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XÂY DỰNG QUY TRÌNH ĐỊNH LƯỢNG ĐỒNG THỜI IMIDACLOPRID VÀ AZOXYSTROBIN BẰNG PHƯƠNG PHÁP HPLC

Page 1

KHOA

-

QUY IMIDACLOPRID AZOXYSTROBIN U CAO

Sinh Th.s. MSSV: 12D720401175 7B

2017


KHOA

Qua 5

-

khoa cho em

em

t

trong gia em xin

em

n

,

Ths. Ng

em trong em trong em

khai dung

trong ra

QUY IMIDACLOPRID AZOXYSTROBIN CAO

em ,

em

em

xin nh ng

em

cho C

, Sinh

Sinh Th.s. MSSV: 12D720401175 7B

2017

6

2017


Hai u

imidacloprid tin d

hai ,

azoxystrobin

cao trong

imidacloprid

azoxystrobin

trong .......................................................................................................................i

ra hai

N azoxystrobin

cao (HPLC). RP- C18 (250 x 4,6mm, 45%), trong azoxystrobin l

trong pha

gian 15 0,0048 ppm

acetonitril 250 nm. Gi quan

0,997.

RSD < 2%. V imidacloprid azoxystrobin

n

1.1.

......................................................................................................1

c (55%:

1.2.

...........................................................................................................1 ......................................................................2

hai

pic

imidacloprid trong

0,9978 98-102%

quy

: imidacloprid, azoxystrobin,

u

a imidacloprid

0,048 ppm.

quan azoxystrobin

.................................................................................................1

quy

imidacloprid

hai

ng, HPLC.

nhanh trong

2.1.

.................................................2

2.1.1.

...............................................................2

2.1.2.

.................................................................2

2.1.3. Imidacloprid ...................................................................................................3 ................................................................................................3 2.1.3.

.....................................................................................................3 .....................................................................3

2.1.3.4

.......................................................................4 ...........................................................4

2.1.4. Azoxystrobin ..................................................................................................5 ................................................................................................5 .....................................................................................................5 .....................................................................5 .......................................................................5 .......................................................................................6 ..........6 ....................................................6 .... 6 ...8 2.1.6.3. Ph azoxystrobin. ........................................................................................................8 i


.......9

........................................................................19

.......................................................................................................9

...........................................................................20

.........................................................................................................9

.......................................................21

........................................................9

..............................................................................21

..............................................................................10

.......................................................................................................21

....................................................................................10

.............................................................................................21

............................................................................................11

...............................................................................................21

..................................................................................11

........................................................................................21

..............................................................................................11

....................................................................................................21

...................................................................................................11

.................................................................................................22

..............................................................................11

...............................................................................22

..................................................................................12

......................................................................23

....................................................12 2.2.5.

-

3.3.1. Chu

......................................................................................23

......................................................................12

.............................................23

............................................................13

.................................................................................................23

....................................................................................13

................................................................23

....................................................................................14

.................................................23

.............................................................................14

..............................................................24

...........................................................................................15

.............................................................................25

......................................................................................15

..............................................................................25

....................................................................................15

...........................................................................25

...................................16

.........................................................................................26

.............................................................16

...........27

..................................................................................17

......................................................................................27

................................................................................17

..........................................................................................28

............................................................................19

...................................................................28

..............................................................19

.......................................................29

................................................................19

..................................................................30

ii

iii


...30

DANH

......................................................................................30 4.1.2 K

..............................................................30 ...................................................................30

ACN

:

As

: :

..............................................................31 ..............................................................................35

Acetonitrile

HPLC

:

Nam High Performance Liquid Chromatography cao)

...............................................................35

LC50

:

Lethal concentration (

.....................................................................................35

LD50

:

Lethal dose

..........................................................................35

LOD

:

Limit of quantitation

LOQ

:

Limit of detection

PDA

:

Photo Diode Array

..........................................................................................38 ......................................................39 ......................................................................................40

50% ) 50%)

diod quang)

ppm

:

Part per million (p

Rs

:

Resolution

RSD

:

Relative Standard Deviation

SD

:

Standard Deviation

..........................................................................................43 .................................................................................................45 .....................................................................................................47 .................................................................48

Speak

:

.........................................................................................................48

UV

:

...........................................................................................................49

Vis

:

pic

...........................................................................................50

iv

v


DANH

DANH

2.1.

...............................................................................3

2.2.

zoxystrobin ...............................................................................5

3.

.........................................................................9

2.1.

....................................................4 ...................................................6

3

................................................................18 m

................................................14 D

................31

..............................21 -

......................21

..............................................................32

......................................24

5%:5%). ................................................................32

............................36

...............................................................33

..................................37

..............................................................33 ...............................................................................40

..............................................................34

...........41

..............................................................34

...........41

.........................................36

..............................43

................38

..................................43

....................39 ..................39

8.

..............................44 ......................................44

...42

....................................45

..42

...........45 ...................46

azoxystrobin...................................................................................................................48

vi

vii


1.

2.

1.1. Trong

2.1. 2.1.1.

nguy n m...

t

vai

QUAN

QUAN

h

quan nay, khi

hay cho

bao ng

gian

con

trung mong

d trong azoxystrobin

imidacloprid

mua

ng hai

hay

trong trong

c trong

imidacloprid

VIS,

sinh

azoxystrobin : quang UVcho

cao (HPLC)

HPLC cao

nay

sau khi thu

trong

do cho

,

so

Cao

2.1.2. theo ba

khoa

-

t

:

.

an ch

.

trong hai hai

ng

nhanh,

nguy

thay

imidacloprid

.

sinh

con

trong

(Saed Mousa Diab Ali, 2012).

trong

cao (HPLC). imidacloprid

bao

imidacloprid, dichlopropene, methyl isocyanate, chloropicrin, methyl bromide aldicarb, dazomet metham natri, qua

trong

nhanh

Trong

sau:

t

cao (HPLC)

azoxystrobin

2015).

