CHNG I TNG QUAN V TI

1.1. 1.1.1.

Mt s khi nim c bn

Thuc tr su (TTS) Thuc tr su hay ho cht bo v thc vt (HCBVTV) xut pht t thut ng ting anh Pesticide c ngha l cht dit loi gy hi. Dch sang ting Vit cc tc gi s dng nhiu thut ng khc nhau nh: thuc tr su, thuc bo v thc vt, ho cht tr su, ho cht bo v thc vt. Nh vy HCBVTV l danh t chung ch mt cht hoc mt hp cht bt k c tc dng d phng, tiu dit hoc kim sot cc sinh vt gy hi, gy bnh cho ngi v ng vt, cc loi cn trng khc hay ng vt c hi trong qu trnh sn xut.Thuc tr su l cht ho hc hoc hp cht t nhin chit xut t cy c dng tr su hi cy.

1.1.2. Cht c L nhng cht khi xm nhp vo c th sinh vt mt lng nh cng c th gy bin i su sc v cu trc hay chc nng trong c th sinh vt, ph hu nghim trng nhng chc nng ca c th, lm cho sinh vt b ng c hoc b cht . Theo t in bch khoa Vit Nam: Cht c kh nng c ch, ph hu hoc lm cht c th sng: khi a mt lng nh cht c vo c th (qua ming, d dy, th ht qua phi, thm qua da) hoc khi c hp th vo mu trong nhng iu kin nht nh, gy ra nhng ri lon sinh l ca c th, lm nguy hi cho sc kho hoc gy nguy him n tnh mng con ngi. 1.1.3. c tnh c tnh l kh nng gy c ca mt cht i vi c th sinh vt mt lng nht nh ca cht c . Theo t in Bch khoa Vit Nam: c

tnh l tnh gy c ca mt cht i vi c th sinh vt. c tnh c chia ra cc dng:

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c cp tnh: cht c xm nhp vo c th gy nhim c tc th,

k hiu LD50 (letal dosis 50), biu th lng cht c (mg) i vi 1 kg trng lng c th c th gy cht 50 % c th vt th nghim (thng l chut hoc th).. Nu cht c ln vi khng kh (hi c, hay trong nc) th c k hiu LC50 (letal concentration 50) biu th lng cht c (mg) trong 1 m3 khng kh hoc 1 lt nc c th gy cht 50 % c th th nghim. LD50 v LC50 cng thp chng t c cp tnh cng cao.
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c mn tnh (c trng din) ch kh nng tch lu cht c

trong c th, kh nng gy t bin, gy ung th hoc qui thai, d dng. 1.1.4. c c biu th mc ca tnh c, l liu lng nht nh ca cht c cn c gy c mt tc ng no trn c th sinh vt khi chng xm nhp vo c th sinh vt. Theo t in Bch khoa Vit Nam: c lng cht c ti thiu git cht mt kilgam sinh vt sng. 1.1.5. D lng D lng l lng cht c cn lu li trong nng sn hoc trong mi trng khi phun thuc BVTV.D lng c tnh bng g (microgram) hoc mg (milligram) lng cht c trong 1kg nng sn hc th tch khng kh t nc..Trng hp d lng qu nh, n v c tnh bng ppm hoc ppb. MRL (Maximun Residue Limit) : Mc d lng ti a cho php

lu tn trong nng sn m khng nh hng n con ngi vt nui.

ADI (Acceptable Daily Intake) : lng cht c chp nhn hp thu

vo c th, khng gy hi cho ngi hay vt nui trong 1 ngy, c tnh bng mg hoc g hp cht c cho n v th trng.

1.2. 1.2.1.

Cc h thuc tr su thng gp trong nng nghip Cc hp cht clo hu c 1.2.1.1. u im v nhc im ca cc hp cht clo hu c

u im - Quy trnh sn xut tng i n gin, gi thnh ca ch phm thp, d ch bin hot cht thnh nhiu ch phm khc nhau ( BTN, ND, BR). Cc thuc ny c ph tc ng rng, hiu lc kh cao, thi gian

hiu lc di thch hp cho vic phng tr ngoi ng, nht l i vi cc loi cy cng nghip. bn ha hc ln trong nhng iu kin thng thng nn d bo qun ton tr. Nhc im di.
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Do bn ha hc ln nn thuc d lu b trong t ai, cy trng,

nng sn, thc phm.Chng lm cho mi trng b nhim trong thi gian

C kh nng gy trng c tch ly mnh.Qua s tip xc vi

thuc nhiu ln hay qua chui thc n hm lng thuc trong c th.

Khi s dng mt loi thuc Clo hu c trong nhiu nm ti mt a

phng d gy ra hin tng cn trng khng thuc. Do nhng hn ch trn nn thuc tr su Clo hu c b cm hay hn ch s dng nhiu nc. 1.2.1.2. Mt s tnh cht,c im khc

- Cu to ha hc : Trong phn t ca cc hp cht ny u c cha nguyn t Clo v cc vng benzen hay d vng - Tnh cht vt l: Thuc k ngh u dng rn, khng tan hay tan t trong nc, tan nhiu trong dung mi hu c v thng c mi kh chu. Tnh cht ha hc : Cc thuc tr su Clo hu c thng c bn

ha hc ln, tn ti lau di ngay c trong mi trng ngoi ng, phn ln u b phn hy trong mi trng kim. 1.2.1.3. Mt s thuc tr su Clo hu c thng dng

Bng 1.1 Mt s TTS clo hu c thng dng

1.2.2. Cc thuc tr su gc ln hu c STT Tn gi thng dng Cu trc ha hc 1 DDT (Dichlodiphenyl Tricloetane)

Tnh c LD50 (chut) 113 mg/kg =

BHC (benzene hexa clorit)

LD50 = 125 mg/kg

Toxaphen

Andrin

LD50 (chut) 40 mg/kg = 70

Diendrin

LD50 (chut) 25 mg/kg

= 30

1.2.2.1. Mt s tnh cht chung - Cng thc ha hc cha : P, C, O, H,S, ..

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- Ph rng dit c nhiu loi su hi - Tc ng rt nhanh: tip xc ,v c, xng hi rt mnh. - Khng tn ti lu trong mi trng ,hiu lc dit su nhanh. - Gy c cp tnh cao do tc ng h thn kinh rt mnh. - D phn hy bi axit v mi trng kim. - t tan trong nc nhng d tan trong dung mi hu c. 1.2.2.2. Mt s thuc tr su gc ln thng gp

Hnh 1.1 : Mt s thuc tr su gc ln thng gp 1.2.2.3. Cng dng v cch dng

Do c tnh c LD 50(chut) = 25 -50 mg/kg, c cp tnh cao nhng khng tch ly trong c th ngi v ng vt.An ton vi cy trng lng khuyn co.Thuc b cm s dng. Thuc c tc dng v c xng hi ,thm su, ph tc ng mnh,ch yu phng tr su nhai gm v mt s cn trng chch ht.Ngoi ra cn c tc dng trn mt s loi nhn gy hi cho cy n qu.Tc ng dit dch hai nhanh, nht l khi nhit mi trng cao.Thi gian c hiu lc ngoi ng l 2 3 ngy.
1.2.3.

Thuc tr su h carbamate Mt s tnh cht chung Cng thc ha hc: Cht dn xut t acid cacbarmin ( NH2COOH ) Ph tc ng hp hn gc ln v chlo hu c, bt u chuyn tnh

1.2.3.1.
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vi nhm cn trng chch ht 1.2.3.2.

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Gy c tnh kh cao, tc ng ln h thn kinh. D phn hy bi acid v mi trng kim t tan trong nc ,nhng d tan trong dung mi hu c. Mt s thuc tr su gc Carbamate thng gp Thuc tr su carbamate: R1 l nhm metyl (CH3 ). V d:

Aldicarb, Methomyl, Oxamy, Isoprocarb

Hnh 1.2 Mt s thuc tr su gc Carbamat thng dng

1.2.3.3.

Cng dng v cch dng

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Aldicard:

2-Metyl-2-2(metylthiopropionandehit)0-metyl

cacbamely-doxim, tc dng ni hp, dng tr su ming chch ht, tuyn trng. Lng dng 0,34-11,25 kg a.i./ha. Khng dng cho cy lng thc v thc phm v thuc rt c. Thuc nhm c I, LD50 qua ming: 0,93 mg/kg; ADI: 0,05 mg/kg.
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Oxamyl : N,N - dimetyl - 2 metyl cacbamoyl oximino - 2 -

(metylthio) - acetamid, c tc dng ni hp, tip xc, dng tr cn trng, nhn v tuyn trng. Lng dng 0,28-1,12kg a.i/ha. Thuc nhm c I, LD50 qua ming: 5,4- 8,9mg/kg; ADI 0,03mg/kg.
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Methomyl : S-metyl N-(metylcacbanoyl) oxithioaxetimidat, c tc

dng ni hp v tip xc. Dng tr rp, su xanh, su keo, nhn . Thuc nhm c I, LD50 qua ming: 17-24mg/kg. Thuc hn ch s dng.

