Možnosti evropského výzkumu v Možnosti evropského výzkumu v oblasti související s fytosanitární oblasti související s fytosanitární

problematikouproblematikou

Jana HajšlováJana HajšlováÚstav chemie a analýzy potravin, VŠCHT PrahaÚstav chemie a analýzy potravin, VŠCHT Praha

Jana.hajslova@vscht.czJana.hajslova@vscht.cz

Národní fytosanitární workshopNárodní fytosanitární workshop3.3.2009, Praha3.3.2009, Praha

Activity 2.1: Sustainable production and management of biological Sustainable production and management of biological resources from land, forest and aquatic environmentresources from land, forest and aquatic environment

WORK PROGRAMME 2010

COOPERATION

THEME 2

FOOD, AGRICULTURE AND FISHERIES, AND BIOTECHNOLOGY

DRAFT

17.02.2009

KBBE-2010-1-2-06: Deepened and enlarged cooperation between phytosanitary (statutory plant health) research programmes - ERA-NET

Funding schemeFunding scheme: Coordination and Support Action : Coordination and Support Action (supporting action). The requested European Community (supporting action). The requested European Community contribution shall not exceed EUR 1 000 000.contribution shall not exceed EUR 1 000 000.

The aim is to prevent the introduction into the community of organisms harmful to plants or plant products or their spread within the Community. Due to climate changes and globalization, new threats are continuously emerging and the number and complexity of plant health problems increases every year, requiring enhanced research efforts.requiring enhanced research efforts.

KBBE-2010-1-2-05: KBBE-2010-1-2-05: Integrated pest management in Integrated pest management in farming systems of major importance for Europe farming systems of major importance for Europe

Funding scheme: Collaborative Project (large scale integrating project). The requested European Community contribution shall not exceed EUR 9 000 000.

better targeted use of all available pest control measures, including preventive measures, observation and pest monitoring, pesticides and biocontrol agents,

modelling and experimentation, including physiological and molecular understanding of plant-pathogen interactions and development of symptoms

targeting farming systems in which reduction of pesticide use and better control of pests and pathogens is expected to have major effects at the European scale

Jaká jsou rizika pro konzumenty?

Je lepší volit bioprodukty?

Samples from Czech marketSamples from Czech market

23 extra virgin olive oils from organic farming (O1 – O23)

23 extra virgin olive oils from conventional farming (C1 – C23)

Method of analysisMethod of analysis

GCx GC/TOF MSGCx GC/TOF MS

EXTRA VIRGIN OLIVE OILEXTRA VIRGIN OLIVE OIL The oil obtained from the fruit of the olive tree solelyThe oil obtained from the fruit of the olive tree solely by by

mechanical or other physical means under mechanical or other physical means under tthe he cconditions. onditions. particularly thermal conditions, that do not lead to alternations particularly thermal conditions, that do not lead to alternations

in the oil, and which has not undergone any treatment other in the oil, and which has not undergone any treatment other than washing, decantation, centrifugation and filtration.than washing, decantation, centrifugation and filtration.

OLIVE OILOLIVE OIL

Sum of 16 PAHSum of 16 PAH = Sum of B[a]A, Chr, CP[cd]P, 5-MeChr, B[c]Fln, B[b]F, B[k]F, B[j]F, = Sum of B[a]A, Chr, CP[cd]P, 5-MeChr, B[c]Fln, B[b]F, B[k]F, B[j]F, B[a]P, DB[ah]A, B[ghi]P, I[cd]P, DB[ah]P, DB[al]P, DB[ai]P, DB[ae]PB[a]P, DB[ah]A, B[ghi]P, I[cd]P, DB[ah]P, DB[al]P, DB[ai]P, DB[ae]P

0

10

20

30

C1 C2 C3 C4 C5 C6 C7 C8 C9 C10

C11

C12

C13

C14

C15

C16

C17

C18

C19

C20

C21

C22

C23

O1

O2

O3

O4

O5

O6

O7

O8

O9

O10

O11

O12

O13

O14

O15

O16

O17

O18

O19

O20

O21

O22

O23

0,00

0,20

0,40

0,60

0,80

1,00

1,20

1,40

1,60

1,80

2,00

2,20

2,40

Conventional Organic

Su

m o

f 16

PA

H

B[a

]P (

µg

/kg

)Sum of 16 PAH

B[a ]P

PAHs in olive oilPAHs in olive oil

Benzo(a)pyrene)

PESTICIDEPESTICIDE RESIDUES in olive oilsRESIDUES in olive oils

AcephateAcrinathrinAldrinAzinfos ethylAzinfos methylBeta cyfluthrin BifenthrinBromophos ethylBromophos methylBrompropylateBupirimateCaptanCarbarylCarbophenothionCyhalothrinCypermethrinCyprodinylDeltamethrinDiazinonDieldrinDifenoconazole DifenylaminDichlofluanidDichlorvosDimethoateEndosulfan alfa

