ng th
nitrofuran trong mt s loi thc ph
si b
sng khi ph LC/MS/MS Trn Th Hng i hc Khoa hc T 
Lu ThS.  ; : 60 44 29
ng dn:  Hng Ho
o v: 2012 Abstract. Gii thiu v ng c
c ph
dnh ng thi 4 cht chuya nitrofuran trong thc
ph  ng bng LC-MS/MS. ng d       t
chuya nitrofuran trong thc ph







 , ,
u kt qu c.

Keywords. ; ; Thc phm

ng tht s loi thc phng
i bng khi ph LC/MS/MS. NG QUAN

1.1. Gii thiu v 
t ng h

 -nitroc s
d ng, ch yi vm,
lnng thy sn (, ) 

u tr 
nhilmonella spp, gia cm
bnh t u [12].
 itrofuran  Furazolidone, furaltadone, furazolidone,
nitrofurazone, 





 








sinh sc nng tt vi vi khung th
ng c vng vt, mt s cht 






[7].


 thp, nitrofuran  ng dit
khun.
Theo nhing rt t





 salmonellosis,
ng. Trn nitrofuran vi thc
 l 

ng nhanh.
Nitrofuran n sc kht bin [7], khi





. , 
















 



 [9u gii hn nh
nht cn thc hii vt chuy
29].




c phm
T  

, 1 





1 



) 





ng 

0  3,55 ng/g. 2008 


1 



















 , :








 , 

 ( ), 
/MS. U

2.ng, mu
ng, mu
t chuya nitrofuran, bao gm: AOZ, AMOZ,

2.2. u
2.2.1. ng khi ph
 u
 
2.2.3. Thnh 


  liu
ng c u
2.3.1. Dng c t b
2.3.1.1. Thit b
- H thng s  ng khi ph khi ph LC/MS/MS bao gm: HPLC 20 AXL ca
i ph ABI 5500 QQQ ca Aplied Biosystem: b ph
lo phu nhit.
- Ct s
18
.
- Ct chit pha rn SPE (Oasis HLB).
- c vortex VELP.
- ng nht mu.
- 
-  
-  c 0,01g).
- 
2
u nhit.
- B u nhit
2.3.1.2 Dng c
- C 
- 

- Amoni acetate (Merck ho tinh khit 98%)
- 2-NBA (Merck ho tinh khit 99%)
- K
2
HPO
4
.3H
2
O (Merck ho tinh khit 99%)
- NaOH (Merck ho tinh khit 99,9%)
- HCl 37% (Merck ho tinh khit 99,9%)
- nh mnh mn vch bc
ct.
- 2--nh mng methanol.
Dung dch chun b h
- K
2
HPO
4

2
HPO
4
.3H
2
nh mng
c ct 2 ln.
- nh mnh mc 1000 ml bc ct
2 ln.
- CH

- Chunh mc bng
nh mc 10 ml.
- Chun SEM 1000 nh mc b
nh mc 10 ml. Bo qun  - 20
0







- Hn hp chun 4 cht 1 ml chun AOZ 10
nh mc bng methanol vnh
mc 10 ml. Bo qun  - 20
0







- Hn hp chun 4 chn hp chun 4
chnh mc bnh mc 10 ml. Bo
qun  - 20
0






- d
5
-
5
-nh mc b
nh mc 10 ml. Bo qun  - 20
0







- d
5
-
5
-nh mc bnh
mc 10 ml. Bo qun  - 20
0










- Hn hp ni chun (d
4
-AOZ, d
5
-AMOZ, d
2
-AHD.HCl,
13
C
15
N
2
-n
n hp ni chun (d
4
-AOZ, d
5
-AMOZ, d
2
-AHD.HCl,
13
C
15
N
2
-SEM.HCl) 1 mg/l
nh mc 10 ml. Bo qun  - 20
0

nh NPAOZ, NPAMOZ, NPAHD, NPSEM bng k thu
n t ESI vi ch  b tu kin khi ph
  ng chu   c dn xut bng 2-
 khn ch  kh i
vi tng cht, tng ion mu kin chy ngu bng 3.1.
Bảng 3.1: Điều kiện chạy nguồn ion hóa ESI

(CUR)
20 psi
m (CAD)
8 psi
Th  (IS)
5000 V
Nhi mao qun (TEM)
400
0
C
u phun (GS1)
50 psi
t ca lu
40 psi
Trong k thun t vi ch  b 
dng (MH)
+


= (M+1) 




d
5
-AMOZ
339
340
13
C-
15
N
2
SEM.HCl
211
212
D
2
-AHD.HCl
250
251
3.1.1.2. Khu kin b





ion con
Detector s d

 i ph 2 l 
chc la chc ion con t quan trng. Ion con phu g
nht 10 ln so vi ion m c mu cao cn phi chc mc

335
291
80
15
16
262
80
21
16
NPSEM
209
192
80
11
18
166
80
13
14
NPAHD
249
134
80
15
18
104
80
27
14
d

251
134
80
15
14




:
Theo qui 



2002/657/EC, 





(IP) 










 [4]. 
, .










