TIU CHUN VIT NAM TCVN 8319 : 2010

RAU QU - XC NH A D LNG THUC BO V THC VT PHNG PHP SC K KH Vegetables and fruits Determination of pesticide multiresidues Gas chromatographic method Li ni u TCVN 8319:2010 do Cc Bo v thc vt bin son, B Nng nghip v Pht trin nng thn ngh, Tng cc Tiu chun o lng Cht lng thm nh, B Khoa hc v Cng ngh cng b. RAU QU - XC NH A D LNG THUC BO V THC VT PHNG PHP SC K KH Vegetables and fruits Determination of pesticide multiresidues Gas chromatographic method 1. Phm vi p dng Tiu chun ny quy nh phng php xc nh ng thi d lng thuc bo v thc vt (BVTV) trong rau qu, bao gm acephate, chlorpyriphos, diazinon, dimethoate, fenitrothion, profenophos, methidathion, trichlorfon, -endosulfan, -endosulfan, iprodion, cyfluthrin, cypermethrin, fenvalerate, -cyhalothrin, permethrin, difenoconazole, propiconazole, chlorothalonil, fipronil v indoxacarb bng sc k kh. 2. Ti liu vin dn Cc ti liu vin dn sau rt cn thit cho vic p dng tiu chun ny. i vi cc ti liu vin dn ghi nm cng b th p dng phin bn c nu. i vi cc ti liu vin dn khng ghi nm cng b th p dng phin bn mi nht, bao gm c cc sa i, b sung (nu c). TCVN 4851 (ISO 3696), Nc dng phn tch trong phng th nghim Yu cu k thut v phng php th. 3. Nguyn tc D lng cc thuc BVTV trong mu th c chit bng hn hp dung mi axeton v c xc nh bng thit b sc k kh vi detector quang ha ngn la (FPD) v detector cng kt in t (ECD). 4. Thuc th Ch s dng cc thuc th loi tinh khit phn tch, nc loi 3 ca TCVN 4851 (ISO 3696), tr khi c quy nh khc. 4.1. Axeton, tinh khit phn tch. 4.2. Diclometan, tinh khit phn tch. 4.3. Ete du m, nhit si t 40 oC n 60 oC. 4.4. Toluen, tinh khit phn tch. 4.5. Hn hp dung mi, cha -hexan v toluen vi t l th tch 9 : 1. 4.6. Natri sulfat khan, hot ha 130 oC trong 8 h, ngui trong bnh ht m, bo qun trong bnh kn. 4.7. Cc cht chun acephate, chlorpyriphos, diazinon, dimethoate, fenitrothion, profenophos, methidathion, trichlorfon, -endosulfan, -endosulfan, iprodion, cyfluthrin, cypermethrin,

