TP CH KHOA HC, i hc Hu, S 59, 2010

XC NH D LNG HA CHT BO V THC VT NHM LN HU C TRONG NHO NINH THUN

Ng Vn T Trng i hc S phm, i hc Hu Nguyn Th Thy Uyn Trng Cao ng ngh KTCN Dung Qut

TM TT
Bi bo ny m t phng php xc nh d lng ha cht bo v thc vt nhm ln hu c (OPs) trong nho bng phng php sc k kh (GC). tin cy ca phng php c nh gi qua gii hn pht hin, thu hi v lp li. Vi quy trnh phn tch v iu kin sc k thch hp c la chn, gii hn pht hin ca phng php i vi Dimethoate, Diazinon, Metylparathion, Fenitrothion v Malathion ln lt l 18, 19, 12, 19 v16 ppb; lp li v thu hi tt. Phng php c p dng xc nh OPs trong mt s mu nho Ninh Thun.

1. M u Nho ti l loi tri cy giu vitamin v cht khong. Gi tr dinh dng trung bnh ca nho tnh cho 100g nho ti l: 75 calo, 0,4 g protein, 0,3 mg lipit, 6 mg Ca, 24 mg P, 0,4 mg Fe, 83UI vitamin A, 0,1 mg vitamin B1, 0,06 mg vitamin B2, 0,2 mg vitamin PP [9]. Ngoi tc dng cung cp dinh dng, nho cn c tc dng cha bnh, c bit l cc bnh lin quan n ri lon trao i cht cng cc bnh lin quan n tim mch [9]. Cy nho li l i tng tn cng ca nhiu loi su v bnh hi [3]. Do vy, vic lm dng ha cht bo v thc vt (HCBVTV) s tr thnh vn nn cn c quan tm. Tuy nhin, nhng thng tin v d lng OPs trong nho Ninh Thun cha c tm thy. GC l phng php hu hiu xc nh HCBVTV trong nng sn thc phm. c bit, vi trng hp khng c chun ri, sc k kh detector quang k ngn la knh lc photpho (GC/FPD P) phi hp vi sc k kh detector khi ph (GC/MS) l phng php tt nh lng OPs [4]. Phng php c p dng xc nh d lng OPs trong mt s mu nho Ninh Thun.

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2. Thc nghim 2.1. Thit b, dng c v ha cht - H thng GC Agilent 7890A, detector FPD P; h thng GC Perkin Elmer, detector MS; my li tm MIKRO 220R, UNIVERSAL 320R ca hng Hettich (c); h thng my c quay lnh li tm chn khng hiu LABCONCO (M); my lc REAX top ca hng Heidolph (c); cn phn tch Sartorius (sai s cho php 0,1 mg); t lnh, l nung, my xay. - Micropipet, pipet, ng li tm 50ml c np vn, chai ng mu phn tch (vial) 2ml c np vn, cc dng c thy tinh cn thit, - Hn hp chun OPs ca hng Supelco (M), PSA Bonded Silica (axit propylsulphonic trn nn SiO2) ca hng Supelco (M), axetonitril (Merck c), than hot tnh (GCB), MgSO4 khan (sy 150oC trc khi s dng), Na2SO4 khan (sy 450oC trc khi s dng), natriaxetat (NaAc) khan, axit axetic (HAc) bng, nc ct, kh He tinh khit 99,99%, kh H2 tinh khit phn tch. 2.2. Quy trnh phn tch 2.2.1. Ly mu v bo qun mu Mu n c ly ngu nhin theo hnh ch X theo cc mt ct ca l sn phm. S mu n c ly ng vi mi l khong 5 10 mu. Tt c cc mu n c trn u thu c mu nghin cu phn tch (mu hin trng) [1]. Mu hin trng c ly vo 2 t (gm 2 loi nho ph bin Ninh Thun l nho Cardinal v nho xanh NH.01-48): - t 1 (v ng Xun): 6 mu nho c chn ly cc vn nho ca 6 h gia nh nhng vng khc nhau (Phc Thun - Ninh Phc, Nha H Ninh Sn, Phan Rang Thp Chm, Ninh Hi) khi thu hoch v c k hiu: NH1.1, NH1.2, NH1.3, Card1.1, Card1.2 v Card1.3. - t 2 (v H Thu): 5 mu nho c chn ly cc vn nho ca 5 h gia nh ( cc vng Phan Rang Thp Chm, Ninh Sn, Ninh Phc, Ninh Hi) khi thu hoch v c k hiu: NH2.2, NH2.4, Card2.1, Card2.2 v Card2.3. Lng mu hin trng c ly khong 1 kg v cho vo ti PE sch bo qun. Mu phn tch d lng HCBVTV c bo qun 0oC 5oC khng qu 3 ngy trc khi phn tch [5], hoc c th bo qun trong thi gian 3 thng 18oC [1]. Mu phng th nghim c ly t mu hin trng theo quy tc ng cho khi phn tch [5].
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2.2.2. X l mu [5], [6], [7], [8] Mu c x l theo tiu chun chu u CEN/TC 275/WG 4 N 204 [5]. Cn chnh xc 10 g mu xay trn cn phn tch (sai s cho php 0,1 mg) vo ng li tm 50ml, thm vo 15 ml HAc 1% trong axetonitril, lc mnh khong 1 pht. Sau thm 1 g NaAc, lc mnh khong 1 pht. Li tm 4oC, tc 4000 vng/pht trong 15 pht. Ly 12 ml dch chit sau li tm cho vo ng li tm khc cha sn 0,25 g PSA v 1g hn hp GCB/MgSO4 (t l 1 : 19), lc mnh trong 2 pht. Thm vo ng li tm 1 g Na2SO4 khan, lc mnh trong 1 pht. Li tm 4oC, tc 4000 vng/pht trong 15 pht. Ly 10 ml dch chit c lm sch em c quay chn khng 35oC cho n kh. nh mc bng 1ml HAc 1% trong axetonitril v chuyn vo vial. Dch chit c lm sch, lm giu v axit ha c chuyn vo trong vial ca b bm mu t ng xc nh OPs bng GC. 2.2.3. Phn tch trn h thng GC - iu kin lm vic: ct mao qun ZB-5, di 30 m, ng knh trong 0,32 mm, lp phim 0,25 m; kh mang He, 25 psi; khng kh 100 mL/pht; kh H2 70 mL/pht; nhit bm mu 250oC; nhit detector 190oC i vi FPD P (280oC i vi detector MS); th tch bm mu 1 L; kiu bm mu khng chia dng. Chng trnh nhit (vi c 2 h thng GC/MS v GC/FPD P):

