Professional Documents
Culture Documents
inh ng Huy
H Ni - Nm 2009
I HC QUC GIA H NI
TRNG I HC KHOA HC T NHIN
_____________________
inh ng Huy
Chuyn ngnh
: Ho phn tch
M s
: 60 44 29
H Ni - Nm 2009
LI CM N
Bn lun vn ny c thc hin v hon thnh ti Trung tm Cht
lng Nng lm thy sn vng 4, 30 Hm Nghi Qun 1, Thnh ph HCM
vi s hng dn ca GS.TS. Phm Hng Vit.
Vi lng bit n su sc, ti xin trn trng cm n GS.TS. Phm Hng
Vit hng dn v gip ti hon thnh lun vn ny.
Ti xin chn thnh cm n TS. Dng Hng Anh hng dn, gip
ti trong qu trnh nghin cu.
Xin chn thnh cm n Ban lnh o Trung tm Cht lng Nng lm
thy sn vng 4, Tp th phng kim nghim Trung tm vng 4, bn b v
ng nghip quan tm gip ti trong thi gian va qua.
Xin chn thnh cm n Ban lnh o Trng, Khoa Ha hc, cng cc
thy c Trng i hc Khoa hc T nhin tn tnh ch dy v hng dn
trong sut qu trnh hc v lm lun vn.
inh ng Huy
MC LC
M U..........................................................................................................................1
Chng 1 - TNG QUAN...............................................................................................4
1.1. Hin tng thy triu ...................................................................................4
1.2. c t sinh hc bin trong thy sn..................................................................6
1.3. i cng v axt domoic (DA)........................................................................8
1.3.1. Tnh cht ha l. .......................................................................................8
1.3.2. Ngun tch t DA trong nhuyn th: ........................................................8
1.3.3. c tnh ca DA .......................................................................................9
1.4. Mt s phng php phn tch DA ...................................................................9
1.4.1. Phng php sinh ha trn chut............................................................10
1.4.2. Phng php sc k lng (LC-UV, LC-DAD, LC-FLD, LC-MS/MS)..11
1.5. u v nhc im ca cc phng php dn n vic s dng phng
php LC-MS/MS trong phn tch DA.............................................................12
1.5.1. Phng php sinh ha trn chut............................................................12
1.5.2. Phng php sc k lng ........................................................................12
1.6. i cng v sc k lng hiu nng cao ghp khi ph [2], [5] ........................12
1.6.1. Mt s nh ngha v phng trnh c bn .............................................13
1.6.2. Nhng thnh phn c bn ca h thng LC-MS/MS (Waters) ..............15
1.6.3. Loi hp cht ph hp phn tch bng sc k lng ................................23
1.6.4. Cc yu t nh hng n qu trnh tch ca cc cht trong ct............23
1.7. K thut chun b mu cho phn tch sc k...................................................26
1.7.1. Chit lng - lng: ....................................................................................27
1.7.2. Chit pha rn SPE: ..................................................................................27
Chng 2 NI DUNG V PHNG PHP NGHIN CU ..................................29
2.1. Ni dung nghin cu ca ti.......................................................................29
2.2. M hnh thc nghim ......................................................................................29
2.3. Thit b, dng c, ho cht: .............................................................................29
2.3.1. Thit b, dng c: ....................................................................................29
2.3.2. Thuc th, ha cht: ...............................................................................30
2.4. Thng tin v mu nghin cu:.........................................................................30
