54

Tp ch Ha hc, T. 41, s 3, Tr. 54 - 60, 2003

KHO ST NG HC PHN NG DEAXETYL HA CHITIN
THANH CHITOSAN NHIT THONG
n Ta son 2-12-2002
L Th Hi Yn, Nguyn Th Ngc T
Vin Ha hc, Trung tm KHTN&CNQG

summary
The influence of NaOH solution concentration and reaction duration on the deacetylation
reaction of chitin to form chitosan has been investigated. The chitosan product obtained from
the technic treated chitin has high deacetylation degree (DD 97%), while those from the non
technic treated chitin has lower deacetylation degree at the same reaction condition.
13
C-NMR
spectra of chitosan in solid form has been studied.
I - t vn
Chitosan l sn phm deaxetyl ha chitin
(thnh phn chnh ca cc d phm thy sn
nh: v tm, v cua, sng lng mc ng...) *
c nghin cu v s dng rng r*i trong
nhng nm gn y [1, 2]....
Cu trc ha hc ca chitin c cc mch
cao phn t an xen nhau nn kh bn vng, v
vy, thng thng thc hin phn ng
deaxetyl ha thnh chitosan ngi ta phi s
dng dung dch kim c v nhit 90 -100
o
C
[3]. c bit, thu c chitosan c
deaxetyl ha (DD) cao khong trn 90%, c tc
gi phi thc hin phn ng deaxetyl ha chitin
trong mi trng kh tr trnh s oxi ha vi
k thut tng i phc tp [4].
Phn ng eaxetyl ha ca chitin thnh
chitosan cng xy ra trong mi trng kim c
vi thi gian phn ng ko di t 5 - 6 ngy
nhit thng, cho sn phm c deaxetyl
ha khng cao (DD < 85%) [5].
y, chng ti * nghin cu ng hc
qu trnh phn ng eaxetyl ha chitin sau khi
x l k thut (XLKT) v chitin khng qua
XLKT thnh chitosan trong mi trng kim,
nhit bnh thng tm ra iu kin ti u
cho phn ng iu ch chitosan.
II - Thc nghim
1. Nguyn liu
- Chitin t v tm do phng Polyme dc
phm - Vin Ha hc, Trung tm KHTN&
CNQG cung cp.
- NaOH tinh sch.
2. Thit b, dng c
Bnh phn ng thy tinh, cc thy tinh, ng
ong, a thy tinh,
My o nht SNB -1/Shangping ca i
Loan,
My o ph hng ngoi (IR) FIR-Impact
410 ca h*ng Nicolet,
My o ph cng hng t ht nhn JEOL
JNM CMX300 ca Nht Bn.
3. Phn ng deaxetyl ha chitin thnh
chitosan
Cn khong 10 g chitin cho mi mu, sau
khi x l k thut (XLKT), chitin c ngm
ngp trong dung dch NaOH c cc nng
khc nhau trong bnh phn ng, thnh thong
55
khuy o u. Phn ng c thc hin
nhit phng.
Theo di qu trnh deaxetyl ha bng th
ho tan ca sn phm trong dung dch axit
axetic 1%, kt hp vi xc nh DD bng ph
IR.
Kt thc phn ng, loi b dung dch
NaOH v ra sch phn khng tan vi nc ct
nhiu ln n pH = 7 v sy kh sn phm 40
- 50
o
C.
Sn phm c ng trong l thy tinh, y
np kn, dn nh*n.
Mu i chng dng chitin khng qua
XLKT cng iu kin phn ng nh nhau.
4. Xc nh khi l)ng phn t trung bnh
Xc nh khi lng phn t trung bnh ca
chitosan theo phng php o nht vi
phng trnh Mark-Houwink [6] v cc thng
s k, thch hp.
[] = k . M
v

