mu sc ph thuc vo nhm
th v thay i t mu cam n mu hoa c). Thc hin phng php so mu tnh hm lng
hydroxyproline.
> Tin hnh
Chun b dung dch chun: ly 25 mg bt hydroxyproline tinh khit ha tan vo 250 ml
nc (100 Lig/ml). Ke tip, ly 50, 75, 100 v 150 \x\ ca dung dch ny v thm nc ct vo
t c tng th tch l 1 ml.
S oxy ha hydroxyproline xy ra khi cho 1 ml Chloramine B vo mi ng nghim, hn
hp c lc v gi nhit phng trong 20 pht. Lng d Chloramine T c phn hy
bng cch cho 1 ml dung dch HC 1 4 3,15 M vo, hn hp c lc trn. Sau 5 pht, cho 1 ml
dung dch p-dimethylaminobenzaldehyde 20 % vo mi ng nghim v tin hnh lc mt ln na.
Cc ng nghim c gi trong b iu nhit 60 c trong 20 pht v sau c lm mt vi
nc lnh trong vng 5 pht. Bc k tip, 5 ml Ethyl Cellosolve c thm vo mi ng
nghim t tng th tch l 10 ml. Ta thu c cc dung dch cha 0,5, 0,75, 1 v 1,5 |Lig
hydroxyproline. hp thu c o bc sng 557 nm (mu sm). Nc ct c dng lm
mu trng. Ta xy dng c ng chun biu din hp thu theo cc hm lng ca
hydroxyproline.

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

Gii thiu khng gian mu CELAB

e tun li cho vic tnh ton v so snh cc mu vi nhau, nm 1976 CE gii thiu mt h
thng sp xp mu sc CIELAB. Trong s dng 3 thng s:
L*: sng
a1: ta mu trn trc - lc b*: ta mu trn trc vng - lam Trong h thng
CIELAB, s khc nhau gia cc im c biu th trong khng gian mu tng ng vi nhng
1AL*=
-

c,p/, -

: s khc nhau v sng gia 2 mu

Nu AL* mang du +: s lch mu theo hng sng

ln, muth sng

hn mu chun
Nu L* mang du s lch mu theo hng ti hn, mu th ti hn mu chun

Aa* =

asampk - a*Umdard: s khc nhau v ta trn trc - lc

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.
-

Nu b* mang du +: s lch mu theo hng vng hn, mu th c sc vng hn

mu chun
-

Nu Ab* mang du s lch mu theo hng lam hn, mu th c sc xanh lam hn

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

Ac* = c* - c* : s khc nhau v bo ha gia hai mu Ah* =

h* - h*2 : s khc nhau v tng gia hai mu

* Ch
Mc ch chnh ca qu trnh ty mu l lm cho da trng, sng ln. Do , trong qu trnh
ny trc mu chnh cn quan tm l trc mu trng - en (). Trc a* v b*
c xem l 2 trc mu ph kho st s thay i ca nh mu ph trong qu trnh x l.
4.3.2.2

Phng php phn tch sn phm

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

30 pht cho nhit ca dung dch collagen cn bng vi nhit b

iu nhit. Th nghim c tin hnh trong khong nhit t 30 c n 50 c. Nhit c

gia tng tng bc, mi bc ng vi 2,5 c v mi mc nhit c gi trong 10 pht.
nht ca dung dch collagen c xc nh tng ng cc mc nhit . nht tng i
c tnh nh sau:
nht tng i = ^min
;

7max - ;7min

Trong :
rt :

nht o c ti mi mc nhit

Tmax :

nht ln nht

Tmin : nht nh 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

Hm Iig tro tng (Total ash content)

4.3.2.2.5
t tiu chun o lng cht lng 3

Hm lig bo (Fat content)

4.3.2.2.
t tiu chun o lng cht lng 3

Hm lng m (Protein content)

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

Quy trnh th nghim

4.3.3.1

Quy trnh loi bo

Da c Basa

0,5% LasNa Dd NaOH 0.2%

H,0

X3

ln

X3

ln

X3

ln

Ra

5% LasNa Dd H7SO4 0.2%

H,0

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

oe thu c collagen khng c mi, da c nguyn liu cn phi c loi loi b cc

cht bo. Ta p dng phng php ca Gudmundsson v Hafsteinsson (1997), phng php
c trnh by nh sau:
Da c Basa sau khi c ra s b vi nc s c tip tc x l vi ln lt 3 loi dung
dch (NaOH 0,2%, H2SO4 0,2% v Citric Acid 0,7%) vi t l da : dung dch l 1:10 (g/ml)
nhm mc ch thu c collagen khng mi. Vi mi loi dung dch, da c c x l qua 3
ln, mi ln tng ng vi thi gian l lh. Sau mi ln x l, da phi c ra sch di vi
nc n khi t pH = 7 trc khi tin hnh ngm ln tip theo. 0,5% cht hot ng b mt
(LasNa) c thm vo tt c cc dung dch s dng trong quy trnh nhm nng cao kh nng
loi mi.
Tuy nhin, theo l thuyt quy trnh ny c th lm mt i mt lng collagen. Ta tin hnh
thm mt quy trnh da trn nguyn tc ca quy trnh trn nhng khng s dng acid ra. 2
mu da ca 2 quy trnh x l (c dng acid v khng dng acid) s c i chng vi nhau.
4.3.3.2