2.5. Xc nh cc thng s ti u:..........................................................................31
2.5.1. Xc nh cc thng s ti u cho MS .....................................................31
2.5.2. Ct:..........................................................................................................31
2.5.3. Pha ng v ch gradient: ..................................................................32
2.5.4. Dung mi chit:.......................................................................................32
2.5.5. Thit lp bng mu: ................................................................................32
2.5.6. Tnh ton : ...............................................................................................33
2.5.7. Kho st khong tuyn tnh: ...................................................................33
2.5.8. Gii hn pht hin ca phng php: .....................................................33
2.5.9. lp li ca phng php: ...................................................................34
2.5.10. thu hi ca phng php:................................................................34
2.5.11. Thc nghim xc nh DA trn mu nhuyn th..................................35
Chng 3- KT QU V BN LUN........................................................................36
3.1. Xc nh cc thng s ti u:..........................................................................36
3.1.1. Xc nh cc thng s ti u ca MS/MS ..............................................36
i
ii
DANH MC CC CH VIT TT
Ch vit tt
DA
DAD
EU
FA
FLD
Ting Anh
Axt domoic
Diot Array Detector
European
Formic acid
Fluorescence Detector
Ting Vit
Axt Domoic
u d Diot Array
Lin minh Chu u
Axt Formic
u d hunh quang
HPLC
LC-MS/MS
LOD
LOQ
NPLC
ND
RPLC
SPE
TFA
UV
VIS
iii
Sc k lng ghp 2 ln
khi ph
Gii hn pht hin
Gii hn nh lng
Sc k lng pha thun
Khng pht hin
Sc k lng pha o
Chit pha rn
Axt Trifluoro axetic
Cc tm
Nhn thy
DANH MC CC BNG
Bng
Din gii
Trang
iv
DANH MC CC HNH V, TH
Hnh
Din gii
Trang
M U
Theo c tnh ca T chc nng lng (FAO), tng kim ngch xut
nhp khu cc sn phm thy sn trong nm 2008 ca th gii ln u tin
trong lch s vt 100 t USD. Mt na xut khu thy sn trn th gii
bt ngun t cc nc ang pht trin trong khi 80% nhp khu thuc v cc
nc pht trin. Xut khu rng t cc nc ang pht trin t mc 25,4 t
USD trong nm 2008. Cc sn phm t thy sn l mt ngun thu ngoi t
quan trng ti cc nc ang pht trin. Vit Nam, thy sn ngy cng
ng vai tr thit yu vo kim ngch xut khu ca Vit Nam. Sau hn 1 nm
gia nhp WTO, ngnh thy sn Vit Nam c mt bc tin nhy vt trong
cng tc xut khu thy sn, ch trong nm 2007 sn lng thy sn c nc
c t 3,9 triu tn vi kim ngch xut khu 3,75 t USD, trong sn
phm nhuyn th hai mnh v cng chim mt t trng ng k. n nm
2008, kim ngch xut khu thy sn nc ta vt ngng 4 t USD.
Mt trong cc th trng nhp khu ln ca ngnh thy sn Vit Nam
l Lin minh Chu u (EU). Theo quy nh ca y ban lin minh Chu u,
mt nc ngoi khi EU c php xut khu thy sn vo EU phi m
bo cc yu t: (i) H thng vn bn quy pham php lut v nng lc c quan
qun l an cht lng v sinh ton thc phm ca nc xut khu v EU l
tng ng. (ii) H thng phng kim nghim tham gia vo cng tc kim
tra, chng nhn cht lng m bo v sinh an ton thc phm ca nc xut
khu v EU tng ng nhau. (iii) Bt buc phi thc hin cc chng trnh
gim st d lng c hi trong thy sn nui v gim st iu kin m bo
v sinh vng thu hoch nhuyn th 2 mnh v. (iv) ng thi thy sn phi
c phn tch cc ch tiu theo quy nh ca EU trc khi xut khu cng
vi cc i hi nghim ngt v k thut phn tch.
Nh vy, thc hin chng trnh gim st iu kin m bo v sinh
vng thu hoch nhuyn th 2 mnh v l mt iu kin tin quyt gip Vit
Nam c php xut khu nhuyn th hai mnh v vo EU. Hai ni dung lin
1
quan n k thut ng vai tr chnh trong vic thc hin chng trnh ny l
nh danh, phn loi to c (cc loi to c c kh nng sinh c t) v
phn tch c t sinh hc bin (ASP- c t gy mt tr nh, DSP c t gy
tiu chy, PSP c t gy lit c).