(1)
Do cc sn phm chitosan c deaxetyl
ha khc nhau nn chng ti chn h dung mi
o CH
3
COOH 0,2 M/CH
3
COONa 0,1 M 30
o
C
vi cc thng s k v theo [6] nh sau:
DD = 69%, [] = 0,104.10
-3
M
1,12
(ml/g)
DD = 84%, [] = 0,1424.10
-3
M
0,96
(ml/g)
DD = 91%, [] = 6,589.10
-3
M
0,88
(ml/g)
DD = 100%, [] = 16,80.10
-3
M
0,81
(ml/g)
5. Xc nh deaxetyl ha(DD)
S dng phng php ca A. Baxter v
cng s (phng php ny cho php o c
chitosan c DA t 0 - 55%) xc nh
deaxetyl ha ca chitosan.
Trn ph IR ca chitosan, xc nh din
tch hp th ca cc nh ti s sng xc nh
(khng tnh chiu cao pic) bng cch ly tr s
ti 3 s sng (gm hai tip tuyn v 1 im cc
i). Xc nh A
1655
v A
3450
, tnh deaxetyl
ha (%N-acetylation) theo cng thc 2 [7]:
DA = (A
1655
/ A
3450
) 115 (2)
DD (%) = 100% - DA (%)
III - Kt qu v( tho lun
Qua nghin cu phn ng deaxetyl ha
chitin thnh chitosan trong mi trng kim,
iu kin nhit bnh thng, vi chitin c
qua XLKT v chitin khng qua XLKT, kt qu
c trnh by bng 1.
Bng 1: Kt qu kho st qu trnh eaxetyl ha t chitin thnh chitosan nhit thng
Khi lng phn t trung bnh
M
eaxetyl (xc nh theo IR),
DD, %
TT
Nng
NaOH,
%
Thi
gian
phn
ng, gi
Mu 1
(c XLKT)
Mu 2
(khng XLKT)
Mu 1
(c XLKT)
Mu 2
(khng XLKT)
1 2 3 4 5 6 7
1 20 48 KX KX KX KX
2 30 48 KX KX 61 KX
3 40 48 450 600 465 700 77 63
4 50 48 415 360 420 650 82 67
5 60 48 364 530 390 100 83 68
6 40 0 KX KX KX KX
7 40 6 KX KX KX KX
8 40 12 KX KX KX KX
9 40 18 KX KX KX KX
56
50
55
60
65
70
75
80
85
20 30 40 50 60
Nng NaOH, %
mu 1
mu 2
D
D
,
%
1 2 3 4 5 6 7
10 40 24 KX KX 60 KX
11 40 36 455.600 KX 67 KX
3 40 48 450.600 465.700 77 63
12 40 72 434.350 455.600 84 65
13 40 96 385.500 450.100 90 68
14 40 120 380.300 430.000 95 69
15 40 144 369.200 405.500 97 71
*KX: Khng xc nh c theo phng php o.

Phng php tnh khi lng phn t trung
bnh v deaxetyl ha ca chitin/chitosan p
dng trong bi ny, khng xc nh c vi
sn phm c DD < 45% v khng tan trong h
dung mi CH
3
COOH 0,2M/CH
3
COONa 0,1M
(trong bng 1 cho kt qu khng xc nh c
theo phng php o). Tuy nhin phng php
ny li hu dng vi sn phm c DD > 45% v
tan trong h dung mi CH
3
COOH 0,2 M/
CH
3
COONa 0,1 M.
Kt qu nghin cu cho thy,
- Khi lng phn t trung bnh ca
chitosan gim khi nng NaOH tng v thi
gian phn ng tng, iu ny l hp l v nng
kim cao v thi gian phn ng di s gy
ct mch chitosan nhiu hn. Tuy nhin, khi
lng phn t trung bnh ca chitosan thu c
khc nhau khng ng k gia hai mu c
XLKT v khng XLKT. iu ny cho thy qu
trnh XLKT khng nh hng n khi lng
phn t ca chitin/chitosan.
- Chitin c XLKT chuyn ha thnh
chitosan d dng hn loi chitin khng XLKT
v c th cho DD kh cao (97%). Chitin
khng qua XLKT kh chuyn ha thnh
chitosan trong iu kin phn ng nhit
thng v cu trc mng tinh th ca n bn
vng, kh nng deaxetyl ha thp DD 71%.
Mc deaxetyl ha chitin thnh chitosan
theo nng kim hai mu c trnh by
trn hnh 1.
Chitin khng qua XLKT (mu 2) cng b
kh axetyl thnh chitosan cng iu kin
phn ng, nhng mc thp hn so vi chitin
* qua XLKT (mu 1), mu khng qua
XLKT c DD t t 63% - 68% trong khi mu
qua XLKT c DD bng 77% - 83%.