Quy trnh loi mu

a Kolodziejska et al. (1999)

Ngm

aa 170 m2U2 Ngm, ty trong NaOH 0,0IM mu

Da c x l

Nu a* mang du +: s lch mu theo hng ln,

mu th

c sc

hn mu chun
-

Nu Aa* mang du s lch mu theo hng lc hn, mu th c sc xanh lc

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

hay citric 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

Collagen thnh phm

Hnh 4.3 Quy trnh trch ly

Q u trnh trch ly collagen c thc hin nhit 4 c trong 24, 48 v 72 h vi dung

dch c nng 0,5 M ca cc acid citric v acetic. Vi mi loi acid, collagen c trch theo
phng php ton phn (A) v tng phn (B).
- Phng php ton phn (A): da c c trch trong dung dch acid mt ln duy nht
ng vi cc mc thi gian 24, 48 v 72 h.

- 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

Cc thng s kho st trong qu trnh th nghm

4.3.4.1 Trong qu trnh x l nguyn liu

-

Nng dung dch H202 (%)

Thi gian ty mu (h)

ncHA ) =

f(0,5; 0,75; 1; 1,25; 1,5)

f(r)

f(l; 2; 3; 4; 5; 6; 24)

T l da/dung dch (g/ml)

f(F)

f(l:10; 1:12,5; 1:15; 1:17,5;

1:20)

4.3.4.2 Trong qu trnh trch collagen

-

Loi acid
Citric v Acetic acid

Phng php trch

Phng php A (tng phn) v phng php B (ton phn)

 Thi gian trch (h)

f(r) =
4.3.5

f(24; 48; 72)

p ng

4.3.5.1 Trong qu trnh x l nguyn liu

p ng c chn lm p ng chnh l gi tr lch sng gia da c nguyn liu ban
u v da c sau qu trnh x l AL* theo cc thng s nng dung dch H 202, thi gian ty
mu v t l da/dung dch. Cc gi tr C* v h* l cc p ng ph nh gi s thay i
mu sc ca da c trong qu trnh x l.
4.3.5.2 Trong qu trnh trch collagen
p ng chnh l hiu sut trch ly theo cc yu t nh loi acid, phng php trch
v thi gian trch.CHNG 5KTQUTHNGHIM
5.1 nh gi nguyn liu
5.3.1
-

Ngun nguyn liu

Xut x: da c Basa c mua t Cng ty TNHH Thu Sn Bnh An (Binh An

Seafood Joint Stock Company), L 2.17, Khu Cng Nghip Tr Nc II, TP. cn Th.
-

Da c c tch ra bng phng php c hc nh my. Sau khi mua v, ta ra sch

bng nc, loi b s b cc phn m cn st li trn da v tn tr khong -

20c cho n khi s dng.

5.3.2

Thnh phn

Kho st cc thnh phn ca da c Basa (Ph lc 1)

Bng 5.1 Thnh phn da c Basa nguyn liu
Hm lng (%)

Protein

31,1

Lipid

6,5

58,7

tro

0,1

Cc thnh phn khc

3,6

5.2 nh gi quy trnh loi bo nguyn liu (Ph lc 1)

Bng 5.2 So snh thnh phn gia da nguyn liu v da sau khi x l
ca hai quy trnh

Cm quan

- Hiu sut loi bo

Quy trnh c s dng acid
15 75-3 27 H% =
X100 = 79,24 %
15.75

Quy trnh khng s dng acid

15 75-15 35 H% =

100 = 2,54 %

15.75

- Lng protein mt i sau khi loi bo

Quy trnh c s dng acid
-]<

_ on I o

75.35

X100 = 48,00 %

Quy trnh khng s dng acid

75,35-64,40 x 100 = 14 53 o/o
75.35

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

sch, c th dng lm nguyn liu cho

chn quy trnh x l c mt ca acid

cc

tinh khit, khng mu v khng mi.