Dng tn ti chnh ca c t gy mt tr nh (ASP) l axit Domoic c
cng thc cu to:
Ngnh thy sn hin nay ang rt cn c nhng qui trnh phn tch ph
hp theo yu cu ca cc th trng nhp khu gip c quan chc nng kim
sot cc ha cht c hi ni chung v c bit l cc c t c mi nguy gn
lin vi loi (c t sinh hc bin vi loi nhuyn th hai mnh v, Histamine
i vi h c thu ng). V vy, cn thit phi xy dng qui trnh phn tch
xc nh Axt domoic trong nhuyn nh 2 mnh v. Ngoi mt s nghin
cu ca mt s t chc khoa hc hoc tiu chun nc ngoi vi phng
php c s dng xc nh hm lng Axt domoic bng k thut HPLCUV v LC/MSn, phng php sinh ha trn chut th hin nay Vit nam cha
c tiu chun ring v phng php th cho loi c t ny. V vy vn
nghin cu, ci tin phng php c nghin cu trn th gii c th
p dng xc nh hm lng DA trong nhuyn th hai mnh v, ph hp vi
iu kin ca Vit nam (nn mu phn tch, thit b, ha cht mi trng )
l rt cn thit. N gip cc phng th nghim ng dng thc t gip c quan
chc nng kim sot cht ch d lng c t ny p ng yu cu xut
khu.
Vi khun kh lun vn thc s chuyn ngnh ha phn tch, chng ti
tp trung tm ra iu kin phn tch ti u trn thit b LC-MS/MS hin c ti
2
NSP
(Neurotoxic
Shellfish
Poisoning)
do
cc
loi
edulis), razor clams (Siliqua patula) and ip (Pecten maximus) [Wekell et al.,
1994; Rhodes et al., 1998; Vale and Sampayo, 2001].
DA b tch t trong nhuyn th do c ch n kiu mng lc nn c th
trc tip cc loi to c hoc cc vi sinh vt c hm lng DA b tch t
trong NT2MV, hm lng c t h thng tiu ha ca nhuyn th cao hn
so vi c tht, tc tch t DA trong nhuyn th cng rt khc nhau gia
cc loi nhuyn th [Vale and Sampayo, 2001; Hay et al., 2000], Kh nng
chuyn ha DA trong nhuyn th l rt thp.
Khi nhuyn th b tch t c t sinh hc bin, ta c th nui lu
(nui trong mi trng nc bin sch) th c th lm gim hm lng c
t. V d, theo nghin cu ca (Novaczek, 1992): 50% hm lng DA trong
loi vm xanh c ra gii trong vng 24 h nui lu, trong khi phi mt
86 ngy ra gii 50% DA trong razor clams (Siliqua patula) (Horner et al.,
1993)
1.3.3. c tnh ca DA
LD50 ca DA khong 2,3 mg/kg khi lng c th theo nghin cu ca
Iverson v cc cng s (1989).
Quy nh ca cc th trng nhp khu (Canada, M, Chu u, c...) v
Vit Nam quy nh mc gii hn ti a cho php trong nhuyn th l 20 g
DA/g tht nhuyn th (20ppm), khi kt qu phn tch vt mc trn s b nh
ch vng thu hoch v ly mu tng cng kim tra ch tiu ny.
1.4. Mt s phng php phn tch DA
Tuy nhin ti Vit Nam, hin cha c mt tiu chun hoc phng php
no cp n vic phn tch DA ngoi tr mt s qui trnh t xy dng ca
mt s phng kim nghim.
Sau y l mt s phng php in hnh, ph hp p ng c phn
tch d lng DA trong thy sn v sn phm thy sn:
1.4.1. Phng php sinh ha trn chut
Pha long dung dch chun ASP mt s nng tng dn bng nc
v trng v tim vo mng bng mt s con chut, tim mi con 1 ml cho n
khi xc nh c nng gy cht chut trong khong thi gian t 5 n 7
pht. Lu : kim tra pH ca cc dung dch trc khi tim. pH ca dung dch
phi nm trong khong t 2-4, khng c ln hn 4,5.
T nng c chn, pha long thm hai nng mi bng cch
thm hay bt 1 ml nc v trng so vi th tch ban u. Th d nu 10ml
chun c pha long vi 25ml nc v trng th khi pha thm hai dung dch
mi s dng th tch nc v trng l 24ml v 26ml.