fenvalerate, -cyhalothrin, permethrin, difenoconazole, propiconazole, chlorothalonil, fipronil v indoxacarb, bit tinh khit. 4.8. Dung dch chun gc, nng 1 000 g/ml Dng cn phn tch (5.7) cn 0,01 g tng cht chun (4.7), chnh xc n 0,01 mg, cho vo cc bnh nh mc dung tch 10 ml (5.1), thm toluen n vch v trn. 4.9. Dung dch chun trung gian 4.9.1. Dung dch chun trung gian 1 Dng micropipet (5.3) ly 500 l cc dung dch chun gc acephate, chlorpyriphos, diazinon, dimethoate, fenitrothion, profenophos, methidathion, dng pipet (5.2) ly 2 ml dung dch chun gc trichlorfon cho vo bnh nh mc dung tch 50 ml (5.1), thm hn hp dung mi (4.5) n vch v trn. Trong dung dch chun trung gian 1, cc hot cht acephate, chlorpyriphos, diazinon, dimethoate, fenitrothion, profenophos, methidathion c nng 10 g/ml, hot cht trichlorfon c nng 40 g/ml. 4.9.2. Dung dch chun trung gian 2 Dng micropipet (5.3) ly 500 l cc dung dch chun gc difenoconazole v permethrin; 400 l cc dung dch chun gc cyfluthrin, cypermethrin, fenvalerate, indoxacarb v iprodion; 200 l cc dung dch chun gc chlorothalonil, -cyhalothrin, -endosulfan, -endosulfan, fipronil, propiconazole cho vo bnh nh mc dung tch 50 ml (5.1), thm hn hp dung mi (4.5) n vch v trn. Trong dung dch chun trung gian 2, hot cht difenoconazole v permethrin c nng 10 g/ml, cc hot cht cyfluthrin, cypermethrin, fenvalerate, indoxacarb v iprodion c nng 8 g/ml, cc hot cht chlorothalonil, -cyhalothrin, -endosulfan, -endosulfan, fipronil v propiconazole c nng 4 g/ml. 4.10. Dung dch chun lm vic Pha long lin tc dung dch chun trung gian 1(4.9.1) v dung dch chun trung gian 2 (4.9.2) thu c 6 dung dch chun lm vic vi cc mc nng nh sau: - Dung dch chun lm vic 1: cc hot cht acephate, chlorpyriphos, diazinon, dimethoate, fenitrothion, profenophos, methidathion c nng 1 g/ml, hot cht trichlorfon c nng 4 g/ml; - Dung dch chun lm vic 2: cc hot cht acephate, chlorpyriphos, diazinon, dimethoate, fenitrothion, profenophos, methidathion c nng 0,5 g/ml, hot cht trichlorfon c nng 2 g/ml; - Dung dch chun lm vic 3: cc hot cht acephate, chlorpyriphos, diazinon, dimethoate, fenitrothion, profenophos, methidathion c nng 0,02 g/ml, hot cht trichlorfon c nng 0,08 g/ml; - Dung dch chun lm vic 4: cc hot cht difenoconazole v permethrin c nng 2 g/ml, cc hot cht cyfluthrin, cypermethrin, fenvalerate, indoxacarb v iprodion c nng 1,6 g/ml, cc hot cht chlorothalonil, -cyhalothrin, -endosulfan, -endosulfan, fipronil v propiconazole c nng 0,8 g/ml; - Dung dch chun lm vic 5: cc hot cht difenoconazole v permethrin c nng 0,4 g/ml, cc hot cht cyfluthrin, cypermethrin, fenvalerate, indoxacarb v iprodion c nng 0,32 g/ml, cc hot cht chlorothalonil, -cyhalothrin, -endosulfan, -endosulfan, fipronil v propiconazole c nng 0,16 g/ml; - Dung dch chun lm vic 6: cc hot cht difenoconazole v permethrin c nng 0,025 g/ml, cc hot cht cyfluthrin, cypermethrin, fenvalerate, indoxacarb v iprodion c nng 0,02