-

quy

Hormon trong

azoxystrobin mong ra

tan g

ph

1.2. M

trong t

2,4,5-T; 2,4-D;

cao

bao

trong dintrophenols, xianophenols, pentachlorophenol

Paraquat (Saed Mousa Diab Ali, 2012). hai imidacloprid HPLC

- Th

sau: azoxystrobin

detector UV-VIS. quy

h

l

imidacloprid

hay

vi

nhau nhau.

azoxystrobin.

carbamat

benomyl. vi sinh

trong t 1

gian

(Saed Mousa Diab Ali, 2012). 2

t trong


hai

Trong imidacloprid

-

azoxystrobin. Theo

trong

pH

5-11, khi

nh

n

cao, sinh ra

Imidacloprid (Raihanah, 2016)

2.1.3.4.

azoxystrobin

imidacloprid D

0.

m. Theo

ra

, imidacloprid trong

. 2.1:

2.1.3. Imidacloprid 2.1.3.1. Imidacloprid

tin

t

imidacloprid

trong

LD50

trong Pyridin

Clo

cao

sau:

ng Azo 5

LD50 > 5000 mg/kg

da

(Sacramento, 2002).

130 mg/kg

da

[31]

2.1.3.2.

LC50 > 5,33 mg/l

t, run

2.1. chung

-

imidacloprid

imidacloprid

-

Trong

C9H10ClN5O2

-

255,662 g/mol

- Danh IUPAC: N-[1-[(6-chloropyridin-3-yl)methyl]-4,5dihydroimidazol-2-yl]nitramide -

HCBVTV:

-

(Spivastava, 2004).

imidacloprid

Neonicotinoid

tinh

-

1.54 g/cm3

cho 0,06 mg/kg

Imidacloprid

be,

Imidacloprid

o

1.00 x 10-7 mm Hg (20 C) trong tan trong gian

Neonicotinoid nicotin. acetylcholin,

receptor

144 C

-

n

kinh o

-

(ADI) trong (Spivastava, 2004). imidacloprid (Spivastava, 2004)

2.1.3.5.

(Sprivastava, 2004)

-

-

imidacloprid trong

t

2.1.3.3.

-

Imidacloprid bao

cao

g

kinh

qua da ng

0,61 g/l (20oC) o

30

(25 C

tan: Tan trong isopropanol, toluene 20oC

dung 3

do kinh trung

cho

pH 7) dchloromethane,

qua

nhanh trong trong

48 4

.


2.2.

2.1.4. Azoxystrobin (Bursic Vojislava and Lazic Sanja, 2012)

tin

azoxystrobin

2.1.4.1. Azoxystrobin b nh

m t lo i thu c di t n m ph r ng

nhi u

qu

ho t

ch ng l i m t s LD50 > 2000 mg / kg.

c nh nh m

Kh 2.1.4.2.

LD50 > 2000 mg / kg da

LC50

0,38 mg /l trong

da 2.2. -

chung

da

: Azoxystrobin

-

C22H17N3O5 403,4 g/mol

- Danh

da

azoxystrobin

L

cho

azoxystrobin

IUPAC: Methyl (E)-2-[2-[6-(2-cyanophenoxy)pyrimidin-4-

m

(ADI) trong

0-0,2 mg/ kg

(T.Nageswara Rao

(T.Nageswara Rao

2.1.4.5.

2012).

2012).

yl]oxyphenyl]-3-methoxyprop-2-enoate. -

HCBVTV:

-

Azoxystrobin

m

methocyacrylate. strobilurin

methoxyacrylates ty

trong

(Rao Nageswara, 2012)

2.1.4.3. -

tinh

-

1.25 g/cm3 ( 25 trong

-

ra.

tan: Tan trong

- Danh

imidacloprid

azoxystrobin

imidacloprid: Confidor 100 SL, Actador 100WP,

Javidan 100 WP, Anvado 100WP, Conphai 10WP, Kola 700WO, Abamix 1,45SP, dung

hexane, methanol, toluen, acetone, 20oC.

ethyl acetat, acetonitril, dichloromethane, -

th

2.1.5.

116 oC

khi

cao, sinh ra

Aba- plus 100EC

[32]

- Danh

nitrogen oxide (N2O)

azoxystrobin: Amistar Top 250 SC, Amistar Top

325 SC, Mi stop 350 SC, Ohho 3255SC, Neoamistagold 360SC, 400SC, 450SC, 500SC, Ammisdotop 400SC, Dovatop 400SC , Paramax 400SC[34].

2.1.4.4.

a azoxystrobin (Sh.A.Ashorkr, 2006) C

Theo

T.Nageswara Rao

2012

azoxystrobin qua

trong .

sau: 2.1.6. T

trong

2.1.6.1.

ng imidacloprid 1(J.Serb, 2009)

5

6

HPLC


C18 (250 cm x 4,6 mm, 5

-

azoxysrobin

2.1.6.2.

UV-Vis

1 (Bursic Vojilava and Lazic Sanja, 2012):

: acetonitril / 0,01 M dung

- Pha 25%: 75%

phosphate (pH 3.0) -

-

UV-Vis

- Pha -

25oC

-

C18 (250 cm x 4,6 mm, 5

-

1

-

270 nm 20

: acetonitril 80% : n 1

2 (Sacramento, 2002): -

20

-

gian

C18 (25 cm x 4,6 mm, 5

azoxystrobin: 11,07

.