Hnh 1: Mc deaxetyl ha chitin thnh
chitosan theo nng NaOH
Mu 1: chitosan t chitin * qua XLKT
Mu 2: chitosan t chitin khng qua XLKT
Hnh 2 cho thy s bin i chitin * qua
XLKT thnh chitosan nhit thng theo
nng NaOH.
Kh nng chuyn ha chitin thnh chitosan
tng theo nng NaOH v thi gian phn ng
(DD tng). Vi thi gian 48 gi kho st, nng
NaOH t 30% - 40% th phn ng xy ra
mnh (DD tng t 61% - 77%), nhng khi nng
NaOH tng (60%) th DD tng chm v ch
t n 83%. Mt khc, khi nng NaOH cao
(60%) th khi lng phn t chitosan gim
( M= 364.530) do b ct mch mt phn. Nng
NaOH chng ti la chn l 40% v cho sn
phm chitosan t c c DD = 77%, M =
450.000.
57
60
65
70
75
80
85
30 40 50 60 70
D
D
,
%
50
55
60
65
70
75
80
85
90
95
100
24 36 48 72 96 120 144
mu 1
mu 2
D
D
,
%
Nng NaOH, %
Hnh 2: S bin i chitin c qua XLKT thnh chitosan theo nng NaOH
deaxetyl ha chitin thnh chitosan theo thi gian phn ng hai mu trnh by hnh 3.
Thi gian, gi

Hnh 3: Mc bin i chitin thnh chitosan theo thi gian phn ng nhit phng
Mu 1: chitosan thu c t chitin * qua XLKT; Mu 2: chitosan thu c t chitin khng qua XLKT

Hnh 3 cho thy, deaxetyl ha ca chitin
thnh chitosan tng theo thi gian phn ng
c hai mu nghin cu, tuy nhin DD mu 2
thp hn mu 1. Kt qu bin i chitin thnh
chitosan nhit thng (khng qua XLKT)
trong 6 ngy thu c sn phm c DD = 71%
ph hp vi nghin cu ca Pius Thomas v
cng s [5].
S bin i deaxetyl ha chitin * qua
XLKT thnh chitosan theo thi gian phn ng
nhit thng trong NaOH 40% c trnh
by trn hnh 4.
Chitin trong t nhin c deaxetyl ha
khong 10%, xc nh DD ca chitin
ngi ta phi s dng phng php ph cng
hng t ht nhn
1
H-NMR hay phn tch
nguyn t.... Thng thng, t khi ngi ta xc
nh eaxetyl ha ca chitin, m ch xc
nh eaxetyl ha ca chitosan. Trong
nghin cu ny, chng ti s dng phng php
ph hng ngoi IR xc nh deaxetyl ha
ca chitosan (cho kt qu c sai s thp) [7].
Tuy nhin, phng php ny c nhc im l
khng xc nh c DD ca chitin m ch
xc nh c DD ca chitosan (DA t 0 -
55% hay DD t 45 - 100%). Chnh v vy, trn
hnh 4 th hin s bin i deaxetyl ha ca
chitosan t chitin * qua XLKT theo thi gian
phn ng nhit thng trong NaOH 40%
t 0 - 24 gi c xc nh bi ngoi suy.
48 gi u ca phn ng chuyn ha
chitin (qua XLKT) thnh chitosan nhit
58
0
10
20
30
40
50
60
70
80
90
100
0 50 100 150
D
D
,
%
Thi gian, gi
Hnh 4: S bin i chitin * qua XLKT thnh chitosan theo
thi gian phn ng nhit phng

