LUAN VAN TOT NGHIEP

LI CAM N
Hn bon nam ngoi tren ghe giang ng ai hoc, va ac biet la
khoang thi gian lam luan van tot nghiep va qua a giup cho em rat
nhieu trong viec phat trien kha nang hoc tap nghien cu cung nh la
ren luyen nhan cach cua mnh. e co c nhng ieu nay, em xin
gi li cam n chan thanh nhat en tat ca cac quy thay co trong
trng ai hoc Bach khoa, va ac biet la cac thay co trong Bo mon
Cong nghe thc pham, nhng ngi a giang day va ch bao tan tnh
cho em trong suot thi gian qua.
Em cung xin gi li cam n chan thanh va sau sac nhat en thay
Ngo Manh Thang va thay Hoang Minh Nam a tan tnh hng dan va
ch bao e em co the hoan thanh tot luan van nay.
Tran trong cam n Ths.oan Tan Vinh, giam oc cong ty VIPESCO
a ho tr hoat chat Dimethoate. Trung tam nong dc thuoc cong ty
VIPESCO a phan tch oi chng mot so mau Dimethoate t dch
trch rau.
Con xin cam n bo me a nuoi dng cham lo, ong vien cho con
e con co the hoan thanh tot nhiem vu cua mnh.
Toi xin gi li cam n en tat ca ban be toi, nhng ngi luon gan
bo, giup toi rat nhieu trong hoc tap cung nh trong cuoc song.
Tp HCM, ngay 05 thang 01 nam 2008
Sinh vien
inh Th Thu Hang

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LUAN VAN TOT NGHIEP

TOM TAT LUAN VAN

Luan van a nghien cu quy trnh phan tch Carbaryl, Dimethoate, Vitamin C
trong cai bap vi thiet b HPLC. Cu the:
oi vi Carbaryl, Dimethoate:
Tien hanh quet pho hap thu bc song UV -VIS cua Carbaryl, Dimethoate trong
dung dch.
La chon pha ong vi ty le Axetonitril/nc thch hp e phan tch Carbaryl,
Dimethoate.
Xay dng ng chuan xac nh ham lng Carbaryl, Dimethoate trong dung dch
vi bc song hap thu cc ai va ty le Axetonitril/nc a chon.
Khao sat, cai thien quy trnh trch ly Carbaryl, Dimethoate trong cac mau rau cai
bap.
ng dung quy trnh trch ly x ly va phan tch ham lng Carbaryl, Dimethoate e
xac nh d lng trong mau rau mua ch va hieu suat phan huy Carbaryl trong cai
bap sau qua trnh chieu xa.
oi vi Vitamin C:
Tien hanh quet pho hap thu UV cua Vitamin C trong dung dch.
Xay dng ng chuan xac nh ham lng Vitamin C trong dung dch vi bc
song hap thu cc ai a xac nh.
Khao sat o thu hoi cua quy trnh trch ly Vitamin C trong cai bap.

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LUAN VAN TOT NGHIEP

MUC LUC
Chng 1: M AU................................................................................................................1
Chng 2: TONG QUAN......................................................................................................... 3
2.1. Thuoc bao ve thc vat: [1, 4, 7].................................................................................... 3
2.1.1. nh ngha: .............................................................................................................. 3
2.1.2. Phan loai:................................................................................................................. 3
2.1.3. Cach tac ong cua thuoc len dch hai: .................................................................. 3
2.1.5. D lng thuoc tren cay trong va nong san: [7].................................................. 4
2.1.6. Tnh hnh s dung thuoc BVTV: [9, 20] ................................................................ 4
2.1.7. Tnh hnh ngo oc thuoc BVTV: [20] .................................................................... 5
2.2. Thuoc tr sau Carbaryl: ............................................................................................. 5
2.2.1. c tnh sinh hc. ng dng. [1]............................................................................ 5
2.2.2. Tnh chat ly hoa: [4] ............................................................................................... 5
2.2.3. oc tnh: [4] ............................................................................................................ 6
2.3. Thuoc tr sau Dimethoate:........................................................................................... 7
2.3.1. ac tnh sinh hoc. ng dung. [1] ........................................................................... 7
2.3.2. Tnh chat ly hoa: ..................................................................................................... 7
2.3.3. oc tnh: [4] ............................................................................................................ 9
2.4. Cai bap: .......................................................................................................................... 9
2.5. Vitamin C: [15, 18, 17, 19]........................................................................................... 11
2.5.1. Gii thieu chung ve Vitamin C: [15] ................................................................... 11
2.5.2. Tnh chat vat ly: [18] ............................................................................................ 11
2.5.3. Tnh chat hoa hoc: [18] ........................................................................................ 12
2.5.4. Nguon goc: [15]..................................................................................................... 12
2.5.4. Vai tro cua Vitamin C : [17] ................................................................................ 12
2.6. Phng phap phan tch HPLC: [8]............................................................................. 13
2.6.1. Phng phap sac ky long cao ap:......................................................................... 13
2.6.2. He thong thiet b sac ky long cao ap:.................................................................. 15
2.7. Cac phng phap phan tch Vitamin C: [10, 11, 12] ................................................. 18
2.7.1. Phan tch Vitamin C bang phng phap HPLC ................................................. 18
2.7.2. Phan tch Vitamin C bang phng phap chuan o:............................................ 18
2.7.3. Phng phap so mau:............................................................................................ 19

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LUAN VAN TOT NGHIEP

2.7.4. Phng phap s dung enzim: ............................................................................... 20
2.8. Phng phap phan tch thuoc tr sau: [7]................................................................. 21
Chng 3: MO TA THC NGHIEM .................................................................................. 22
3.1. Hoa chat va thiet b s dung....................................................................................... 22
3.1.1. Hoa chat: ............................................................................................................... 22
3.1.2. Thiet b: ................................................................................................................. 22
3.2. Khao sat pho hap thu UV-VIS Carbaryl, Dimethoate, Vitamin C. ......................... 23
3.3. Thiet lap cac thong so cho he thong HPLC............................................................... 23
3.4. Khao sat thanh phan pha ong phan tch hai chat Carbaryl va Dimethoate......... 24
3.5. Xay dng cac ng chuan cua cac chat phan tch .................................................. 24
3.6. Xac nh o thu hoi mau qua cot cua moi chat ng vi ieu kien chon phan tch. 24
3.7. Khao sat cac bac trch ly Carbaryl, Dimethoate va xac nh hieu suat thu hoi qua
cac bac................................................................................................................................. 25
3.7.1. La chon quy trnh trch ly oi vi Carbaryl va Dimethoate............................ 25
3.7.2. Khao sat hieu suat trch ly dch Carbaryl va Dimethoate trong nc qua cac
bac trch ly....................................................................................................................... 27
3.7.3 Khao sat o thu hoi mau cua quy trnh trch ly Carbaryl va Dimethoate tren
mau cai bap ..................................................................................................................... 27
3.8. ng dung quy trnh phan tch Carbaryl e khao sat s phan huy Carbaryl trong
cai bap sau khi chieu xa lieu thap. .................................................................................... 30
3.9. Khao sat hieu suat thu hoi cua quy trnh trch ly Vitamin C trong cai bap. ........... 30
3.9.1. Quy trnh trch ly Vitamin C trong cai bap......................................................... 30
3.9.2. Khao sat hieu suat trch ly oi vi quy trnh trch ly Vitamin C trong cai bap 30
Chng 4: KET QUA VA BAN LUAN ............................................................................. 33
4.1. Kt qu khao sat pho hap thu cua Carbaryl, Dimethoate, Vitamin C:................... 33
4.1.1. Pho hap thu UV cua Carbaryl trong nc. ......................................................... 33
4.1.2. Pho hap thu UV cua Dimethoate trong nc. .................................................... 33
4.1.3. Pho hap thu UV cua Vitamin C trong em:........................................................ 34
4.2. Ket qua khao sat pha ong oi vi Carbaryl va Dimethoate .................................. 34
4.2.1. Ket qua khao sat pha ong cua Carbaryl:.......................................................... 34
4.2.2. Ket qua khao sat anh hng thanh phan pha ong phan tch Dimethoate ..... 35
4.3. Ket qua xay dng ng chuan Carbaryl, Dimethoate, Vitamin C........................ 36

LUAN VAN TOT NGHIEP

4.3.1. ng chuan cua carabryl trong dung dch nc: ............................................. 36
4.3.2. ng chuan Dimethoate trong dung dch nc: .............................................. 37
4.3.3. ng chuan dung dch Vitamin C trong em:.................................................. 38
4.3. Ket qua o thu hoi mau Carbaryl, Dimthoate va Vitamin C: ................................. 39
4.3.1. o thu hoi mau Dimethoate qua cot: .................................................................. 39
4.3.2. o thu hoi qua cot cua Carbaryl: ........................................................................ 39
4.3.3.o thu hoi qua cot cua Vitamin C: ....................................................................... 40
4.4. Ket qua khao sat hieu suat trch ly dch Carbaryl va Dimethoate trong nc qua
cac bac trch ly.................................................................................................................... 41
4.5. Ket qua khao sat o thu hoi mau cua quy trnh trch ly Carbaryl va Dimethoate
tren mau cai bap: ............................................................................................................... 42
4.5.1. Ket qua o thu hoi Carbaryl cua cai bap ........................................................... 42
4.5.2. Ket qua o thu hoi Dimethoate cua cai bap:...................................................... 44
4.6. Ket qua khao sat s phan huy Carbaryl di tac dung cua chieu xa: .................... 47
4.7. Ket qua khao sat o thu hoi cua quy trnh trch Vitamin C t cai bap: ................. 48
4.7.1. o thu hoi Vitamin C khi bo sung them chuan vao rau: ................................... 48
4.7.2. o thu hoi Vitamin C khi bo sung vao dch rau: ................................................ 50
Chng 5: KET LUAN VA KIEN NGH ........................................................................... 52
5.1 Ket luan: ....................................................................................................................... 52
5.2. Kien ngh:..................................................................................................................... 52
TAI LIEU THAM KHAO .................................................................................................... 53
PHU LUC ............................................................................................................................... 55

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DANH MUC CAC BANG
Bang 2.1: Mot so tnh chat vat ly cua Carbaryl......................................................................... 6
Bang 2.2: o tan cua Dimethoate trong mot so dung moi 250C. ........................................... 8
Bang 2.3: o tan trong nc cua Dimethoate 200C ............................................................... 9
Bang 2.4: Thanh phan cac chat dinh dng co trong 100 g cai bap: [20] .............................. 10
Bang 2.5: Gii han d lng thuoc tr sau trong cai bap ........................................................ 11
Bang 2.6: Thanh phan Vitamin C trong mot so nguyen lieu thc pham ................................. 12
Bang 4.1: Dien tch peak va thi gian lu cua Carbaryl cac pha ong ................................ 35
Bang 4.2: Dien tch va thi gian lu cua Dimethoate cac pha ong .................................... 35
Bang 4.3: o thu hoi qua cot cua Dimethoate: ....................................................................... 39
Bang 4.4: o thu hoi qua cot cua Carbaryl ............................................................................. 40
Bang 4.5: o thu hoi qua cot cua Vitamin C........................................................................... 40
Bang 4.6: Ket qua khao sat cac bac trch ly Dimethoate ........................................................ 41
Bang 4.7: Ket qua khao sat cac bac trch ly Carbaryl ............................................................. 41
Bang 4.8: Ket qua o mau xac nh o thu hoi cua Carbaryl ................................................. 42
Bang 4.9: Ket qua o mau xac nh o thu hoi Carbaryl them vao dch ................................ 44
Bang 4.10: Ket qua o mau xac nh o thu hoi cua Dimethoate........................................... 45
Bang 4.11: Ket qua o mau Dimethoate bo sung vao dch sau loc ......................................... 45
Bang 4.12: Ket qua o mau bo sung Vitamin C ...................................................................... 49
Bang 4.13: Ket qua o mau bo sung Vitamin C vao dch ....................................................... 50

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LUAN VAN TOT NGHIEP

DANH MUC HNH VE
Hnh 2.1: Cong thc cau tao cua Carbaryl ................................................................................ 5
Hnh 2.2: Cong thc cau tao cua Dimethoate ........................................................................... 7
Hnh 2.3: Cai bap..................................................................................................................... 10
Hnh 2.4: (a)-Acid Ascorbi, (b)-Acid Dehydroascorbic....11
Hnh 2.5: Cot sac ky ................................................................................................................ 16
Hnh 2.6: He thong may sac ky long cao ap ........................................................................... 17
Hnh 2.7: Ong dan FIA s dung e nh lng acid ascorbic bang phng phap quang pho. 20
Hnh 3.1: S o quy trnh trch ly Carbaryl va Dimethoate.26
Hnh 3.2: S o trch ly 3 bac..28
Hnh 3.3: Quy trnh trch ly Vitamin C .................................................................................... 31
Hnh 4.1: Pho hap thu UV cua dung dch Carbaryl trong nc ............................................... 33
Hnh 4.2: Pho hap thu UV cua dung dch chuan Dimethoate trong Axetonitril: nc............. 33
Hnh 4.3: Pho hap thu UV cua Vitamin C trong dung dch em..34
Hnh 4.4: Sac ky o o chuan Carbaryl nong o 10 ppm (a) va 5 ppm (b) .......................... 36
Hnh 4.5: ng chuan phan tch Carbaryl trong nc ........................................................... 36
Hnh 4.6: Sac ky o o chuan Dimethoate nong o 5 ppm (a) va 1 ppm (b) ....................... 37
Hnh 4.7: ng chuan phan tch Dimethoate trong dung dch Axetonitril: nc .................. 37
Hnh 4.8: Sac ky o o chuan Vitamin C 1ppm va 2 ppm....................................................... 38
Hnh 4.9: ng chuan Vitamin C trong dung dch em ........................................................ 38
Hnh 4.10: Quy trnh trch ly cai bap..43
Hnh 4.11: Hieu suat phan huy tren cai bap cac lieu chieu khac nhau................................ 48