2 (Burket and Sapiests, 2005):

UV-Vis

- Pha -

: acetonitril 30% : 1

-

-

70%

gian

UV-Vis : acetonitril 55% : n

-T -

20

-

RP C18 (25 cm x 4,6 mm, 5

- Pha

270 nm

-

imidacloprid: 5,5

-

20

-

gian

C18 (75 cm x 4,6 mm, 3,5 QuEChERS

- Pha

: acetonitril 20%:

-

C18 (250 cm x 4,6 mm, 5

-

80%

-

t

UV-Vis

- Pha -

: 270 nm

-

20

: acetonitril 70% : acid formic 30% 1

4 (Raihanah, 2004): - Pha

: acetonitril 20% : n

240 nm

-

C18 (25 cm x 4,6 mm, 5 UV-Vis

-

azoxystrobin: 8,3 3 (Rao Nageswara, 2012):

1

-

20 hai

2.1.6.3. Ph 80% trong imidacloprid ng

270 nm

-

t imidacloprid

azoxystrobin.

1

-

45%

1 207 nm

3 (Sprivastava, 2004):

10 (LOQ),

hai

azoxystrobin hai ng

p (LOD), gian),

20%

255 nm

-

-

HPLC

HPLC. N HPLC.

imidacloprid

azoxystrobin hay

trung cho

quan cao,

trong

cao hay

h cho

2015

Cao

trong

trong 7

c

. Trong 8


ra trong

imidacloprid

ph

Trong

azoxystrobin bao .

1-

dung

2.2.

3-

methanol,

Nam (

Cao

2015).

QUAN

CAO

HPLC di trao

pha

cao.

i ion hay

pha

h

(pha

6-

detector

hay

b

(HPTLC) Trong ng

in

2.2.3.1. HPLC bao

hai

cao

HPLC, i ta chia

pha

-

pha trao

ion

pha

trong

hay pha

tan

4 dung

dung ta

3 hai pha

HPLC 4

n

theo

tan ng.

dung

ta

cao (HPLC).

-

-

5-

8-

2.2.2.

-

syringe hay auto sampler).

7trong 2016)

t

cao

4-

2.2.1. Kh

pha

pha

2-

trong ngh

acetonitril,

dung

khi

4

4

2

ng

cho mong

theo kinh ta

i

c pha

pha

ng

tinh

t

nh. dung cho HPLC cho HPLC hay dung tinh

ghi pha

.

pha

0,2

hay

2013)

dung

m

qua Duy

100%. Tuy

pha

nhau.

o.

HPLC (

cao theo

ng dung

cho

p ion.

2.2.3.

dung m 1

ra

tinh 0,45

peak

c trong

2.2.3.2. i trong dung pha

trong

dung sau

-

pha

a peak thay

pha

y ra: dung

cho

i gian

i.

- Trong Pump

dung

e) khi

ng. Trong 2.3:

c

cho

HPLC 9

10

sai.


pha

t tan ra t (0

pha 0,5 2

m trong

ghi

+ Trong

cao

2.2.3.3.

qua

200 bar)

gian

ra

peak,

+

cho

peak trong

ng

theo

2.2.3.4. 2.2.3.8.

theo dung

a loop

5

100

2

trong

ng syringe)

in

Sau khi

li xong

pha

coi

tim

pha trong 1- 10 mm,

5 - 10

u

cao.

ng 10

30 cm,

tan trong

do

ra

theo

ra

Pha

(trong

t). c silan

USP 23, 24

ta

di

nhau

oxid, polyme

pha

Pha

qua

pha

theo pha

pha

nhau,

silicagel (pha (pha

bao

2016)

cho qua pha trong hay

tan

silicagel

in

trong

2.2.4.

2.2.3.5.

ra

ta

(auto sampler).

tan.

di cho

r

nh trao

Pha

theo

t

ion. cao

p

4

trao

trong

Trong .

T

2.2.5.

2.2.3.6. (hay cho

detector)

khi detector

-

chia

2

pha

Pha

theo

ghi

Thu, 2007)

pha

pha

g detector

cao khi

2.2.3.7.

pha

theo ta

An

cao

ra

ghi trong

nay, detector

ion

trong

cho

-

khi

quang cho

hay UV-Vis ko

Pha

pha

bao

mang silica 3 Pha

10

th

50 n theo

trong

ghi do detector 11

sang.

pha

Pha

c 12

h

mang

trong pha


-

Trong pha silica qua

silanol.

pha Pha Pha

trong ta dung

dung

thay

pha 2

pha

-

hay

dung

pha

thay

pha

,

ta chia

pha

pha

bao

pha pentan, hexan,

pha

Dung m

p

pha

pha

hydrocacbon pha

i

theo

2.4.

trong -

pha

pha tan

pha

2.2.6.2.

pha trong

trong dung

Dung

methanol, trong

pha

tan trong pha

tan qua pha

gian

Cs

mol

tan trong pha

CM

mol

ch tan trong pha

K ra

khi

cho

n khi

di

gian gian

K

(mol/l)

pha

pha

tan qua pha

g

nhau

h

tan.

K

trong ra

ng

n.

nhau t.

2.2.6.3.

gian

dung

dung

cho

trong hai

pha

pha

tan trong pha

pha

tan trong pha trong

tan. Trong

gian

pH

t

Trong hao gian

pha

Trong

peak

1-5

a ta

13

t R t0 t0

tR

gian dung

.

K'

.

ut

peak thay

(mol/l)

tR gian

theo

CS CM

An, 2007)

trong HPLC

gian

peak

K:

K

quan Trong

t

nhau gian

B

o.

2.2.6. 2.2.6.1.

A

K di

. pha

i gian

tR 14


2.2.6.4. (hay

gian

g

hai

A

RS

B)

hai

Trong hai

tRA, tRB:

hai

KA KB

A, B

tRA tRB

gian

Trong

nhau

1

2.2.6.5. trong trong H.

pha

c u

hai

pha

ng.

trung

t

B

ra

nhau

ta

AF Trong

a: n

(

sau peak

a

AF

b

1/10

1, peak

2

5,54

tR W0,5

peak. trong

2500

-

a

-

AS

5500

2.2.7.

ra hai peak

b

1/20

nhau

peak

peak trong

AS

1,5).

cho m:

Pha

sau: 15

0,8 - 1,5 (0,8

p ta bao

nhau theo

cao peak 1, peak

-

RS

g

a b 2a

As

gian

W0,5

cho.

cao peak)

AS

N

2.2.6.6.

b a peak

b: n

sau:

N

:

AF:

nh.