1.2.4. 1.2.4.1.

Thuc tr su gc Pyrethroids( Gc cc tng hp) Mt s tnh cht chung

Hot cht Pyrethrin c trch ly t cy hoa cc Pyrethrum cinerariaetrifolium vo nhng nm 1960. Cng thc ha hc c nhiu ng phn nn rt phc tp. - Cu trc ha hc:

- Ph tc dng rt rng, chuyn bit vi cn trng ming ht chch v cn trng ming nhai ,c bit l u trng b cnh vy. - Tc ng nhanh: tip xc,v c, xng hi yu v khng c tnh ni hp. - Khng tn ti lu trong mi trng, d phn hy bi nh sng v nhit . - S dng liu thp so vi thuc gc Clo, Ln, Cacbamat - Gy c cp tnh yu.Tc ng h thn kinh gy thiu oxy, ngoi ra cn tc ng ln h thn kinh ngc lm cn trng mt thng bng khi bay. - Cht c thi ra ngoi theo ng nc tiu, gii c ging nh gc ln , cacbarmat. - t c vi mi trng v ng vt mu nng, gy tnh khng nhanh khi s dng nhiu. - t tan trong nc, d tan trong m v dung mi hu c. 1.2.4.2.

Mt s thuc tr su pyrethroid thng dng

Cypermethrin

- Lch s : Cypermethrin c tng hp u tin nm 1974. - Tn gi khc l : Sherpa ,Ambush C, Cymbush, Peran, Cyperan.

Tn ha hc: (RS)--cyano-3-phenoxybenzyl (1RS,3RS;1RS,3SR)-

3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropanecarboxylate hoc (RS)-cyano-3-phenoxybenzyl(1RS)-cis-trans-3-(2,2-dichlorovinyl)-2,2dimethylcyclopropanecarboxylate.

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Cng thc ha hc :

C22H19Cl2NO3.

- Cu trc ha hc :

- Phn t lng : 416,3

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c tnh : Thuc k thut dng c st ( 60 0C chuyn thnh dng

dung dch lng), hu nh khng tan trong nc, tan nhiu trong dung mi hu c, tng i bn trong mi trng trung tnh v axit nh, thy phn trong mi trng kim, quang gii km, khng n mn kim loi , thuc nhm c II.
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S dng : Cypermethrin tc dng tip xc v v c, c ph tc

ng rt rng, tr c nhiu loi cn trng v nhn hi, c bit l cn trng thuc b Lepidoptera. Lng dng t 25-200 g a.i./ha ty thuc vo loi cy trng ngoi ng. Cypermethrin cn c dng tr ve, bt, chy, rn cho vt nui ,tr rui, mui trong nh.Cypermethrin c gia cng thnh dng sa,dng hn hp vi nhm Ln hu c (nh Polytrin-P440).... Cypermethrin c th hn hp c vi nhiu loi thuc nhm Clo hu c, Carbamate v Ln hu c s dng.

Deltamethrin
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Tn gi khc : Decis, Decamethrin, K-Obiol, K-Othrin. Tn ha hc : (S) - - Cyano - 3 - phenoxibenzyl (1R, 3R) - 3 -

(2,2 - dibromovinyl) - 2,2 - dimetylxiclopropancacnoxylat.

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Cng thc ha hc : C22H19Br2NO3

- Cu trc ha hc :

- Phn t lng : 505,2

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c tnh : Thuc k thut (>98% Deltamethrin) dng bt, khng

tan trong nc, tan nhiu trong dung mi hu c nh acetone (500 g/l), benzene (450 g/l), dioxane (250 g/l); bn vng trong mi trng acid hn mi trng kim, tng i bn di tc ng ca khng kh, nhng di tc ng ca nh nng mt tri hoc tia t ngoi b phn hu, khng n mn kim loi.
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S dng : Deltamethrin tc dng v c v tip xc, tr c

nhiu loi cn trng v nhn hi rau mu, cy cng nghip, tr c nhiu loi su mt hi nng sn bo qun v nhiu loi cn trng y t ,tr c ve, bt, chy, rn hi vt nui.. Deltamethrin d gy tnh khng i vi cn trng chch ht nh ry nu hi la.

Cyhalothrin

- Tn gi khc : Karate, PP 321.

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Tn ha hc : ( RS ) - - Cyano-3-phenoxibenzyl ( Z ) -

(1RS,3RS)-(2-clo-3,3,3-triflopropenyl)-2,2-dimetylxiclopropalcacboxylat.
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Cng thc ha hc : C23H19ClF3NO3.

- Cu trc ha hc :

- Phn t lng : 449,9 - c tnh : thuc k thut (90%) th du nu, khng tan trong nc, tan trong acetone v nhiu dung mi hu c; thy phn mnh pH > 9.
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S dng : Cyhalothrin c tc dng tip xc, v c, c dng tr

su cho rau, mu, cy cng nghip, tr k sinh trng cho vt nui (ve, bt, rui, mui, chy, rn).. Thuc c pha nc nng 0,05-0,07% phun tr nhn , b xt mui, ry xanh hi tr.
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Fenvalerate Tn gi khc : Esfenvalerate, Pydrin, Belmark, Sumicidin,

Furitrothion, Phenvalerate, Sumifleece, Aqmatrine, Insectral.

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Tn

ha

hc

(RS)--cyano-3-phenoxybenzyl

(2RS)-2-

(chlorophenyl)-3-methylbutyrate. Hoc cyano(3-phenoxyphenyl)methyl 4chloro--(1-methylethyl)benzeneacetate.

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Cng thc ha hc : C25H22ClNO3.

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- Cu trc ha hc :

Phn t lng : 419.9 c tnh : Thuc k thut th lng, khng tan trong nc, tan

trong nhiu dung mi hu c nh acetone, xylene, tng i bn vng di tc ng ca nhit v nh nng mt tri, bn vng trong acid hn trong kim (bn vng pH=4); khng n mn kim loi. Nhm c: II. Khng dng cho cy lm thuc.
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S dng : Thuc tc dng tip xc v v c, tr c nhiu loi

cn trng v nhn, tr ve, bt cho vt nui, dng tr su t, su khoang, su xanh, rp, b nhy, nhn hi rau, ry xanh, b xt mui, b cnh t, nhn hi tr, nhn , su v ba hi cam v tr rp bp.
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Permethrin Tn gi khc : Transpermethrin, Permethrine, Biopermethrin,

Ambush,
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Tn ha hc : 3-phenoxybenzyl (1RS,3RS;1RS,3SR)-3-(2,2

dichlorovinyl)-2,2-dimethylcyclopropanecarboxylate.
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Cng thc ha hc : C21H20Cl2O3.

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- Cu trc ha hc : - Phn t lng : 391

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c tnh : thuc k thut th lng v c chiu hng kt tinh

nhit trong phng, hu nh khng tan trong nc, tan trong nhiu dung mi hu c nh hexane, xylene bn trong mi trng acid, thy phn trong mi trng kim; khng n mn nhm. Nhm c: III..
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S dng : Thuc k thut tc dng tip xc v v c, tr c

trng ca cn trng, c ph tc ng rt rng, c dng tr su hi bng ,su hi cy n tri ,su hi nho, thuc l ,su hi rau ,tr rui, mui, k sinh trng gia sc,tr gin v kh trng kho tng. 1.2.4.3.
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Mt s loi thuc pyrethroit khc Allethrin (Pynamin, Pallethrin): 3-Allyl-2 metyl-4-oxoxiclopentc

tenyl-2,2-dimetyl-3-(2-metylprop-1-enyl)-xiclopropancacboxylat,c

ng tip xc,v c, dng tr rp rau, cy n qu,tr cn trng y t v th y. Thuc c nhm III, LD50per os = 685-11000 mg/kg, LD50 dermal: 2500mg/kg.
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Cyfluthrin (Baythroit): thuc c kh nng tip xc v v c, tr

c nhiu loi su hi rau, cy n qu ,bong ay, ma..Thuc dng 2,5 v 5 SL . Dng 40-60g a.i/ha tr su xanh, su hng,rp hi bong, ai,lc.

1.2.4.4.