thng vi nng NaOH bng 40%, DD ca
sn phm tng nhanh (mu 1 c DD = 77%),
sau tc phn ng deaxetyl ha chm li.
iu ny c l gii nh sau: cc nhm -
NHCOCH
3
ban u nhiu v d dng tham gia
phn ng nn tc deaxetyl ha din ra
nhanh. V sau cc nhm -NHCOCH
3
gim dn
v v tr kh tip xc (do mch cao phn t
an xen v cun xon ln ln che lp cc nhm
chc) nn kh b loi hon ton trong phn ng
kh axetyl. iu ny cng ph hp vi vi
phng php deaxetyl ha ca Keisuke Kurita
v cng s [4], khi mun thu c sn phm c
DD cao (> 95%), phn ng phi thc hin
trong iu kin nit v lp i lp li nhiu ln
loi hon ton nhm axetyl cn li trong
mch chitin/chitosan. Chitin * qua XLKT c
mng phn t linh ng nn phn ng xy ra
d dng hn so vi chitin khng qua XLKT,
nhng thu c chitosan c DD cao chng
ti phi lp li qu trnh XLKT nhiu ln v
ko di thi gian phn ng n 144 gi mi thu
c sn phm c eaxetyl ha bng 97%.
c bit, sn phm chitosan sau qu trnh
deaxetyl ha c mu trng v khng b nh
hng bi nhit , khc vi chitosan dng
nhit cao thng c mu vng nht v trng
lng phn t gim. Theo nghin cu ca S. Z.
Rogovina v cng s, qu trnh eaxetyl ha
chitin nhit cao d b oxi ha, sn phm b
vng v khi lng phn t trung bnh gim [8].
Cn qu trnh deaxetyl ha ca chng ti thc
hin nhit thng v chitin c XLKT
cho sn phm chitosan c khi lng phn t
trung bnh gim cht t ( M = 369.200) so vi
chitin khng qua XLKT ( M= 405.500), tuy
nhin s khc bit ny khng ln lm m vn
thu c sn phm c kh axetyl cao.
Ph
13
C-NMR dng rn ca chitosan (hnh
5 v 6) cng th hin deaxetyl ha trong sn
phm. Trn ph
13
C-NMR hnh 5 c cc nh
23,76 ppm (CH
3
) v 174,43 ppm (CO), chng
t trong i phn t chitosan vn cn nhm
axetyl do s deaxetyl ha xy ra khng hon
ton, khc vi ph hnh 6 ca chitosan c
deaxetyl ha cao phn ng xy ra gn nh hon
ton.
IV - Kt lun
Qua kho st ng hc ca phn ng
eaxetyl ha ca chitin thnh chitosan nhit
thng, chng ti * xy dng c quy
trnh sn xut chitosan t cht lng cao, tn t
nng lng v d thc hin Vit Nam.
Sn phm chitosan thu c c eaxetyl
ha nh mong mun. c bit qua qu trnh x
59
Hnh 5: Ph
13
C-NMR ca chitosan c DD = 77%

Hnh 6: Ph
13
C-NMR ca chitosan c DD = 97%

l k thut, c th iu ch chitosan c DD
cao trn 90% m cc phng php thng
thng kh thc hin c.
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st
Century: a review;
Pharma. Sci., Vol. 10, No. 1, P. 5 - 22
(2000).
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Arachchi and You-Jin Jeon. Trends in Food
Sci. & Tech., Vol. 10, P. 37 - 51 (1999).
3. ng Vn Luyn. Chitin/chitosan. Cc bi
ging v bo co chuyn , tp 2, Tr. 27 -
60
35 (1995).
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Tada, Shigeru Ishii, Shin - Ichiro Nishimura
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13, P. 281 - 285 (1991).
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Anthony Taylor, and George A. F. Robets.
Int. J. Biol. Macromol, June, Vol.14, P. 166
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