Trong

ra

pic pic

cao H

tR

RS hai peak

quan

trong

Trong

hai

t

pha

i

ra xa nhau, Do

H pha

theo

A

o

khu n

peak hai peak

cho (pha

2.2.6.7. thu

pic

RS = 1,5

R

hai

trong

nhau

peak

xa dung

nhau

tan trong pha

hai peak

RS = 1,0. Trong

nhau

nhau

cao

t RB t RA WB WA

. WB, WA:

KA KB

2

16


pha

-

pha

-

t.

Pha

u sau:

- Pha

ng:

pha

( pH,

isocratic. -

u

ch

pH,

- Pha

i

tan khi thay

phosphat

ngay

pha

hay

n

ta

pha

pha

-

l ta pha

2

pha

(NP)

ng (NP): Silicagel trung hay trung .

c OH

pha

alkyl

Pha

OH

ng (C8 hay C18 phenyl).

2.2.7.2.

thay

H

RP-8 hay RP-18) hay hay

c

pha

tinh

t

c

Trong pha ACN, MeOH, acid hay base

dung

trong

pha

(RP u

trong

RP- HPLC ra

ion carbon t t.

cho HPLC,

quang.

dung

ion.

alkyl

u.

amin hay acid amin -

c,

gian: gia

trong

dung

t

o (RP): Silicagel

m

i detector: detector UV-Vis dung acid acetic c < 220 nm). Detector

kinh HPLC), pha

c.

pha

carbon (phenyl-

quang

g

trong t.

cao: dung

nhanh

quang

(RP).

Pha

tinh

-

o

hay pha

theo trong

pha trong

dung

ACN hay MeOH

- Pha

u

pha

trong

dung

i gradient. 2.2.7.1.

cho pha

pH: 2,5 < pH < 8,5).

Khi

dung

pH. ta

v

E

dung

theo

sau: 2.3.

dung

trong u t

ta trong

pha

Dung

cho peak

i gian

dung

u,

. Pha

+ +

gian

+

n-Hexan

0,1

Tetrachlorua carbon

1,6

Toluen

2,4

Methanol

5,11

Acetonitril

10,20

gian thay

c

E

i:

t. i.

+ +

th t trong

peak.

Do

trong

pha

ta

pha

khi

p t

ta 17

18

nhanh

ta


cao.

c

tuy t

trong

gian, kinh

2.2.8.4.

. nhau 2012)

2.2.8. 2.2.8.1.

c

HPLC

-

theo

t,

cho

dung theo

dung khi cho

theo pha

c

ra. u

C

tra

theo

quy S

pha

c.

pha ch.

cho

hay khi

nghi

sau

dung

pha

ng a

NP-HPLC: R

RP-HPLC: khi pha l ACN hay methanol:

methanol; 50%: 50% cho

i trong

khi 100% sau

gian

c, N

dung

2.2.8.2. dung

pha

cho

tinh

tinh dung

t

theo

gian theo HPLC

2.2.8.3. X

theo

- Dung

, quy

tan

dung

pha

hay - Ph ra

h cho HPLC.

Pha dung

tan trong pha

theo

c sau:

trong

u

tan u. tan trong pha

t ly

qua

c 0,2

0,45 t.

t.

Pha dung

trong

dung ra

19

c sau: trong

t.

-

do

u

HPLC. Tuy

khi

20


3.

PH

STT

3.1.

Glassico

India

8

Neuburger

Germany

9

S100H Elmasonic

Germany

10

Balances and Precious USA Metal scales

7

dung

3.1.1. 3.1.1.1.

n

-

imidacloprid

azoxystrobin.

-

Sigma Aldrish

-

g: imidacloprid: 99,82% azoxystrobin: 99,60%

3.1.1.2. Dung 3.1. Danh m

11 dung

tinh

cho HPLC

cho HPLC 3.1.2.2.

STT

dung

1

Acetonitril

cho HPLC

2

Methanol

cho HPLC

-

3

hai

T

tinh 50 ml, 100 ml.

-

tia.

-

cho HPLC

vial

3.2. 3.1.2.

100 ml, 50 ml, 25 ml, 10 ml. 1 ml, 2 ml, 5 ml, 10 ml, 20 ml, 25 ml.

- Pipet

DUNG

-

imidacloprid

3.1.2.1

cao

3.2. Danh

-

cho -

STT 1

cao

2

cho dung

Pump

Germany

Vacuum Degasser

Germany

pha

pha

quy

3

Syringe

Hamilton

-

4

Injector

USA

-

5

Columns

Germany

hi nh

peak

-

RP-18, 250x4,6 mm,

6

ghi

azoxystrobin

sau:

UV/Vis Detector 21

Germany 22


imidacloprid

azoxystrobin

3.3. 3.3.1.

-

dung dung

pha

- Qua tham

-

, ,

ra dung

-

pha dung

pha

.

pha

dung c

RP

azoxystrobin,

i gian: 0

1

3.3.2.4.

Qua tham

ra dung

dung ra

ng

xu

pha

C8

C18

Tuy

m

danh

trong kinh

nay, p

m

so

tra,

ng.

ta

250 x 4,6 mm cho

RP

3.3.2.2.

18,

quan Do

u

detector UV-Vis

-

ng HPLC

imidacloprid

ta

t

: RP

18, 5

azoxystrobin

250 x 4,6 mm. 50 ppm.