5.3 Kho st cc yu t nh hng n qu trnh ty mu nguyn liu
5.3.1
5.3.1.1

Nng H202 v thi gian ty mu

Diu kin kh st

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)

C2 (%) = 0.5,0.75,1,1.25,1.5 r (h) =

1,2,3,4,5,6,24

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

Thi gian (h)

30

hng
gian

0.5%
0.75%
1%
1.25%
1.5%

ca nng H202 v thi

0

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

5.3.2.1 iu kin kho st

iu kin c nh
C22(%)= 1
T (h)

tAC)

=4

30

Kho st
AL* =f(F)
AC* =f(F)
Ah* =f(F)

F(g/ml) = 1:10, 1:12.5, 1:15, 1:17.5, 1:20

5.3.2.2 Kt qu kho st (Ph lc 3)

Bng 5..4 Kt qu lng collagen trch bng lactic acid (ph lc 6)

T l da/dung dch (g/ml)

Hnh 5.2 th biu th nh hng ca t l da/'dung dch ln qu trnh ty mu

5.3.2.3

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

Theo phng php nh lng hydroxyproline (Ph lc 4)

Hnh 5.3 th biu din ng chun ca hp th theo nng

hydroxyproline
Bng 5.3 Tng lng collagen trong da c (xt trn 30 g da c) (Ph lc 5)
Ln Hydroxyproline
th
(g/ml)
Loi da c Hm lng
nghim

Da ban u
Da sau kh
x l

Bng 5..4 Kt qu lng collagen trch bng lactic acid (ph lc 6)

> Da c ban u
-

Hm lng collagen (%) tnh trn khi lng t (wet weight)

-X100 = 15,53 %
30

Hm lng collagen (%) tnh trn khi lng kh (dry weight)

100 = 37,64 %
12,38

Da c sau khi x l
-

Hm lng collagen (%) tnh trn khi lng t (wet weight)

4 37
-^-x 100 = 14,57 %
30

Hm lng collagen (%) tnh trn khi lng kh (dry weight)

4 37
-^-x 100 = 39,26 %
11,13

5.4.2

Theo phng php trch ly bng lactic acid (Ph lc 6)

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

- Lng collagen tng cng:

3,32 + 0,89 + 0,19 = 4,4 (gram)
- Phn trm collagen theo khi lng t (wet weight)
% collagen = X100 = 14,67 %
30
- Phn trm collagen theo khi lng kh (dry weight)
% collagen = X100 = 35,54 %
12,38
5.4.3

chnh lch gia 2 phng php

4 4-4 37
X100 = 0,69 %
4,37

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(Phng php trch)

=

f(r)

Acetic v Citric acid

tng phn v ton phn
r (h)
=
24, 48, 72
5.5.2

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

Thi gian trch: hiu sut trch collagen

ng bin theo thi gian i vi c 2 loi acid.

-

Phng php trch: mi loi acid, lng

collagen thu c theo phng php trch tng

phn cao hn so vi phng php trch ton

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

nh gi tnh cht sn phm

5.6.1 Xc nh s bn nhit ca collagen

(Thermal stability)
5.6.1.1
8)

Phng php o nht (Ph lc

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)

Hnh 5.5 ng cong bin tnh theo nhit ca colagen

Phng php DTA/TG (Ph lc 9)

5.6.1.2
DTA /(uVAng)
DDTA J[uVAngAnin
)

ng phn tch nhit DTA/TG ca collagen trch being acetic aci

d
DTA 4uV/tag)

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

irngphn tch nhit DTA/TG ca collagen trch bang citric acid

ng phn tch nhit DTA/TG ca colagen trch bng lactic acd

Hnh 5.6 ng phn tch DTA/TG ca collagen
Nhn chung, ng phn tch nhit ca collagen trch bng 3 loi acid c xu hng ging
nhau. ng biu din ca collagen (trch bng acetic acid) c 2 peak gn st vi collagen
(trch bng lactic acid, xem nh l mu i chng). sch ca cc collagen sau khi tinh ch
l tng i cao, nhit phn hy vo khong 119,2 c.

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

Thi gian (pht)

Thi gian (pht)Collagen ha tan

0,0087
0,0095
0,0105
0,0114
0,0136
0,0141
0,0149
0,0149
0,0149
0,0149
0,0149
0,0149
0,0149
0,0149
0,0149
0,0149
Thi gian (pht)

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

5..4.2 So snh vi collagen thu c t cc nghin cu khc

trch ly t da c Basa (s dng lactic acid) Hnh 5.9 Wavenumber (cm'1)
Ph IR ca
collagenVng

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
a
t
h

e
o
p
h

n
g
p
h

p
tr

c
h
l
y
b
a
n
g

a
c
i
d
t
h

h
i

n
k

t
q
u

p
h

I
R

g
i

n
g
v

i
p
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t
u

c
t

n
h


n
g
l
o

i
c
o
ll
a
g
e
n
k
h

c
(
s
o
s


n
h
v

i
c
o
ll
a
g
e
n
t
h
u

c
t

n
h

n
g
q
u
y
tr

n
h

c
c

n
g

).
V

n
g
a
m
i
d
e
I,
I
I
v

I
I
I
c

li

n
q
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a
n
tr

c
ti

n
h

n
h
d

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g
c

a
c
h
u

i
p
o
l
y
p
e
p
ti
d
e
.
N

a
m
i
d
e
A
(
3
4
0
0
T
3
4
4
0

c
m
'1
)
li

n
h

i
m

i
k

o
N
H
.