g/ml, cc hot cht chlorothalonil, -cyhalothrin, -endosulfan, -endosulfan, fipronil v propiconazole c nng 0,01 g/ml; Cc dung dch chun lm vic c bo qun 4 oC v c thi hn s dng l 6 thng. 4.11. Kh ni t, c tinh khit khng nh hn 99,999 %. 4.12. Kh hydro, c tinh khit khng nh hn 99,999 %. 4.13. Khng kh nn dng cho thit b sc k kh. 5. Thit b, dng c S dng cc thit b, dng c ca phng th nghim nh sau: 5.1. Bnh nh mc, dung tch 10 ml, 50 ml v 100 ml. 5.2. Pipet, dung tch 2 ml, 5 ml v 10 ml, chia vch n 0,1 ml. 5.3. Micropipet, dung tch t 50 l n 200 l v t 200 l n 1 000 l. 5.4. Cc ly tm, dung tch 250 ml. 5.5. ng ong, dung tch 50 ml. 5.6. ng nghim, dung tch 15 ml. 5.7. Xyranh, dung tch 10 l, c chia vch n 1 l. 5.8. Cn phn tch, c th cn chnh xc n 0,01 mg. 5.9. Cn k thut, c th cn chnh xc n 0,01 g. 5.10. Thit b ng ha Ultra-Turrax, tc khng nh hn 13 500 r/min. 5.11. Thit b thi kh nit. 5.12. My nghin mu. 5.13. My ly tm, tc khng nh hn 2 000 r/min, c ng ly tm dung tch 250 ml. 5.14. Thit b sc k kh 1, c trang b nh sau: - Bung bm mu chia dng v khng chia dng; - Detector quang k ngn la, knh lc phospho (FPD/P); - Ct mao qun DB-5, c chiu di 30 m, ng knh 0,32 mm, chiu dy pha tnh 0,25 m, hoc loi tng ng; - My vi tnh. 5.15. Thit b sc k kh 2, c trang b nh sau: - Bung bm mu chia dng v khng chia dng; - Detector cng kt in t (ECD); - Ct mao qun DB-5, c chiu di 30 m, ng knh 0,32 mm, chiu dy pha tnh 0,25 m, hoc loi tng ng; - My vi tnh. 6. Ly mu Vic ly mu khng c quy nh trong tiu chun ny. Nn ly mu theo TCVN 5139:2008 (CAC/GL 33-1999) Phng php khuyn co ly mu xc nh d lng thuc bo v thc vt ph hp vi cc gii hn d lng ti a (MRL). 7. Cch tin hnh

7.1. Yu cu chung Ton b quy trnh phn tch cn c thc hin trong ngy. Nu khng phn tch c trong ngy th phi bo qun dch mu 4 oC. 7.2. Chun b mu Mu th c nghin trong my nghin mu (5.12) n khi ng nht. 7.3. Chun b phn mu th Dng cn (5.9) cn khong 20 g mu th (m) c ng nht (xem 7.2), chnh xc n 0,01g, vo cc ly tm dung tch 250 ml (5.4). Thm vo cc 40 ml axeton, ng ha trong 30 s bng thit b Ultra-Turrax (5.10) vi tc 13 500 r/min, thm ln lt 40 ml ete du m, 40 ml diclometan, 5 g natri sulfat khan, ng ha trong 30 s bng thit b Ultra-Turrax (5.10) vi tc 13 500 r/min, sau ly tm bng thit b ly tm (5.13) vi tc 2 000 r/min trong 10 min, thu ton b dch chit v ghi li th tch (V1). Dng pipet dung tch 10 ml (5.2) ly chnh xc 8 ml (V2) dch lng thu c cho vo ng nghim (5.6) v thi kh hon ton bng thit b thi kh nit (5.11). Ha tan phn cn vi 2 ml (VE) hn hp dung mi (4.5) thu c phn mu th. 7.4. Chun b phn mu trng Mu trng l mu c bit khng c d lng acephate, chlorpyriphos, diazinon, dimethoate, fenitrothion, profenophos, methidathion, trichlorfon, -endosulfan, -endosulfan, iprodion, cyfluthrin, cypermethrin, fenvalerate, -cyhalothrin, permethrin, difenoconazole, propiconazole, chlorothalonil, fipronil v indoxacarb, c chun b theo quy trnh trong 7.3. 7.5. Chun b phn mu kim tra hiu sut thu hi Dng cn (5.9) cn khong 20 g mu trng c ng nht theo 7.2, chnh xc n 0,01 g, vo cc ly tm dung tch 250 ml (5.4). Dng micropipet (5.3) thm 200 l dung dch chun trung gian 1 v 200 l dung dch chun trung gian 2, yn trong nhit phng ti thiu 15 min. Tip tc thc hin theo 7.3. 7.6. iu kin phn tch 7.6.1. Thit b sc k kh 1 Nhit bung bm mu: Th tch bm mu: Nhit ct tch: 240 oC; 2 l, khng chia dng; Nhit ban u 80 oC gi trong 1 min, tng 40 oC/min n nhit 160 oC, tng 3 oC/min n 250 oC, tng 10 oC/min n nhit cui 280 oC v gi trong 10 min; 2 ml/min; 220 oC; 75 ml/min; 100 ml/min; 60 ml/min. 240 oC; 1 l, khng chia dng; Nhit ban u 80 oC gi trong 1 min, tng 40 oC/min n nhit 160 oC, tng 3 oC/min n 250 oC, tng 10 oC/min n nhit cui