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Chng 1: M AU
Rau xanh la nhu cau khong the thieu trong c cau ba an hang ngay cua con
ngi tren khap hanh tinh. ac biet khi lng thc va cac thc an giau am a c
am bao th yeu cau ve so lng va chat lng rau lai cang gia tang. Rau khong ch
cung cap mot lng chat x, cac sinh to A, B, C ma con cung cap cac nguyen to vi
lng va a lng rat can thiet trong cau tao te bao. Co nhieu chung loai rau nh rau
an la (cai bap, cai xanh), rau an qua (da leo), rau an cu [9].
Hien nay, do muon thu li nhuan cao nen ngi nong dan s dung rat nhieu cac
loai thuoc tr sau, thuoc tr benh goi chung la thuoc bao ve thc vat (BVTV) len cac
loai rau e tang nang suat cay trong. Cac loai thuoc tr sau c s dung rong rai
trong nong nghiep nham bao ve mua mang khoi s pha hoai cua con trung, tr cac
benh tren thc vat do con trung hoac cac vat k sinh ben ngoai gay ra. Tuy nhien, viec
s dung qua lieu va phun gan ngay thu hoach khien cho lng thuoc ton d cao tren
san pham sau khi thu hoach. Theo thong ke, hang nam mien ong Nam nc ta,
tong lng thuoc bao ve thc vat s dung len en con so 1000 tan, trong o thuoc bao
ve thc vat dung tren rau rat ln chiem t 50 80% tong lng thuoc dung cho cac
loai cay trong. ieu o gay ra nhng moi nguy cho sc khoe con ngi. Mot so ni
ngi s dung rau b ngo oc xay ra ngay cang nhieu va pho bien.
V vay e am bao an toan cho ngi tieu dung rau, can thiet phai co bien phap
quan ly, kiem soat cac nguon rau cung cap. Van e xac nh d lng thuoc tr sau
con ton ong trong rau qua co chnh xac th mi co the ket luan ung ve o an toan
cua rau em i kiem tra. Nh vay phng phap phan tch d lng thuoc tr sau trong
rau cung can nghien cu e co the phan tch c nhanh chong va chnh xac d lng
thuoc co trong rau. T o van e kiem soat nguon rau sach cung n gian va de dang
hn.
Moi phng phap, qui trnh phan tch d lng thuoc tr sau trong rau eu bao gom
2 phan chnh: Tach d lng thuoc tr sau can phan tch, va xac nh d lng nay vi
thiet b phan tch phu hp. Vi thiet b sac ky long hien co bo mon Cong nghe thc
pham, bc au a co mot vai nghien cu kha nang ng dung e phan tch d lng

LUAN VAN TOT NGHIEP

Carbaryl, Dimethoat [5, 6] trong rau ma khong phai ton kem nhieu ve tien thue thiet
b phan tch. Tuy nhien, ket qua at c con han che do o thu hoi mau cua giai oan
trch ly t rau va giai oan phan tch qua cot HPLC con thap. Muc tieu cua luan van
nay la khao sat kha nang cai tien qui trnh trch ly Carbaryl, Dimethoat t rau cai bap
bang cach nang so bac trch ly nham at o thu hoi cac hoat chat o t rau cao hn, t
o lam tang o chnh xac, o tin cay cua ket qua phan tch d lng hai chat nay trong
rau. Ben canh o cai bap co cha mot lng Vitamin C ang ke nen cung can co
phng phap nh lng chnh xac ham lng Vitamin C trong o.

LUAN VAN TOT NGHIEP

Chng 2: TONG QUAN

2.1. Thuoc bao ve thc vat: [1, 4, 7]
2.1.1. nh ngha:
Thuoc bao ve thc vat (BVTV) con goi la thuoc tr dch hai hoac san pham nong
dc, bao gom nhng che pham dung e phong tr cac sinh vat gay hai tai nguyen
thc vat, cac che pham co tac dung ieu hoa sinh trng thc vat, cac che pham co
tac dung xua uoi hoac thu hut cac sinh vat gay hai hoac tai nguyen thc vat en e
tieu diet.
2.1.2. Phan loai:
Theo oi tng phong tr, ngi ta phan loai thuoc BVTV vao cac nhom sau:
-

Thuoc tr sau

Thuoc tr benh

Thuoc tr co

Thuoc tr chuot

Thuoc tr nhen

Thuoc tr oc sen

Thuoc ieu tiet sinh trng cay trong

2.1.3. Cach tac ong cua thuoc len dch hai:

Cach tac ong la ng xam nhap gay hai cua thuoc vao c the dch hai. Thuoc
BVTV co cach tac ong chu yeu la:
-

Tiep xuc: thuoc tr sau tiep xuc xam nhap vao c the sau qua bieu b

V oc: La tac ong cua thuoc khi xam nhap vao bo phan tieu hoa cua ong vat
(con trung, chuot, chim). Chat oc an qua ng mieng vao trong ruot, hoa tan
vao trong dch v da day va ruot gia, tham qua thanh ruot va di chuyen en
cac c quan trong c the e gay hai.

Xong hi: thuoc co the sinh ra kh, khoi, mu co tac dung diet con trung, nam, vi
khuan, chuot. Thuoc co tac ong xong hi co the dung phun len cay, xong hi
trong nha , nha kho, kho tang, nha knh, hang hoa hoac trong at e tieu diet
cac vi sinh vat gay hai. Hi thuoc oc xam nhap qua lo th hoac trc tiep tieu
diet dch hai.

LUAN VAN TOT NGHIEP

-

Noi hap (lu dan): la kha nang cua thuoc co the xam nhap va di chuyen trong
cay e e diet dch hai bang cach tiep xuc hay v oc.

Tham sau: thuoc co kha nang tham qua cac lp te bao bieu b cay e giet dch
hai nam di lp bieu b, ma khong co kha nang di chuyen trong cay.

2.1.5. D lng thuoc tren cay trong va nong san: [7]

Thuoc bao ve thc vat ton tai tren cay trong va nong san tren mot thi gian la ieu
can thiet e bao ve cay trong va nong san chong lai s gay hai cua dch hai tren
ruong, trong qua trnh van chuyen va bao quan. Nhng d lng thuoc co tren nong
san se la nguon gay hai cho ngi tieu dung.
D lng thuoc co the gom nhng chat tan c trong lipit, nhng khong tan c
trong nc, thng ton tai lp bieu b nen co ten la d lng bieu b. D lng gom
nhng chat tan trong nc nhng khong tan trong lipit, ton tai di lp bieu b goi la
d lng noi b. Con d lng la nhng chat khong tan ca trong lipit va nc, nam
phan ngoai bieu b mang ten d lng ngoai b. D lng thuoc thng co trong tc
pham va chung ch gay hai khi vt ngng cho phep.
D lng thuoc BVTV c tnh bang mg thuoc co trong 1 kg nong san.
D lng toi a cho phep (Maximum Residue Limit, viet tat MRL): la d lng
thuoc BVTV toi a c phep ton tai trong nong san ma khong gay oc cho ngi va
vat nuoi khi s dung nong san o lam thc an.
2.1.6. Tnh hnh s dung thuoc BVTV: [9, 20]
V muc ch bao ve cay trong, thuoc BVTV c s dung ngay cang rong rai va
nhieu. Theo mot so ieu tra gan ay, nc ta nong dan s dung khoang 28 loai thuoc
tr sau, 19 loai thuoc tr benh khac nhau. Hau het nong dan xem bien phap hoa hoc la
cach chu yeu e phong tr sau benh tren rau. Co ti 43,8% nong dan s dung thuoc co
oc tnh cao a cam s dung tren rau, 48,42% lai khong am bao thi gian cach li an
toan, co khi ch sau mot hoac hai ngay phun a mang ra ch ban.
Ket qua nghien cu khao sat ve d lng thuoc bao ve thc vat va chat bao quan
trong rau qua nam 2002 2003 cua TS Ha Th Anh ao (Vien Dinh dng quoc gia)
va cong s [18] cho thay: Vi 240 mau rau thu thap tai cac khu vc buon ban rau qua
ln Ha Noi va TP. Ho Ch Minh, t le rau qua co d lng hoa chat la 30,4%, vi cac
chat Dimethoat, Diazinon, Cypermethin Trong o d lng Dimethoat, Carbendazin,
Cypermethin vut qua gii han cho phep.

LUAN VAN TOT NGHIEP

2.1.7. Tnh hnh ngo oc thuoc BVTV: [20]
Tren the gii, hang nam co tren 40000 ngi chet v ngo oc rau ten tong so 2
trieu ngi b ngo oc (da theo thong ke cua to cha Lao ong Quoc te ILO). Tuy
cha co thong ke chnh thc ve so ngi ngo oc do thuoc tr sau nc ta, nhng
theo nhng thong bao t nhng nam qua ve cac trng hp ngo oc le te hay hang
loat tng benh vien, tng a phng cho thay so ngi ngo oc la con so ang ke.
T nam 2000 en thang 6 nam 2003, co nhieu thong bao ve ngi b nhiem oc do
an phai rau qua con d lng thuoc tr sau, nang nhat van la cac tnh ong bang song
Cu Long. Nam 2005, co 13000 ngi nhiem oc va trong o co en 354 ngi chet.
Nguyen nhan ngo oc chu yeu la an phai thc pham co lng thuoc tr sau cha phan
huy het hoac thc pham co s dung chat oc e bao quan, lam hap dan ve hnh thc
va mui v
2.2. Thuoc tr sau Carbaryl:
2.2.1. c tnh sinh hc. ng dng. [1]
Carbaryl la thuoc tr sau tiep xuc, v oc va noi hap nhe, vi pho tac ong diet tr
sau benh kha rong. No la mot trong nhng thuoc tr sau ho Carbamat lau i nhat (bat
au s dung t nam 1956) nhng c s dung nhieu nhat va thay the thuoc tr sau
clo hu c (Hien nay thuoc tr sau nhom bong cuc s dung thay the dan nhom
Carbamate). Thng dung e tr cac loai sau, ray hai lua, cay cong nghiep, cay an
qua ..No c anh gia co o oc trung bnh, LD50 (cap tnh oi vi chuot c) 850
mg/kg, LD50 (man tnh) > 2000 mg/kg. Tuy nhien, san pham gia cong co the oc hn
nhieu, v du Tecyl 85 WP co o oc loai I. oc vi ong. Khong tch luy trong m.
2.2.2. Tnh chat ly hoa: [4]
Cong thc phan t: C12H11NO2.
Cong thc cau tao:

Hnh 2.1: Cong thc cau tao cua Carbaryl

Khoi lng phan t: 201,22.
Ten hoa hoc: 1-naphtol-N-metyl carbamate hoac 1-naphtyl-N-metyl carbamine.
Ten thng mai va cac ten khac: Sevin, Atoxan, Caprotin, Gamonil, Panam, Sevidol.
5

LUAN VAN TOT NGHIEP

Carbaryl 99% la tinh the mau trang co iem nong chay 142oC, ap lc hi 0,002
mmHg (40oC). Khong tan trong nc, tan trong phan ln cac dung moi phan cc nh
Axeton, Cyclohexanon. Ban than ben vi anh sang va o am, nhng b thuy phan
trong moi trng kiem manh thanh - naphtol va metylamin.
Trong c the con trung, Carbaryl b oxy hoa thanh 4-hydroxyCarbaryl, sau o b
thai ra ngoai di dang lien ket vi axt glucuronic. [4]
Bang 2.1: Mot so tnh chat vat ly cua Carbaryl
Phan t lng

201.2 vC

Nhiet o nong chay

138 0C (o tinh khiet 99.1%)

Nhiet o hoa hi

1930C

T trong 200C

1.232

o hoa tan trong nc

40 mg/l 300C

Hoa tan trong cac dung Acetone (200 - 300 g/kg), Methanol (79.6 g/kg),
moi hu c
Dicloromethan (242.6 g/kg), Hexan (0.214g/kg).
o on nh

- On nh trong moi trng trung tnh va acid yeu

nhng de dang b thuy phan trong moi trng kiem
tao thanh 1-naphthol
- On nh vi nhiet o len en 700C va anh sang.
Khi b gia nhiet e phan huy no tao ra khoi oc
NOx.

2.2.3. oc tnh: [4]

Thuoc nhom oc loai II, lieu lng Carbaryl tieu diet 50% so sinh vat tiep xuc
(LD50): LD50 qua mieng 246-283 mg/kg, LD50 qua da > 2000 mg/kg; nong o Carbaryl
tieu diet 50% so sinh vat tiep xuc (LC50): LC50 xong hi > 6,08 mg/l.
Carbaryl de dang c hap thu qua ng ho hap va ng mieng, nhng t b hap
thu hn qua tiep xuc vi da. Khi tiep xuc trc tiep co the gay bong da hoac mat. Ht
vao hay nuot phai Carbaryl vi lng nhat nh co the gay ngo oc cho he than kinh,
he ho hap, gay ra cac trieu chng non ma, co that da day, au au, hon me, roi loan
than kinh, suy ho hap, Dau hieu nhiem oc tang nhanh sau khi hap thu va cung

LUAN VAN TOT NGHIEP

nhanh chong ket thuc sau khi khong con tiep xuc na. Khi b hap thu vao c the ngi,
no se nhanh chong c bai tiet ra theo nc tieu (bai tiet khoang 91,5% trong vong
24h). Carbaryl rat co hai cho con ngi neu vao mieng (LD50 = 614 mg/kg the trong)
va ht vao (LC50 = 2,43 mg/l). Carbaryl co o oc qua da thap (LD50 > 5000 mg/kg the
trong).
2.3. Thuoc tr sau Dimethoate:
2.3.1. ac tnh sinh hoc. ng dung. [1]
Dimethoate la thuoc tr sau co tac dung tiep xuc va lu dan vi pho tac ong rong.
No con co tac dung tr nhen. o oc thap oi vi ong vat mau nong, LD50 185- 245
mg/kg, khong gay chay la. Do vay co the dung Dimethoate trong nhieu lnh vc: rau
qua, ngu coc, chan nuoi.
Tai Viet Nam, Dimethoate van c s dung nhieu. Thng dang hon hp cac
thuoc BVTV khac. Tuy nhien, thuoc co mui hoi nen anh hng en moi trng.
2.3.2. Tnh chat ly hoa:
Dimethoat la thuoc BVTV thuoc nhom lan hu c, co ten goi theo danh phap quoc
te la O,O-dimethyl Smethylcarbamoylmethyl phosphorodithioate, so CAS: 60-51-5.
Cong thc hoa hoc: C5H12NO3PS2,
Khoi lng phan t: 229,2
Cong thc cau tao:

Hnh 2.2: Cong thc cau tao cua Dimethoate

Dimethoate dang ran, iem nong chay 51-52oC; ap lc hi 1,1 mPa (25oC). Tan
trong nc va cac dung moi hu c chu yeu, kho tan trong hydrocacbon. B thuy phan
trong moi trng kiem, nhng tng oi ben trong moi trng trung tnh (2< pH < 7).
B phan huy khi un nong, au tien tao dan xuat cua O,S-dimetyl. [4]

LUAN VAN TOT NGHIEP

Bang 2.2. o tan cua Dimethoate trong mot so dung moi 250C.
Dung moi

o hoa tan (g/100mL)

Aceton

140

Acetonitril

140

Cyclohexane

120

Dodecane

0.043

Etanol

150

Ethyl acetate

120

Hexane

0.03

2-propanol

120

Methanol

160

Dicloromethane

150

1-octanol

52

Toluene

100

Xylene

31

1,2-dicloroethane

120

n-heptane

0.024

Dimethoate phan huy nhanh khi gia nhiet en nhiet o >800C. S phan huy nay
tao ra cac kh oc hai dimethyl sulfic, carbon monoxide, methyl mercaptane,
pentoxide phospho.