Endosulfan betaEndosulfan sulfatEndrinEthionEtrimfosFenamiphosFenarimolFenchlorphosFenitrothionFenoxycarbFenthionFenvalerate FludioxonilFolpetFormothionHCBHeptachlorHeptenophosHexythiazoxHCH alfaHCH betaHCH deltaChlorfenviphos ChlorophamChlorothalonilChlorpyrifos

Chlorpyrifos-methylImazalilIprodioneIsofenphosKresoxim methylLindaneMalathionMetalaxylMethacrifosMethamidophosMethidathionMevinphosMonocrophosDDDDDEDDTOmethoateOxyfluorfenOxychlordaneParaoxon ethylParathionParathion methylPenconazolePermethrin PhenothrinPhenthoate

PhosalonePhoshamidone PhosmetPirimicarbPirimiphos methylProcymidoneProphamPyridabenQuinalphosSulfotepTebuconazoleTetraconazoleTetradifonThiobendazol Tolclofos-methylTolylfluanidTriadimefonTriazamateTriazophosTrifloxystrobinTrichlorfonVinclozolin

Endosulfan sulphate

0,0

100,0

200,0

300,0

C1

C2

C3

C4

C5

C6

C7

C8

C9

C1

0

C1

1C

12

C1

3C

14

C1

5C

16

C1

7

C1

8C

19

C2

0C

21

C2

2C

23

O1

O2

O3

O4

O5

O6

O7

O8

O9

O1

0O

11

O1

2O

13

O1

4O

15

O1

6O

17

O1

8

O1

9O

20

O2

1O

22

O2

3

(µg

/kg

) Conventional Organic

Fenthion and chlorpyrifos

0,0

100,0

200,0

300,0

C1

C2

C3

C4

C5

C6

C7

C8

C9

C1

0

C1

1C

12

C1

3

C1

4C

15

C1

6C

17

C1

8C

19

C2

0C

21

C2

2C

23

O1

O2

O3

O4

O5

O6

O7

O8

O9

O1

0

O1

1O

12

O1

3

O1

4O

15

O1

6O

17

O1

8O

19

O2

0O

21

O2

2O

23

Fenthion

Chlorpyrifos

(µg

/kg

)

PPESTICIDESESTICIDES DETECTEDDETECTED ININ OLIVEOLIVE OILSOILS

PesticidesPesticides

RESIDUERESIDUE

CONVENTIONALCONVENTIONAL ORGANICORGANIC

nmean (µg/kg)

Min - Max (µg/kg)

% of positive samples n

mean (µg/kg)

Min - Max (µg/kg)

% of positive samples

EndosulfanEndosulfan sulphatesulphate

2222 55.255.2 26.9 - 210.226.9 - 210.2 9696 88 33.833.8 24.5 - 43.524.5 - 43.5 3535

Fenthion Fenthion 88 127.1127.1 58.0 - 31958.0 - 319 3535 00 -- -- 00

Chlorpyrifos Chlorpyrifos 33 40.540.5 37.8 - 41.537.8 - 41.5 1313 22 31.731.7 99

n - number of positive samples

komoditakomodita produkční produkční systémsystém

počet počet detekovanýdetekovaných reziduích reziduí

mrkev

organické

0

0

0

konvenční

2

4

1

jablka

organické

0

0

0

konvenční

6

3

6

komoditaprodukční

systém

počet detekovanýc

h reziduí

rajčata

organické

0

1

0

konvenční

5

11

9

papriky

organické0

1

konvenční

3

5

3

9

Rezidua pesticidů ve vzorcích z ekologického a Rezidua pesticidů ve vzorcích z ekologického a konvenčního zemědělství na českém trhukonvenčního zemědělství na českém trhu

22882 pesticides in 2 pesticides in LC- MS/MS multiresidue methodLC- MS/MS multiresidue method

Column: Supelco DiscoverySupelco Discovery C18 ,Mobile phase: MeOH (A), 0.01M amm. acetate (B)Flow: 0.3 ml/min

PERFORMANCE OF METHOD EMPLOYING PERFORMANCE OF METHOD EMPLOYING QQQQQQ:: for all analytes 4 identification points at concentration levels 0.01 mg/kg available requirements of

2002/657/EC met

Kovalczuk T., Lacina O., Jech M., Poustka J.,

Hajšlová J.: Novel approach to fast

determination of multiple pesticide residues

using UPLC-MS/MS. Food Addit. Cont. 25(4),

444–457 (2008).

Various contaminants may occur in cereal grainsVarious contaminants may occur in cereal grains

Is it possible to examine Is it possible to examine the presence of these the presence of these toxins in a single run?toxins in a single run?