N
h

a
m
i
d
e
I
(
1
6
0
0
T
1
6
6

0
c
m
'1
)
b
i

u
t
h

s
tr
e
t
c
h
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n
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v

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b
r
a
ti
o
n
c

a
c

c
g

c
c
a
r
b
o
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l
tr
o
n
g
c

c
p
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ti
d
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,
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t
h


n
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p
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n
q
u
a
n
tr

n
g
n
h

t
tr
o
n

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v
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c
k
h

o
s

t
c

u
tr

c
b

c
h

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i
c

a
c

c
p
r
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t
e
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n
.
N
h

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i
d
e
I
I
(
~
1
5
5
0
c
m

"'
)
b
i

u
t

N
H

b
e
n
d
i
n
g
v

C
N

s
tr
e
t
c

h
i
n
g
.
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h

a
m
i
d
e
I
I
I
(
1
2
2

0
*

1
3
2
0
c
m
'1
)
b
i

u
d
i

n
C
N

s
tr
e
t
c
h
i
n
g
v

N
H
,
c

li

n
q
u

a
n

n
c

u
tr

c
c
h
u

i
x
o

a
c
o
ll
a
g
e
n
.
T
r
o
n
g
p
h

g
n
g
o

i
t
h
u

c
t

c
o
ll
a
g
e
n

a
c

B
a
s
a
,
t

t
c

n
h

n
g

n
g
v

i
c

c
d

i
a
m
i
d
e

tr

c
b
i

u
d
i

c
b

c
s

n
g
t

n
g
t

n
h

i
v

i
c

o
ll
a
g
e
n
t

l
o

i
c

c
n

g
h
i

n
c

n
(
c

n
g

).

 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:
-

Collagen trong da c Basa c t cc lin kt lin phn t nn d dng c ha tan bang

acid hn c r v c ng.

Acetic l dung mi trch ly hu hiu nht trong 3 loi acetic, citric v lactic acid.

CHNG 6 NHN XT- BN LUN

6.1 Nhn xt trn ngun nguyn liu
C Basa l loi c tng trng nhanh, giu dinh dng v c gi tr xut khu cao.
nc ta, c bit l vng ng bng sng Cu Long, vic nui c Basa rt pht trin. Mt
hng xut khu ch yu l fillet c, lng xut khu khong 30.000 -T- 35.000 tn llet/nm.
Trong qu trnh ch bin c, da l ngun ph phm chim khi lng khong 0,5%. Nh vy,
ta c khong 1.500 -5-1.750 tn da/nm. Neu c tn dng lm ngun nguyn liu dng
trong vic sn xut collagen th gi tr kinh t ca da c Basa, vn d trc y ch lm thc
n gia sc hay phn bn, s c nng ln. v mt cu to, da c Basa cng c lp biu b
(epidermis) v lp b (dermis) nh da ngi nn collagen c ngun gc t da c c ng
dng rng ri trong nhiu lnh vc ca i sng.
6.2 Nhn xt trn quy trnh th nghim

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

x l, da c tr trong t ng nhit khong - 20 c cho n khi c s dng cho cc

bc tip theo.
- Khu 2: Trch chit collagen, s dng acetic acid lay collagen t da c x

- Khu 3: Tinh ch collagen, collagen c kt tinh v ly ra khi dch trch bng

phng php ly tm.
Quy trnh th nghim c th tm tt bng s khi nh sau

Lun vn tt nghip

Trang 1

:Da nguyn liu

1 Loi bo%

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

Hnh 6.1 Quy trnh

sn xut colagen
t da c

Mt im bt li cn phi cp n ca collagen trch ly t da c l s lu tn dai dng

ca mu v mi hi ca c. Do , cn phi c giai on x l da c loi b cc cht bo gy
mi hi v cc sc t c th thu c collagen khng mu v khng mi.
6.2.1
Quy trnh loi bo
p dng quy trnh ca Gudmundsson v Hafsteinsson (1997), da c c x l 3 ln vi mi
loi dung dch sau 0,2 % NaOH, 0,2 % H2SO4 v 0,7% citric acid. Da sau khi x l khng cn
mi hi. Tuy nhin, nhc im ca phng php ny l quy trnh kh phc tp v tn sc. Da c
c x l tri qua mt lot dung dch, sau mi ln x l da phi c ra sch n khi pH t
bng 7. y chnh l phn tiu tn nhiu thi gian nht ca quy trnh. ng thi, mt lng
collagen trong da b mt i sau khi tri qua x l. Tuy nhin, nu loi b cc hp cht noncollagen ch vi dung dch NaOH th da vn cn rt hi v nhp nhp. Tt c cc dung dch dng
trong qu trnh x l c thm vo 0,5% cht hot ng b mt LaSNa nhm lm tng hiu qu
ca qu trnh loi mi.
6.2.2
Quy trnh ty mu
p dng quy trnh ca Kolodziejska et al. (1999), nhn chung cc sc t c loi mt
cch kh tng i. trng ca da c sau khi x l uc ci thin ng k so vi da c ban u.
Hai thng s quan trng trong qu trnh loi pigment ca da c l nng H2O2 v thi gian ty
mu. Mt im cn ch l mu ph trong qu trnh x l. Neu ngm da c qu lu th da s b
ng mu vng.
6.2.3

Quy trnh trch ly

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.