LUAN VAN TOT NGHIEP

Bang 2.3. o tan trong nc cua Dimethoate 200C
pH

o tan (g/L)

23,3

23,8

25,0

Dimethoate phan huy trong thi gian ngan trong at, nc va cay trong. Thi gian
ton tai trong at cua Dimethoate la 4 16 ngay, ngan hn trong at am. Trong nc
song, sau 18h 8 tuan th b giam mot na. Trong ong vat cung nh trong thc vat, c
che bien oi cua Dimethoate la giong nhau. No b oxi hoa thanh O,O-dimethylphosphorothioate va hydro hoa thanh O,O-dimethyl-phosphorodithioate, phosphorothioate, -phosphat. S oxi hoa Dimethoate tao nen hp chat omethoate, mot
chat oc va la chat c che enzyme cholinesterase manh, no cung phan huy trong moi
trng nhanh nh Dimethoate. [4]
2.3.3. oc tnh: [4]
Dimethoate thuoc nhom oc II. oi vi ngui, LD50 qua mieng la 235mg/kg, qua da
>400mg/kg. ADI: 0.02mg/kg the trong / ngay. Nong o cao nhat c chap nhan cua
Dimethoate trong nc uong la 0,02mg/L.
Cac nghien cu mi ay tai Vien nghien cu hat nhan a Lat a phat hien bang
chng bien loan nhiem sac the tren te bao limpho ngi mot nhom dan c san xuat
nong nghiep Lam ong gay ra bi thuoc tr sau BAI 58. Thanh phan chnh cua BAI
58 la Dimethoate 40EC va 60% chat tao nhu.
Dimethoate la oi tng nghien cu oc chat moi trng cua the gii t nhng
nam 60. Mac du, BAI 58 la loai thuoc tr sau lan hu c ang c xem xet lai
nhieu nc tren the gii, song Viet Nam van ang c cap phep s dung va ang
c s dung pho bien trong nong nghiep Viet Nam, nhat la nhng vung cay cong
nhiep nh ca phe, dau tam
2.4. Cai bap:
Ten khoa hoc: Brassica oleracea
var. capitata.
Ho: Cruciferae.

LUAN VAN TOT NGHIEP

Hnh 2.3: Cai bap

ac iem bap cai trang
La ngoai co nhiem vu quang hp nen co mau xanh. La trong khong tiep xuc trc
tiep vi anh sang, co mau trang nga. Chung co nhiem vu d tr chat dinh dng va la
bo phan dung lam thc pham. Ham lng Vitamin la gia cao hn.
Thu hoach va bao quan:
Cai bap c trong t thang 9 en thang 10, vu muon t thang 11 en thang 12.
Thi gian sinh trng t 80 90 ngay. Khi bap cai cuon chat th co the thu hoach. Ta
dung dao cat va e lai vai la gia ben ngoai. Nhiet o thch hp e bao quan cai bap la
0 10C.
S dung cai bap
Cai bap la loai rau nhieu chat dinh dng, co the che bien c nhieu mon an nh:
an song, luoc, xao vi tht, muoi chua
Theo ong y, cai bap v ngot, tnh bnh, co cong dung bo cot tuy, khoe gan cot. No
co tac dung cat cn au va thuc cho mau lanh oi vi chng loet da day.
Bang 2.4: Thanh phan cac chat dinh dng co trong 100 g cai bap: [20]
Protein

Lipit

Carbs

Ca

1.1 g

0.2 g

3.4 g

32mg

Fe

Caroten

Vit B1

Vit B2

Vit B3

Vit C

0.3mg 0.02mg

0.04mg

0.04mg

0.3mg

30mg

Nc ep t cai bap la loai "may loc" tuyet vi va giup giam can v cha t calo.
Chat sulfur va chlorine trong cai bap giup tay sach mang nhay bam bao t va ruot.
Ngoai ra, nc cai bap cung cha u lng iot can thiet cho c the ngi, ong thi rat
tot cho viec cha viem da day, ung loet da day, viem mieng va viem li. Mot so
nghien cu cho thay chat isothiocyanates cha nhieu trong cac loai rau nha ho cai co
tac dung tieu diet cac khoi u gay ung th phoi.

10

LUAN VAN TOT NGHIEP

Bang 2.5: Gii han d lng thuoc tr sau trong cai bap
Thuoc tr sau

Gii han d lng

MRL(ppm)

Carbaryl

Dimethoate

2.5. Vitamin C: [15, 18, 17, 19]

2.5.1. Gii thieu chung ve Vitamin C: [15]

(a)

(b)

Hnh 2.4: (a)-Acid Ascorbic; (b)-Acid Dehydroascorbic

Vitamin C la ten goi thong dung cua acid ascorbic. Cac ten goi hoa hoc la LAscorbic acid, L-xyloascorbic acid, 3-oxo-L-gulofuranolactone (dang enol), L-3ketothreohexuronic acid lactone. Vi cong thc phan t C6H8O6. Vitamin C co khoi
lng phan t 176,1. Hnh 2.4 cho thay cong thc cau tao va mo hnh cau truc phan t
Vitamin C.
2.5.2. Tnh chat vat ly: [18]
Acid ascorbic tinh khiet ton tai dang tinh the, khong mau hoac co mau trang.
Acid ascorbic thng khong co mui, co v chua acid, tan vo han trong nc, t tan trong
ethanol, khong tan trong ether va chloroform. Nhiet o nong chay 190oC.
Acid ascorbic b hoa en khi e trc tiep ngoai anh sang mat tri, tuy nhien s
bien mau nhe co the khong lam mat hoat tnh cha benh cua acid ascorbic. Tham ch
khi khong co mat anh sang th acid ascorbic van de dang b bien tnh khi tiep xuc vi
o am trong khong kh va s phan huy nay cang nhanh khi nhiet o moi trng cang
cao.

11

LUAN VAN TOT NGHIEP

2.5.3. Tnh chat hoa hoc: [18]
Acid ascorbic co cac gia tr pKa la 4,2 va 11,6. Trong dung moi nc, dung dch
acid ascorbic 5% co pH = 2,2 2,5.
Trong dung dch, acid ascorbic nhanh chong b oxi hoa khi co mat khong kh va
pH vung kiem. Acid ascorbic ben trong moi trng trung tnh, acid, khong ben trong
moi trng co cac ion kim loai nh Cu2+, Fe2+, Enzym ascorbatoxydase
2.5.4. Nguon goc: [15]
Acid ascorbic co ca nguon goc t nhien va nhan tao. Trong t nhien, acid
ascorbic c tm thay trong cac loai rau qua ti. Nguon thc an cha Vitamin C rat
tot la nhng trai cay thuoc ho citrus hay nc ep t trai ho citrus, qua mong, ho tieu
xanh hay o, ca chua, bong cai, rau cai bo xoi.
Bang 2.6: Thanh phan Vitamin C trong mot so nguyen lieu thc pham
STT

Nguyen lieu

Vitamin C (mg/100g)

Da

17

Cai bap

30

Me

75

Vai

167

Hat ieu

Da leo

12

Nhan

Chanh

46

Cai xanh

70

10

250

2.5.4. Vai tro cua Vitamin C : [17]

Vitamin C c Albert Szent-Gyorgyi phan lap nam 1928. Gan 70 nam sau, cac
nha nghien cu a kham pha ra nhieu li ch cua Vitamin C tren sc khoe. ay la loai
12

LUAN VAN TOT NGHIEP

Vitamin ma c the can nhieu nhat. oi vi ngi bnh thng can khoang 80 100mg
Vitamin C trong 24 gi. Tuy thuoc vao he di truyen va cac yeu to trong loi song, mot
so ngi oi hoi lng Vitamin C nhieu hn lng trung bnh cua mot ngi bnh
thng nham ngan chan s gian oan cua cac phan ng sinh hoa quan trong. Chang
han nh ngi ln tuoi, uong ru, ngi b benh tieu ng, ngi hut thuoc la
thng b thieu Vitamin C.
Khong nh hau het cac loai ong vat khac, c the ngi khong the t san xuat
Vitamin C. Mot benh do thieu hut Vitamin C pho bien nhat la benh scorbus (scurvy).
2.6. Phng phap phan tch HPLC: [8]
2.6.1. Phng phap sac ky long cao ap:
Sac ky long cao ap (High pressure liquid chromatography_HPLC) hay sac ky long
hieu nang cao la mot phng phap phan tch sac ky long hien ai, c s dung rong
rai trong ky thuat phan tch e phan tch cac chat co ham lng nho nh: Vitamin, acid
amin, oc to cua vi sinh vat, d lng thuoc tr sau trong thc pham...
Nguyen tac:
Sac k long la ky thuat phan tch sac ky dung e tach cac ion hoac cac phan t hoa
tan trong dung dch. Khi dung dch mau tiep xuc vi mot pha ran hay mot pha long th
hai, cac chat hoa tan khac nhau trong dung dch mau se tng tac vi cac pha o vi
mot mc o khac nhau da tren s khac nhau ve kha nang hap phu, kha nang trao oi
ion, kha nang phan bo hay da tren s khac nhau ve cau truc phan t cua cac chat
(sac k loc gel). S khac nhau nay se tach cac cau t ra khoi hon hp. Cau t nao co ai
lc vi pha ong manh se i ra trc, cau t nao co ai lc vi pha ong yeu hn se i
ra sau. Cac cau t c xac nh thong qua thi gian lu cua cau t trong cot sac k.
e nh danh cac cau t o, chung ta phai c ho tr bi mot phng phap khac, v
du nh s dung au do hong ngoai IR, au do t ngoai UV
ac iem:
- S dung cot sac ky: kch thc cua cot sac ky nham muc ch phan tch thng be
(Chieu dai: 10-25cm, ng knh: 2-4mm).
- Pha ong la chat long.
- Pha tnh a dang tuy vao kieu cot sac ky nh sac ky hap phu, phan bo, loc gel, trao
oi ion. Do o kha nang ng dung cua phng phap nay rat ln va a dang.
- Hat vat lieu nhoi co kch thc rat be, khoang 1 4 m. Do o, toc o chay cua pha
ong se rat cham. V vay phai s dung ap suat cao (100 400 bar), toc o dong khi o
13

LUAN VAN TOT NGHIEP

at khoang 0,1 en 10 ml/phut. Thi gian phan tch mau thng dao ong trong
khoang ti 30 phut.
Phan loai: Co the chia phng phap HPLC ra lam 2 loai:
Sac ky pha thuan (Normal-phase HPLC): la phng phap sac ky co pha
ong la dung moi khong phan cc nh hexane, con pha tnh la cac hat chat mang bang
silicagel tren o co gan cac goc phan cc nh _CN hoac _NH2.
Sac ky pha ao (Reversed-phase HPLC): la phng phap sac ky co pha
ong la dung moi phan cc nh nc hoac nc cung vi cac dung moi phan cc khac
nh methanol hay Axetonitril, con pha tnh la cac hat chat mang bang silicagel tren o
co gan cac goc khong phan cc nh octadecyl- hay octylsilane.
Thng s dung sac ky pha ao cho sac ky long cao ap:
pha ong phan cc
pha tnh kem phan cc
Nguyen nhan la do pha ong phan cc cho phep s dung nhieu loai pha ong a
dang, ngoai ra co the bo sung them cac thanh phan khac nhau vao pha ong e tao ra
cac o phan cc khac nhau. Thanh phan chnh cua pha ong la nc, la mot dung moi
re, t gay o nhiem moi trng va khong oc hai.
Pham vi ng dung cua phng phap:
Phan tch cac thanh phan hoa hoc trong thc pham nh cac thanh phan dinh
dng, phu gia, oc to, theo doi s bien oi cua thc pham trong qua trnh bao quan
va che bien.
Cu the:
Vitamin
Acid amin
Thuoc, khang sinh, dc lieu.
oc to (aflatoxin).
Thuoc tr sau.
Thuoc tr co.
Phu gia cong nghiep va thc pham.
Acid hu c

14

LUAN VAN TOT NGHIEP

Cac loai ng
Chat kch thch.
2.6.2. He thong thiet b sac ky long cao ap:
Nguyen tac hoat ong:
Pha ong sau khi c bai kh c bm a vao he thong sac ky. He thong t
ong bm mau vao cot (hoac chung ta t bm mau vao). Pha ong khi i ngang qua se
cuon mau can phan tch i theo. Khi vao trong cot sac ky, chat nao co ai lc manh vi
pha ong se theo pha ong i ra trc, chat nao co ai lc yeu hn se i ra sau. au do
phat hien cac cau t i ra khoi cot, chuyen tn hieu cho may ghi ket qua.
Cau tao:
Bm :
La bo phan co tnh quan trong hang au trong he thong HPLC. Bm ap lc cao
c s dung e ay dung moi i xuyen qua pha tnh. Kch thc vat lieu chat nhoi
cot cang nho th ap suat phai cang cao.
Yeu cau: tao c dong pha lien tuc, on nh va khong co xung.
Bm thng s dung la bm pittong.
Cac loai bm hien ai co nhng thong so sau:
Van toc dong: 0,1 10 ml/phut.
o on nh cua dong lu lng: Dao ong khong ln hn 1%.
Ap lc ln nhat: len en 5000psi.
(He thong HPLC trong phong th nghiem Hoa sinh cua khoa s dung bm LC10AS).
Bo phan nhap mau:
Cho phep lay chnh xac lng mau a vao cot sac ky mot mot cach t ong va
c chng trnh hoa:
The tch mau bm vao.
Thi gian thay oi mau.
Thi gian ra bo phan nhap mau.
Nhiet o bm mau.