1 C: acetonitril (CH3CN); D: n gian: 0 -20

3.3.

c

= 250 nm

thay

n pha

pha

acetonitril

Sau

HPLC

ng 220 nm, 250 nm, 255 nm, 260nm. UV-VIS SHIMADZU cho

hai

200-400nm

imidacloprid

thu

ra

(%)

Acetonitril (%) 95

10

90

5

85

15

80

20

azoxystrobin. tham

p 3.3.2.3.

nh

HPLC c 3.3.2.3 ta 23

t

20

pha sau:

ta

tham

gian

pha

- Detector: UV- Vis

i

c a

thay

nhau:

- Pha

g

thay

nhau

-

cho detector

Trong

thay

-

.

a HPLC, pha

pha kh

-

trang

ra pha

Khi pha thay

dung qua

ng

Trong

cho

2013)

t

t RP 18

ng

pha

pha

cho

t5

220 nm, 250 nm, 255 nm, 260nm. 20

.

3.3.2.1.

20 55%C: 45%D. 50 ppm.

-

dung

sau:

18, 5 m, 250 x4,6 mm.

- Detector: UV- Vis

dung imidacloprid

3.3.2.

HPLC

C: acetonitril (CH3CN); D:

-

cho

n

dung

t

- Pha

50 ppm

24

n

(gradient)

ng


(%)

Acetonitril (%)

peak (S), 60

40

55

45

50

50

40

(N) khi

6

gian

-

peak

-

3.3.2.5.

tan gian

peak, cho

pha

peak

trong

ra

0,8 - 1,5 (0,8

peak

trong

RP

50

dung

c

. 1

1,2

pha

tan

hay pha

dung

n imidacloprid

dung

azoxystrobin hai

.

:

= 250 nm. -

imidacloprid v

(ICH, 1996)

3.3.3.

1,5).

theo quy

dung

t; 0,8

- Detector: UV- Vis 20 -

AS

trong quy

sau 5

- Pha C: acetonitril (CH3CN); D: gian: 0 20 55%C: 45%D. 0,5

2,0%

3.3.3.2.

nhau.

18, 250 x 4,6 mm,

t

peak

1,5 (RS 1,5).

quy

ra

-

n

nhau

peak

-

1,0%

chung

HPLC quy

pha Tuy

(AS),

RSD > 2%, trong

C

pha

-

(RS),

2000.

RSD

-

(tR),

. t

-

60

gian

n

minh quy

u

Khi

t

y

-

pic

trong

sau dung

trong

-

y

2009)

3.3.3.1

6 25

ta

theo quy

pha

ng

.

tan

imidacloprid

dung

azoxystrobin hai

:

azoxystrobin cho

ch

hay pha

dung

dung n imidacloprid 50 ppm,

pic

2009) trong

tin

gian trong

gian ch

quy

sai

cho peak

dung

gian 1,0%, so

quy

azoxystrobin

gian

imidacloprid 26

azoxystrobin


Ghi

gian

peak

tinh

trong

x

- Ghi

.

peak trung

u.

Microsoft Excel 2003 v

quy

pic theo -

trong dung

cho peak peak

t

: peak

nhau

dung gian

gian 1,0%, so

0,997 (hay R2

ng quan (r)

nhau

i

gian

trong peak

trong peak

0,995).

r < 0,997

3.3.3.5.

peak

trung gian.

trong

- Peak

tinh

3.3.3.3.

LOD v

Trong quy (LOQ) hai

a ph

LOQ

c a h th ng s c

(LOD) quan

tuy n

h th ng s c K t qu

(LOD) cho h

c kh o

m t m u chu n h n h p imidacloprid

so

hay

v i

c peak

(RSD)

b ng

l pl i6l n

azoxystrobin

n ng

u ki n t i qua

S peak

c kh o

l ch chu n (SD), gian tR. RSD

(LOQ)

xem

RSD

trong 3 LOQ

5-7

- Pha 10 ppm -T

ta

imdacloprid dung

azoxystrobin RSD

pha

RSD 6 2,0%.

suy ra

3

ba pha ba dung

LOQ =

trong

80%, 100%

dung quy suy ra

10 Ã&#x2014; LOD 3

quan

theo

R%

quy

theo

hay

khi

20-160% i5

Mt 100 Ms

Trong

trong g

40%, 80%, 100%, 120%, 160%),

n

pha 5 3

- Ms : N

khi pha ( g/ml)

- Mt: N

( g/ml)

quy

cho -

HPLC

3.3.2 27

-

thu

6

trong

khi pha

ra

120% So

3.3.3.4.

dung

nhau

%)

3.3.3.6.

ghi k cao peak LOD

-

2,0%.

sau:

chu

HPLC

i

trung gian

hay LOD

m c 3.3.2. l ch chu n

2%,

so tr

n m trong kho ng

: 98,0% 102% cho quy 28


2,0 %

- RSD

4.

s

4.1.

3.3.4.

4.1.1.

dung

trong

Theo gian

dung

cao peak

quan

tan

Trong ta

quan

sau:

Hi = k1 . Ci = f(C)

(1)

Si = k2 . Ci = f(C)

(2)

pha

u

cao

n

Ci

peak

i

gian

Dung 10 mg, cho

i.

10 ml acetonitril,

ml,

400 ppm.

6 ml dung

- Dung xi

thu

qua

i 1

x1 x

imidacloprid Dung

2

n 1

RSD %

s .100 x

-T -L

khi

azoxystrobin.

p:

1 ml dung azoxystrobin

i 1

dung

0,45 50 ppm.

Dung -

SD

cho

acetonitrile

n

n

imidacloprid

pha

- H

theo p

1 n

ch

Dung

(1) (2) ta hay x

hai

azoxystrobin. Dung - Dung dung

tRi.

25 ml.

10

acetonitril

t pha

trung

dung

-

k1 , k2

a

dung

trong pha

Si

trong acetonitril

acetonitril dung

Trong Hi

pha

dung

imidacloprid 50 ml.

5 ml dung

acetonitril qua

0,45

imidacloprid

khi

azoxystrobin

Dung 50 ppm.