.2.3.2 Cc thng s quan trng trong qu trnh trch ly

C rt nhiu yu t nh hng n qu trnh trch ly chang hn nh loi acid, phng php
trch ly, thi gian trch, t l nguyn liu : dung mi trch ly,... Kch c nguyn liu v t l nguyn
liu : dung mi c xem l hai thng s quan trng bi v qu trnh tip xc gia hai pha rn lng cng tt khi b mt tip xc pha cng ln. Nguyn liu c kch thc nh s lm tng din
tch tip xc pha lm qu trnh trch ly xy ra hon ton, ngc li th trch ly s b hn ch do
khng c s tip xc tt gia hai pha. T l da : dung dch acid ln th da s phn tn u trong
dung dch, nu lng acid qu t th b mt da khng c tip xc hon ton vi dung dch, trch
ly khng hon ton. Thng s thi gian quyt nh hiu sut trch ly. Neu thi gian ngn th
collagen ha tan vo acid cha nhiu nn hiu sut thp. Tuy nhin, thi gian qu di cng khng
tt v mch collagen c th b ct su xung hnh thnh gelatin.
Trong ti lun vn ny, quy trnh trch ly cn tng i n gin, ch c s h tr ca s
khuy gin on. Do , hiu sut thu c vn cha cao lam, collagen cha ho tan hon ton.
Ta c th nng cao hiu sut trch ly collagen bng cch tin x l c hc, ha hc hay enzyme.
Nguyn liu cn c xay nh tng kh nng tip xc vi dung dch. ng thi trong qu trnh
trch ly cn kt hp vi s khuy trn c hc. S khuy nhm gip cho s ho trn gia nguyn
liu v dung mi c tt hn v cung cp nng lng cho qu trnh trch ly. Mt phng php
na lm tng lng collagen trch ly c l p dng s tin x l vi enzyme. Cc enzyme
thng c s dng nh pepsin, trypsin, pancreatin, fcin, bromelin hay papain. Cc enzyme ny
ch ct b nhng phn u mt khng xon c (telopeptide) ca collagen. Chnh nh s ct t
cc telopeptide ny lm t cc lin kt gia cc phn t do lm tng kh nng ho tan ca
collagen. Mc gia tng ca s ho tan collagen tu thuc vo tng loi c. Collagen trong da
ca mt s loi c c th ho tan hon ton trong dung dch acetic acid sau khi x l enzyme. Tuy
nhin, mt s loi khc, hiu sut trch ly sau khi x l vi enzyme ch tng khong 5%. S
khc bit ny c gy ra bi s khc nhau v v tr cng nh loi lin kt lin phn t ca cc
loi c.
6.3 nh gi kt qu

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

c. Sau , trng c khuynh

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

Thi gian trch ly

72 h

Phng php trch ly

Tng phn

Nng acid

0,5M

Nhit trch ly

4 c

T l da : acid

1:6 g/ml

nh gi tnh cht sn phm

6.3.3.1
Nhit bin tnh (Dnaturation temperature)
Nhit bin tnh ca collagen t da c Basa l khong 37,5 c, cao hn so vi collagen tu
c t nhng loi c c nghin cu nh c tuyt (15 C), c thu (26,1 C), c chp (32,5
C), c r (36,5 C). Nhit bin tnh cao ca collagen t da c Basa l do cu trc da ca n c
cha nhiu thnh phn imino acid (proline v hydroxyproline).
Hm lng imino acid, c bit l hydroxyproline, ph thuc vo nhit mi trng sng
ca c v n c nh hng ln s bn nhit ca collagen. Collagen c ngun gc t cc loi c
sng trong mi trng nc lnh c thnh phn hydroxyproline thp hn so vi nhng loi sng
vng nc m. Cc imino acid, proline v hydroxyproline, c trong cc lin kt hydrogen gia
cc chui. Cc lin kt ny gp phn lm n nh cu trc chui xon c ca collagen.
C Basa l loi c sng vng nc m nn c cha nhiu hydroxyproline nn nhit bin
tnh cao.
.3.3.2 Cc yu t nh hng n tnh cht ca collagen
tui ca ng vt
tui ca c nh hng n cc m lin kt cng nh s lng, cht lng ca cc protein