- Xc nh thi gian lu: dng hn hp chun OPs 0,4 ppm xc nh th t i ra khi ct ca OPs bng h thng GC/MS, t xc nh thi gian lu ca OPs bng h thng GC/ FPD P. - Kho st khong tuyn tnh v xy dng ng chun (trn h GC/FPD P): khong tuyn tnh ca detector c kho st qua cc dung dch chun OPs c nng 0,04 ppm; 0,1 ppm; 0,2 ppm; 0,4 ppm; 1 ppm; 2 ppm; 4 ppm, iu kin k thut sc k nh trn. ng chun xc nh OPs c xy dng trn c s kho st khong tuyn tnh. - nh gi phng php qua gii hn pht hin, lp li, thu hi [10]. - p dng quy trnh phn tch xc nh OPs trong mt s mu nho Ninh Thun (trn h GC/FPD P).

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3. Kt qu v tho lun 3.1. Thi gian lu ca OPs - Dng hn hp chun OPs 0,4ppm xc nh th t i ra khi ct ca OPs bng h thng GC/MS. - Tin hnh th nghim trn h thng GC/ FPD P i vi cc dung dch chun cha 5 ch tiu (5ct) ca OPs c nng 0,04 ppm; 0,1ppm; 0,2 ppm; 0,4 ppm; 1 ppm; 2 ppm; 4 ppm. Kt qu c trnh by bng 3.1.

Hnh 3.1. Sc dung dch chun 5ct ca OPs 0,4ppm Bng 3.1. Thi gian lu ca OPs

STT 1 2 3 4 5

Cht chun Dimethoate Diazinon Metyl parathion Fenitrothion Malathion

Thi gian lu (pht) (P = 0,95) 6,477 0,116 7,383 0,053 8,775 0,078 9,941 0,078 10,554 0,029

lch chun (S), pht 0,0127 0,0058 0,0085 0,0086 0,0032

lch chun tng i RSD (%) (n = 21) 0,20 0,08 0,10 0,09 0,03

3.2. Khong tuyn tnh v ng chun ca OPs Khong tuyn tnh ca detector c kho st qua cc dung dch chun OPs c nng 0,04 ppm; 0,1 ppm; 0,2 ppm; 0,4 ppm; 1 ppm; 2 ppm; 4 ppm. Kt qu cho thy, detector cho kt qu tuyn tnh tt i vi tt c OPs trong khong nng kho st t 0,4 4 ppm (hnh 3.2). Trong khong ny, ng chun xc nh OPs c h s tng
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quan R t 1,0000 (ring Malathion cho h s tng quan R = 0,9998 trong khong 0,4 4 ppm v R = 1,0000 trong khong t 1 4 ppm), (hnh 3.3 v bng 3.2).
7000

6000

Din tch pic

5000

D im e th o a te D ia z in o n M e ty lp a ra th io n F e n itro th io n M a la th io n