C ch tc ng ln c th ca TTS h cc tng hp

Pyrethroid l nhng cht c vi t bo thn kinh. Cc cht ny lin kt vi cc cht thnh phn ca mng si trc thn kinh (l Protein v Lipid), cn tr s vn chuyn ca Ion (ch yu l Na+ v K+) qua mng, lm
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mt in th to nn s dn truyn xung ng thn kinh, dn n thn kinh b t lit, su cht. C ch gy c ca pyrethroid trong c th ngi rt nhanh, ch yu l qu trnh thy phn v hydro ha este ca pyrethroid,cc phn t lin hp ca chng v cui cng b o thi qua ng nc tiu. Trong cc cht chuyn ha tng ng trong nc tiu l cis- and trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane carboxylic acid (cisDCCA and trans-DCCA), cis-3-(2,2-dibromovinyl)-2,2dimethylcyclopropane carboxylic acid (-DBCA), 3-phenoxybenzoic acid (3PBA), and 4-fluoro-3-phenoxybenzoic acid (FPBA).Chu k bn hy sinh hc ca cc pyrethroids trong mu,huyt tng l khc nhau t 2 12 gi.Chu k bn hy ca cc cht cis-DCCA, trans-DCCA, v FPBA c tm thy sau qu trnh bi tit nc tiu, khi tip xc bng ming hoc ht phi bi cc tnh nguyn vin cho th nghim ny l 6,44 gi.Trong s cc cht chuyn ha th 94% cc cht c lc thi qua thn sau 48 gi sau khi tip xc.Cc thuc tr su h hoa cc nh pyrethrin t nhin hay (S)bioallethrin cng b chuyn ha trong c th bi qu trnh thy phn,oxy ha v cc phn t lin hp cui cng cng b loi tr qua ng nc tiu.

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Hnh 1.3 Qu trnh trao i cht ca pyrethroid : cyfluthrin, cypermethrin, detalmethrin v permethrin ngi. Qu trnh chuyn ha ch yu l -(E)-chrysanthemumdicarboxylicacid (trans-CDCA).Kt qu cho thy l trans-CDCA cng nh trans-chrysanthemic acid khng tm thy trong c th ngi.Do khi b phi nhim (S)bioallethrin, qu trnh bi tit trans-CDCA cao nht l 24 gi sau khi b phi
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nhim.V sau 72 gi th nng ca trans-CDCA l di gii hn pht hin( 5 g/l).

Hnh 1.4 Qu trnh chuyn ha ca (S)-bioallethrin trong c th ngi ngi cc triu chng c th b o ngc khc nhau,chng hn nh cm gic khc thng,, d ng da nim mc ,nhc u, chng mt,bun nn.Vi nhng nh hng bt li i vi sc khe , pyrethroid/pyrethrin gc v qu trnh bi tit kh c khng phi l nguyn nhn gy nn cc nh hng bt li i vi sc khe .Do theo quan im ca y hc nu
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b phi nhim pyrethroid liu lng cao th b ng c cp tnh .Do qu trnh bi tit th sau 24 gi nng pht hin c gim mt cch nhanh chng.Trong mi trng ngh nghip ngi tip xc vi pyrethroid cng nh ngi tip xc vi mi trng c cha pyrethroid c th hm lng pyrethroid khng th tm thy trong mu nhng hm lng cc cht chuyn ha ca chng c th tm thy trong nc tiu. 1.3. Phng php xc nh DLTTS h cc tng hp trong mu nng

sn thc phm bng phng php sc k kh 1.3.1. Nguyn tc ca phng php D lng thuc tr su h cc trong tri cy v rau qu c chit vi dung mi ,cc nn mu ht c chit bng hn hp ACN : H2O.Cc cht phn tch c chuyn sang dung mi n hexan v chit ngc lai bng ACN.Dch chit c lm sch bng ct Florisil c gim hot tnh v ra gii vi 6% Ethylether trong n hexan .Hm lng cht phn tch c nh lng bng sc k kh u khi ph. 1.3.2. Quy trnh xc nh Giai on chit mu
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i vi mu c hm lng nc ln(>75%).Cn 50g mu vo

erlen 500ml ,thm 120 ml acetone ng nht mu.

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i vi mu kh hoc c hm lng nc nh.Cn 20 g mu vo

erlen 500ml , thm 150 ml hn hp dung mi ACN : H2O (2 :1).ng nht mu.
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Lc mu v trng ra erlen bng acetone hoc ACN : H2O (2:1) 3

ln ,mi ln vi 25 ml dung mi.Ton b dch lc c thu vo phu chit 500ml

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Giai on chit lng lng Thm 60 ml n hexan vo phu chit.Lc mnh 5 pht Thm 200ml dung dch NaCl 4% ,lc nh trong 30 giy.X b

phn nc
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Chuyn lp n hexan vo bnh cu 250ml thng qua phu lc v

15g Na2SO4 khan.Ra phu chit vi 20ml n hexan, gp vo bnh cu.C gn cn trn my c quay 400C Ho tan cn vi 10ml n hexan v chuyn vo phu chit 125ml,

trng ra bnh cu 2 ln ,mi ln vi 5 ml n hexan. Thm 30ml hn hp ACN bo ha trong n hexan ,lc mnh 5 pht. yn,chuyn pha ACN vo bnh cu 250ml.Chit tip vi 30ml ACN
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C cn dch chit v ha tan cn vi 5ml n hexan Giai on lm sch Ct lm sch c chun b bng 1 lp Na2SO4 khan ( khong 1

cm) ,y ct c lt lp bng thy tinh.Cho n hexan n khong 50ml.Sau cho 10 g Florisil gim hot ha vo ct.Cho tip Na2SO4 vo bn trn ct khong 1cm.X lp dung mi cho ti khi lp dung mi va tip xc vi ct. Chuyn dch chit vo ct.trng bnh 2 ln mi ln vi 10ml n

hexan.M kha cho dung mi chy qua ct vi tc 3ml/pht. Ra gii bng 120 200 ml dung dch ra gii 6% ethykether trong n hexan.Ch : trnh lm kh ct Ton b dch c thu vo bnh cu .C quay ti va cn Ha tan cn vi 2ml n hexan.Chuyn vo vial sc k.

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Phn tch bng GC-MS hoc GC ECD

1.3.3 Tnh ton kt qu

X ( m g / kg= )
Trong : Ss,Sst Cst F M Vdm

S s C st Vdm S st m )g (

ln lt l din tch peak mu v chun nng chun ( g/ml) H s pha long khi lng mu(g) th tch nh mc ( ml)

1.4. Nguyn l ca phng php sc k kh ghp khi ph ( GC MS ) 1.4.1. Gii thiu chung Sc k kh ghp khi ph (Gas Chromatography Mass Spectometry) l mt trong nhng phng php phn tch hin i nht hin nay vi nhy v c hiu cao c s dng trong cc nghin cu v phn tch kt hp. Chc nng: phn tch v xc nh cu trc phn t ca cc hp cht hu c d bay hi, bn nhit. Xc nh hm lng a lng, vi lng ca cc hp cht hu c. Lnh vc ng dng: ngy nay k thut GC/MS c s dng rng ri trong cc ngnh nh y hc, mi trng, nng sn, kim nghim thc phm, v sinh mi trng, nc ung, nc thi cng nghip - sinh hot, cc cht c, thuc tr su, thuc dit c, v cc d lng khng sinh 1.4.2. Nguyn l hot ng Thit b GC/MS c cu to gm hai phn: phn sc k kh (GC) dng tch cc cht cn phn tch ra khi hn hp cht ban u, phn khi ph (MS) l mt u d xc nh cht cn phn tch bng cch ct t cu t
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cht phn tch to thnh cc ion, qua b phn tch khi n u d. Ti y din ra qu trnh ion ha. Ion sinh ra c tp trung v c a vo b phn tch khi. Mi ion phn tch s c mt gi tr m/z xc nh, nh chng c tch chn lc ra khi b phn tch khi v to tn hiu c trng ti h thng pht hin ion. S d c th thc hin kh thun li lin hp sc k khi ph l v c hai phng php ny u c cng nhng c tnh chung sau y: Mu c nghin cu trong trng thi kh. C hai phng php u c nhy pht trin cao. Tc phn tch ca c hai phng php tng t nhau. C th ni hai phng php trn c tng thch cao. S khc bit duy nht ca c hai phng php ny l trong ct sc k lun tn ti mt p sut ln hn p sut mi trng (do chnh lch p sut u v cui ct) trong khi bung ion ca my khi ph hot ng mt chn khng cao (khong 10-6 mmHg). c th ghp ni gia ct tch sc k vi ngun ion, gii php duy nht l phi tch kh mang ra khi trc khi vo bung ion ha. C nh vy th chn khng ca ngun ion mi khng b nh hng. Ton b h thng GC MS c ni vi my tnh iu khin t ng, x l s liu v ghi ph.