15

LUAN VAN TOT NGHIEP

Ap suat bm mau (Chu y ap suat bm mau phai nho hn 70bar th mau
mi tap trung c vao cot, kha nang phan tch mau mi tot).
Ngoai ra, ta co the bm mau thu cong bang cach dung xilanh bm mau vao cot.
Cot sac ky :
Hnh 2.5: Cot sac ky
La bo phan chnh cua he thong thiet b HPLC, o chnh la ni dien ra s phan
tach cac cau t trong hon hp nh kha nang hap phu hay phan bo cua cac cau t gia
pha ong va pha tnh. Pho bien nhat la cot ODS (Octadodecyl siloxane).
Rat ngan, khoang 10 25 cm, ng knh trong 2 4 mm.
Vat lieu nhoi cot:
oi vi sac ky hap phu: chat mang ran thng la silicagel hay oxyt
nhom.
oi vi sac ky phan bo: chat mang ran la dan xuat cua silicagel (cac hat
silicagel c gan vi cac nhom chc khac nhau tao ra cac hat co chc nang
phu hp vi mau can phan tch). V du nh: Siloxanes (C18, C8).
Yeu cau oi vi chat mang:
Co tnh chon loc can thiet oi vi thanh phan mau nghien cu.
Ben nhiet.
Ben c hoc.
Ben va tr ve mat hoa hoc vi cac cau t khac trong mau phan tch.
au do:
Ngay nay cac au do quang hoc c s dung pho bien trong cac he thong sac ky
long. Nhng au do nay a mot tia sang xuyen qua dong chay ra khoi cot. S thay
oi ve cng o anh sang do s hap thu tia UV, s phat xa huynh quang t cac thanh
phan se c ghi nhan lai dang thay oi cua ien the au ra. Nhng ien the nay
c bieu dien tren mot o th va a vao may tnh e hien th thi gian lu va peak.
au do trong HPLC rat a dang:
au do UV.
au do UV VIS.

16

LUAN VAN TOT NGHIEP

au do huynh quang.
au do dan nhiet.
au do khuc xa ke vi sai (RI).
He thong HPLC trong phong th nghiem Hoa sinh cua khoa s dung au do
quang pho UV-VIS loai SPD - 6VA.
Cac ac trng quan trong cua mot au do la:
o nhay.
Gii han do tm.
Quan tnh va pham vi phu thuoc tuyen tnh gia cng o tn hieu vi
nong o.
May ghi :
Co the ve sac ky o, ghi lai thi gian lu, tnh dien tch peak, ghi lai phng phap
xac nh, ong thi co the s dung may ghi e ieu khien, chng trnh hoa qua trnh
chay sac ky.
He thong HPLC trong phong th nghiem Hoa sinh cua khoa s dung may ghi
Chromatopac CR 6A.
S o he thong:

Cot sac ky

au do

May tron
May ghi

Pha ong

Valve phun

Bm

Hnh 2.6: He thong may sac ky long cao ap

17

LUAN VAN TOT NGHIEP

2.7. Cac phng phap phan tch Vitamin C: [10, 11, 12]
Vitamin C co the c phan tch bang nhieu phng phap, tuy thuoc vao ac iem
cua mau can phan tch, thi gian phan tch va thiet b co the tiep can c. Cac
phng phap thng c s dung e phan tch Vitamin C la:
-

Phng phap sac ky long cao ap (HPLC)

Phng phap chuan o.

Phng phap so mau.

Phng phap s dung enzim

2.7.1. Phan tch Vitamin C bang phng phap HPLC

Phng phap nay c gii thieu trong nhieu tai lieu tham khao va a tr thanh
mot bai th nghiem trong PTN Hoa sinh, B/m CN thc pham. Cot sac ky s dung la cot
pha ao ODS C18. Pha ong s dung la dung dch em phosphate pH = 2,8 tron vi
methanol theo ty le the tch 90/10, lu lng pha ong qua cot 1 mL/phut. S dung au
do UV o o hap thu bc song 245 nm. Pha ong c anh sieu am e uoi kh
hoa tan trc khi s dung, va c loc lien tuc trong qua trnh chay sac ky bang mang
loc 450 nm. Xay dng ng chuan vi cac mau acid ascorbic tinh khiet vi nam nong
o khac nhau. Mau rau qua c nghien trong dung dch em phosphate pH = 2,8 e
trch Vit.C ra, pha loang theo ty le thch hp roi phan tch nh vi cac mau chuan.
Ham lng Vitamin C trong mau c xac nh bang cach noi / ngoai suy tn hieu cua
mau t ng chuan. [10]
2.7.2. Phan tch Vitamin C bang phng phap chuan o:
Chuan o vi 2,6- dichloroindophenol (DCP)
ay la mot phng phap hu dung e nh lng Vitamin C trong rau qua. C s
cua phng phap nay la phan ng oxi hoa acid ascorbic trong dung dch bi DCP
thanh dang dehydroascorbic acid. San pham cua phan ng oxy hoa kh nay eu khong
mau.
Phng trnh phan ng:
(pH=3)
C6H8O6 (khong mau) + C12H7O2NCl2(o) C6H6O6 (khong mau) + C12H9O2NCl2

(khong mau)

S chuan o nay ac biet thuan li bi v DCP cung ong vai tro nh chat ch th
mau. Khi nho dung dch DCP vao mot dung dch co cha Vitamin C, th hon hp phan
ng khong mau cho en khi tat ca lng Vitamin C co trong hon hp chuyen het thanh
18

LUAN VAN TOT NGHIEP

dang dehydroascorbic acid. Giot dung dch DCP tiep theo them vao neu xuat hien
mau o chng to o la giot DCP con d va phan ng a ket thuc. Do o, iem tng
ng c xac nh (ch th) khi dung dch chuan o t khong mau chuyen sang mau
o cua DCP.
Yeu iem cua phng phap nay la dung dch DCP rat kem ben nen chung ta ch
pha dung dch ngay trc khi s dung chung.
Trc khi tien hanh chuan o, can x ly mau thc pham vi metaphosphoric
acid. Bc x ly vi acid nay nham lam bien tnh va ket tua protein trong mau e
tranh anh hng en ket qua phan tch, ong thi cung nham giup on nh acid
ascorbic tranh b phan huy. Acid hoa en pH nho hn 4 cung nham lam giam phan ng
cua DCP vi cac hp chat khac.
Chuan o bang iode
ay la qui trnh chuan o ngc, lng ion I 3 d trong dung dch sau khi a
phan ng vi Vitamin C c xac nh bang cach cho phan ng vi dung dch
thiosulfate. C s cua phng phap chuan o Vitamin C bang Iod la phan ng oxi hoa
Vitamin C thanh dehydroascorbic acid, C6H6O6 theo phng trnh sau :
I 3 C 6 H 8 O6 H 2 O C 6 H 6 O6 3I 2 H

Lng ion I 3 trong dung dch sau khi Vitamin C a phan ng het se c xac
nh bang phep chuan o vi dung dch thiosulfate S 2 O32 theo phng trnh sau:
I 3 2 S 2 O32

3 I S 4 O62

Co mot kho khan la phai pha che dung dch I 3 t dung dch KIO3 va muoi ran KI
trong ieu kien acid nh phng trnh sau:
6 H (aq ) KIO3 (aq ) 8 KI ( s ) 9 K (aq ) 3I 3 (aq ) 3H 2 O(l )

Lng I 3 tao thanh co the c tnh toan thong qua lng KI va KIO3 a s
dung. Lng nay tr i lng d trong chuan o vi thiosulfate se thu c lng I 3
a phan ng vi acid ascorbic.
2.7.3. Phng phap so mau:
C s cua phng phap la phan ng kh Fe(III) thanh Fe(II) bi acid ascorbic.
San pham Fe(II) co the c o de dang sau khi ket hp vi tac nhan mau nh 1,10phenanthroline hoac neocuproline. ay, mono (1,10-phenanthroline)-Fe(III) b kh

19

LUAN VAN TOT NGHIEP

trc tiep thanh tri(1,10-phenanthroline)-Fe(II) bi acid ascorbic, san pham tao thanh
hap thu anh sang bc song 510 nm.
Phng phap nay rat nhay, nhanh, va a c nghien cu t ong hoa bi ky
thuat tiem dong (FIA), cho hieu suat phan tch 200 mau/gi. ng chuan cua acid
ascorbic tren day 0 - 50ppm trong ieu kien ly tng c ghi nhan. RSD = 0,5%(
n=15).
Mono(1,10-phenanthroline)-Fe(III) cung co the c s dung e nh lng
adrenaline.

Hnh 2.7: Ong dan FIA s dung e nh lng acid ascorbic bang phng phap
quang pho.
C s cua phng phap la phan ng kh muoi tetrazolium MTT [3-(4,5dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide] thanh mot Formazan bi acid
L-ascorbic (L-ascorbate) khi co mat cua chat mang ien t PMS (5methylphenazinium methosulfate) tai pH = 3,5. MTT-Formazan c xac nh thong
qua o o hap thu anh sang thay c tai bc song 578 nm.
Trong th nghiem vi mau th tong lng chat kh se c tnh toan theo:
L Ascorbate ( x H ) MT Dehydroascorbate ( x) MTT Formazan H
2
e nh lng L-ascorbate, s dung ong thi mot mau trang. Mau trang nay
c cho phan ng vi enzym ascorbate oxidase (AAO) vi s co mat cua O2 e loai
i s co mat cua L-Ascorbte (2.5). Dang dehydroascorbate khong phan ng vi
MTT/PMS.
L Ascorbate 1 / 2 O2 Dehydroascorbate H 2 O

S khac nhau ve o hap thu gia mau th va mau trang se chnh la lng LAscorbate co trong mau.
2.7.4. Phng phap s dung enzim:
Phng phap enzyme s dung ascorbate oxidase e xuc tac phan ng oxi hoa
acid L-ascorbic bang O2 trong moi trng acid theo phan ng sau:
Ascrobate oxydase
2 L-ascorbate + O2 + 2H+
2 dehydro-L-ascorbate + 2H2O

20

LUAN VAN TOT NGHIEP

Nong o cua acid ascorbic c xac nh gian tiep thong qua mc o tieu thu O2,
xac nh bi ien cc oxy. ng chuan cua phng phap co dang ng thang trong
khoang nong o 1,2.10-4 en 1,0.10-3 mol/L acid ascorbic.
2.8. Phng phap phan tch thuoc tr sau: [7]
Phan tch d lng thuoc BVTV la mot lnh vc ac biet. D lng co trong nong
san, at ai va moi trung co ham lng rat thap (ppm hay ppb) thng c phan tch
bang phng phap sac ky long cao ap hay sac ky kh hien ai vi cac detector cong
ket ien t. Qua trnh chuan b mau tach chiet phai lam rat can than va chnh xac e
tranh sai so, phai la chon dung moi thch hp vi ac tnh ly hoa tng loai thuoc
BVTV e co the tach chiet c d lng toi a va tach c d lng nay khoi hp
chat khac trong cay nh glucosit, clorophyl
Phan tch d lng Carbaryl:
Hien nay co nhieu tai lieu tham khao mo ta phng phap e phan tch Carbary: sac
ky kh, sac ky long, quang pho so mau. Tuy nhien, cac hp chat thuoc ho Carbamate
la nhng chat khong on nh khi gia nhiet, de b phan huy khi nhiet o cao nen a so
s dung sac ky long HPLC e phan tch Carbaryl. [11]
Phan tch d lng Dimethoate:
Cac phng phap phan tch ham lng Dimethoate sau c tm thay trong tai lieu
tham khao:
- Phng phap sac k kh.
- Phng phap sac k long cao ap.
- Phng phap sac k ban mong.
- Phng phap dung bo KIT enzym

21

LUAN VAN TOT NGHIEP

Chng 3: MO TA THC NGHIEM

3.1. Hoa chat va thiet b s dung

3.1.1. Hoa chat:

- Carbaryl_ o tinh khiet > 98%
- Dimethoate_ o tinh khiet > 98%
- Axeton_Trung Quoc
- NaCl_Trung Quoc
- Axetonitril_Merk_HPLC
- Methanol_Merk_HPLC
- H3PO4
- Acid ascorbic_Trung Quoc
- KH2PO4
- Nc cat 2 lan
Dung dch chuan goc Carbaryl, Dimethoate: Can chnh xac lng 0,1000 g moi
chat, hoa tan trong Axetonitril va nh mc len 100 ml bang Axetonitril thu c dung
dch chuan goc nong o 1000 ppm, bao quan trong tu lanh e s dung cho cac th
nghiem.
Dung dch chuan goc Vitamin C 1000 ppm moi lan th nghiem se c pha mi.
Can 0,1000 g Acid ascorbic nh mc len 100 ml bang dung dch em KH2PO4 pH=
2,8.
3.1.2. Thiet b:
- May sac ky long_ hang Shimazdu SPD 6AV_ au do UV VIS
- Be anh sieu am_ Decon
- May co ac chan khong_ Heidolph WB 2000
- May o quang pho vung UV VIS.
- May xay sinh to Gali
- May khuay t
- He thong loc chan khong

22

LUAN VAN TOT NGHIEP

- Phieu chiet 250 ml
- Mang loc 0,45 m, giay loc 0,6 m
- Bnh nh mc 10, 25, 50, 100 ml
- Ong ong 100, 500 ml
3.2. Khao sat pho hap thu UV-VIS Carbaryl, Dimethoate, Vitamin C.
Chuan b cac dung dch Carbaryl, Dimethoate trong nc va dung dch Vitamin C
trong em pH = 2,8 vi nong o khoang 3ppm. S dung cuvet thach anh e o. Tien
hanh quet pho bang may quet pho t ong co bc song t 200 nm en 350 nm. Ghi
nhan so lieu o c va ve cac o th bieu dien pho hap thu vung UV cua 3 chat tren.
Da vao o th pho thu c chon bc song hap thu cc ai cho moi chat. Sau o
at bc song cho may HPLC tng ng khi phan tch tng chat tren.
3.3. Thiet lap cac thong so cho he thong HPLC
Luan van nay s dung he thong may sac ky long cua hang Shimazdu: au do UVVIS SPD 6AV, bm LC 10AS, may ghi sac ky o C R6A.Cot phan tch Gemini
pha ao C18: 250 x 4,6 mm x 5 m x 110 Ao.
Cac thong so chnh cua he thong:
- at gia tr ap suat cc ai (P.MAX): 300 kgf/cm2. Muc ch la e bao ve cot va cac
he thong khac, khi ap suat vt P.MAX bm se t ong tat.
- at gii han di cua ap suat (P.MIN): thng la 0. Muc ch la khong cho khong
kh vao he thong khi co s co h tren he thong va an toan khi o.
- Ap suat at c khi he thong on nh: 98 110kgf/cm2
- Toc o dong pha ong 1ml/phut
- The tch bm mau 20 l.
- Atten: co gia tr t 0 en 10, gia tr cang nho th khoang bieu dien tren sac ky o
cang nho, dung atten nho khi o nong o thap ma khong anh hng ti dien tch peak
o c, thng s dung atten =10.
- Speed (toc o giay): thng dung la 5, muon peak sac nhon th giam Speed
- Bc song cua au do: - oi vi Carbaryl: 220 nm
- oi vi Dimethoate: 204 nm
- oi vi Vitamin C: 245 nm