7000

6000

5000

D im e th o a te D ia z in o n M e ty lp a ra th io n F e n itro th io n M a la th io n

4000

Din tch pic

4000

3000

3000

2000

2000

1000

1000

0 0 1 2 3 4

0 0 .0 0 .5 1 .0 1 .5 2 .0 2 .5 3 .0 3 .5 4 .0 4 .5

Nng chun, ppm

Nng chun, ppm

Hnh 3.2. th biu din kt qu kho st khong tuyn tnh ca OPs

Hnh 3.3. ng chun xc nh OPs

Bng 3.2. Phng trnh ng chun ca OPs

STT 1 2 3 4 5

Cht chun Diazinon Dimethoate Fenitrothion Malathion

Phng trnh ng chun Y = ( 197,252 7,694) + (1296,370 2,733)X Y = ( 319,113 6,302) + (1450,594 2,255)X Y = ( 212,872 6,560) + (1509,967 3,242)X Y = ( 270,067 6,686) + (978,003 1,752)X

R 1,0000 1,0000 1,0000 1,0000 1,0000

Metylparathion Y = ( 249,478 3,696) + (1555,592 1,636)X

(Ghi ch: Y l din tch pic; X l nng , ppm)

* Thit b cho lp li tt (RSD ca tn hiu din tch pic khi xy dng ng chun t 0,99 6,60% v cc RSD ny u nh hn 0,5RSD tnh theo hm Hortwiz) [10]. * Kt qu tnh ton gii hn pht hin (LOD) v gii hn nh lng (LOQ) ca thit b theo "quy tc 3" c trnh by bng 3.3 [10].
Bng 3.3. Gi tr LOD v LOQ ca thit b

Gi tr LOD (ppb) LOQ (ppb)

Diazinon Dimethoate Fenitrothion Malathion Metylparathion 8 26,67 15 50,00 15 50,00 3S y b

; LOQ =

14 46,67
10LOD ) 3

8 26,67

(Ghi ch: LOD c tnh theo cng thc

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3.3. nh gi phng php 3.3.1. lp li v thu hi ca phng php

Tin hnh phn tch mu Card1.1 theo quy trnh trn vi nng thm chun nh sau: (Dimethoate 0,972 ppm; Diazinon 0,979 ppm; Metylparathion 0,786ppm; Fenitrothion 0,704 ppm; Malathion 0,982 ppm). lp li ca phng php c nh gi qua lch chun tng i (RSD). Kt qu phn tch 3 ln lp li v lp li ca phng php c nu ra bng 3.4. thu hi ca phng php cng c xc nh bng cch tin hnh phn tch 3 ln lp li i vi mu Card1.1 theo quy trnh trn vi nng thm chun l 0,8 ppm. Kt qu thu hi ca phng php c nu ra bng 3.4.
Bng 3.4. Kt qu xc nh lp li v thu hi (Re) ca phng php

Hot cht

Hm lng OPs thm chun, ppm

0,972 0,979 0,786 0,704 0,982

Hm lng OPs xc nh c, ppm ln 1 ln 2 ln 3

0.940 0,914 0,726 0,668 0,946

TB
0.961 0,901 0,733 0,656 0,936

RSD (%) (n=3)

1,86 1,50 1,03 1,58 1,85

Re (%)
98,9 92,0 93,3 93,2 95,3

Dimethoate Diazinon Metylparathion Fenitrothion Malathion

0,971 0,971 0,901 0,887 0,732 0,741 0,650 0,650 0,946 0,916

Nhn xt: Kt qu tnh ton cho thy, phng php cho ng tt vi thu hi t 92,0 98,9% 3.3.2. LOD v LOQ ca phng php
xc nh LOD v LOQ ca phng php, tin hnh phn tch 3 ln lp li i vi cc mu nho Card1.1 c thm chun ln lt 0,4 ppm; 0,6 ppm; 0,8 ppm; 1,0 ppm. Gi tr LOD, LOQ c xc nh theo quy tc "3". Kt qu tnh ton LOD v LOQ ca phng php c trnh by bng 3.5.
Bng 3.5. LOD v LOQ ca phng php xc nh OPs trong nho

Gi tr
LOD (ppb) LOQ (ppb) R

Diazinon Dimethoate Fenitrothion

19 63,33 0,9999 18 60,00 0,9999 19 63,33 0,9998

Malathion Metylparathion
16 53,33 0,9999 12 40,00 0,9999

(Ghi ch: R l h s tng quan gia din tch nh v nng khi xc nh LOD) 168

3.4. Xc nh OPs trong mt s mu nho Ninh Thun

Quy trnh phn tch trn c p dng nh tnh (da vo thi gian lu) v nh lng OPs trong mt s mu nho Ninh Thun. Kt qu cho thy d lng OPs trong nho rt t. Do vy, cc mu phn tch u c lm giu trc khi xc nh. Cc kt qu thu c khi nh lng OPs trong mu sau lm giu u ln hn LOQ tng ng bng 3.5. Sau khi thc hin nhng php tnh cn thit, kt qu phn tch OPs trong mt s mu nho c dn ra bng 3.6.
Bng 3.6. Hm lng OPs trong mt s mu nho Ninh Thun, ppm