Hnh 1.5

S khi GC-MS
21

1.4.3. Sc k kh ghp khi ph (GC MS) Sc k kh l mt phng php chia tch trong pha ng l mt cht kh (c gi l kh mang) v pha tnh cha trong ct l mt cht rn hoc cht lng ph trn b mt cht mang tr dng rn ph u ln thnh pha trong ca ct. GC gm cc b phn chnh: b phn cp kh mang, b phn tim mu, ct tch, ct v detector. GC c th ghp vi cc u d: detector dn nhit (TCD), detector ion ha ngn la (FID), detector cng kt in t (ECD), detector nit photpho (NPD), detector quang k ngn la (FPD) v detector khi ph (MS) 1.4.3.1. H thng cung cp kh Ta thy rng gim p sut qua ct tch t l thun vi nht ca kh mang. Khi la chn kh mang ta phi ch n tnh cht ca ct, detector s dng, tinh khit ca kh mang, yu cu tch... Kh mang c dng phi khng c thay i trng thi l, ho hc khi i qua my sc k kh. Cc loi kh mang thng s dng: kh hidro, kh heli, kh Ar, kh N2 1.4.3.2. B tim mu Ca tim mu (injection port): hn hp cc cht s c tim vo h thng ti ca ny. Mu sau c dn qua h thng bi kh tr, thng l helium. Nhit ca tim mu c nng ln (thng l 300oC) mu tr thnh dng kh. Trc khi vo ct tch, thng thng mu c i vo bung bay hi ri dng dng kh mang a vo ct. Trong sc k kh mao qun, lng mu rt t v lu lng dng kh mang cng rt nh (0,5 5 mL/pht) nn mu c th c a vo ct theo cc hnh thc nh: bm mu gin tip v chia dng, gin tip v khng chia dng, bm mu trc tip.

22

1.4.3.3. Ct sc k kh L ni xy ra qu trnh tch cc cht da trn hai qu trnh hp ph v gii hp xy ra lin tc trn pha tnh v pha ng. Ct (column): Ct trong GC c lm bng thy tinh, inox hoc thp khng r c kch thc, kch c rt a dng, chiu di c th l 25m, 30m, 50m, 100m v c ng knh rt nh (nh mt ng mao dn), mt bn trong c trnh bng mt lp polymer c bit. Thng thng ct c s dng l semivolatile, hp cht hu c khng phn cc nh PAHs. Ngi ta thng chia ct sc k thnh cc loi ct tch : Ct nhi, ct mau qun. Ct nhi : c s dng trong giai on u mi pht trin sc k kh, ct thng lm bng thp, i khi lm bng thy tinh, ct di t 1 3m, ng knh t 3 6mm. Ct c nhi vi mt cht rn c l xp ngp y hoc ghp vi mt pha tnh chn lc. Thnh bn trong ca ct c x l tn hiu ng vi cc xc tc trn mu phn tch lu lng kh mang i qua thng t 10 40ml/pht.

Ct mao qun: c bt u s dng t 1970, ban u thng

c lm bng thp v ngy nay bng thy tinh silic, di t 10 100m, ng knh t 0,05 0,35mm. Nay ngi ta sn xut cc ct ny bng cch ko nhit cao, silic c t inh khit 99,999%. chc chn hn bn ngoi ct thy tinh c ph mt lp polyimide hoc mt lp mng aluminium. Ct c cun trn v nh trn mt gi kim loi to thnh mt lng hnh tr. 1.4.3.4. u d khi ph Khi ph gm 4 b phn chnh: - B to ion (ion source). - B tch ion ra phn tch (Mass analyzer). - o m/z ca mi loi ion (detector).
23

- B x l kt qu, in khi ph.

Hnh 1.6

S khi ca khi ph

B to ion (ion source). Mu cht cn phn tch s c chuyn thnh trng thi hi sau mi bt u qu trnh o khi ph. o c c tnh ca cc phn t c th my khi ph s chuyn chng thnh cc ion ,kim sot chuyn ng ca chng bng in trng ngoi.Qu trnh c thc hin trong mi trng chn khng. Ion ha cng (hard ionization) - Ion ha in t (electron ionization EI)
-

Electron c nng lng 70ev c bn vo cc phn t ha cht

trong chn khng.

AB + e(70ev) AB + + 2e

(ion phn t) (mnh ion)

AB + e(70ev) A + + B + 2e

in t ion mang nng lng 675kj/mol Nng lng ni t 100 n 1000 kj/mol Do kh nng t ni rt ln.

24

Hnh 1.7 Cu to b to ion Ion ha mm (soft ionization) Ion ha ha hc (chemical ionization) Ion ha hc dng PCI Phn ng vi kh tc cht (metan)
CH 4 + e CH 4+ , CH 3+ , CH 2+ CH 4+ + CH 4 CH 5+ + CH 3 CH 3+ + CH 4 C 2 H 5+ + H 2
+ + CH 2 + CH 4 C 2 H 4 + H 2

m / z 16,15, 14 m / z 17* m / z 29*

m / z 28 m / z 28

C2 H

+ 3

+ H2 +H

C 2 H 3+ + CH 4 C3 H 5+ + H 2

m / z 41

Chuyn proton
M + CH 5+ [ M + H ]+ + CH 4 M + C 2 H 5+ [ M + H ]+ + C 2 H 4 [ M + 1]+ [ M + 1]+

Kh hydro
M + CH 5+ [ M H ]+ + CH 4 + H 2 M + C 2 H 5+ [ M H ]+ + C 2 H 6 [ M 1]+ [ M 1]+

To hp cht cng

25

M + C 2 H 5+ [ M + C 2 H 5 ]+ M + C3 H 5+ [ M + C3 H 5 ]+

[ M + 29]+ [ M + 41]+

Ion ha ha hc m NCI Tc cht : Metan

CH 4 + e CH 4+ + e(Nng lng E ln) + e (nng lng E nh)

Do va chm, nng lng in t gim dn v tr thnh in t nhit c nng lng nh, cc phn t c cng nhiu nguyn t hay nhm c m in ln cng d bt cc in t ny.

B tch ion phn tch Khi ph n tiu

Mt dng in mt chiu thng ng vung gc vi trc xy c dng phn tch ion khi cht phn tch i qua bung chn khng.p sut trong bung khong 10-5 10-8 mmHg.Ion sau khi c to thnh s c phn tch bng gia tc v tp trung chng thnh mt dng tia m sau s b un cong bi mt t trng.Nguyn tc ca b ph khi n tiu : gi r khng i, i V hay B hay c hai cho tt c cc ion c m/z khc nhau u hi t ti mt im trn b phn thu ion.

Hnh 1.8 S ph khi n tiu . - B khi ph t cc

26

Xung quanh t cc c t trng c tn s dao ng RF lm cho cc ion i qua t cc s dao ng, nhng ion no b cng hng do t tng ca t cc pht ra s c nh hng i thng n detector, cn cc ion khng cng hng th b lch ra khi qu o v khng i n detector.

Hnh 1.9
-

S khi ca t cc

By ion

Hnh 1.10 S khi ca ion trap Cc bc c bn by ion: a ion vo by; gi ion trong by; tng ion ra chn lc.Tng ion ra khi by mt cch chn lc theo khi lng: tng in th RF (V) ln lt tng ra khi by t m/z nh n m/z ln. zV = 1/2mv2 Detector

27

m s lng cc ht c cng khi lng. Thng tin ny sau c chuyn n b phn x l s liu.

Hnh 1.11

Detector ion

Detector nhn c tn hiu ri thc hin cc php tnh ton v xut ra kt qu gi l khi ph. Khi ph l mt biu phn nh s lng cc ion vi cc khi lng khc nhau i qua b phn tch khi. nh tnh v nh lng bng GC-MS

.4.

nh tnh

Trong qu trnh sc k ngi ta s ghi c sc . T peak sc ngi ta s ghi c cc tn hiu tng ng vi tng cu t gi l peak sc k. Thi gian lu ca pick l c trng nh tnh cho cht cn tch. Ngi ta so thi gian lu ca mu th vi thi gian lu ca mu chun ghi cng iu kin. i khi ngi ta a cht chun vo trong mu phn tch, ghi sc k . So snh chiu cao v din tch peak ca sc k trc v sau khi cho mu chun vo mu. Vic tng chiu cao v din tch ca cc peak chng t s c mt gi nh trong mu. Tuy nhin phng php ny cho kt qu cng khng hon ton tin cy v thi gian lu ca nhiu cht kh gn nhau.