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-

1600 cm'1), ch yu da vo ni i c = o trong lin kt peptide (khong 80 %). Ph hng ngoi

ca collagen trch ly t da c Basa bang acetic acid phn nh s hin din ca di amide A (3449
cm'1), amide B (2925 cm"1), amide I (1619 cm"1), amide II (1560 cm'1) v amide IT (1241 cm'1).
Di amide I c lin i vi cu trc bc hai ca protein v amide chng minh s c mt ca cu
trc chui xon c. kt qu ph R cho thy c cu trc xon c trong collagen thu c t da c
Basa.
.3.3.5 Sn phm ph ca quy trnh sn xut collagen t da c Basa
Sn phm chnh ca quy trnh l collagen ho tan trong acid (acid soluble collagen ASC).
Phan d cn li cn cha mt lng collagen v c xem nh l collagen khng ho tan
(insoluble collagen). Thay v lm tan r v chuyn thnh vt liu ha tan, collagen vi nhiu lin
kt ngang c th c phn tn thnh nhng huyn ph si mu trng sa bng nhng tc nhn
bin tnh nh hoc lm vn bng phng php c hc iu kin pH acid. Si collagen bn hn
vi s thy phn so vi nhng thnh phn khc trong m khng phi l collagen, c tch khi
trong sut qu trnh thy phn v ra. Trong nhng bc x l c b sung thm vo, vt liu
collagen c th c dng cho nhng ng dng ha hc nh s acetyl ha, s methyl ha hoc c
th dng gn kt vi cc polymer khc.

Sizeimicions)

Hnh 6.2 Kt qu o kch thuc ht ca insoluble collagen x l bng acetic acid

Hnh 6.3 Kt qu SEM ca insoube coagen x l bng 3 loi acid

A. Acetic acd, B. Ciric acid,

c. Lactic acid

6.4 Ket lun - kin ngh

6.4.1 Kt lun
Collagen l mt loi protein cu trc chnh ca c th, c rt nhiu chc nng trong c th
con ngi, n c phm vi ng dng rng ri trong nhng ngnh quan trng nh y hc, cng
nghip v trong m phm. Trong nhng nm gn y, c nhiu nghin cu trong vic tm ra
cc ngun thay th nhm p ng nhu cu collagen ca th trng. Nhng ph phm t c
c chng minh l ngun thay th l tng nht cho vic cung cp collagen.
C Basa l
loi c ph bin Vit Nam, c gi tr kinh t cao visnlng nui
v xut khau ln. Da l mt ngun ph phm ca ngnhcng nghip ch bin thy,
hi

sn vi mt khi lng khng l. Vic tn dng ngun ph phm ny iu ch collagen gp

phn em li hiu qu kinh t.
Trong phm vi kho st ca lun vn ny, c th rt ra iu kin
ti un cho qu
trnh
trch chit nh sau: acetic acid l loi acid tt nht cho qu trnhtrch ly collagen
t da c Basa. Cc mc gi tr cho kt qu kh quan nht ca cc thng s nh nng acid, t l
da : dung dch, nhit , thi gian ln lt l 0,5 M, 1 : 6 g/ml, 4 c v 72 h. Phng php trch ly
tng phn cho hiu sut cao hn.
6.4.2 Kin ngh
V thi gian thc hin ti c hn cng nh my mc, thit b cn thiu thn nn khng th
kho st ht tt c cc yu t nh hng n qu trnh trch ly collagen t' da c Basa. Do , ti
ngh mt s nghin cu cn kho st nh sau :
-

Kho st

nng acid, t l da : acid

Kho st

nhit trch ly

Kho st qu trnh trch ly collagen t xng c Basa

Kho st tnh nng, thm d ng dng ca sn phm collagen

TI LIU THAM KHO

1.

Wolfgang Friess, U.O.E., Erlangen, Germany, Collagen - biomaterial for drug

delivery. 1997.

2.

Fratzl, p., Collagen Structure and Mechanics. 2008.

3.

Frederic M.Richards, D.S.E., John Kuriyan, Advances in Protein Chemistry. 2005.

Fibrous Proteins Coiled-Coils, Collagen and Elastomers, Volume 70

4.

A.de Meijere, K.N.H., Collagen Primer in Structure, Processing and Assemblyics.

2005. Topics in Current Chemistry.
5.

Garg, F.H.S.a.A.K., Handbook of Biodegradable Polymers. Collagen:

Characterization, Processing and Medical Applications.

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

dnaturation temperature and Hydrodynamic properties. 1967.

8.

Nagai, T., Collagen from diamondback squid. 2003.

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

backed toadfish (Lagocephalus gloveri) skin. 2005.

11.

Food and Agricultural Organisation - Food and Nutrition Paper. 14/7/1986.

12.

N.Yu. Ignat'eva, N.A.D., Determination of Hydroxy proline in Tissues and the

Evaluation of the Collagen Content of the Tissues. 2005.

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

adult Nile perch (Lates niloticus). 2003.

15.