3.1.2. La chn c
 ng thi theo khuy
sn xut thit b khi ph  ng s di thit b 
cn 1%, amoni
acetate (0 ts da chn ct
o C18 









:
- Ct C18 :
- 

Gradient 1, 



3.1 :
Th
0,01
12,00
14,00
15,00
20,00
% MeOH
20
90
90
20
20 3.1: 









1 20 ng/ml


2 20 ng/ml
c) 

3, 



3.3 :
Th
0,01
8
10
13
13,01
% MeOH
20
90
90
20
20  3.3: 












4 20 ng/ml
e) 

5, 



3.5: Th
0,01
12,00
13,00
13,01
16,00
% MeOH
20
90
90
20
20  3.5: 












 -NH
2
 -
NH
3
+
. 

, 









 , 

3.1.4. Kht mu
Qua tham kho mt s u [12, 13, 14, 16, 17, 18, 19, 21, 22, 23, 26] t s
c ng d m : quy
 u bng chit lng  l u bng chit pha rn. Do 
t mu 







.
3.1.4.1. La ch u bng chit lng  lng
D ki 




-NBA 10mg/ml



n hp ni chun i ng



ch chuu



2
O:MeOH (60:40)



Chuyn m LC/MS/MS

3.1.4.2. La ch u bng chit pha rn




:





-NBA 10mg/ml



l hn hp ni chun i ng



ch chuu




O



 t SPE



Ra gii 



bng nghim



Thng N
2

tan cn bng 1 ml H
2
O:MeOH (60:40)



Chuyn m LC/MS/MS


AMOZ 20 ng/g
92,7
45,0
AHD 20 ng/g
73,3
33,7
SEM 20 ng/g
70,5
30,6





:  thu h
vt lng lnh c
u tip theo
3.2. 






3.2.1 












 1 



2 ion


4 

, 

 













0,9984

Nhận xét: t l dii chun (IS) trong khong 0,5  5 ng/ml
 h s 
2
> 0,99.
3.2.1.2. Đường chuẩn của các chất nhóm nitrofuran không sử dụng nội chuẩn (để đánh giá
hiệu suất thu hồi)
Bảng 3.8: Phương trình hồi quy của 4 nitrofuran không sử dụng nội chuẩn
TT
Cht
i qui
H s R
1
AOZ

0,9994
2
AMOZ

0,9989
3
AHD

0,9993
4
SEM

0,9993
Nhận xét: Dii n trong khong 0,5   s 



. .
3. lp l thu hi
Thc hiu tht la nitrofuran  c n
i vi mi chng/g; 1,5 ng/g; 2 ng/g. p li 6
ln mi mu.
: 



 80,5  106%, 





(%RSD <
13,7%) 





 2002/657/EC. 









 ng d 
mt s loi thc pht qu  bng 3.16
Bảng 3.16: Kết quả phân tích mẫu thực theo phương pha
́
p thêm chuâ
̉
n

STT
Mu
Nitrofuran



thy (ng)
R%
1
Tht ln ch
Nguyn Cao
AOZ
0
KPH
-
1
0,87
87
AMOZ

AMOZ
0
KPH
-
1
1,02
102
AHD
0
KPH
-
1
0,95
95
SEM
0
KPH
-
1
0,89
89
3
Th Mai
ng
AOZ
0
KPH
-
1
0,99

88
AMOZ
0
KPH
-
1
0,97
97
AHD
0
KPH
-
1
0,75
75
SEM
0
KPH
-
1
0,69
69
5
Tht ln ch Cu
Giy
AOZ
0
KPH

1

1
0,92
92
AMOZ
0
KPH

1
0,88
88
AHD
0
KPH

1
0.79
79
SEM
0
KPH

1
1,06
106
7
Tht ln ch Ngc

AOZ
0
KPH

KPH

1
0,76
76
AMOZ
0
KPH

1
0,82
82
AHD
0
KPH

1
0,97
97
SEM
0
KPH

1
0,73
73
KPH: không phát hiện < LOD của phương pháp,
R %: Phần trăm độ thu hồi.
:
Qua t s mu thn mu