Tim mi nng chun pha long thu c bc trn vo mt
nhm gm 10 chut (10 chut/ mt nng ), mi con chut 1ml dung dch.
Xc nh thi gian gy cht chut tnh t thi im tim xong n thi im
tim chut p ln cui cng.
Lp li qui trnh trn 1 hoc 2 ngy sau vi dung dch chun lm
vic mi c chun b.
Tnh thi gian gy cht trung bnh ca mi nhm 10 chut ng vi mi
pha long. Loi b cc pha long c thi gian gy cht chut trung bnh
nh hn 5 pht hoc ln hn 7 pht. T thi gian gy cht chut thu c, tra
bng Sommers, xc nh hm lng c t tnh theo n v MU trong
1ml dung dch (MU/ml) ng vi mi pha long. Tnh h s chuyn i CF
(h s chuyn i gia lng c t tnh bng g v tnh bng MU) cho mi
pha long bng cch chia nng ca dung dch (g/ml) vi s n v c
t tnh theo MU va tra (MU/ml). n v ca CF s l g/MU
10
vi u d khi ph cu t ra khi ct c a vo u d MS v y cu
t c ion ha, phn mnh v c pht hin da vo thng s m/z. Trn c
s ny nn u d MS c th nh danh v nh lng mt cch r rng,
khng nh v mt hp cht no .
1.5. u v nhc im ca cc phng php dn n vic s dng
phng php LC-MS/MS trong phn tch DA
12
k=
tr tm
(0.2)
tm
Cs
(0.3)
Cm
t
t
n = 5,545 r = 16 r
Wb
Wh
(0.4)
t r'
t r'
N = 5,545 = 16 (0.5)
Wb
Wh
n : tng s a l thuyt
L : chiu di ca ct (mm)
14
L
(0.6)
n
1.6.1.6. H s tch ()
Cn c gi l chn lc, cho bit khong cch gia 2 pic.
=
k2
(0.7)
k1
(0.8)
15
16
18
19
Mao qun
Bung ha hi
Cht lng
B phn phn tch
Bm chn khng
Gia nhit
Khng kh
kh
Kh nebulizer
20
Vng va chm
Kh nebulizer
Block lm bay hi
bng gia nhit
Vng phun in
(plasma)
u kim dn mu
(hiu in th cao)
21
u d
Ion cng hng
Ion khng cng hng
Ngun
Hiu in th mt chiu
v xoay chiu
22
B mt photon
va chm
Ion s cp
Ion th cp
23
25
Dung mi
Kh nng tan
trong nc
Khng phn cc
Hexan
Khng
Iso-octan
Khng
Petroleum ete
Khng
Cyclo-hexan
Khng
Cacbon tetra-hydrochlorit
Khng
Cloroform
Khng
Dicloro metan
Khng
Tetra-hydrofuran
Phn cc
Dietyl ete
Khng
Etyl axetat
Axeton
Axetonitril
Isopropanol
Metanol
Nc
Axt axetic
[A]hc
KA=
(0.9)
[A]n
27
28
Xy dng qui
trnh chy mu
Xy dng
phng php
Xy dng qui
trnh chit mu
Chn dung mi
chit
Kim tra
lp li
Kim tra
thu hi
Tm gii hn
pht hin
Xc nh
c hiu
Phn tch mt
s mu tht
Tn mu
1.
Nghu Bn Tre
2.
3.
S mu
03
Nghu Bn Tre
03
Tu hi
03
Ni ly mu
Giao Thy Nam nh
Tin Hi Thi Bnh
Vn n - Qung Ninh
30
Thi gian ly
mu
20/7; 18/8;
09/11
20/7; 18/8;
09/11
20/7; 17/8;
09/11
Thi gian ly
mu
ip
03
28/7; 18/8;
4.
Phan Thit Bnh Thun
10/11
S anti
03
28/7; 18/8;
5.
Tuy Phong Bnh Thun
10/11
Nghu Bn Tre
03
28/7; 18/8;
6.