Phanat Kittiphattanabawon, S.B., Wonnop Visessanguan, Takashi Nagai,

Munehiko Tanaka, Characterisation of acid-soluble collagen from skin and bone

of bigeye snapper (Priacanthus tayenus). 2004.

16.

Jin-Wook Woo, S.-J.Y., Seung-Mock Cho, Yang-Bong Lee, Seon-Bong Kim,

Extraction optimization and properties of collagen from

yellowfin tuna (Thunnus albacares) dorsal skin. 2007.

17.

V. Vijayasundaram, V.R. and P.R. Palaniappan, The study of the changes in the
thermal properties ofLabeo rohita

bones due to arsenic exposure. 2009.

18.

Yun Shana, Y.Z., Yong Caoa,Qunhua Xua, Huangxian Jub, Zonghan Wuc,
Preparation and infrared emissivity study of

collagen-g-PMMA/In203 nanocomposite. 2003.

19.

HaiYing Liu, D.L., ShiDong Guo, Studies on collagen from the skin of channel
catfish (Ictalurus punctaus). 2006.

20.

S.Rama, G.C., Physico - chemical characterisation and molecular organisation of

hte collagen from the skin of an air - breathing fish (Ophiocephalus striatus). 1983.

MC LC
CHNG 1 TNG QUAN V COLLAGEN.......................1
1.1

Gii thiu s lc v Collagen.....................................................................................1

1.2

Cu trc ca collagen....................................................................................................1

1.2.1

Cu trc phn t ca collagen ...............................................................................1

1.2.2

Cu trc si ca collagen ......................................................................................4

1.3

Phn loi collagen.........................................................................................................7

1.4

ng dng ca collagen................................................................................................10

1.4.1

ng dng trong cng nghip................................................................................10

1.4.2

ng dng trong y hc...........................................................................................10

1.4.3

ng dng trong m phm.....................................................................................13

1.5

Cc ngun cung cp collagen.....................................................................................17

1.6

Cc dng collagen nguyn liu thu c t da c......................................................18

1.6.1.

Collagen dng bt................................................................................................18

1.6.2.

Collagen dng gel.................................................................................................18

1.7
1.7.1

Mt s sn phm collagen...................................................................................19
Collagen EX.........................................................................................................19

1.7.2

Collagen update (nc ung b dng)..............................................................19

1.7.3

Ediva collagen, Hp 30 gi X 2.2g (bt)..............................................................20

1.7.4

Viospring collagen active essence.......................................................................20

1.7.5

Collagen essential mask sheet..............................................................................21

CHNG 2 MT S QUY TRNH TRCH CHIT

COLLAGEN C CNG B....................................22
2.1

Quy trnh trch ly collagen t da heo .........................................................................22

2.2

Quy trnh trch ly collagen t gn chn g ................................................................23

2.3

Quy trnh trch ly collagen t da mc ..........................................................................24

2.4

Quy trnh trch ly collagen t da c tuyt (Baltic cod)................................................25

2.5

Tnh hnh sn xut collagen hin nay...........................................................................26

2.6

Collagen c ngun gc t' c........................................................................................26

2.7

So snh collagen t c v collagen t cc loi gia sc.................................................28

2.8

Ngun nguyn liu c cho vic sn xut collagen Vit Nam....................................29

CHNG 3 TNG QUAN V CBASA........................30

3.1

Gii thiu v c Basa....................................................................................................30

3.2

c im sinh l, sinh thi ca c Basa.......................................................................31

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

nh hng ca mt s yu t mi trng i vi c Basa..................................33

3.3

H vi sinh vt trong c..................................................................................................34

3.4

Cc yu t nh hng n qu trnh sn xut, cht lng sn phm c Basa.34

3.4.1

Anh hng ca thc n.........................................................................................34

3.4.2

Anh hng ca mi trng...................................................................................35

3.5

Gi tr thc phm ca c Basa......................................................................................35

3.6

Cc sn phm c ch bin t c Basa.......................................................................36

CHNG 4 PHNG PHPNGHIN

cu...................38

4.1

Mc ch ti..............................................................................................................38

4.2

Ha cht - Thit b........................................................................................................38

4.2.1

Ha cht................................................................................................................38

4.2.2

Thit b..................................................................................................................38

4.3

Phng php nghin cn...............................................................................................39

4.3.1

Phng php tin hnh th

4.3.2

Phng php phn tch.........................................................................................39

4.3.3

Quy trnh th nghim.............................................................................................48

4.3.4

Cc thng s kho st trong qutrnh th nghim.................................................51

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

Ngun nguyn liu................................................................................................53

5.1.2

Kho st cc thnh phnca da c Basa...............................................................53

5.2

nh gi quy trnh loi bo nguyn

liu ................................................................53

5.3

Kho st cc yu t nh hng nqu trnh tay mu nguyn liu..............................55

5.3.1

Nng H202 v thi gian

ty mu.............................................................55

5.3.2

T l da/dung dch.................................................................................................57