28

 khc phc kh khn ny ngi ta thc hin sc k mu trn cc ct vi cc cht hp ph khc nhau. Nu ta thc hin c cc kt qu ging nhau trn cc ct vi cc cht hp ph khc nhau, s tng tin cy ca kt qu phn tch.

nh lng

Phn tch nh lng da vo vic o cc tham s khc nhau ca cc peak sc k nh chiu cao, rng din tch, th tch lu hay t s ca th tch lu vi chiu cao ca peak. Cc i lng ny v nguyn tc l t l vi nng cc cu t trong hn hp. Khi iu kin lm vic ca qu trnh sc k cng nh vic d tm (s lm vic ca detector) n nh. Vic tnh ton kt qu c th da vo chiu cao ca peak. Vic tnh ton kt qu cng c th da vo din tch ca peak cho php gim bt yu cu v n nh ca iu kin chy sc k so vi vic tnh ton da vo chiu cao pick.. Khi lm vic vi sc k ny ta s dng cch tnh c bit ch yu da vo chiu cao ca peak. Cc phng php sc k kh nh lng ch yu l: Phng php chun ha, phng php chun ha theo h s hiu chnh, phng php ng chun tuyt i, phng php ni chun.

29

CHNG II
2.1.

NI DUNG NGHIN CU

S lc v phng php nh tr

2.1.1. nh ngha v mc ch nh tr l s khng nh thng qua vic kim tra v cung cp cc bng chng rng cc yu cu qui nh ca mt phng php c c tha mn. nh tr phng php c thc hin nhm m bo rng ti mi thi im v trn phng din chnh xc, mc ch s dng v sai s th phng php l ph hp vi cc yu cu phn tch t ra. 2.1.2. Khi no thc hin vic nh tr Ta tin hnh nh tr mt phng php trong cc trng hp sau:

30

Phng php mi hoc phng php c xy dng da trn phng php c ph duyt. Kim tra li phng php c ph duyt hoc khi c s thay i trong phng php c ph duyt. Khi c s thay i kim nghim vin, thit b s trong phng php c ph duyt hoc khi phng php c ph duyt c p dng phng th nghim khc. Khi h thng kim sot cht lng (QC Quality control) pht hin s khng n nh ca phng php c ph duyt. Khi cn chng minh s tng ng gia hai phng php (phng php mi v phng php chun). nh tr phng php phi c thc hin mt cch nh k. Khi c bt k thay i nh no trong h thng phn tch cng u phi tin hnh nh tr li. Tt c cc phng php m mt phng th nghim ang thc hin u phi c phng th nghim nh tr theo nh k. 2.1.3. Mt s i lng c xc nh trong qu trnh nh tr chnh xc ca phng php: mc gn nhau ca kt qu o v gi tr thc ca i lng o. lp li ca kt qu o (Repeatability): mc gn nhau gia kt qu cc php o lin tip cng mt i lng o trong nhng iu kin nh nhau. iu kin lp li l cc iu kin khi cc kt qu c lp nhn c vi cng mt phng php trn cc iu kin th ging nhau trong cng mt phng th nghim, do cng mt nhn vin s dng cng thit b trong khong thi gian ngn.
-

ti lp ca kt qu o (Reproducibility): thng s gn vi kt qu ca php o, c trng cho s phn tn cc gi tr c coi l hp l ca i lg o. ti lp l cc iu kin khi cc kt qu th nhn c vi

31

cng mt phng php trn cc i tng th ging nhau trong cc phng th nghim khc nhau do nhng nhn vin khc nhau s dng nhng thit b khc nhau tuyn tnh: l kh nng m phng php s em li cc kt qu th nghim t l thun vi nng cu t trong mt khong cho trc. Khong tuyn tnh c th hin thng qua ng chun: Vic chun b thit lp ng chun phi c tin hnh nhiu hn mt ln. Phi xc nh t nht 5 im dng ng chun. H s tng quan l tng l cao hn 0,995; c th chp nhn gi tr thp hn nu qu trnh chun b thit lp ng chun c lin quan n vic to dn xut hoc phn ng c xc tc enzyme. Gii hn pht hin (LOD): gi tr nh nht ca khong pht hin ca mt phng php. Gii hn nh lng (LOQ): Thng bng 10 ln LOD v c xem l mc nng thp nht c th o c chnh xc. Hiu sut thu hi: t l % gia gi tr xc nh c v gi tr thc ca i lng o. 2.2. Phng php phn tch DLTTS trong mu nng sn thc phm bng Quechers 2.2.1. Gii thiu chung v phng php Quechers Quechers c pht trin bi United States Department of Agriculture ( S nng nghip Hoa K) vo nm 2003.Quechers l t vit tt ca Quick Easy, Cheap, Effective, Rugged and Safe.Phng php ny n gin v kh hiu qu trong vic chit v lm sch mt s mu c nn phc tp.Quechers l mt k thut c p dng cho vic phn tch a - d lng TTS. Association of Official Analytical Chemists (AOAC) ban hnh v tiu

32

chun ha phng php vo nm 2007.Trong khi theo tiu chun chu u ( EN 15662 2007) cng s dng phng php tng t. u im ca phng php - Chun b mu nhanh d dng,c th x l mu nhanh cho mt lng mu ln. - Chi ph thp khng cn cc thit b cu k tinh vi. - S dng lng t ha cht, dung mi.Gim c nh hng ti sc khe i vi kim nghim vin. Nhc im ca phng php:
-

Khi hydrat ha khi cho mui khan vo c th dn ti s ta nhit

v lm mt mu do nhit hoc lm bay hi cu t cn phn tch.

-

Mt s TTS b phn hy trong mi trng acetonnitil nn axit ha

ACN trnh s phn hy .

-

Trong tt c cc loi thc phm, thnh phn nn mu bao gm cht

bo, carbohydrate, protein, nc v s lng khong cht t, vitamin v mt lot cc sn phm t nhin ty thuc vo loi mu.Thnh phn nn mu ny lm nh hng ti qu trnh chit cng nh phn tch.
-

Mt s TTS tin hnh theo phng php Quechers thng b mt

hay phn hy bi pH. 2.2.2. C s l thuyt ca phng php Mu sau khi ng nht c chit vi dung mi ACN (Acetonitrile) vi cc tc nhn loi nc, to pH nh MgSO4, NaCl, Na3Citrat2H2O,
2

Na

HCitrate 1 H 2 O.Dch chit thu c tip tc c chit v lm sch vi

33

hn hp mui PSA v MgSO4.Dch chit cui cng c dng phn tch bng LC MS/MS hoc GC MS . Bng 2.1 Lng nc thm vo cho mt s loi mu khc nhau Nhm mu Tn mu Khi lng mu(g) Ng cc Rau qu hm Ng cc c Tri cy sy kh: to, 5g 10g hm lng nc thm 10g X X = 10g hm lng mu 10g - Thm 600l nc cha trong 10 g Ch thch

lng mn,o,chui. Ti,u,ht c ph nc t 25 80 % Rau hm qu Tr ,cy c tho mc kho. c Chanh, nho,qu mn

lng xi.. Xoi,c rt,l da, ci nc ln hn xoong, rau dip, 80% Mt ong gia v 5g 2g 10g 10g

NaOH 5M S dng GCB trong giai on lm sch

2.2.3. Mt s ch trong qu trnh xc nh DLTTS bng phng php Quechers

-

Trong qu trnh chun b mu,trc khi xay nh th mu ct nh

cn c bo qun trong t lnh ( -180C). nhit phng mt s cu t ca TTS c th b mt i nhng mt khc mc ng nht khng c tt khi lnh.

34

Mu ng nht nh hoa c th cn trc tip vo ng ly tm sau

thm lng nc cn thit trnh s phn hy ca mt s TTS mu cn c gi trong iu

kin nhit thp c th. Vi mt s tri cy kh vic ng nht mu khi nghin nh c th

gp kh khn.Do thm nc trc khi nghin nh. Lng dung mi v mui thm vo cn phi t l vi khi lng

mu, th tch dch chit.