Bnh i Bn Tre
10/11
Nghu Bn Tre
03
28/7; 18/8;
7.
Ba Tri Bn Tre
10/11
S huyt
03
28/7; 18/8;
8.
Thnh Ph Bn Tre
10/11
Nghu Bn Tre
03
28/7; 18/8;
9.
Cn Gi - H Ch Minh
10/11
Nghu Bn Tre
03
28/7; 18/8;
10.
Tn Thnh Tin Giang
10/11
S Lng
03
20/7; 17/8;
11.
B La Kin Giang
09/11
Nghu la
03
20/7; 17/8;
12.
Hip Thnh Tr Vinh
09/11
o
Tt c cc mu c bo qun ng t nht -20 C cho n khi x l.
TT
Tn mu
S mu
Ni ly mu
2.5. Xc nh cc thng s ti u:
32
nh nht trong dy chun c din tch pic trung bnh cao gp 10 ln dn tch
pic ca mu trng.
- Trong trng hp mu trng khng c pic ti vng ln cn thi gian
lu ca pic DA th tin hnh xc nh nhy ca phng php bng cch
dng dy mu thm chun (spike) gim dn vi nmg phn tch trong 3
ngy v kim tra nhy ca pic thng qua tnh nng signal-to-noise ratio
ca chng trnh. Gii hn pht hin (LOD) l nng nh nht trong dy
chun c t l pic:nhiu trung bnh ca pic DA t nht gp 3 ln so vi nhiu
nn. Gii hn nh lng (LOQ) l nng nh nht trong dy chun c t l
pic:nhiu trung bnh ca pic gp 10 ln so vi nhiu nn.
2.5.9. lp li ca phng php:
th nghim lp li ca phng php, tin hnh phn tch trong 3
ngy, mi ngy 7 mu nhuyn th thm chun DA 2ppm. Tnh ton lp li
ca kt qu kt qu thu c thng qua lch chun Sr :
Sr =
Trong :
(X
X) 2
n 1
n 1
(0.1)
Sr = lp li
Std: lch chun
xi : kt qu thu c trn mu th i
Rm =
Trong :
C C uns
C spike
(0.2)
c
tnh nh sau: C = xi/n
34
35
Chng 3- KT QU V BN LUN
3.1. Xc nh cc thng s ti u:
Capillary
Cng tn hiu
1.
1 kV
2,93 e5
2.
2 kV
1,00 e6
3.
3 kV
8,13 e5
4.
4 kV
7,1 e5
36
Cone volt
Cng tn hiu
1.
10 V
9,74 e4
2.
20 V
4,78 e5
3.
30 V
4,89 e5
4.
40 V
1,36 e5
5.
50 V
5,81 e5
37
Nng lng
va chm
10 eV
15 eV
20 eV
25 eV
30 eV
35 eV
40 eV
184,51
0
0
52348
0
21304
0
0
38
293,78
0
21573
0
0
0
0
0
- H2O
184
Hnh 17. Cch phn mnh ion ca DA
Kt qu kho st: chng ti chn c cc mnh Ion th cp v cc
iu kin ti u ca Collision energy tng ng nh sau:
Bng 06. Cc Ion th cp v cc iu kin ti u ca nng lng va chm
STT
Ion
1.
184,51
20 eV
2.
219,31
20 eV
3.
247,64
15 eV
4.
266, 25
15 eV
5.