CHNG 6 NHN XT - BN LUN................................75

6.1

Nhn xt trn ngun nguyn liu..................................................................................75

6.2

Nhn xt trn quy trnh th nghim...............................................................................75

6.2.1

Quy trnh loi bo.................................................................................................77

6.2.2

Quy trnh ty mu.................................................................................................77

6.2.3

Quy trnh trch ly...................................................................................................77

6.3

nh gi kt qu............................................................................................................79

6.3.1

Qu trnh ty mu..................................................................................................79

6.3.2

Qu trnh trch ly...................................................................................................80

6.3.3

nh gi tnh cht sn phm.................................................................................80

6.4.1

Ket lun - kin ngh

Kt lun..84
6.4.2

Kin ngh

84
85

DANH MC CAC HINH

Hnh 1.1 Cu trc ca phn t collagen (triple

helix).......................................................................2
Hnh .2 S hnh thnh chui xon c ni phn t v gia cc chui polypeptide....................2
Hnh 1.3 Cu trc ca procoagen v s chuyn t procoagen sang tropocoagen 4
Hnh 1.4 Cu trc si ca co!lagen.............................................................................................4
Hnh 1.5 Qu trnh t hp to si ca cc phn t collagen........................................................5
Hnh 1.6 S sp xp ca phn t collagen trong mt si..........................................................6
Hnh 1.7 Hnh nh ca b si (a) v si (b) collagen................................................................7
Hnh 1.8 Mt vi dng sinh hc ca collagen...............................................................................8
Hnh 1.9 Keo dn collagen.........................................................................................................10
Hnh 1.10 Collagen dng trom phn thut thm m..............................................................11
Hnh 1.11 Collagen p dng trong vic iu tr vt bng.........................................................12
Hnh 1.12 S lo ha da.............................................................................................................14
Hnh 1.13 Tc dng chong lo ha ca collagen.......................................................................16
Hnh 1.14 Tc dng tr so mn ca collagen..........................................................................16
Hnh 1.15 Bt co!lagen sn xut t da v vy c.....................................................................18
Hnh . 16 Collagen dng gel.....................................................................................................18
Hnh 1.17 Collagen EX..............................................................................................................19
Hnh 1.18 Collagen Update........................................................................................................19
Hnh 1.19 Ediva collagen...........................................................................................................20
Hnh 1.20 Viospring collagen active essence............................................................................20
Hnh 1.21 Collagen essential mask sheet....................................................................................21
Hnh 2.1
Quy trnhtrch ly collagen t da heo..................................................................22
Hnh 2.2
Quy trnhtrch ly collagen t gn chn g.........................................................23
Hnh 2.3
Quy trnhtrch y collagen t da mc.................................................................24
Hnh 2.4
Quy trnhtrchly c oil agen t da c tuyt...........................................................25
Hnh 2.5 th so snh kh nng hp th vo mu ca collagen trch ly t c v t
ln................................................................................................................................................29
Hnh 3.1 C Basa.......................................................................................................................30
Hnh 4.1 Quy trnh loi bo......................................................................................................48
Hnh 4.2 Quy trnh ty mu.......................................................................................................49
Hnh 4.3 Quy trnh trch y........................................................................................................50
Hnh 5.1 th biu th nh hng ca nng H202 v thi gian ln qu trnh ty
mu..............................................................................................................................................56
Hnh 5.2 th biu th nh hng ca t da/dung dch ln qu trnh ty mu.............58
Hnh 5.3 o th biu din ng chun ca hp th theo nng hydroxyproine

......................................................................................................................................................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

DANH MC CAC BANG

Bng 1.1 S phn bo cc amino acid trong chui

poypeptide............................................................3
Bng 1.2 Ket cu chui v s phn b ca cc oi coagen trong c th ngi.............9
Bng 2.1 Thnh phn coagen trong tht c...........................................................................27
Bng 3.1 Hai v chnh th c giong vo b.........................................................................33
Bng 3.2 Sc chu ng ca c Basci trong mi trng nc c mn khc nhau. 33
Bng 3.3 Nhng iu kin mi trng c Basa song v pht trin...................................34
Bng 5.1 Thnh phn da c Basa nguyn liu........................................................................53
Bng 5.2 So snh thnh phn gia da nguyn liu v da sau khi x ca hai quy
trnh.............................................................................................................................................53
Bng 5.3 Tng lng coagen trong da c..............................................................................59
Bng 5.4 Ket qu lng collagen trch bng lactic acid..........................................................61
Bng 5.5 Lng coagen thu c trong qu trnh trch ly..................................................63
Bng 5.6 V tr cc nh (peak) trong ph R ca coagen ha tan trong acd t da c
Basa.............................................................................................................................................72
Bng 5.7 So snh v tr cc nh trong ph hng ngoi ca coagen trch ly t da c Basa,
c r (Lates
noticus) v c ng (Thunnus abacares).............................................73