23

LUAN VAN TOT NGHIEP

3.4. Khao sat thanh phan pha ong phan tch hai chat Carbaryl va Dimethoate.
T dung dch chuan goc Carbaryl va Dimethoate 1000 ppm pha san, tien hanh pha
loang bang nc c dung dch co nong o10 ppm cua Carbaryl va Dimethoate trong
nc.
Chuan b pha ong khao sat: ay chung ta tien hanh khao sat kha nang tach
Carbaryl va Dimethoate cua cac pha ong co thanh phan Axetonitril va nc theo
nhng t le 85/15, 70/30, 55/45, 40/60.
Tien hanh pha cac pha ong vi t le ve the tch Axetonitril: nc = 85/15, 70/30,
55/45, 40/60. ng vi moi pha ong nay ta thc hien phan tch cho ca Carbaryl va
Dimethoate nong o 10 ppm a pha tren. Moi pha ong khi phan tch vi tng chat
se cho so lieu ve thi gian lu va dien tch peak tng ng.
3.5. Xay dng cac ng chuan cua cac chat phan tch
Chuan b cac dung dch chuan Carbaryl, Dimethoate, Vitamin C vi cac nong o 1,
2, 5, 10, 20 ppm bang cach pha loang ra t dung dch chuan goc 1000 ppm.
Pha ong chon e xay dng ng chuan da vao ket qua cua th nghiem muc 3.4.
oi vi Carbaryl va Dimethoate chon pha ong la Axetonitril: nc theo t le the tch
55: 45. Con oi vi pha ong phan tch Vitamin C s dung la methanol: he em
KH2PO4 co pH = 2,8 theo the tch 10 : 90.
He em c pha bang cach hoa tan 6,5g KH2PO4 trong 1l nc cat 2 lan, chnh pH
ve 2,8 bang H3PO4 roi loc bang mang 0,45m.
Tien hanh o cac dung dch Carbaryl va Dimethoate tren may sac ky vi cac thong
so nh muc 3.3 tren. Moi mau chuan phan tch 3 lan, lay gia tr trung bnh cua dien
tch peak. Thiet lap ng chuan phu thuoc cua dien tch peak thu c vao nong o
chat phan tch trong dung dch mau chuan.
3.6. Xac nh o thu hoi mau qua cot cua moi chat ng vi ieu kien chon phan
tch.
e xac nh o thu hoi mau cua cac chat qua cot phan tch ta tien hanh phan tch
cac nong o t 1 en 20 ppm.
che o co i qua cot: moi nong o phan tch 3 lan theo che o ng vi chat a
neu tren, tnh dien tch peak trung bnh (Xc).
che o khong cho qua cot: c khoang 1 phut ta tiem mau vao cot mot lan vi
the tch 20 l, sau 5 -6 phut th dng (cac lan bm t lan 2 tr i khong nhan phm
START). Tnh gia tr trung bnh cua dien tch peak moi nong o (X0).
24

LUAN VAN TOT NGHIEP

Gia tr hieu suat thu hoi qua cot Hc (%) c tnh theo cong thc:
Hc (%) =

X
X

100

[Cong thc 1]

3.7. Khao sat cac bac trch ly Carbaryl, Dimethoate va xac nh hieu suat thu hoi
qua cac bac.
3.7.1. La chon quy trnh trch ly oi vi Carbaryl va Dimethoate.
Tai lieu tham khao t AOAC va cac luan van tot nghiep ai hoc nam trc cho
thay, quy trnh phan tch da tren cac bc c ban: trch ly, lam sach, phan tch tren
thiet b. Trong quy trnh trch ly cac chat Carbaryl va Dimethoate th quan trong nhat
phai chon c dung moi trch ly tot nhat. ay chung toi chon dung moi trch ly la
Axeton. Cac bc tien hanh cua qua trnh trch ly Carbaryl va Dimethoate c thc
hien nh sau:
-

Bc 1: Can 100g mau rau cho vao may xay Magic bullet, them 200 ml Aceton
vao, nghien trong thi gian 1 phut.

Bc 2: Chuyen hon hp sau khi nghien vao trong bercher 500 ml, trang sach
may xay bang Axeton, sau o em i khuay t trong thi gian 15 phut

Bc 3: Tien hanh loc chan khong qua giay loc tren pheu loc Buchner, thu dch
loc vao Bercher 500 ml. Ba gi lai tren pheu. Can khoang 20g muoi NaCl (tuy
vao lng nc co trong mau) cho vao dch trch, khuay ca t e hoa tan hoan
toan muoi

Bc 4: Chuyen dch a bo sung muoi vao trong phieu chiet, tach pha hu c ra
khoi pha nc.

Bc 5: Lay pha hu c pha tren em i co quay chan khong cho en khi can va
dch khong con thay oi the tch na th dng.

Bc 6: Chuyen phan dch sau co quay vao bnh nh mc 100 ml, trang bnh co
quay bang 25 ml Axetonitril vao bnh nh mc tiep tuc nh mc en vach.

Bc 7: Dch sau khi nh mc em i loc vi mang loc 0,45 m sau o anh

sieu am loai bot kh va tien hanh phan tch tren may HPLC.

Sau ay la s o khoi cua quy trnh tren: (xem hnh 3.1, trang 26)

25

LUAN VAN TOT NGHIEP

100g rau
200 ml
Axeton

Xay

Lac

Loc chan
khong
20 g NaCL

Tach pha

Ba

Pha
nc

Co quay chan khong

120 v/p, 45oC
Trang bnh

nh mc 100
ml
Sieu loc 0,45 m

Dch phan
tch

Hnh 3.1: S o quy trnh trch ly

Carbaryl va Dimethoate.

26

LUAN VAN TOT NGHIEP

3.7.2. Khao sat hieu suat trch ly dch Carbaryl va Dimethoate trong nc qua cac
bac trch ly.
Cach tien hanh nh sau: cho 25 ml dung dch moi chat Carbaryl va Dimethoate 20
ppm a pha san trch ly trong 50 ml Axeton. Sau o cung tien hanh khuay t e hoa
tan Carbaryl va Dimethoate vao trong Axeton. Tien hanh giong cac bc t 4 en 7
nh quy trnh muc 3.7.1 tren. Pha hu c tach ra lan th nhat sau khi tien hanh cac
bc tiep theo ta c dch trch bac 1 em i phan tch, tiep tuc lay pha nc bo sung
them Axeton va muoi em i khuay t tiep thu c pha hu c va nc lan 2. Pha
hu c tiep tuc quy trnh pha sau thu dch trch bac 2, pha nc thu c lan 2 lai cho
them Axeton va muoi em khuay t va tach pha lan 3. Pha hu c thu c lan 3 tiep
tuc qua trnh e co dch trch bac 3. Ch khao sat 3 bac v vay pha nc lan 3 c loai
bo.
Dch trch ly bac 1, 2, 3 c em o tren may HPLC e xac nh dien tch cac
peak. Moi mau tien hanh o 3 lan, lay dien tch peak trung bnh.
e tnh c hieu suat thu hoi cac chat qua cac bac trch ly ta tien hanh ong thi
o dien tch peak cua dung dch Carbaryl, Dimethoate 20 ppm em i trch e so sanh.
S o quy trnh trch c the hien hnh 3.2: (trang 28)
3.7.3 Khao sat o thu hoi mau cua quy trnh trch ly Carbaryl va Dimethoate tren
mau cai bap
S o quy trnh trch da tren quy trnh s o hnh 3.1 va ket qua chon so bac se
c trnh bay phan ket qua va ban luan.
Tien hanh khao sat o thu hoi Carbaryl va Dimethoate tren oi tng phan tch la cai
bap. Cai bap c mua ch em ve bao quan trong ngan mat cua tu lanh.
e anh gia mc o hoan thien cua mot quy trnh phan tch chung toi da vao o
thu hoi mau. o thu hoi mau la mc o thu hoi lng hoat chat a them vao trong
mau rau khi phan tch. V vay ta phai biet chnh xac lng thuoc them vao tren rau.
Va tnh hieu suat thu hoi da tren lng thuoc them vao nay.
Ta tien hanh khao sat o thu hoi mau cac nong o bo sung tng ng vi lng phun
them 2,5 ; 5 ; 10 ml dung dch 100 ppm Carbaryl, Dimethoate vao cai bap va lng
them cung 2,5 ; 5; 10 ml dung dch chuan Carbaryl, Dimethoate 100 ppm vao dch
trch a c loc chan khong.

27

LUAN VAN TOT NGHIEP

25mL Carbaryl , Dimethoate

20ppm

Khuay t

50mL Axeton

5 7g NaCl

Tach pha

Pha hu c

Pha nc

2-3g NaCl

Co quay chan khong

2nl Axetonitril

Khuay

Axeton

Tach pha

Trang bnh

nh mc 50mL

Pha hu c

Pha nc
2-3g NaCl

Dch phan
tch 1

Co quay chan khong

Khuay
t
Tach pha

2mL Axetonitril

Trang bnh
Pha hu c

nh mc 10mL

Dch phan
tch 2

Hnh 3.2: S o trch ly 3 bac

28

Dch phan
tch 3

Axeton

Nc

LUAN VAN TOT NGHIEP

Xac nh o thu hoi cua Carbaryl, Dimethoate cua cai bap khi bo sung trc
tiep dung dch chuan len rau:
Vi moi chat ta se chuan b 4 mau rau 100g trong o:
-

Mau 1 khong bo sung g ca

Mau 2 bo sung 2,5 ml dung dch chuan 100 ppm

Mau 3 bo sung 5 ml dung dch chuan 100 ppm

Mau 4 bo sung 10 ml dung dch chuan 100 ppm

Sau o tien hanh trch mau theo quy trnh a chon. Phan tch bang HPLC nh a mo ta
phan trc.
Xac nh dien tch peak cac mau khong bo sung va co bo sung Carbaryl, Dimethoate
tren rau nh tren, ong thi o mau Carbaryl, Dimethoate trong nc nong o tng
ng vi lng chat chuan bo sung sau khi a nh mc e em phan tch tren may
HPLC. o thu hoi mau H(%) moi nong o bo sung c tnh:

H(%) b=
(X

(X
b

100

chuan

[Cong thc 2]

Vi Xb: dien tch peak trung bnh cac lan o cua mau co bo sung them chan
X: dien tch peak trung bnh cac lan o cua mau khong bo sung
Xchuan: dien tch peak trung bnh cac lan o cua cac chuan tng ng nong o a
bo sung.
Xac nh o thu hoi mau Carbaryl, Dimethoate khi bo sung vao dch cai bap
sau khi a loc ba:
oi vi moi chat khao sat ta cung tien hanh chuan b 4 mau rau cai bap, moi mau
100g. Moi mau cung c trch ly 2 bac cho ca ba va tach pha. Nhng trong th
nghiem nay ta khong bo sung dung dch chuan Carbaryl, Dimethoate vao trong rau ma
bo sung vao dch qua loc chan khong sau khi a trch 2 lan tren rau. Nh vay o thu ho
lng chat chuan them vao trong th nghiem nay so sanh vi o thu hoi khi phun chat
chuan trc tiep len rau trong th nghiem tren se cho ta thay s mat mat lng chat van
con nam trong ba va s ton that qua khau loc chan khong.
Tien hanh trch ly 8 mau cai bap, moi mau 100g thc hien theo quy trnh tren
nhng khau dch loc trc khi khuay t 1 se co 6 mau c bo sung cac lng tng
ng 2,5; 5; 10 ml dung dch chuan 100 ppm cho moi chat Carbaryl, Dimethoate, 2 mau

29

LUAN VAN TOT NGHIEP

con lai khong bo sung g them. Trong qua trnh lay mau, cac mau bo sung cung mot
chat va mau trang oi chng phai lay cung mot trai cai bap e tranh sai so.
Phan tch tren may HPLC moi mau 3 lan, lay ket qua dien tch peak trung bnh o
c.
3.8. ng dung quy trnh phan tch Carbaryl e khao sat s phan huy Carbaryl
trong cai bap sau khi chieu xa lieu thap.
Vi quy trnh phan tch Carbaryl a c khao sat va chon nh tren, chung toi thc
hien trch ly cac mau cai bap co bo sung them 5 ppm Carbaryl a c x ly chieu xa
cac lieu chieu 0,2 ; 0,4; 0,6; 0,8; 1 KGy. e xem xet s anh hng cua chieu xa en
lng Carbaryl trong cai bap la nh the nao.
X ly mau rau em i chieu xa: ng vi moi lieu chieu xa chung toi dung nguyen trai
cai bap tam vao mot lng dung dch chuan Carbaryl vi ham lng 5 ppm tng ng
vi bo sung lng 5 mg Carbaryl vao 1kg cai bap. Sau khi tam them Carbaryl vao mau
cai bap c e qua em trong ngan mat cua tu lanh. Chuan b them mot mau cai bap
oi chng ngha la mau nay cung c tam them 5 ppm Carbaryl bao quan trong ieu
kien giong cac mau tren nhng khong em i chieu xa. Sau o mau c gi i chieu
xa Vien cong nghe hat nhan a Lat.
Mau sau khi chieu xa xong tien hanh lay moi mau 100g em i trch ly Carbaryl va
phan tch tren HPLC.
3.9. Khao sat hieu suat thu hoi cua quy trnh trch ly Vitamin C trong cai bap.
3.9.1. Quy trnh trch ly Vitamin C trong cai bap
Da vao tai lieu tham khao Giao trnh phan tch thc pham [10], chung toi xay
dng quy trnh trch ly Vitamin C nh sau: Lay mau rau khoang 25g em xay trong
trong dung dch em KH2PO4 pH = 2,8 e trch ly Vitamin C co trong cai bap. Sau o
tien hanh loc bang he thong loc chan khong. Dch loc thu c c nh mc len 100
ml, sau o tiep tuc loc qua mang loc 0,45 m thu c dch chiet Vitamin C cho phan
tch. Dch thu c tiep tuc pha loang 10 lan bang dung dch em. Sau o tien hanh
phan tch vi dch a pha loang nay tren may HPLC. S o quy trnh trch ly Vitamin
C t cai bap nh hnh 3.3 (trang 31).
3.9.2. Khao sat hieu suat trch ly oi vi quy trnh trch ly Vitamin C trong cai bap
e co the tnh c hieu suat thu hoi ta phai them lng chat chuan biet trc ham
lng vao mau can phan tch va co mau trang oi chng e so sanh.