293,78
15 eV
39
A%
B%
10,0
10,0
50,0
90,0
90,0
10,0
90,0
90,0
50,0
10,0
10,0
90,0
40
Tc dng
(mL/pht)
0,400
0,400
0,400
0,400
0,400
0,400
Curve
1
1
1
1
1
1
A%
B%
Tc dng
Curve
(mL/pht)
0,00
50,0
50,0
0.400
5,00
50,0
50,0
0.400
6,00
90,0
10,0
0.400
10,00
90,0
10,0
0.400
10,01
50,0
50,0
0.400
41
42
Curve
1
1
1
1
1
1
A%
B%
Tc dng
Curve
(mL/pht)
0,00
50,0
50,0
0,400
5,00
50,0
50,0
0,400
6,00
90,0
10,0
0,400
10,00
90,0
10,0
0,400
10,01
50,0
50,0
0,400
43
A%
B%
Tc dng
Curve
(mL/pht)
0,00
50,0
50,0
0,400
5,00
50,0
50,0
0,400
6,00
90,0
10,0
0,400
10,00
90,0
10,0
0,400
10,01
50,0
50,0
0,400
44
45
46
2,07 e5
47
Dung mi 2
2,03 e5
Dung mi 3
1,74 e5
H s hi quy
H s gc a
H s gc b
tuyn tnh R2
219,31
0,9984
346,635
-133,465
184,51
0,9846
314,745
-196,319
293,78
0,9972
1733,17
-784,847
266,25
0,9995
16764
-5244,29
247,64
0,9969
1812,02
-902,768
48
Ion
Blank
0.5 ppm
2 ppm
4 ppm
7 ppm
10 ppm
184,51
2,4
7,9
53
144
259
439
266,25
20,7
373
2372
5368
8850
9189
Ngy 2
Ngy 3
2,63
2,61
2.40
2,43
2,93
2,58
2,45
2,58
2,53
2,46
2,25
2,36
2,72
2,73
2,20
2,60
2,42
2,74
2,43
2,61
2,60
2,53
2,59
2,49
SD (1 ngy)
0,12
0,22
0,18
2,53
49
TT
SD (3 ngy)
Ngy 2
0,17
CV%
6,72 %
thu hi (3 ngy)
101,2 %
50
Ngy 3
Cng
Nng
(pht)
pic
(ppm)
1.
5,07
1016
0,37
2.
5,10
1018
0,37
3.
5,10
997
0,37
4.
5,13
933
0,37
5.
4,93
775
0,36
6.
5,10
854
0,36
7.
5,07
852
0,36
8.
4,97
260
0,33
9.
4,70
20
0,31
10.
5,17
811
0,36
11.
5,13
894
0,37
12.
5,13
762
0,36
13.
4,77
39
0,32
14.
5,13
559
0,35
15.
4,80
52
0,32
16.
5,10
808
0,36
17.
5,07
580
0,35
18.
5,03
734
0,36
19.
5,07
739
0,36
STT
51
20.
4,73
28
0,31
21.
5,03
515
0,34
22.
5,03
429
0,34
23.
5,03
376
0,34
24.
5,03
485
0,34
25.
5,07
494
0,34
26.
5,07
482
0,34
27.
4,93
90
0,32
28.
5,03
474
0,34
29.
5,00
444
0,34
30.
5,00
462
0,34
31.
5,07
484
0,34
32.
5,00
560
0,35
33.
5,07
355
0,33
34.
5,13
486
0,34
35.
5,10
277
0,33
36.
5,00
346
0,33
Tn mu
Kt qu
(HPLC-UV)
Kt qu
(LC-MS/MS)
1.
Mu nghu Nam nh
ND
ND
2.
Mu Tu hi Vn n
ND
ND
3.
ND
ND
4.
Mu S lng B La
ND
ND
5.
S huyt Thnh Ph
ND
ND
6.
Mu Spiked 2 ppm
2.12 ppm
2.06 ppm
7.
2.64 ppm
2.53 ppm
8.
Mu CRM
DA: 41 2 ppm
C5 Epiaxt domoic:
3.2 0.3
42.5 ppm
48.12 ppm
DA
53
54
56
A%
B%
(Pht)
Tc dng
Curve
(mL/pht)
0,00
50,0
50,0
0,400
5,00
50,0
50,0
0,400
6,00
90,0
10,0
0,400
10,00
90,0
10,0
0,400
10,01
50,0
50,0
0,400
Axt Domoic
ion s
ion th
Thi gian
in th
Nng
cp
cp
Dwell (s)
cone (V)
lng
(m/z)
(m/z)
312
cone (eV)
184,51
0,1
30
20
266,25
0,1
30
15
59
Th t tim mu:
-Tim cc dch mu xy dng ng chun.
-Tim dch mu trng.