.
n c 
Trong bn luu kin ch  
ng LC/MS/MS trong mt s mu thc phng
(, ,  c m rng
dng nn mu sinh hc, mu sy rt c
nhu ti n 











bng chit pha rn vi ct chit oasis HLB.
3.  cm:
- Khong tuyng t 0,5  i h s 
2

> 0,996.
- Gii h
      u v  nh  




nitrofuran.
-  thu h



80,5  106% cho th  tt.
-  lch chui thp (RSD% < 13,7%) cho th lp li
.
4. ng d u thc phm t 
i, cho th ng d 




nitrofuran trong thc phng.

bi.
8. NguyGiáo trình Các phương pháp tách, i hc
Khoa hc T i hc Qui.
9. S - vic cấm sản xuất, nhập khẩu, lưu
thông và sử dụng một số loại kháng sinh hóa chất trong sản xuất và kinh doanh thức ăn chăn
nuôi ca B 
10. 28 TCN 194:2004,Các chất chuyển hóa thuộc nhóm nitrofuran trong thủy sản và sản
phẩm thủy sản – Phương pháp định lượng bằng sắc ký lỏng khối phổ - khối phổ, B Thy
Sn.
11. Trang web (2008), http//vietbao.vn/Kinh-te/My-kiem-tra-thuy-san-nuoi-Trung-Quoc-
Viet-Nam-lo-ngai/55154852/93. Vi

Ting Anh
12. Alexander Leitner, Peter Zollner, Wolfgang Lindner (2001)  
metabolites of nitrofuran antibiotics in animal tissue by high-performance liquid
chromatographyJournal of Chromatography A, Volume 939,
pp. 49-58.
13. Caroline Douny, Joelle Widart, Edwin De Pauw, Frederic sivestre, Packtrick Kestemont,
Huynh Thi Tu, Nguyen Thanh Phuong (2012), Development of an analytical method to
detect metabolites of nitrofurans: Application to the study of furazolidone elimination in
Vietnamese black tiger shrimp (Penaeus monodon), Aquaculture, Volume 376-379, pp. 54-
58.
14. C. Bock, C. Stachel, P.Gowik (2007), Validation of a confirmatory method for the
determination of residues of four nitrofurans in egg by liquid chromatography-tandem mass
spectrometry with the software interval, Analytica Chimica Acta, Volume 586, pp. 348-358.
15. C. Chang, D.P. Peng, J.E. Wu, Y.L. Wang, Z.H. Yuan (2008), Development of an indirect
competitive ELISA for the detection of furazolidone marker residue in animal edible tissues.
Journal of Agricultural and Food Chemistry, Vol.53, pp 8934-8939.
16. Chung-Wei Tsai, chuan-ho tang and Wei-Hsien wang (2010) 
Determination of Four Nitrofurans and Corresponding Metabolites in the Fish Muscle by

 Veterinarni Medicina, Vol.53, pp. 469500.
26. Pascal Mottier, Seu-Ping Khong, Eric Gremaud, Janique Richoz, Thierry Delatour, Till
Goldmann, Philippe A.Guy (2005), Quantitative determination of four nitrofuran metabolites
in meat by isotope dilution liquid chromatography-electrospray ionization-tandem mass
spectrometry, Journal of chromatography A, Volume 1067, pp. 85-91.
27. P. Vinas, N. Campillo, L. Carrasco, M. Hernandez-Cordoba (2007), Analysis of
nitrofuran residues in animal feed using liquid chromatography and photodiode-array
detection, Chromatographia, Vol.65, pp. 85-89.
28. Randox laboratory (2008) itrofuran metabolites with elisa kits
using multi-Randox life sciences.
29. Sujittra Phongvivat, Bangkok, Thailand (2004),    
, Joint FAO/WHO Technical Workshop on, Residues of Substances without
ADI/MRL in Food.
30. Tao Ding, Jingzhong Xu, Chongyu Shen and Kefei Wang (2007)
level nitrofuran metabolites in crawfish meat by electrospray LC-MS/MS on the TSQ
quantum discovery MAX, Thermo fisher scientific, San Jose, CA, USA.
31. W. Lui, C. Zhao, Y. Zhang, S. Lu, J. Liu, R. Xi (2007), Preparation of polyclonal
antibodies to a derivative of 1-amonihydantoin (AHD) and development of an indirect
competitive ELISA for the detection of nitrofurantoin residue in water. Journal of
Agriculture and Food Chemistry, Vol.55, 6829-6834.
32. 2002/657/EC implementing Council Directive 96/23/EC concerning the performance of
analytical methods and the interpretation of results.