4.1.2 K Sau khi

theo

3.3.2, ta thu

sau: cho detector

4.1.2.1.

detector

quan nh sao cho

ta

200-400 nm sau: 29

30

thu

50


1.500

azoxystrobin

1.000

Abs.

imidacloprid 0.500

0.000

-0.500 200.00

4.1:

250.00

th

4.1 cho

300.00 nm.

a azoxystrobin

350.00

imidacloprid trong acetonitrile.

azoxystrobin

thu

imidacloprid

c

270 nm. Hai

nhau hai

400.00

250 nm.

207 nm, thu giao

ACN/

(90%:10%).

4.3.

ACN/

(95%:5%).

pha

4.1.2.2. pha 3.3.2 imidacloprid dung

4.2.

250 nm

thay azoxystrobin

c pha pha

pha

Dung dung

chu c 400 ppm.

h

acetonitril

nhau. pha trong

c 3.3.2.4.

t

sau:

31

32


4.4.

4.5.

ACN/

ACN/

(85%:15%) 4.6.

ACN/

h 4.7.

ACN/

(55%:45%).

(60%:40%). (50%:50%).

4.2; 3.3; 4.4; 4.5; 4.6; 4.7, ta

pha

gian ACN

(80%-95%)

pic

gian , peak

nhau cao

nhau, .

, n.

peak imidacloprid

pha nhau (55%:45%) cho 33

a peak. hai

h dung

ACN theo.

34

ra khi ta

t

ng


4.1.2.3 c imidacloprid

azoxystrobin

nhau:

0,5 0,8 1 ml 1,2 1,5 sau: -

t

RP

18,

-

250 x 4,6 mm.

u: 20

t 50 ppm - Detector UV-Vis: 250 nm. - Pha

pha

3.3.2.4. theo

c 3.3.2.5. Theo

ta

qua 4.8:

gian -

0,6

0,8

cho

i gian

4.1.

n, peak

ng

imidacloprid

nhau -

1

tR

cho

nhau

,

-

Do

ta

4.2.

hao gian d

dung peak

1

ng

NG

4.2.1.

As

Rs imi

N

1

3,954

626144

1,140

0,941

0,00

39194,789

2

3,949

634100

1,126

0,955

0,00

39100,101

3

3,969

638942

1,138

0,967

0,00

39103,700

4

3,947

642210

1,156

0,942

0,00

38679,280

5

3,974

643648

1,162

0,951

0,00

39004,980

6

3,960

660550

1,128

0,960

0,00

38243,829

Trung

3,957

640932

1,142

0,953

0,00

38887,780

%RSD

0,309

1,797

1,282

0,011

0,00

0,933

SD

0,012

11517

0,015

0,0101

0,00

362,722

ra

imidacloprid.

IMIDACLOPRID

x

.

(> 1

azoxystrobin

Speak

HAI AZOXYSTROBIN quy

4.2.1.1. Pha dung

azoxystrobin 6

35

imidacloprid

50 ppm.

36


4.2.

ng azoxystrobin Speak

tR

x

As

Rs azo

4.2.1.2 N

1

12,058

1805578

1,013

2,012

2,056

76407,704

2

12,081

1814130

1,009

2,212

2,059

75540,833

3

12,165

1824734

1,007

2,411

2,065

75960,530

4

12,072

1832351

1,021

2,254

2,059

76049,748

5

12,215

1835474

1,014

2,211

2,074

76593,872

6

12,144

1828874

0,999

2,231

2,067

73276,171

Trung

12,122

1823524

1,010

1,889

2,063

75638,143

%RSD

0,513

0,631

0,731

21,12

0,315

1,605

SD

0,012

11498

0,0073

0,399

0.0065

1213,992

hai gian (AS)

(tR), trong

4.1

4.2 cho

peak (S), ng 0,8-1,5,

4.9:

2%, (RS)

imidacloprid

azoxystrobin

cho

1,5, -

theo

pha

gian c

dung

imidacloprid azoxystrobin. - Trong

pha

azoxystrobin trong trong

dung

a

hai peak

gian

imidacloprid

pic azoxystrobin.

1 peak

uk

gian imidacloprid

pic c

a imidacloprid

imidacloprid gian

nhau

azoxystrobin n n trong s

peak

peak azoxystrobin

pic

peak imidacloprid

2,606 trong azoxystrobin

azoxystrobin:

. Peak imidacloprid

to cho

dung azoxystrobin

rang

hai peak imidacloprid gian

3,959

37

12,210 peak

38


gian

imidacloprid

-

azoxystrobin

,

cho hai peak

azoxystrobin

imidacloprid

g

dung

T

a imidacloprid

- LOD peak

suy ra:

gian

- LOQ

a azoxystrobin =

10 10 LOD = x 0,048 = 0,16 ppm. 3 3

- LOQ

a imidacloprid =

10 10 LOD = x 0,0048 = 0,016 ppm. 3 3

quy

4.2.1.3.

LOD

LOQ

quy Trong (LOQ) hai

h quan

LOD chu dung

4.2.1.4. a imidacloprid

ta

imdacloprid pha

cao peak = 3

(LOD)

ng.

LOQ

sau: pha

azoxystrobin

5

10 ppm

Sau y

pha

t

dung

n

khi

pha

LOD

n

- Pha

dung

imidacloprid

50 ml

1,6 ppm:

- Pha

azoxystrobin

0,048 ppm

sau: 0,4

azoxystrobin 40 ml dung

ng acetonitril cho dung 10 ml

cho 4.3: khi

n

40%, 80%, 100%, 120%, 160%.

sau:

50 ml,

p imidacloprid

azoxystrobin

0,4-1,6 ppm

ppm; 0,8 ppm; 1 ppm; 1,2 ppm; 1,6 ppm

HPLC.

cao

7

2 ppm

4.10:

0,0048 ppm.

pha

ch 1,6 ppm

ch

thu ng acetonitril.

dung HPLC

Vx ml sau:

imidacloprid

azoxystrobin

C(ppm)

0,4

0,8

1

1,2

1,6

Vx ml

2,5

5

6,25

7,5

10

Vacetonitrile

c

Sau

cho

.

n

HPLC

c

sau:

-

t

RP

18, 250 x 4,6 mm,

- Pha

C: ACN 55%; D: N

c: 45%

pha ng: 1 - Detector UV-Vis: = 250 nm. 4.11.

imidacloprid 4.10

imidacloprid - LOD

0,0048 ppm a azoxystrobin

azoxystrobin

4.11 cho

khi

cao = 3 0,048 ppm 39

u: 20

0,0048 ppm azoxystrobin

n

0,048 ppm

pic S

C

.

pic S thu

quy

n

n.