-

i vi mt s cu t thuc tr su c tnh acid

(nh phenoxyalcanoic) s tng tc vi PSA .Do vic phn tch cn tin hnh trn LC/MS/MS.Dch chit s c s c tim trc tip vo vial ( 200 l).Khng qua qu trnh lm sch bng PSA.
-

Khi to h m bng cch thm mui th pH ca mu nm trong

khong 5 5,5.khong pH thch hp cho vic nh lng thuc tr c c tnh acid ,lm cho cc TTS c kim khng bn c bo v.( captan, folpet, tolylfluanid). Mt s nn mu c tnh acid pH < 3 ( nh chanh.) th c th

dng NaOH 5N iu chnh pH

-

Trong nn mu MgSO4 c th ht m to thnh cc, nn sau khi

thm mui vo cn vortex lin.

-

Vi loi mu nh ng cc hay tri cy sy kh khi x l c th b

nh hng bi cht bo hay nht ti qu trnh phn tch bng GC.C th loi b cht bo v nht bng cch dch chit trong t lnh nhit thp( hn 1 gi hoc qua m).Hoc cng c th loi bng cch s dng C18 cng vi PSA v MgSO4

35

i vi mu c hm lng sc t ( nh c rt, t ) hoc cht dip

lc (nh rau dip,rau ci..)Qu trnh chit lm sch s dng PSA, MgSO4 kt hp vi GCB( than hot tnh).Nhng c mt s loi TTS phn cc c i lc ln i vi cu trc phn cc ca GCB c th lm cho hiu sut thu hi b gim i.Qua mt s qu trnh kho st cho thy hiu sut thu hi gim i khng ng k.V sau khi chit lm sch bng GCB vn cn mt lng nh sc t v chlorophyll. Lng GCB c th s dng : + 2,5mg cho c rt ,rau dip, v cc loi tng t.T l

MgSO4 : GCB (60:1) + 7,5 mg cho ht tiu ngt,rau chn vt,tht cu v cc loi tng t.t l MgSO4 :GCB ( 20 :1)

36

2.2.4. So snh cc phng php Quecher

37

hod 2007.01

EU prEN-15662 (CEN)

Na 2 HCitrate 1 H 2 O

1g Na3Citrate.2H2 O + 0.5g

4g MgSO4 + 1g NaCl,

10 g mu/10 mL MeCN,

rong ACN ,

50mg PSA

1 mL dch chit + 150mg MgSO 4 + 25mg PSA

(pha ACN)

5 5.5 ( pha nc ) ~ 8 ( pha ACN ) ~ 8 (pha ACN )

34

38

Bng 2.2

Bng so snh cc phng php Quechers

39

2.3.

Thc nghim nh tr h cc tng hp trn nn mu c chua bng

phng php Quechers 2.3.1. Ha cht v thit b Ha cht

-

Acetonitrile ( Merck)

- Acetone ( Merk) - Methanol ( MeOH) Merk - Natri clorua ( NaOH )

-

Disodium hydrogencitrate sesquihydrate (C6H8Na2O8) Trisodium citrate dehydrate (C6H5Na3O7.2(H2O)) Magnesium sulphate anhydrous ( MgSO4) Merck IS( Internal standards) Triphenyl phosphate (TPP) 10 ppm, 50 ppm

T dung dch chun gc TPP 2537 ppm ly 985 l vo bnh nh mc 50 ml. Sau nh mc bng MeOH. Ta c dung dch chun TPP 50ppm.Pha long dung dch chun 50ppm trong MeOH ta c dung dch 10ppm Chun gc thuc tr su ( 5 cu t).Cc cht chun phi c chng nhn v cn hn s dng.
-

40

Pha chun mix 5 cu t 100ppm trong 10ml. Chun gc Permethrin 1235,2 ppm : Ly 809,58 l chun gc Permethrin vo bnh nh mc 10 ml. Chun gc Cypermethrin 1056,2 ppm : Ly 3787 l chun gc Cypermethrin vo bnh nh mc 10 ml. Chun gc Fenvalerate 1130,6 ppm : Ly 884,48 l chun gc Fenvalerate vo bnh nh mc 10 ml. Chun gc Deltamethrin 1263,2 ppm : Ly 3166 l chun gc Deltamethrin vo bnh nh mc 10 ml. Chun gc Cyhalothrin 1105,8 ppm : Ly 904,32 l chun gc Cyhalothrin vo bnh nh mc 10 ml. nh mc 10ml bng acetol v lc u.Dung dch chun mix cc 5 cu t 10ppm c pha long t dung dch 100ppm bng acetol.Chun c bo qun trong t lnh nhit thp. -

n hexan ( Merck) PSA (primary secondary amine) 100mg GCB (graphitized carbon black) C18 silica

Tt c cc ha cht u phi t tinh khit ha hc Thit b ng ly tm loi 15ml, 50ml ( cn sn lng mui cn thit cho qu trnh lm vic)
-

Micropipet (10-100L, 200-1000l)

41

Pipet bu 1ml,2ml,4ml,10ml My xay mu My nh vortex My ly tm H thng thi kh bng kh N2 Thit b sc k kh ghp khi ph ( GC-MS ) Cc loi kh s dng cho GC Cn phn tch cp chnh xc 0.1mg - Vial sc k kh 1.5 ml

2.3.2. Quy trnh xc nh, tnh ton kt qu

Mu QC

Mu Blank

Mu phn tch

100 l TPP 50ppm

Spike 2,0 mg/kg cypermethin, deltamethrin v 0,5 mg/kg cc cu t khc vo mu QC

Khng spike cho mu phn tch v Blank

Lc u bng tay

42

Thm 10ml ACN,lc mnh 1 pht

Thm 4g MgSO4 + 1g NaCl + 1g Na3Citrate.2H2 O + 0.5g Na 2 HCitrate.1H 2 O

nh Vortex 1 pht. Ly tm vi tc 4000 vong/phut trong 5 pht

Ly 6ml dch chit vo ng ly tm cha sn 150mg PSA + 900mg MgSO4

nh Vortex 1 pht. Ly tm vi tc 3000 vng/pht trong 5 pht

Ly chnh xc 4ml dch chit thi va ti kh bng kh N2

nh mc bng 1ml n hexan

Chuyn vo vial. Phn tch bng GC - MS

Hnh 2. S tm tt quy trnh xc nh bng phng php Quechers nh lng bng GC MS iu kin phn tch trn GC

43

Ct sc k kh :

DB 5MS

Tc dng: 2,5 ml/pht Th tch tim mu: 2,0l Chng trnh nhit l ct: Tc tng (0C/pht) Nhit (0C) 100 15 300 Thi gian gi ( pht) 0.5 15

- iu kin phn tch trn khi ph Nhit Source : 200 0C Nhit interface : 260 0C Tn hp cht M khi lng phn t m/z M1 Ion nh danh (m/z) Th detector: 350V Th ion ha : 70 eV M2 Ion nh lng (m/z) RT (pht:giy)

Cyhalothrin Trans permethrin Cypermethrin Fenvalerate Deltamethrin IS_TPP

449.8 391.3 416.3 420.0 505.2

197 163 163 167 253

181 183 163 125 181 326

13:36 14:56 16:37 18:41 21:18 13:00

44

Tnh ton kt qu

Da vo phng trnh hi quy tuyn tnh y = aC + b ca tng cu t ta tnh c nng da trn phng trnh ng chun. Phng trnh hi quy t khi R2 > 0.99. Da vo khi lng cn v th tch nh mc, hm lng thuc tr su trong mu c tnh theo cng thc sau: X(%) = Ctnh*F*V/m Trong : X: hm lng cc cht c trong mu (mg/kg) Ctinh: hm lng tnh c theo ng chun (mg/kg) F: h s pha long V: th tch nh mc (ml) m: khi lng mu cn (g)

45

CHNG 3 3.1.

KT QU V THO LUN

Phng php nh tr h cc tng hp trong mu nng sn thc

phm bng phng php Quecher 3.1.1. Kho st gii hn pht hin (LOD), gii hn nh lng ( LOQ) xc nh gii hn pht hin v gii hn nh lng, ta tin hnh thc hin trn 7 mu blank c thm chun.Trn 7 sc k thu c, ta tin hnh ct cc peak c thi gian tng ng thi gian lu ca 5 cu t.

140000

120000

100000

80000

60000

40000

20000

Time (min:sec)

6:40.00

326

10:00.00

183

13:20.00 163

16:40.00 125

20:00.00 181

23:20.00

Hnh 3.1 Sc k ca mu blank c ni chun Din tch cc peak ny c gi l noise. Nh vy, ng vi mi cht ta s c cc noise ring bit. Da vo cc noise ny ta s tnh c LOD v LOQ theo cng thc: - Cng thc tnh LOD: Cng

LOD =

C 3 Noise S LO = Q

thc tnh LOQ:

C 10 Noise S
46

Trong

C: nng chun thm vo. S: din tch peak ng vi nng C.