30

LUAN VAN TOT NGHIEP

Cach thc tien hanh: ay chung toi khao sat hieu suat thu hoi chung cua ca quy
trnh trch va khao sat s mat mat rieng cua khau loc ba va cua ba sot lai.
Khao sat o thu hoi cua ca quy trnh trch ly Vitamin C trong cai bap
Chuan b mau trch ly: lay 4 mau cai bap cung t mot trai, moi mau can 25g. Sau o
tien hanh x ly mau nh sau:
-

Mau 1: khong bo them chuan Vitamin C

Mau 2: bo sung them 5ml dung dch chuan 1000 ppm Vitamin C vao rau

Mau 3: bo sung them 10 ml dung dch chuan 1000 ppm Vitamin C vao rau

Mau 4: bo sung them 15 ml dung dch chuan 1000 ppm Vitamin C vao rau.
25g rau

75 mL dd em

Xay

Loc chan khong

nh mc 100mL

Sieu loc

Dch chiet
Vitamin C

Hnh 3.3: Quy trnh trch ly Vitamin C

31

LUAN VAN TOT NGHIEP

Tien hanh trch ly cac mau a x ly tren theo quy trnh trch ly nh muc 3.9.1 thu
c dch loc chuan b cho phan tch HPLC. ong thi cung pha loang t dung dch
goc 1000 ppm thanh cac dung dch co nong o 5 ppm, 10 ppm, 15 ppm.
Dch loc thu c cua cac mau tien hanh pha loang len 10 lan bang cach lay moi
mau dch 1ml cho vao bnh nh mc 10 ml va nh mc ti vach bang dung dch em
pH =2,8. anh sieu am cac mau trc khi phan tch tren may.
o mau tren may HPLC. Moi mau o lap lai 3 lan, lay gia tr trung bnh dien tch
peak o c.
Khao sat s mat mat Vitamin C trong ba va qua trnh loc.
Lay 4 mau cai bap moi mau 50g t cung mot trai. Sau o tien hanh quy trnh trch ly
nh muc 3.9.1. Dch loc thu c sau khi loc chan khong tien hanh x ly nh sau:
-

Dch loc mau 1: khong bo sung chuan Vitamin C

Dch loc mau 2: bo sung them 5 ml dung dch chuan Vitamin C

Dch loc mau 3: bo sung them 10 ml dung dch chuan Vitamin C

Dch loc mau 4: bo sung them 15 ml dung dch chuan Vitamin C.

Tiep tuc quy trnh trch va thu c dch trch sau sieu loc qua mang 0,45 m. Pha
loang dch trch moi mau len 10 lan, anh sieu am tat ca cac mau va tien hanh phan
tch tren may HPLC. Moi mau o lap lai 3 lan, lay gia tr trung bnh dien tch peak o
c.

32

LUAN VAN TOT NGHIEP

Chng 4: KET QUA VA BAN LUAN

4.1. Kt qu khao sat pho hap thu cua Carbaryl, Dimethoate, Vitamin C:
4.1.1. Pho hap thu UV cua Carbaryl trong nc.

1.6

Ph h p thu ca carbaryl

h p thu

1.2
0.8
0.4
0
200

220

240 260 280 300

Bc sng (nm)

320

340

Hnh 4.1: Pho hap thu UV cua dung dch Carbaryl trong nc
Da vao o th pho hap thu UV cua Carbabyl hnh 4.1 ta thay Carbaryl co 2 nh
hap phu bc song 220 va 280 nm. Nhng bc song 220 o hap thu at c cao
hn. So sanh vi ket qua cua nam trc ket qua nay hoan toan giong. Nh vay co the
chon bc song 220 nm e at che o cho dau do UV khi o Carbabryl cua may
HPLC.
4.1.2. Pho hap thu UV cua Dimethoate trong nc.
3

Ph hp thu ca Dimethoat

h p thu

2.5
2
1.5
1
0.5
0
-0.5 200

220

240

260

280

Bc sng (nm)

Hnh 4.2: Pho hap thu UV cua dung dch chuan Dimethoate trong Axetonitril: nc

33

LUAN VAN TOT NGHIEP

Pho hap thu cua Dimethoate hnh 4.2 thu c khi chnh pH ca dung dch
Dimethoate pH = 6. o th co mot nh hap thu bc song 204 nm cho ket qua phu
hp vi nam trc. ong thi chung toi nhan thay trong qua trnh o quang pho cua
Dimethoate trong nc co s anh hng ln cua pH dung dch. Khi chnh pH cua dung
dch Dimethoate gia tr 9 10, chung toi thay pho hap thu khong co nh hap thu cc
ai va hnh dang ng pho dao ong zch zac. ieu nay co the giai thch la moi
trng pH = 9 -10 mang tnh kiem th Dimethoate khong on nh va b phan huy nhanh
lam cho gia tr o hap thu khi o dao ong nhieu.
4.1.3. Pho hap thu UV cua Vitamin C trong em:

Ph Vitamin C trong dung dch m

0.7
0.6

hp thu

0.5
0.4
0.3
0.2
0.1
0
-0.1210

240

270

300

330

360

390

Bc sng (nm)

Hnh 4.3: Pho hap thu UV dung dch Vitamin C trong em

Pho quet c cho thay nh hap thu cc ai cua Vitamin C 245 nm, ket qua nay
hoan toan phu hp vi tai lieu phan tch Vitamin C cua bo mon. Nh vay luan van nay
cung at gia tr bc song au do UV cua may HPLC la 245 nm khi o viamin C trong
suot luan van.
4.2. Ket qua khao sat pha ong oi vi Carbaryl va Dimethoate
4.2.1. Ket qua khao sat pha ong cua Carbaryl:
Ket qua ve dien tch peak va thi gian lu khi khao sat cac pha ong khac nhau
cho thay khi tang dan lng Axetonitril len th dien tch peak tang theo va thi gian
lu giam dan.ieu o chng to t le Axetonitril cang ln th kha nang ra giai cua pha
ong cang cao.V Axetonitril la dung moi phan cc dung e ra giai cot pha ao trong
phan tch cac chat co o phan cc trung bnh en manh. Luan van nay khao sat kha
nang ra giai cua cac pha ong va t o chon ra pha ong phu hp e tiep tuc khao
sat Carbaryl trong cac phan sau luan van.

34

LUAN VAN TOT NGHIEP

Bang 4.1: Dien tch peak va thi gian lu cua Carbaryl cac pha ong
He pha ong Thi gian lu trung Dien tch peak trung
bnh, phut
bnh
40/60

11,374 0.038

1270426 60726

55/45

6,291 0.084

1507740,7 7791

70/30

5,502 0,166

1775789,7 18823

85/15

3,359 0.806

2016552 52632

e chon c mot pha ong chay HPLC ta khong ch da vao kha nang ra giai cao
ma con phai xem xet ve thi gian lu phu hp.Va con tnh en viec tiet kiem dung
moi s dung v dung moi chay HPLC la dung moi co ch ph at. Thi gian lu chon
phai lam sao khong qua dai e rut ngan thi gian phan tch cung nh khong qua ngan
v thi gian lu ngan de trung vi peak dung moi ra, luc o dien tch peak o c se
khong chnh xac. Da tren Bang 4.1 tren ta thay pha ong vi t le Axetonitril : nc
la 55/45 la co the chap nhan c. Neu tang t le nay len 70/30 th thi gian lu giam
xuong khong ang ke ma lai tang len mot lng khong t dung moi nen chon pha ong
nay khong kinh te lam.
4.2.2. Ket qua khao sat anh hng thanh phan pha ong phan tch Dimethoate
Bang 4.2: Dien tch va thi gian lu cua Dimethoate cac pha ong
He pha Thi gian lu Dien tch peak trung
ong
trung bnh, phut
bnh
40/60

6,549 0,100

184982,3 961,9

55/45

3,973 0,063

219896 7476

70/30

3,654 0,048

342067,7 15324

85/15

2,971 0,004

364542 17498

Tng t khao sat pha ong cua Carbaryl, oi vi Dimethoate ta cung tien hanh
khao sat e chon ra pha ong phu hp nhat. Da tren Bang 4.2 ta thay Dimethoate
cung vay khi tang t le cua Axetonitril th thi gian lu giam ong thi dien tch peak

35

LUAN VAN TOT NGHIEP

tang len. pha ong t le 70/30 va 85/15 dien tch peak kha cao so vi t le 55/45 va
40/60. Nhng pha ong 55/45 cho sac ky o co hnh dang can oi hn so vi cac pha
ong khac. Chung toi thay pha ong 55/45 cho thi gian lu va dien tch peak tng
oi trung bnh gia cac pha ong. V luan van nay ong thi khao sat oi vi hai chat
nen chung toi chon pha ong chay Dimethoate cung la 55/45 e tien viec phan tch
ong thi cho hai chat. Muc ch la tiet kiem thi gian khi chuyen t phan tch chat
nay sang chat kia v thi gian thay pha ong e phan tch mot chat rat lau mi co the
phan tch on nh.
4.3. Ket qua xay dng ng chuan Carbaryl, Dimethoate, Vitamin C
4.3.1. ng chuan cua carabryl trong dung dch nc:
Pha ong cho phan tch Carbaryl va Dimethoate dung la Axetonitril: nc theo t le
55/45.
6,198

( a)

(b)

6.297

Hnh 4.4: Sac ky o o chuan Carbaryl nong o 10 ppm (a) va 5 ppm (b)
4000000

Dien tch peak

y = 185795x
R 2 = 0,9932

3000000
2000000
1000000
0
0

10

15

20
25
Nong o Carbaryl, ppm

Hnh 4.5: ng chun phan tch Carbaryl trong nc

36

LUAN VAN TOT NGHIEP

Ket qua o cac mau chuan cua Carbaryl cac nong o cho dien tch peak va thi gian
lu co o lap lai tot, vi o lech chuan chap nhan c cho sai so thc nghiem < 5%.
ng chuan xay dng cho khoang nong o t 1 ppm en 20 ppm co o tuyen tnh cao
vi he so tng quan R = 0,996.
4.3.2. ng chuan Dimethoate trong dung dch nc:
Tng t Carbaryl, oi vi Dimthoate ket qua thu c cung co o lap lai cao gia
cac lan o. ng chuan trong khoang nong o o t 1ppm en 20 ppm co o tuyen
tnh cao R = 0,997. ay ta thay thi gian lu cua Dimethoate khoang gan 4 phut
ngan hn so vi thi gian lu cua Carbaryl la hn 6 phut co the giai thch c la do
o phan cc cua Dimethoate ln hn so vi o phan cc cua Carbaryl. V vay pha
ong se loi keo Dimthoate ra nhanh hn so vi Carbaryl.

3,937

(a)

(b)
3,968

Hnh 4.6: Sac ky o o chuan Dimethoate nong o 5 ppm (a) va 1 ppm (b)
Dien tch peak

400000

y = 16006x
2

R = 0,995
300000

200000

100000

0
0

10

15

20
25
Nong o Dimethoat, ppm

Hnh 4.7: ng chuan phan tch Dimethoate trong dung dch Axetonitril: nc
37

LUAN VAN TOT NGHIEP

4.3.3. ng chuan dung dch Vitamin C trong em:
Ket qua ng chuan cua Vitamin C trong khoang nong o o t 1 ppm en 20 ppm
nh hnh 4.7. Sai so gia cac lan o < 5 % la chap nhan c, he so tuyen tnh cua
ng chuan cao R = 0,997.
2,397
2,397

Hnh 4.8: Sac ky o o chuan Vitamin C 1ppm va 2 ppm

Dien tch peak

y = 29352x
2

R = 0,9957

500000
400000
300000
200000
100000
0
0

10

15

20

25

Nong o Vitamin C, ppm

Hnh 4.9: ng chuan Vitamin C trong dung dch em

Ngng nh lng cua may HPLC
Do han che cua phan mem ng dung ghi va x ly so lieu nen khong the tnh
c gia tr ly thuyet cua ngng phat hien (LOD) va ngng nh lng (LOQ). Gii
han nh lng (LOQ) cua qui trnh phan tch cho ba chat tren c xac nh qua thc
nghiem bang cach o cac mau chuan co nong o thap. Thc nghiem cho thay ket qua
o on nh va co o lap lai tot (ca dien tch peak lan thi gian lu) nong o 0,1ppm
oi vi Dimethoate. Co the lay ay la ngng nh lng cua Dimethoate. Da vao he
so goc cua ng chuan cua 3 chat, chung toi thay o nhay cua may oi vi Carbaryl
la ln nhat roi ti Vitamin C va Dimethoate. Nh vay da vao t so he so goc ng

38

LUAN VAN TOT NGHIEP

chuan va ngng nh lng cua Dimethoate la 0,1 ppm, chung toi suy ra c ngng
nh lng cua Vitamin C la 0,05 ppm va Carbaryl la 0,008 ppm.
4.3. Ket qua o thu hoi mau Carbaryl, Dimthoate va Vitamin C:
4.3.1. o thu hoi mau Dimethoate qua cot:
Phan tch cac mau Dimethoate co nong o 1 20ppm lap lai 3 lan, lay gia tr trung
bnh cua dien tch peak va thi gian lu. Sau o cung tiep tuc phan tch cac mau nay
nhng e dong pha ong dan mau thang t au do, khong qua cot phan tch. Lay gia
tr trung bnh cua dien tch peak mau khong qua cot. T so gia dien tch peak cua mau
qua cot va mau khong qua cot nong o tng ng la o thu hoi mau qua cot.
Ta co bang sau:
Bang 4.3: o thu hoi qua cot cua Dimethoate:
Nong
o mau,
ppm

Mau qua cot

Mau khong qua cot

o thu hoi
mau qua cot,
%

Dien tch peak

RSD%

Dien tch peak

RSD%

34538,3

2,48

107228

4,31

32,21 1,57

61590,5

3,68

151350

3,24

40,69 1,99

163449,5

4,12

253129,8

4,2

64,57 3,74

10

211424,5

3,84

323055,3

3,09

65,45 3,2

20

463938,3

1,51

584562

3,4

79,36 2,93

Tnh o thu hoi theo cong thc 1 muc 3.6, ta thay o thu hoi mau qua cot cua
Dimethoate tang len khi tang nong o mau. ieu nay co the giai thch la do nong o
thap mot lng nho Dimethoate b gi lai cung lam anh hng ang ke en ket qua.
Nhn chung th ket qua o thu hoi mau cung tng oi cao, chng to he pha ong ra
giai Dimethoate kha tot.
4.3.2. o thu hoi qua cot cua Carbaryl:
Tien hanh tng t nh oi vi Dimethoate ta co bang sau:

39

LUAN VAN TOT NGHIEP

Bang 4.4: o thu hoi qua cot cua Carbaryl
Nong
o
mau,
ppm

Mau qua cot

Mau khong qua cot

o thu hoi mau

qua cot, %

Dien tch peak

RSD%

Dien tch peak

RSD%

195813,5

4,69

236675

1,55

82,74 4,07

313848,8

1,44

372470,8

1,27

84,26 1,61

750989,5

3,9

788714,8

1,42

95,22 3,95

10

1512305

0,74

1543244

0,72

98,00 1,00

o thu hoi mau qua cot cua Carbaryl cao hn rat nhieu so vi cac mau
Dimethoate, ieu nay chng to he pha ong Axetonitril/ nc vi thanh phan 55/45
ra giai Carbaryl ra khoi cot tot hn nhieu so vi ra giai Dimethoate. Tuy nhien e
thuan tien trong qua trnh th nghiem, tao s on nh cho cot phan tch, trong luan van
nay ch dung mot he pha ong 55/45 e phan tch ca 2 chat.
4.3.3.o thu hoi qua cot cua Vitamin C:
Da vao ket qua tren bang 4.5 ta thay o thu hoi qua cot cua Vitamin C kha cao,
tang dan khi nong o mau tang.
Bang 4.5: o thu hoi qua cot cua Vitamin C
Nong o
mau,
ppm

o thu hoi mau

qua cot, %
Mau qua cot

Mau khong qua cot

Dien tch peak

RSD%

Dien tch peak

RSD%

30427

1,56

35170

2,34

86,51 2,43

57752

4,78

65252

4,17

88,51 5,57

143544

5,01

157669

1,34

91,04 4,71

10

281456

4,3

302972

2,45

92,9 4,58

20

559646

1,45

598297

4,3

93,54 4,23

40

LUAN VAN TOT NGHIEP

4.4. Ket qua khao sat hieu suat trch ly dch Carbaryl va Dimethoate trong nc
qua cac bac trch ly.
Theo tai lieu luan van nam trc qua trnh trch ly ch tien hanh mot bac va lng
mau x ly nho nen hieu suat thu hoi t rau cai bap cua qua ch at 70,38% oi vi
Carbaryl va 82% oi vi Dimethoate. V vay chung toi tien hanh khao sat them so bac
trch ly e at c hieu suat thu hoi mau cao hn. Trc het chung toi khao sat tren
dung dch chuan trong nc cua hai chat Carbaryl va Dimethoate e xem so bac trch
ly thc s y ngha la bao nhieu.
Sau khi tach pha ta thu c pha hu c va pha nc. Pha hu c tiep tuc c x
ly nh lan 1, thu c dch 1 em i phan tch. Con pha nc lai tiep tuc cho Axeton
vao, qua cac qua trnh nh trc ta thu c dch 2 em i phan tch. Tiep tuc tach
pha lan 2 va lam nh lan 1 c dch 3. 2 lan x ly sau, dch c nh mc 10mL.
moi mau phan tch 3 lan, lay gia tr trung bnh cua dien tch peak va thi gian lu, ta
thu c bang sau:
Bang 4.6: Ket qua khao sat cac bac trch ly Dimethoate
Bac trch ly

Dien tch peak

RSD%

So bac
trch ly

o thu hoi mau

(%)

1 (nh mc 25mL)

290692,6

1,08

90,81 1,98

2 (nh mc 10mL)

11318,3

3,93

92,22 4,14

3 (nh mc 10mL)

1248

4,52

92,38 5,81

T ket qua dien tch peak cac mau o c nay ta tnh c o thu hoi mau qua cac
bac trch ly tng ng. oi vi trch ly mot bac th o thu hoi mau at c kha cao,
khoang 90,81%. khi tang them mot bac trch ly th o thu hoi tang len la 92,22%, con
khi tang len 3 bac th o thu hoi tang len khong ang ke. Do o e tiet kiem thi gian
va dung moi, ta se tien hanh trch ly 2 bac.
Tng t ta co ket qua oi vi Carbaryl nh bang 4.7 sau:
Bang 4.7: Ket qua khao sat cac bac trch ly Carbaryl
Bac trch ly

Dien tch peak

RSD (%)

So bac trch

o thu hoi mau (%)

1 (nh mc 25 ml)

3383912

0,18

90,05 3,22

2 (nh mc 10 ml)

280926

2,09

94,09 3,89

3 (nh mc 10 ml)

138435

1,18

95,58 4,11

41

LUAN VAN TOT NGHIEP

oi vi carabryl chung toi cung chon 2 bac trch ly v ket qua cho thay bac th 3
o thu hoi mau tang len khong ang ke ma lai ton them thi gian va dung moi e thc
hien.
T ket qua nay chung toi a ra c quy trnh trch ly 2 bac cho Carbaryl va
Dimethoate trong cai bap nh hnh 4.10 sau: (trang 43)
4.5. Ket qua khao sat o thu hoi mau cua quy trnh trch ly Carbaryl va
Dimethoate tren mau cai bap:
4.5.1. Ket qua o thu hoi Carbaryl cua cai bap
o thu hoi mau cua Carbaryl bo sung len rau va e qua em:
Cac mau sau khi c trch ly em i phan tch moi mau t nhat 3 lan, ket qua thu
c bang sau:
Bang 4.8: Ket qua o mau xac nh o thu hoi cua Carbaryl
Mau

Dien tch peak

RSD (%)

Mau trang

40 706

2,78

Bo sung 2,5 ml

381443

5.03

Bo sung 5 ml

865077

4,89

Bo sung 10 ml

1702964

5,01

Chuan 2,5 ml

398339

2,07

Chuan 5 ml

897271

1,96

Chuan 10 ml

1759452

1,50

T ket qua nay chung toi tien hanh tnh o thu hoi mau cua quy trnh phan tch theo
cong thc 2 muc 3.7.3 c ket qua sau:
Mau

o thu hoi (%)

Bo sung 2,5 ml

85,53

Bo sung 5 ml

91,87

Bo sung 10 ml

94,6

42

LUAN VAN TOT NGHIEP

100g Cai bap
200mL Axeton

Xay

Lac
Ba

Loc 1

25 27g NaCl

Khuay t 1

Lac

50mL Axeton

Tach pha 1
Loc 2

Pha hu c

Pha nc

50ml Axeton

Khuay t 2

Tach pha 2

Pha hu c

Pha nc

Co quay chan khong

25mL
Axetonitril

Trang bnh

nh mc 100mL

Sieu am

Sieu loc

43

Hnh 4.10: Quy trnh trch ly cai bap

Dch phan
tch

LUAN VAN TOT NGHIEP

o thu hoi mau carabryl khi bo sung them carabryl vao dch trch rau:
Khac vi viec bo sung them carabryl vao rau nh tren, th nghiem nay chung toi
bo sung them Carbaryl vao ngay dch trch sau qua trnh loc ba e khao sat xem long
Carbaryl b mat mat trong giai oan loc va con sot trong ba la bao nhieu.
Bang 4.9: Ket qua o mau xac nh o thu hoi Carbaryl them vao dch
Mau

Dien tch
peak

RSD (%)

Mau trang

43134

1,08

Bo sung 2,5 ml

399139

4,60

Bo sung 5 ml

900240

3,67

Bo sung 10 ml

1765974

5,10

Chuan 2,5 ml

398339

2,07

Chuan 5 ml

897271

1,96

Chuan 10 ml

1759452

1,50

Ket qua t bang tren chung toi tnh c o thu hoi mau ra ket qua nh sau:
Mau

o thu hoi (%)

Bo sung 2,5 ml

89,17 4,5

Bo sung 5 ml

92,2 3,87

Bo sung 10 ml

95,5 5,15

4.5.2. Ket qua o thu hoi Dimethoate cua cai bap:

o thu hoi mau cua Dimethoate bo sung len rau va e qua em
Cac mau cai bap c bo sung them lng dung dch chuan sau khi tien hanh trch ly,
tien hanh em phan tch moi mau t nhat 3 lan, ket qua thu c bang sau 4.10:
(trang 45)

44

LUAN VAN TOT NGHIEP

Bang 4.10: Ket qua o mau xac nh o thu hoi cua Dimethoate
Mau

Dien tch peak

RSD (%)

Mau trang

26518

3,01

Bo sung 2,5 ml

99846

4,01

Bo sung 5 ml

128588

3,90

Bo sung 10 ml

220272

4,10

Chuan 2,5 ml

51205

1,04

Chuan 5 ml

84256

1,17

Chuan 10 ml

186593

1,50

o thu hoi tnh t ket qua thu c tren nh sau:

Mau

o thu hoi (%)

Bo sung 2,5 ml

84,61 4,31

Bo sung 5 ml

85,65 4,28

Bo sung 10 ml

87,76 4,65

o thu hoi mau carabryl khi bo sung them Dimethoate vao dch trch rau
Cac mau em o tren may HPLC thu c ket qua:
Bang 4.11: Ket qua o mau Dimethoate bo sung vao dch sau loc
Mau
Mau trang
Bo sung 2,5 ml

Dien tch peak

24991
68754

RSD(%)
3.12
4,12

Bo sung 5 ml

101639

2.56

Bo sung 10 ml

195660

3,56

Chuan 2,5 ml

51205

1,04

Chuan 5 ml

84256

1,17

Chuan 10 ml

186593

1,50

45

LUAN VAN TOT NGHIEP

o thu hoi tnh theo cong thc 2 muc 3.7.3, t so lieu cua bang tren cho ket qua
sau:
Mau

o thu hoi(%)

Bo sung 2,5 ml

85 4,42

Bo sung 5 ml

90,9 3,72

Bo sung 10 ml

91,46 4,48

Nh vay ket qua o thu hoi mau cua quy trnh trch ly tng oi cao, oi vi
Carbaryl t 85,53% en 94,6% va oi vi Dimethoate t 84,61% en 87,76%. So vi
ket qua cua nam trc th ket qua nay tng oi cao. o thu hoi cua Carabryl cao hn
cua Dimethoate nhng khong qua ln. ieu o cho thay quy trnh phan tch la chon
co kha nang phan tch kha tot ham lng Carbaryl cung nh Dimethoate trong cai bap.
Ngoai ra ket qua o thu hoi cua mau bo sung them vao dch con cho chung ta thay
rang giai oan loc ba lam mat mat mot lng en khoang 3,64 % oi vi Carbaryl va
khoang 5,25 % oi vi Dimethoate.
T so lieu o c cua mau trang cai bap oi vi Carbaryl va Dimethoate ta co
the tnh ra c ham lng that s cua Carabryl va Dimethoate co trong mau rau a
mua ngoai th trng.
Thiet lap cong thc tnh ham lng Carbaryl va Dimethoate co trong rau:
oi vi quy trnh phan tch Carbaryl va Dimethoate a chon, chung toi a xac nh
c o thu hoi mau qua khau o tren may HPLC va trong qua trnh trch ly Carbaryl
va Dimethoate nh tren. Nh vay khi co mot mau rau cai bap, chung toi tien hanh
trch nh quy trnh trch tren va phan tch tren may HPLC ket qua ra dien tch peak
cua chat can o. T so lieu dien tch peak nay, da vao ng chuan moi chat va o
thu hoi qua tng khau, chung toi co the xac nh c ham lng chat can phan tch.
T ket qua o thu hoi mau qua cot bang 4.3, 4.4 va o thu hoi cua quy trnh trch ly
bang 4.8, 4.10, ket qua ng chuan hnh 4.4, 4.5, thiet lap c cong thc sau:
100 S mau
100 100

F
m 16006 56,45 86

[Cong thc 3]

100 S mau
100
100

F
m 185795 90,05 90,67

[Cong thc 4]

oi vi Dimethoate:

X ( ppm)

oi vi Carbaryl:

X ( ppm)

46

LUAN VAN TOT NGHIEP

Vi m: khoi lng mau em phan tch (g)
F: He so pha loang mau trc khi phan tch tren may HPLC
Smau: Dien tch peak o c bi may HPLC
Cac gia tr 56,45; 90,05 va 86; 90,67 lan lt la o thu hoi qua cot trung bnh va
o thu hoi quy trnh trch trung bnh cua Dimethoate va Carbaryl.
Cac gia tr 16006 va 185795 lan lt la he so goc ng chuan cua dung dch
Dimethoate va Carbaryl.
Xac nh d lng Carbaryl va Dimethoate co trong cai bap:
Mau cai bap o ham lng Carbaryl co dien tch peak 43134 ap dung cong thc 4
tren tnh c ham lng la 0,28 ppm. Nh vay ham lng nay la be hn so vi gia tr
gii han cho phep cua Carbaryl tren rau cai bap la 5 ppm.
Tng t oi vi d lng cua Dimethoate, mau o co dien tch peak 36518 ap
dung cong thc 3 tnh c ham lng Dimethoate co tron cai bap la 4,7 ppm. D
lng nay vt mc gii han cho phep cua Dimethoate tren cai bap la 2 ppm.
Theo thong tin chung toi biet c t Trung tam nghien cu va phat trien nong
dc VIPESCO th hien nay thuoc tr sau Carbaryl lng s dung a t han i. Nhng
Dimethoate th van con s dung pho bien. Nh vay ket qua phan tch tren oi vi cai
bap la phu hp vi thong tin nhan c.
4.6. Ket qua khao sat s phan huy Carbaryl di tac dung cua chieu xa:
ng dung quy trnh trch ly Carbaryl t cai bap vao phan tch mau cai bap co bo
sung them 5 ppm Carbaryl em i chieu xa cac lieu chieu khac nhau e xem xet kha
nang phan huy carabryl trong dch rau cua tia bc xa. Cac mau sau khi trch ly c
em phan tch moi mau t nhat 3 lan, lay ket qua dien tch peak o c so sanh vi
dien tch peak cua mau cai bap bo sung them 5 ppm lam oi chng chung toi ra c
hieu suat phan huy Carbaryl cac lieu chieu khac nhau.
Ket qua hieu suat phan huy Carbaryl theo cac lieu chieu khac nhau c the hien
tren hnh 4.11: (trang 48)

47

LUAN VAN TOT NGHIEP

Hiu sut
(%)

Hiu sut phn hy Carbaryl trn

ci bp
100
80
60
40
20
0

85.73
75.63
61.45
44.0837.75

1 KGy
0,8 KGy
0,6 Kgy
0,4 KGy
0,2 KGy

Liu chiu
1 (KGy)

Hnh 4.11: Hieu suat phan huy tren cai bap cac lieu chieu khac nhau
Ket qua tren cho thay khi tang lieu chieu dan t 0,2 KGy len ti 1 KGy th kha
nang phan huy cua carabryl cang tang,ieu nay cung hp ly v khi tang lieu chieu th
nang lng cung cap cang tang dan en mc o lam phan huy Carbaryl cung tang
theo.Theo ket qua o thu hoi mau cua Carbaryl qua quy trnh trch ly la tng oi cao
nh vay so lieu o c cua mau rau cai bap sau khi chieu xa co the tin cay cao
hn.Nh vay t ket qua nay chung toi thay vi lieu chieu en 1KGy cung co the lam
giam lng Carbaryl en 85,73 %, ket qua nay cho kha nang co the ng dung chieu
xa e lam giam d lng thuoc tr sau tren rau am bao an toan ve sinh cho rau qua
trc khi s dung.
4.7. Ket qua khao sat o thu hoi cua quy trnh trch Vitamin C t cai bap:
4.7.1. o thu hoi Vitamin C khi bo sung them chuan vao rau:
Vi quy trnh trch nh phan tren a gii thieu, chung toi tien hanh trch ly cac mau
cai bap co bo sung them chuan Vitamin C, sau khi pha loang mau em o tren may
sac ky thu c ket qua bang 4.12 sau: (trang 49)

48

LUAN VAN TOT NGHIEP

Bang 4.12: Ket qua o mau bo sung Vitamin C
Mau

Dien tch
peak

RSD(%)

Mau trang

71938

2,78

Bo sung 5 ml

189076

3,56

Bo sung 10 ml

312040

5.23

Bo sung 15 ml

453275

2,67

Chuan 5 ml

141236

1,23

Chuan 10 ml

282400

2,34

Chuan 15 ml

423679

1,23

T bang tren ta co the tnh c o thu hoi cua Vitamin C cho ket qua sau:
Mau

o thu hoi(%)

Bo sung 5 ml

82 3,38

Bo sung 10ml

84,41 5,31

Bo sung 15 ml

86,5 3,49

Ket qua o thu hoi cua Vitamin C at c t 82% en 86,5% tng oi cao. So
sanh vi ket qua at c cua nam ngoai la 89,5% th ket qua o c cua nam nay
co thap hn mot t nhng khong ang ke. Co the la do thao thac th nghiem, sai so chu
quan do ngi thc hien. V Vitamin C la mot chat de phan huy di anh sang nen
thi gian sau khi trch ly xong en khi phan tch cung lam anh hng ln en ham
lng Vitamin C o c. Thi iem tien hanh trch ly va phan tch cung anh hng
en ket qua thu c, th nghiem tien hanh vao buoi toi nhiet o thap va t anh sang
gat th Vitamin C cung t phan huy hn so vi tien hanh th nghiem vao buoi ban ngay
nhat la buoi tra. Nhn chung vi o thu hoi mau at c qua e tai nh vay cho thay
quy trnh trch ly va phan tch Vitamin C a chon co the ap dung tot e phan tch
Vitamin C trong rau vi o tin vay cao.