Do

ra trong ghi

a

Microsoft Excel 2003 40

ng,

, quy.


4.4.

peak

imidacloprid trong

Imidacloprid C (ppm)

Speak

0,4

81789

0,8

152152

1

226200

1,2

291980

1,6

368627

4.5.

peak

azoxystrobin trong

4.12.

peak

imidacloprid

4.13.

peak

azoxystrobin

ch

azoxystrobin C (ppm)

Speak

0,4

185609

0,8

357070

1

506195

1,2

670685

1,6

870305

ta trong

41

4.12, 4.13

4.4

quy sau 4.5.

42


4.6.

quy

azoxystrobin

imidacloprid

4.8.

th gian

quy

quan R2

(ppm)

HPLC

u imidaclopid

imidacloprid

peak

imidacloprid

TT gian

imidacloprid

R2=0,9978

Y=74105x

azoxystrobin

0,4-1,6

2

Y=171865x

R =0,997

Qua

ta

imidacloprid

0,4

1,6 ppm. Do

hay

3,964

trung

2360311

Xtb=3,959

Xtb=2383429,5

quan hai t

3

3,971

4

3,966

imidacloprid trong

5

3,973

6

3,922

2339617

SD=0,0188

2375992

SD=34611,11

ta

ng azoxystrobin

u

2

2375675

trung

peak

c 3.3.3.4.

azoxystrobin

3,959

0,4-1,6

nhau cao

1

.

2419586

RSD(%)=0,47%

2429396

RSD(%)=1,45%

4.2.1.5. Theo p6 p azoxystrobin u

c 3.3.2. (RSD)

imidacloprid

peak

4.7.

qua Speak

HPLC gian

gian

peak

2

12,077

3

12,132

4

12,167

5

12,205

6

12,311

trung Xtb=12,162

6472807

G

6535964

Xtb=6495348.67

6419486

peak 1

2

3

1

2367836

2341037

2352453

2

2301897

2401004

2390451

3

2401211

2390312

2385421

4

2315012

2352789

2410207

5

2342202

2321107

2331652

SD=60348.26

6

2296789

2321775

2314473

Trung

2337491

2354671

2364109

2352090

1,75%

1,44%

1,57%

1,59%

RSD 6592855

43

imidacloprid

trung

6475006

RSD(%)=0,72%

trung gian

azoxystrobin

6475974

SD=0,088

imidacloprid

trung gian 4.9.

gian 12,081

n pic 0,92 % 3.3.3.5. Quy .

(SD), tR .

TT

1

ta

HPLC

u azoxystrobin

azoxystrobin

ng, azoxystrobin

RSD(%)=0,92%

44

1,45 %


4.10.

trung gian

azoxystrobin

2455806

250

248,79

101,00

2505612

250

254,83

98,12

3

2502276

250

254,13

98,25

2927766

300

306.12

98,00

2865312

300

298,53

100,30

2887210

300

301,20

99,60

peak 1

100%

2

1

6399178

6417754

6475441

2

6400153

6438765

6390435

3

6403347

6443221

6485421

4

6421182

6432119

6410379

5

6501122

6421277

6478553

6

6467781

6456423

6455763

120%

ng 4.12.

s Azoxystrobin pic

(%) Trung RSD

%

6432127

6432927

6449332

6438128

x

0,66%

0,23%

0,61%

0,5%

4860132

200

202,03

98,98

4788941

200

201,51

99,25

4888295

200

203,50

98,28

5804669

250

250,52

99,79

5898400

250

259,31

98,41

5870421

250

253,89

98,46

6779319

300

300,51

99,82

6698359

300

296,36

101,22

6700758

300

296,48

101,48

thu

theo

cao (RSD

4.9 peak

4.10 cho

80%

quy

2%).

quy

hai

imidacloprid

azoxystrobin

HPLC

cao, do

100%

quy 4.2.1.6. 4.11.

Imidacloprid

120%

Imidacloprid

t imidacloprid

pic

(%) x

80%

i azoxystrobin

quy

3

azoxystrobin

trong

trong

4.9

4.10 c

ng 98-102%

% quy

2050270

200

199,50

101,00

2082793

200

203,46

98,29

2054821

200

200,06

99,97

45

quy

ng imidacloprid

HPLC

azoxystrobin t

46


4.3.

5. HPLC:

5.1. K trang

Tuy

cao, dung

qu

HPLC

so khi

.

u

trong

cao: t cho

-

,

trong

azoxystrobin trong h cao cao.

-

sai

HPLC

Nam. Nam

Trong s

RP

18; 250 x4,6 mm;

- Pha

acetonitril:

-T

g: 1

-

imidacloprid

(55%:45%)

20 c

g: 250 nm sau: C

cho chung

hai

RSD

quy

chi

Imidacloprid

1,45%

0,92% nh

quan )

quy trong

0,997 (hay R2 0,995) 98-102% quy

hai trong xu , trong

5.1: azoxystrobin

. Trong g

tham hai

azoxystrobin

. gian,

.

azoxystrobin

trong

peak

trong .

c, dung

RSD

hai

azoxystrobin.

t,

cho peak imidacloprid

Qua

Quy

peak

gian

azoxystrobin

HPLC

LOD, LQD

IV

quy

cao, .

n

cho 2

imidacloprid

imidacloprid

azoxystrobin

LOD (ppm)

0,0048

0,048

LOQ (ppm)

0,0016

0,016

Y=74105x

Y=171865x

hai . Do

5

cho

quy

imidacloprid

c detector UV-Vis.