Hm lng OPs Cht

NH1.1* NH1.2 Mu t 1 Card1.1 Card1.2 Card1.3 NH2.4* Mu NH2.2 t Card2.1 2 Card2.2 Card2.3 0,0420,004 0,0610,013 NH1.3

Diazinon Dimethoate Fenitrothion Malathion Metylparathion

0,0590,013

0,0370,004 0,0240,004 0,0260,004

0,0430,004 0,0260,004

(Ghi ch: Du "*": Nho trng theo k thut VietGAP (thc hnh sn xut nng nghip tt); NH: k hiu nho NH.01 48; Card: k hiu nho Cardinal; du "": khng pht hin c)

4. Kt lun
la chn c quy trnh phn tch thch hp xc nh OPs trong nho bng phng php GC/FPD P. Phng php c p dng phn tch OPs trong mt s mu nho Ninh Thun. Kt qu ban u cho thy: vi OPs vn cn tn lu trong mt s mu nho kho st. iu ni ln rng, mt s OPs c s dng cho nho trong qu trnh sn xut. D lng OPs trong nho c th l do vic qu lm dng thuc ca ngi nng dn, nhng li thiu quan tm n vn liu lng v thi gian cch ly khi s dng thuc. Tuy nhin, tt c cc mu nho u c gi tr hm lng OPs thp
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hn so vi gii hn d lng ti a (MRL) theo tiu chun ca B Y t (Diazinon 0,2 ppm; Dimethoate 1,0 ppm; Fenitrothion 0,5 ppm; Malathion 8,0 ppm; Metylparathion 0,5 ppm).

TI LIU THAM KHO

1. B Nng nghip & PTNT (1999), Tiu chun ngnh s 10TCN 386 99, Phng php ly mu kim nh cht lng v d lng thuc bo v thc vt (ban hnh km theo Quyt nh s 116/1999/QBNNKHCN ngy 4 thng 8 nm 1999 ca B Nng nghip v PTNT). 2. B Y t (2008), Quy nh gii hn ti a nhim sinh hc v ha hc trong thc phm (ban hnh km theo Quyt nh s 46/2007/Q BYT ngy 19 thng 12 nm 2007 ca B trng B Y t), Nh xut bn H Ni. 3. Phm Hu Nhng, Nguyn Hu Bnh, L Xun Bnh, L Quang Quyn (2000), K thut trng nho, Nh xut bn Nng nghip. 4. Critina Aprea, Claudio Colosio, Teresa Mammone, Claudio Minoia, Marco Maroni (2002), Biological monitoring of pesticide exposure: a review of analytical methods, Journal of Chromatography B, 769, 191 219. 5. CEN/TC 275/WG 4 (2006), Foods of plant origin Determination of pesticide residues using GC/MS or LC/MS/MS following acetonitrile extraction/partitioning QuEChERS-method (European Standard), the results of discussions at the 13th meeting of CEN/TC 275/WG 4 in Berlin. 6. J.S. Fritz (1999), Analytical Solid Phase Extraction, Wiley VHC, New York. 7. William Horwitz (2005), Official Methods of Analysis of AOAC International, 18th Edition, USA. 8. Somenath Mitra (2003), Sample Preparation Techniques in Analytical Chemistry, John Wiley and Sons, Inc., New York. 9. Nguyen Minh Chau, Pham Ngoc Lieu, Le Thi Thu Hong, Dang Kim Son, Nguyen Van Tinh, Nguyen Man (2002), Fruits in Vietnam, Agriculture Publishing House. 10. J. C. Miller and J. N. Miller (1988), Statistics for Analytical Chemistry, Ellis Horwood Limited.

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DETERMINATION OF ORGANOPHOSPHORUS PESTICIDES RESIDUES IN GRAPE IN NINH THUAN PROVINCE

Ngo Van Tu College of Pedagogy, Hue University Nguyen Thi Thuy Uyen Dung Quat Vocational College of Technique and Technology

SUMMARY
This article describes the method of determining orangnophosphorus pesticides (OPs) residues in Grape using Gas Chromatography. This method was appreciated by means of detection limit, recovery and precision. With the analytical procedure and the working conditions for chromatographic system which was chosen, detection limit of the method for Dimethoate, Diazinon, Metylparathion, Fenitrothion and Malathion was 18, 19, 12, 19 and 16ppb, respectively and its recovery and precision might be accepted. This method was applied to determine OPs residues in some grape samples in Ninh Thuan province.

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