Tc kh mang (N2): Nhit detector FPD: Tc kh hydro (H2): Tc d dng khng kh: Tc nit b tr: 7.6.1. Thit b sc k kh 2 Nhit bung bm mu: Th tch bm mu: Nhit ct tch:

280 oC v gi trong 10 min; Tc kh mang (N2): Nhit detector ECD: Tc nit b tr: 7.7. Dng ng chun Dng ng chun ca cc thuc BVTV (tng quan gia din tch/chiu cao pc v nng cht chun) ti 3 im c nng tng ng trong dung dch chun lm vic 1, dung dch chun lm vic 2 v dung dch chun lm vic 3 trn thit b sc k 1 (5.14). Dng ng chun ca cc thuc BVTV (tng quan gia din tch/chiu cao pc v nng cht chun) ti 3 im c nng tng ng trong dung dch chun lm vic 4, dung dch chun lm vic 5 v dung dch chun lm vic 6 trn thit b sc k 2 (5.15). 7.8. Xc nh Bm ln lt dung dch mu trng (7.4), dung dch mu th (7.3), dung dch mu kim tra hiu sut thu hi (7.5) vo thit b sc k kh (5.14) v thit b sc k kh 2 (5.15). Xc nh nng ca mu th v mu kim tra hiu sut thu hi bng ng chun. Nu nng ca mu th nm ngoi ng chun th iu chnh bng cch pha long dung dch phn mu th (khng pha long lng mu bm). 8. Tnh kt qu D lng tng thuc BVTV (4.7), X, biu th bng miligam trn kilogam (mg/kg), c tnh theo cng thc: X = Xo x Trong : Xo l nng tng thuc BVTV (4.7) trong mu th c xc nh theo 7.8, tnh bng microgam trn mililit (g/ml); VE l th tch phn mu th thu c (xem 7.3), tnh bng mililit (ml); V1 l th tch dch chit thu c (xem 7.3), tnh bng mililt (ml); V2 l th tch dch chit c ly ra thi kh (xem 7.3), tnh bng mililt (ml); m l khi lng mu th, tnh bng gam (g); P l tinh khit ca cht chun, tnh bng phn trm (%). 9. Hiu sut thu hi v gii hn xc nh 9.1. Hiu sut thu hi ca phng php: t 70 % n 110 %. 9.2. Gii hn nh lng ca phng php (LOQ) - i vi acephate, chlorpyriphos, diazinon, dimethoate, fenitrothion, profenophos, methidathion, iprodion, cyfluthrin, cypermethrin, fenvalerate, permethrin, difenoconazole, indoxacarb: 0,05 mg/kg; - i vi chlorothalonil, -endosulfan, -endosulfan, fipronil, lambda cyhalothrin, propiconazole: 0,02 mg/kg. 10. Bo co th nghim Bo co th nghim phi ghi r: a) Mi thng tin cn thit v vic nhn bit y mu th; 2 ml/min; 300 oC; 60 ml/min.

VE V1 P V2 m 100

b) Phng php ly mu s dng, nu bit; c) Phng php th s dng v vin dn tiu chun ny; d) Mi thao tc khng c quy nh trong tiu chun ny, hoc nhng iu c coi l t chn, v bt k chi tit no c nh hng n kt qu. e) Kt qu th nghim thu c.

TH MC TI LIU THAM KHO

[1] British Crop Protection Council 2003, The Pesticide Manual. [2] Food and Drug Aministration of USA, 1994, Pesticide Analysis Manual, Volume I, Section 302, E4.