- Th

HPLC

. Do .

trong

imidacloprid

trong h

t. gian, dung

-T

azoxystrobin n sau:

ch

:

-

imidacloprid cho

cao

quy quan

2

R =0,9978

R2=0,997

0,4-1,6

0,4-1,6 ,

trang 47

cao

, dung 48


, quy

,

THAM

5.2. thu t

ta

gp HPLC cao, imidacloprid azoxystrobin

cho HPLC

trong

.

c ra

Thay

thay

hai

dung

Thay

imdacloprid

azoxystrobin

Y 7

(2009),

2.

sau: gian

1.

hao

dung

Minh

dung 3.

t do

HPLC

h

Hoa (2013).

chung cho

g trong quy

h trong

u

quy

Dung (2014).

4.

Hi trong trong

(2014).

quy

Y

dung

RP - HPLC

ASEAN v 22/2009/TT-BYT Quy

trong

5.

(2015).

fluconazol

.L

ra. 6.

.

Minh. Duy Cao

quang

UV VIS

(2015).

trong

s

8.

An

9. 10.

An

(2007).

. NXB Y Thu (2007). . NXB Khoa

(1995).

11.

(2016).HPLCY

12. trong

TP.

(2003).

cao (HPLC) 7.

Y

,

II, NXB Y , , .

trong

.

Minh. (2013). s

Calci Atorvastatin cao.

Simvastatin

ANH 13. BP 2010. Validation of Analytical Procedures, Supplementary Chapter III E. Printed in the United Kingdom by The Stationery Office, N5977690 C34 8/2009. 14. ICH Topic Q2B (1996). Validation of Analytical Procedures: Text and Methodology. 15. Ferrer Imma. et al (2005). Multi-residue pestiside analysis in fruits and vegetables by liquid Chromatography- time of-flight mass spectrometry. Pesticide Residue Research Group, University of almera, Spain. 49

50


16. J.Serb (2009). A rapid spectrophotometric determination of Imidacloprid in

28. Vojislava Bursic and Sanja Lazic (2012). Dissipation of Fungicide

selected commercial formulation in the presence of 6- chloronicotinic acid.

Azoxystrobin from Cucumber. Faculty of Agriculture. Universit of Novi Sad,

Journal of the Serbian Chemical Society. 74(12). pp.1455-1465. 17. Mastovka Katerina (2006). Azoxystrobin. Agricultural Research Service.

Serbia. WEBSITE

United State Department of Agriculture, USA. 18. M.K. Sprivastava (2004). Analysis of imidacloprid residues in fruits, vegetables, cereals, fruit juices, and baby foods and daily intake estimation in and around lucknow, India. Environmental Toxicology and Chemistry, India. 19. Raihanah. Et al (2016). Ultra hight performance liquid Chromatography technique to determine Imidacloprid residue in rice using QuEChERS method. International Food Research Journal. 23(4), pp.1396-1402. 20. Schonind Ralf and Schmuck Richard (2003). Analytical determination of Imidacloprid and relevant metabolite residues by LC, MS/MS. Bulletin of Insectology. 56(1), pp.41-50. 21. S.R Burket and A Sapiests (1995). Residue analytical method for the analysis of ICIA 5504 and R2 30310 in cereals (grain) and wine (grapes). Center for

29. National pesticide information center. Imidacloprid (10.11.2016). http://npic.orst.edu/factsheets/imidagen.html. Truy 10 03 30.

(2005). Danh

https://nonghoc.com/thuoc-bao-ve-thuc-vat/1/thuocbvtv1.aspx. Truy 19

04

2017.

31. Pubchem open chemistry database. Azoxystrobin (5.2010) https://pubchem.ncbi.nlm.nih.gov/compound/Azoxystrobin. Truy

12

6 2017 32. Pubchem open chemistry database. Imidacloprid (5.2010) https://pubchem.ncbi.nlm.nih.gov/compound/Imidacloprid. Truy th 6 2017

Analytical Chemistry, USA. 22. Sacramento (2002). HPLC Determination of Total Imidaclopridin Vegetation, Center for Analytical Chemistry, California. 23. Saed Mousa Diab Ali (2012). Evaluation of Imidacloprid and Abamectin Residue in Tomato, Cucumber and Pepper by High Performance Liquid Chromatography (HPLC). MSc. Thesis, Faculty of graduate Studies, An-Najah National University. 24. Ahmad Baig Sajad. Et al (2012). Imidacloprid residues in vegetables, soil and water in the southern Punjab, Pakistan. Journal of Agricultural Technology. 8(3), pp.903-916. 25. Sh.A.Ashorkr. et al (2006). Persistence and fate of Carbosulfan and Imidacloprid residue in potato plants. Central Agricultural Pesticides Laboratory. Agricultural Research Centre, Egypt. 26. T. Nageswara Rao. Et al (2012). Development and Validation of HPLC- UV method for Simultanous determination of Strobilurin fungicide residues in tomato fruits followed by matrix solid phase Disperision (MSPD). Department of Analytical Chemistry International Insitiuate of Biotechnology and Toxicology. 3(11). pp, 113-118. 27. USP 34/ NF 29 (2011). Validation of Compendial<1225>. Printed in the United States by United Book, Inc., Baltimore, MD.

51

2017.

(04.12.2005)

52

12


1:

Imidacloprid

1.3. 1.1.

imidacloprid C= 80ppm, 1.1 Speak

5,425

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255nm

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As

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nm


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Azoxystrobin

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3.

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N

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39194,789

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website theo

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dung :

quy

imidacloprid

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quy

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n ng sung dung sung imidacloprid

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trung gian.

azoxystrobin trong pha

dung trong

sung tham

trong


SINH

PGS.TS


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