Bng 3.1 LOD v LOQ ca cc cu t Cc trn nn mu c chua STT Tn cu t C (mg/kg) 1 2 3 4 5 6 Cyhalothrin 0.2 Noise (N) 261.31 189.98 6522.60 323.67 157.29 1981.77 Area (S ) 284030.1 96598.7 508254.2 139513.6 32543.6 161909.3 LOD LOQ

(mg/kg) (mg/kg) 0.0138 0.0295 0.1925 0.0348 0.0725 0.1836 0.0460 0.0983 0.6417 0.1160 0.2417 0.6120

Trans permethrin 0.2 Cypermethrin Fenvalerate I Fenvalerate II Deltamethrin 0.2 0.2 0.2 0.2

Da vo bng LOD, LOQ trn ta thy a s cc cu t c gii hn pht hin kh thp c vi chc ppb.Duy ch c cu t Cypermethrin v cu t Deltamethrin cho gii hn pht hin v gii hn nh lng cao nn trong qu trnh tin hnh th nghim.Pha chun mix ta cn ch ti 2 cu t ny.V d chun mix 10mg/kg ta phi tng nng ca cu t Cypermethrin v cu t Deltamethrin gp 2,3 ln.Trong phm vi bi bo co ny chng ti tin hnh kho st v kt qu cho thy vi t l 1: 4 cho kh nng nh lng tt.Ngha l trong chun mix 10 mg/kg th 2 cu t trn s l 40 mg/kg. 3.1.2. Kho st hiu sut thu hi Tin hnh kho st hiu sut trn nn mu QC c chua ( khng cha DLTTS h Cc tng hp).Chun b 16 mu QC theo quy trnh di y:
-

T QC1 n QC8 spike 0,5 mg/kg t chun mix 5 cu t 10 mg/kg

47

T QC11 n QC16 spike 1 mg/kg t chun mix 5 cu t 10 mg/kg 1 mu Blank ( khng spike chun mix ) Xy dng ng chun 1ppm, 2ppm, 4ppm, 6ppm

ng chun IS (TPP 50ppm) Chun mix 10ppm nh mc ( n hexan)

1 ppm 80 l 1ml

2ppm 80 l 2ml

4ppm 80 l 4ml

6 ppm 80 l 6ml

nh mc bng n hexan ti 10 ml

Bng 3.2

Phng trnh dng chun v R2 ca cc cu t Cc

Tn cu t Cyhalothrin Tran permethrin Cypermethrine Fenvalerate I Fenvalerate II Deltamethrin

7 . 00
12.0

Phng trnh ng chun Y = 0.863086*X 0.978665 Y = 1.37919*X 0.50725 Y = 0.570972*X 0.812922 Y = 0.680581*X 0.211978 Y = 0.213353*X 0.143199
Std 6p pm :2

R^2 0.99941 0.99869 0.99896 0.99929 0.99993 0.99947

S td 6 p p m :2 Std 6ppm :2

Y = 0.184728*X 0.317572
5.00

Std 6ppm :2

4.50 3 . 50 4.00
3 . 00

6 . 00
10.0

3.50

5 . 00
Ratio Ratio

Std 4ppm :2
8.00

Std 4ppm:2

4 . 00

Ratio Ratio

2 . 50 3.00

St d 4 4ppm :2 Std p pm :2

2 200 0 . .5

6.00

3 . 00

2.00 1 . 50 1.50
1 . 00

2 . 00

4.00

Std 2ppm :2

Std 2ppm :2

S td 2 p p m :2 Std 2ppm :2

1.00
2.00 1 . 00

48
S td 1 p p m :2 Std 1ppm :2
1 1 2 2 33 44

Std 1ppm :2 Std 1ppm :2

1
5

0. 5 0 0
10

4 15

yy= + 1.3 79 19x -- 0. 5072 5 = +0.570972x 0.812922 rr = 0. 99 869 = 0.99896

5 20 C o ncen tra t io n Concentration

25

30 8

935

yy = ++ 0.680581x -- 00.211978 = 0. 86 30 8 6x .222279 r r== 00.99929 .99941

5 5 C o noncetntrtaio n n C ce n ra tio

6 6

Hnh 3.2 ng chun ca cc cu t h Cc trn nn c chua

4.00

S t d 6 p p m :2

Std 6ppm :2
1. 0 0

3.50

3.00
0. 80 0

Ratio

0. 60 0

Ratio

Std 4ppm :2

2.50

S td 4 p p m :2

2.00

0. 40 0

1.50

Std 2ppm :2
0. 20 0

1.00

S td 2 p p m :2

0.500

Std 1ppm :2
1 2 3 4

S td 1 p p m :2
5 10 15

y = + 0.213353x - 0.143199 r = 0.99993

5 C o nce n tra tio n

y = + 0.18 4728x - 0 .31 7572 r = 0.9994 7

20 C o n c e n t r a t io n

25

30

35

49

140000

120000

100000

80000

60000

40000

20000

Time (min:sec)

6:40.00

326

10:00.00

183

13:20.00 163

16:40.00 125

20:00.00 181

23:20.00

Hnh 3.3 Peak ca cc cu t Cc trn nn c chua ( QC spike 0.5 mg/kg)

250000

200000

150000

100000

50000

Time (min:sec)

6:40.00 326

10:00.00 183

13:20.00 163

16:40.00 125

20:00.00 181

23:20.00

Hnh 3.4 Peak ca cc cu t Cc trn nn c chua ( QC spike 1 mg/kg )

50

lp li : lp li c nh gi thng qua gi tr lch chun Sr:

Sr =

(x

x) 2

n 1

lp li:
R = 2 Sr t n 1

Vi tin cy

t n 1

gi tr tra bng Student vi n s ln thc nghim v

Bng 3.3

RSD ngh chp nhn i vi lp li cc mc nng khc nhau

Nng cht phn tch (n v) RSD (%) 100g/kg 10g/kg 1g/kg 100mg/kg 10mg/kg 1mg/kg 100g/kg 10g/kg 1g/kg 0,1g/kg 2 3 4 5 7 11 15 21 20 43

T kt qu hiu sut thu hi ,Sr v RSD(%) ca tng cu t Cc .Ta c c bng tng kt v hiu sut thu hi , lch chun, lp li ca tt c cc cu t kho st: Bng 3.3 hiu sut thu hi v RSD ( lp li ) ca cu t Cyhalothrin

51

STT 1 2 3 4 5 6 7 8

m (g) 10.05 10.22 10.38 10.20 10.18 10.24 10.28 10.46

C0(mg/kg) 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Trung bnh

R 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25

Vm 10 10 10 10 10 10 10 10

C tnh (g/mL) 1.88 1.62 1.85 1.87 2.01 1.97 2.08 2.17 1.93

Sr

RSD%

H (%) 93.53 79.26 89.11 91.67 98.72 96.19 101.17 103.73

0.16763

8.67962

94.1724

STT 9 10 11 12 13 14 15 16

m (g) 10.20 10.20 10.16 10.20 10.03 10.57 10.53 10.47

C0(mg/kg) 1 1 1 1 1 1 1 1 Trung bnh

R 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25

Vm 10 10 10 10 10 10 10 10

C tnh (g/mL) 4.14 4.09 4.08 4.08 3.28 3.98 4.15 3.91 3.96

Sr

RSD%

H (%) 101.4706 100.2451 100.3937 100.0000 81.7547 94.1343 98.5280 93.3620

0.2877

7.258

96.236058

Bng 3.1 Bng hiu sut thu hi, lch chun, lp li, cho cc cu t h Cc tng hp
Tn cu t Cyhalothrin C (mg/kg) 0.5 Sr 0.17 RSD 8.68 H (%) 94.17

52

1 Trans permethrin Cypermethrin Fenvalerate I 0.5 1 0.5 1 0.5 1 Fenvalerate II 0.5 1 0.5 1

0.29 0.095 0.12 0.58 0.45 0.13 0.33 0.14 0.26 0.29 0.82

7.26 5.26 3.30 8.64 2.92 7.51 9.28 9.62 7.87 4.12 5.46

96.24 88.59 92.31 8.86 93.63 84.05 86.55 74.98 79.75 86.67 91.39

Deltamethrin

Nhn xt: Qua bng s liu ta thy a s cc cu t cho hiu sut thu hi kh cao. Do cc cu t h Cc tng i bn trong mi trng m citrate.C th bo qun mu trong t lnh qua m nhng lng cu t phn tch b mt i do phn hy l rt nh.i vi phng php nh tr kt qu t c cho php khi -

H s tng quan ng chun (R > 0.99) Mu blank khng c cht phn tch Hiu sut thu hi t c nm trong khong 60% - 120% cho

80% cu t thm vo.Do hiu sut thu hi ny t yu cu cho phng php nh tr h Cc( 5 cu t).