49

LUAN VAN TOT NGHIEP

4.7.2. o thu hoi Vitamin C khi bo sung vao dch rau:
o thu hoi cua Vitamin C khi bo sung vao trc tiep vao dch rau so sanh vi o thu
hoi cua Vitamin C khi bo sung tren rau se cho thay s mat mat va lng Vitamin C
con sot lai trong ba, cac mau rau sau khi bo sung Vitamin C vao dch tien hanh cac
giai oan tiep theo thu c dch phan tch, tien hanh o moi mau t nhat 3 lan thu
c bang:
Bang 4.13: Ket qua o mau bo sung Vitamin C vao dch
Mau

Dien tch peak

RSD(%)

Mau trang

72370

2,56

Bo sung 5 ml

190567

5,1

Bo sung 10 ml

317688

3,45

Bo sung 15 ml

460137

4,57

Chuan 5 ml

141236

1,23

Chuan 10 ml

282400

2,34

Chuan 15 ml

423679

1,23

o thu hoi tnh c nh sau:

Mau

o thu hoi (%)

Bo sung 5 ml

83,99 4,87

Bo sung 10ml

86 4,20

Bo sung 15 ml

88 4,66

So sanh 2 o thu hoi ta thay giai oan loc mau va ba cung lam mat mat mot lng
Vitamin C khoang 1,5%. T ket qua o c mau khong bo sung them Vitamin C ta
co the tnh c ham lng Vitamin C thc co trong cai bap trong mau rau mua ngoai
ch e phan tch.

50

LUAN VAN TOT NGHIEP

Tng t nh thiet lap cong thc tnh ham lng Carbaryl va Dimethoate trong cai
bap, chung toi cung lap cong thc tnh ham lng Vitamin C trong cai bap nh sau:
X ( ppm)

100 S mau 100 100

F
m 29352 90,5 84,3

[Cong thc 5]

Trong cong thc gia tr 90,5 va 84,3 la gia tr o thu hoi qua cot trung bnh va o thu
hoi quy trnh trch Vitamin C trung bnh.
Vi dien tch peak 72 370 oi vi mau trang cai bap o ham lng Vitamin C, ap
dung cong thc 5 tren tnh c ham lng Vitamin C co trong cai bap em phan tch
la 129,2 ppm.
So sanh ket qua nay vi ham lng Vitamin C co trong cai bap theo tai lieu tham
khao [9] la 300 ppm, cho thay ham lng Vitamin C thc te trong mau o c thap
hn nhieu. Co nhieu nguyen nhan dan en s sai lech ln nay, theo chung toi th nhat
co the la do mau rau cai bap mua ngoai ch ve phan tch co thi gian sau thu hoach
lau hn so vi so lieu ma tai lieu tham khao co nen theo thi gian bao quan lng
Vitamin C a b ton that. Th hai trong qua trnh trch ly mau cung b mat mat do thi
gian phan tch keo dai trong khi o mau. Th ba la ham lng Vitamin C co trong mau
rau th kho trch ly ra hn la lng Vitamin C bo sung vao rau e trch ly khi khao sat
o thu hoi mau.

51

LUAN VAN TOT NGHIEP

Chng 5: KET LUAN VA KIEN NGH

5.1 Ket luan:

Ket qua luan van co the c tom tat nh sau:
1. a khao sat c pho hap thu cua Carbarayl, Dimethoate, Vitamin C trong
vung bc song UV t o chon ra c bc song toi u e phan tch cac chat
tren tren thiet b HPLC vi au do UV- VIS.
2. a khao sat o thu hoi cua Carbaryl, Dimethoate va Vitamin C khi qua cot
HPLC. Ket qua cho thay, oi vi Vitamin C va Carbaryl o thu hoi tng oi
cao khoang en 93,54 va 98%. Rieng oi vi Dimethoate do chon pha ong
55/45 nen o thu hoi qua cot HPLC hi thap hn khoang ti 79,36%.
3. a khao sat c quy trnh phan tch cho ca Carbaryl va Dimethoate vi so bac
trch ly co y ngha chon la 2 bac.
4. a khao sat c o thu hoi mau oi vi quy trnh phan tch Carbaryl,
Dimethoate va Vitamin C. Ket qua o thu hoi mau cua cac chat vi quy trnh
chon phan tch la kha cao t 84,61% en 95,5%.
5. a ap dung quy trnh trch e o ham lng Carnaryl, Dimethoate va Vitamin C
tren cac mau rau cai bap mua ch. Ket qua d lng cua Carbaryl la an toan
di mc gii han cho phep. D lng cua Dimethoate thi cao hn d lng cho
phep. Ham lng Vitamin C o c tren rau thap hn so vi gia tr tham khao
trong tai lieu.
6. a ap dung quy trnh phan tch e anh gia anh hng cua chieu xa len s
phan huy Carbaryl trong cai bap. Ket qua o phan huy Carbaryl vi cac lieu
chieu < 1 KGy tang dan theo lieu chieu, vi lieu chieu 1 KGy xac nh o phan
huy cua Carbaryl la 85,73 %.
5.2. Kien ngh:
Do thi gian va ieu kien thc hien luan van co han ma khoi lng oi hoi phai
khao sat khong t nen e cho thuan li va giam thi gian phan tch, chung toi a chon
cung pha ong cho phan tch ca 2 chat Carbaryl va Dimethoate.V vay e cho o thu
hoi qua cot oi vi Dimethoate cao hn th co the chon pha ong Axetonitril : nc t
le 70/30 thay v 55/45 nh chung toi a khao sat.
Dch trch cua Carbaryl va Dimethoate em phan tch tren may HPLC con cha
nhieu tap chat, co the lam anh hng en kha nang tach cua pha ong, sac ky o o
c khong c ep. e loai bo het cac tap chat cua dch trch va lam giau cau t
can phan tch chung toi e ngh dch trch can phai c lam giau mau qua qua trnh
trch ly pha ran (SPE) e ket qua o c chnh xac va t b anh hng cua nen cac
chat co trong rau.

52

LUAN VAN TOT NGHIEP

TAI LIEU THAM KHAO

[1]. ao Van Hoang, Ky thuat tong hp cac hoa chat Bao ve thc vat. Nha xuat ban
Khoa hoc Ky thuat, 2003.
[2]. Phung Vo Cam Hong, Khao sat d lng Cypermethrin va Fenobucarb cai ngot
va au Cove tai huyen Hocmon va tren th trng Tp HCM. Luan van thac s. Trng
ai hoc Bach khoa Tp HCM, 2003.
[3]. Tran Th Ngoc Linh, Nghien cu anh hng cua chieu xa lieu thap len tnh chat
cam quan va ham lng Vitamin C trong cai xanh, cai bap va da leo. Luan van tot
nghiep. Trng ai hoc Bach khoa Tp HCM, 2007.
[4]. Nguyen Tran Oanh, Hoa hoc Bao ve thc vat. Nha xuat ban Nong nghiep, 1997.
[5]. Lu Th Thu Trang, Nghien cu anh hng cua chieu xa lieu thap len d lng
Dimethoate trong cai xanh, cai bap, da leo. Luan van tot nghiep, Trng ai hoc
Bach khoa Tp HCM, 2007.
[6]. Tran Th Mai Tram, Nghien cu anh hng cua chieu xa lieu thap len d lng
Dimethoate trong cai xanh, cai bap, da leo. Luan van tot nghiep, Trng ai hoc
Bach khoa Tp HCM, 2007.
[7]. Bui Cach Tuyen, Giao trnh thuoc bao ve thc vat. Trng ai hoc Nong lam. Tp
HCM, 1996.
[8]. Nguyen Th Thu Van, Phan tch nh lng, Nha xuat ban ai hoc quoc gia
Tp.HCM, 2004.
[9]. Tran Khac Phi, Ky thuat trong rau sach, Nha xuat ban Nong nghiep, 1997.
[10]. Cac tac gia. Giao trnh th nghiep phan tch thc pham Phan 2. Trng ai hoc
Bach khoa Tp HCM.
[11]. A Primer, HPLC for food analysis, Agilent Technologies, 1996 2001, 134 trang
[12]. AOAC Official Method 967.21, Ascorbic Acid in Vitamin Preparations and Juices,
1968.
[13].AOAC Official Method 964.18, Carbaryl Insectiside Residuses. Colorimetric
Method, AOAC Official Method of Analysis, 1995.
[14].AOAC Official Method 975.40, N-MethylCarbarmateInsectiside Residuses. Gas
Chromatographic Method, AOAC Official Method of Analysis, 1995.

53

LUAN VAN TOT NGHIEP

[15].http://www.scielo.br/scielo.php?script=sci_arttext&pid=S151689132002000600003
[16].http://www.speclab.com/compound/c60515.htm
[17].http://www.cimsi.org.vn/Sach/dinhduong&attp/chuong6.htm
[18].http://www.inchem.org/documents/pims/pharm/ascorbic.htm#PartTitle:1.%20%20N
AME [26]
[19].http://www.inchem.org/documents/pims/pharm/ascorbic.htm#PartTitle:1.%20%20N
AME [27]
[20].www.quynhluong.gov.vn
[21].http://www.vnn.vn/suckhoe/2004/06/169567/

54

LUAN VAN TOT NGHIEP

PHU LUC
Bang 1: Dien tch peak va thi gian lu xay dng ng chuan Carbaryl
Nong o, Thi gian Thi gian RSD, %
ppm
lu , phut
lu trung
bnh,
phut
6.313
1

10

20

6.187

Dien tch Dien tch RSD, %

peak
peak
trung
bnh
357834

6.256

1.02

364234

6.267

3409350

6.333

408334

6.198

6.255

1.12

418948

6.233

430958

6.367

977156

6.297

6.284

1.44

964026

6.187

957236

6.363

1995450

6.198

6.291

1.34

1964401

6.313

1955700

6.385

3507441

6.288

6.320

0.89

6.288

3767607
3634308

55

354334.3

3.40

419413.3

2.70

966139.3

1.05

1971850

1.06

3636452

3.58

LUAN VAN TOT NGHIEP

Bang 2: Dien tch peak va thi gian lu xay dng ng chuan Dimethoate
Nong o, Thi gian Thi gian RSD, %
ppm
lu , phut
lu trung
bnh,
phut
4.128
1

10

20

3.987

Dien tch Dien tch RSD, %

peak
peak
trung
bnh
14363

4.031

2.09

15496

3.977

15521

3.975

33184

4.038

4.035

1.45

36145

4.092

35945

3.998

75348

4.212

4.054

3.42

75693

3.952

75968

147474

3.945

3.988

0.97

145644

4.02

142072

4.143

319871

4.005

4.1

2.03

4.155

331201
334231

56

15126.7

4.37

35091.3

4.72

75669.7

0.41

145063.3

1.89

328434.3

2.3

LUAN VAN TOT NGHIEP

Bang 3: Dien tch peak va thi gian lu xay dng ng chuan Vitamin C
Nong o, Thi gian Thi gian RSD,
ppm
lu , phut
lu
trung %
bnh, phut
2.397
1

10

20

2.397

Dien
tch
peak
42426

2.397

0.00

43072

2.397

42778

2.408

78163

2.402

2.404

0.14

79714

2.402

79611

2.407

161823

2.4

2.402

0.17

160171

2.4

158997

2.403

287161

2.402

2.402

0.02

287528

2.402

287268

2.407

582738

2.402

Dien tch peak RSD, %

trung bnh

2.404

0.12

2.402

585951
583404

57

42758.7

0.76

79162.7

1.10

160330.3

0.89

287319.0

0.07

584031.0

0.29

LUAN VAN TOT NGHIEP

2,4
2,397

Mau bo sung 5 ml

Mau trang
2,234

Mau bo sung 10 ml
Hnh 1: Sac ky o o Vitanmin C

6,267

6,333

Mau bo sung 5 ml

Mau bo sung 2,5 ml

6,297
6,233

Mau bo sung 10 ml

Mau trang

Hnh 2: Sac ky o o Carbaryl trong cai bap

58

LUAN VAN TOT NGHIEP

3,923

3,92

Mau trang

Mau bo sung 5 ml

3,937
3,968

Mau bo sung 2,5 ml

Mau bo sung 10 ml

Hnh 3: Sac ky o o Dimethoate trong cac mau cai bap bo sung dung dch chuan
Dimethoate

Hnh 4: Sac ky o o thu hoi qua cot cua Dimethoate 10 ppm

Hnh 5: Sac ky o o thu hoi qua cot cua Carbaryl 5 ppm

59

LUAN VAN TOT NGHIEP

Hnh 6: Sac ky o o thu hoi qua cot cua Vitamin C 10 ppm

60