-Tim cc dch mu th nghim.Tnh Hm lng DA trong mu th
nghim.
3.7.6. m bo cht lng
3.7.6.1. nh tnh: Cht cn phn tch c khng nh l c mt khi
cc yu cu v thi gian lu, mc dung sai cng ion tng i p ng
theo ch th 2002/657/EC ca Cng ng Chu u.
3.7.6.2. nh lng:
- T l tn hiu/ nhiu ca cc ion th cp trong mu phi 3:1
- H s hi qui tuyn tnh ca ng chun R2 phi 0.99
3.7.7. Tnh ton kt qu:
3.7.7.1. Tnh t s ion theo phng trnh:
R=
100 A1
(%)
A2
Trong :
R: t s ion (%)
A1: din tch ca cc pic ion th cp c cng thp.
A2: din tch ca cc pic ion th cp c cng cao
3.7.7.2. Dng th h s tn hiu ca dch mu xy dng ng chun
vi nng cht chun b sung theo phng php hi qui tuyn tnh:
Y = ax + b
Trong :
b: tung gc ca ng chun
a: h s gc ca ng chun
3.7.7.4. Tnh hm lng cht cn phn tch trong mu da theo h s tn hiu
ca tng mu phn tch v ng chun.
60
KT LUN
Qua qu trnh nghin cu thc nghim, chng ti hon thnh cc
nhim v ca lun vn ra vi kt qu nh sau:
- Kho st, ti u ha cc iu kin xy dng quy trnh phn tch axt
domoic trong thy sn trn thit b LC-MS/MS: (i) Phng php chit: la
chn phng php chit lng-lng vi dung mi chit l MeOH:H2O t l 1:1.
(ii) Ti u cc thng s lin quan n HPLC: s dng ct C18, kch thc ct
250 x 4.6 mm, kch thc ht 5 m (Merck- Licrocart 250-4 RP-18); la chn
v s dng pha ng l 0.1% FA trong H20 (A) v 0.1%FA trong ACN (B);
tc dng 0,4 ml/pht. (iii) Ti u ha cc thng s ca u d MS/MS:
hiu in th mao qun (capillary = 2 kV), in th cone (cone volt = 30 V),
nng lng va chm (collission energy = 20 eV v 15 eV tng ng vi 02
ion con l 184,51 v 266,25).
- Kho st gi tr s dng ca phng php trn mu tht v mu chun: (i)
Khong tuyn tnh: khong tuyn tnh c nng t 0.5 ppm n 10 ppm (R2
> 0,98). (ii) nhy ca phng php: Gii hn pht hin LOD: 0,5 ppm.
Gii hn nh lng LOQ: 1,5 ppm. (iii) lp li ca phng php: thu
hi ca phng php: 101.2%. lp li ca phng php SD = 0.17 hay CV
= 6.72%.
- phn tch 36 mu nhuyn th ti 12 vng thu hoch nhuyn th ca
Vit Nam.
- Kim i chng: ly xc sut 1 s mu nhuyn th, mu thm chun
(spiked) v mu vt liu chun (CRM).
- Trong khun kh lun vn, vi thi gian v iu kin hn hp nn
chng ti ch mi ch nghin cu trong phm vi l phn tch DA trong nhuyn
th hai mnh v. V vy mong mun ca chng ti l trong thi gian sp n
cn phi m rng nghin cu phng php ny thnh phng php chung c
th p dng phn tch ng thi nhiu cht nhm c t sinh hc bin khc
nh DSP, PSP.
61
62
(NBD-Cl)
and
fluorescence
detection,
Journal
of
63
2. Nhm DSP
2.1 Dinophysis Toxin
2.3 Pectenotoxin
2.4 Yessotoxin
3. Nhm PSP
4. Nhm NSP
1. Kho st capillary
15
Collision Energy
20
25
30
52348
21304
16651
22530
17764
35
40
184
219
19793
13518
11555
247
40379
17217
25038
266
80749
14042
21573
Bng 14. Bng tng hp tn hiu ca cc mnh Ion s cp
294
3.
DA
DA
DA