3.1.3. Kho st ti lp ca phng php

53

Thc hin vi 16 mu QC ( chun b mu ging nh kho st hiu sut thu hi v lp li).Kt qu ti lp c tnh theo phng php x l s liu bng hm anova.

54

55

STT 1 2 Cyhalothrin 3 4 5 C add(mg/kg) TB SD CV 4 H (%)

NV 1

NV 2

Anova: Single Factor

4.14 4.09 4.08 4.08 3.98

4.0740 0.0581 1.4270 101.8500

3.96 4.12 SUMMARY 3.83

Groups Count 5 5 Sum 20.37 20.21 Average 4.074 4.042 Variance 0.00338 0.02072

4.18 Column 1 4.12 Column 2

4.0420 0.1439 3.56 101.05 ANOVA Source of Variation Between Groups Within Groups

SS 0.00256 0.0964

df 1 8

MS 0.00256 0.01205

F 0.212448133

P-value 0.657127

F crit 5.317655

Total

0.09896

TB

4.0580 S.D. Mean RSD C.V. (RSD%)

Cng ngi 0.1098 4.0580 0.0271 2.7051

Chnh lch 2 ngi 0.0436 4.0580 0.0107 1.0736

Intermediate Precision 0.1181 4.0580 0.0291 2.9103

Kt lun

Thu hi /Overall recovery

101.45

Khng c s chch lch

49

STT 1 trans - pemerthrin 2 3 4 5 C add(mg/kg) TB SD CV 4 H (%)

NV 1

NV 2

Anova: Single Factor

3.94 3.82 3.86 3.79 3.98

3.8780 0.0801 2.0661 96.9500

3.96 4.02 SUMMARY 3.93

Groups Count 5 5 Sum 19.39 19.5 Average 3.878 3.9 Variance 0.00642 0.01575

3.69 Column 1 3.9 Column 2

3.9000 0.1255 3.22 97.50 ANOVA Source of Variation Between Groups Within Groups Total

SS 0.00121 0.08868 0.08989

df 1 8 9 Chnh lch 2 ngi 0.0444 3.8890 0.0114 1.1427

MS 0.00121 0.011085

F 0.109156518

P-value 0.749593

F crit 5.317655

TB

3.8890 S.D. Mean RSD C.V. (RSD%)

Cng ngi 0.1053 3.8890 0.0271 2.7073

Intermediate Precision 0.1143 3.8890 0.0294 2.9386

Kt lun

Thu hi /Overall recovery

97.23

Khng c s chch lch

50

STT 1 cypermethrin 2 3 4 5 C add(mg/kg) TB SD CV 16 H (%)

NV 1

NV 2

Anova: Single Factor

15.73 15.6 15.69 15.32 14.98

15.4640 0.3144 2.0329 96.6500

16.54 16.8 15.94 15.01 15.71

16.0000 0.7070 4.42 100.00 ANOVA Source of Variation Between Groups Within Groups Total SUMMARY Groups Column 1 Column 2 Count 5 5 Sum 77.32 80 Average 15.464 16 Variance 0.09883 0.49985

SS 0.71824 2.39472 3.11296

df 1 8 9 Chnh lch 2 ngi 0.2894 15.7320 0.0184 1.8399

MS 0.71824 0.29934

F 2.39941204

P-value 0.159974

F crit 5.317655

TB

15.7320 S.D. Mean RSD C.V. (RSD%)

Cng ngi 0.5471 15.7320 0.0348 3.4778

Intermediate Precision 0.6190 15.7320 0.0393 3.9344

Kt lun

Thu hi /Overall recovery

98.33

Khng c s chch lch

51

STT 1 Fenvalerate I 2 3 4 5 C add(mg/kg) TB SD CV 4 H (%)

NV 1

NV 2

Anova: Single Factor

3.76 3.52 3.72 3.88 3.81

3.7380 0.1357 3.6308 93.4500

4.01 4.15 3.92 3.61 3.79

3.8960 0.2068 5.31 97.40 ANOVA Source of Variation Between Groups Within Groups Total SUMMARY Groups Column 1 Column 2 Count 5 5 Sum 18.69 19.48 Average 3.738 3.896 Variance 0.01842 0.04278

SS 0.06241 0.2448 0.30721

df 1 8 9 Chnh lch 2 ngi 0.0798 3.8170 0.0209 2.0897

MS 0.06241 0.0306

F 2.039542484

P-value 0.191108

F crit 5.317655

TB

3.8170 S.D. Mean RSD C.V. (RSD%)

Cng ngi 0.1749 3.8170 0.0458 4.5829

Intermediate Precision 0.1923 3.8170 0.0504 5.0368

Kt lun

Thu hi /Overall recovery

95.43

Khng c s chch lch

52

STT 1 Fenvalerate II 2 3 4 5 C add(mg/kg) TB SD CV 4 H (%)

NV 1

NV 2

Anova: Single Factor

3.18 3.38 3.41 2.99 3.19

3.2300 0.1707 5.2859 80.7500

3.33 3.51 3.43 3.28 3.36

3.3820 0.0898 2.66 84.55 ANOVA Source of Variation Between Groups Within Groups Total SUMMARY Groups Column 1 Column 2 Count 5 5 Sum 16.15 16.91 Average 3.23 3.382 Variance 0.02915 0.00807

SS 0.05776 0.14888 0.20664

df 1 8 9

MS 0.05776 0.01861

F 3.103707684

P-value 0.11614

F crit 5.317655

TB

3.3060 S.D. Mean RSD C.V. (RSD%)

Cng ngi 0.1364 3.3060 0.0413 4.1264

Chnh lch 2 ngi 0.0885 3.3060 0.0268 2.6766

Intermediate Precision 0.1626 3.3060 0.0492 4.9184

Kt lun

Thu hi /Overall recovery

82.65

Khng c s chch lch

53

STT 1 deltamethrin 2 3 4 5 C add(mg/kg) 16 TB SD CV H (%)

NV 1

NV 2

Anova: Single Factor SUMMARY Groups Column 1 Column 2 Count 5 5 Sum 77.18 80.9 Average 15.436 16.18 Variance 0.11803 1.3934

15.66 15.02 15.24 15.89 15.37

15.4360 0.3436 2.2257 96.4750

17.67 15.03 14.93 16.47 16.8

16.1800 1.1804 7.30 101.13 ANOVA Source of Variation Between Groups Within Groups Total

SS 1.38384 6.04572 7.42956

df 1 8 9 Chnh lch 2 ngi 0.3544 15.8080 0.0224 2.2421

MS 1.38384 0.755715

F 1.831166511

P-value 0.212984

F crit 5.317655

TB

15.8080 S.D. Mean RSD C.V. (RSD%)

Cng ngi 0.8693 15.8080 0.0550 5.4992

Intermediate Precision 0.9388 15.8080 0.0594 5.9387

Kt lun

Thu hi /Overall recovery

98.80

Khng c s chch lch

54

KT LUN V KIN NGH

Sau thi gian hc tp v thc nghim nh tr d lng thuc

tr su h Cc tng hp trong mu nng sn thc phm bng phng php Quechers da trn GC MS.Kt qu t c c p dng vo quy trnh nh lng trn mt s nn mu tng t.
-

Tuy nhin, phng php t c hiu sut thu hi cao cho nhiu

nn mu v ang c trin khai mt cch rng ri trong cc PTN.Nhng mt iu ng lu l kt qu t c s khng tt khi c s tng tc gia cht phn tch v nn mu.V d nh cc mu c nn phc tp nh gia v..
-

C th s dng LC-MS/MS phn tch th qu trnh chun b

mu n gin hn.Nhng kh nng nh lng thp hn. p dng phng phng php vo thc nghim gip ta gim

c thi gian phn tch trn mt s lng mu nhiu.Gim c chi ph cho vic phn tch.Gim c nh hng ti sc khe cho KNV. Nu c iu kin ti xin a ra mt s kin ngh: - Tip tc kho st, nh tr DLTTS gc Clo hu c, Pht pho hu c trn nn mu c chua v mt s nn mu khc.
-

Nghin cu theo hng m rng ra cho cc nn mu khc ngoi

rau qu nh : thuc th y, PAHs ,khng sinh,huyt tng, tht v t

-

Nghin cu kt hp hi ha hai phng php AOAC v prEN-

15662 t c hiu sut thu hi tt hn.

-

C th dng Canxi clorua(CaCl2) thay thay th Magie

sunfat(MgSO4).
55