PESTICIDESPESTICIDES

MYCOTOXINSMYCOTOXINS

PLANT TOXINSPLANT TOXINS

POPsPOPs

PAHsPAHs

Acephate Cymoxanil Etrimfos Isofenphos Oxamyl Propoxur Thiophanate-methyl

Acetamiprid Cypermethrin Fenamiphos Isoproturon Oxydemeton-methyl Propyzamide Tolylfluanid

Acrinathrin Cyproconazole Fenamiphos-sulfon Kresoxim-methyl Oxyfluorfen Prosulfocarb Triadimefon

Alachlor Cyprodinil Fenamiphos-sulfoxide Lenacil Paclobutrazol Pymetrozine Triadimenol

Aldicarb Deltametrin Fenarimol Linuron Penconazole Pyraclostrobin Triazophos

Aldicarb-sulfone Demeton-S-methyl Fenazaquin Malaoxon Pencycuron Pyrazophos Trichlorfon

Aldicarb-sulfoxide Demeton-S-methyl-sulfone Fenbuconazole Malathion Pendimethalin Pyrethrins Trifloxystrobin

Ametryn Desmetryn Fenhexamid Mecarbam Permethrin Pyridaben Triflumuron

Atrazine Diazinon Fenoxycarb Mefenpyr-diethyl Phenmedipham Pyrimethanil Vamidothion

Azinphos-ethyl Dichlofluanid Fenpropathrin Mepanipyrim phenothrin Pyriproxyfen

Azinphos-methyl Dichlorvos Fenprophimorph Metalaxyl Phenthoate Quinalphos

Azoxystrobin Diclofop-methyl Fenpropidin Metconazole Phorate Quinmerac Aflatoxin B1

Benalaxyl Dicrotophos Fenpyroximate Methamidophos Phorate-sulfon Quinoxyfen Aflatoxin B2

Bifenthrin Diethofencarb Fenthion Methidathion Phorate-sulfoxide Resmethrin Aflatoxin G1

Bitertanol Difenoconazole Fipronil Methiocarb phosalone Rimsulfuron Aflatoxin G2

Bupirimate Diflubenzuron Fluazifop-P-butyl Methiocarb sulfone Phosmet Simazine Altenuene

Buprofezin Diflufenican Flufenoxuron Methiocarb sulfoxide Phosphamidon Simetryn Alternariol

Cadusafos Dimethoate Fluoxastrobin Methomyl Picoxystrobin Spinosad Alternariolmethylether

Carbaryl Dimethomorph Fluquinconazole Methoxyfenozide Piperonyl butoxide Spiroxamin Nivalenon

Carbendazim Dimoxystrobin Flusilazole Metobromuron Pirimicarb Sulfotep Deoxynivelonon-3-Glc

Carbofuran Disulfotone-sulfone Fonofos Metolachlor Pirimiphos-ethyl tau-Fluvalinate Deoxynivalenon

Carbofuran-3-hydroxy Disulfotone-sulfoxid Haloxyfop-acid Metolcarb Pirimiphos-methyl Tebuconazole Fusarenon-X

carbophenothion Diuron Haloxyfop-ethoxy-ethyl Metoxuron Prochloraz Tebufenozide 15-ADON

Chlorfenvinphos DMSA Haloxyfop-methyl Monocrotophos Profenofos Teflubenzuron 3-ADON

Chloroxuron DMST Heptenophos Monolinuron Prometon Terbuthylazine HT2

Chlorpyrifos Dodine Hexaconazole Monuron Prometryn Terbutryn T2

Chlorpyrifos-methyl Epoxiconazole Hexythiazox Myclobutanil Propachlor Tetraconazole Fumonisin B1

Clofentezine Ethiofencarb Imazalil Naled Propamocarb Thiabendazole Fumonisin B2

Clomazone Ethion Imidacloprid Neburon Propaquizafop Thiacloprid Fumonisin B3

Clothianidin Ethofumesate Indoxacarb Norflurazone Propargite Thiamethoxam Zearalenon

Cyanazine Ethoprophos Iodosulfuron-methyl Omethoate Propham Thiodicarb a-zearalenol

Cyazofamid Etofenprox Iprovalicarb Oxadixyl Propiconazole Thiometon b-zearalenol

Pesticides:Pesticides:

Mycotoxins:Mycotoxins:

Pesticides and all regulated mycotoxins Pesticides and all regulated mycotoxins within single run in 8 minutes within single run in 8 minutes

(UPLC- TOF MS)(UPLC- TOF MS)

AMBIENT MASS AMBIENT MASS SPECTROMETRYSPECTROMETRY

NEWNEW

……opens the doors to many challenging applications opens the doors to many challenging applications in various areas including food / feed controlin various areas including food / feed control

- Reduced / minimal sample prep- Reduced / minimal sample prep

- No chromatographic separation- No chromatographic separation

MS[1];0.195..0.225;-1.0*MS[1];0.290..0.327; / ESI+ / 20071116_SMPL13

320 340 360 380 400m/z

0

50

100

150

Area (158063)x103

409.13999327.17349

404.12625

388.10808368.11366

397.38513

355.28702

328.17700

370.30724 383.36849337.27645314.14176

371.31562324.29151 357.30173353.27294

Kresoxim methyl

Dimoxystrobin

Azoxystrobin

Picoxystrobin Pyraclostrobin

Trifloxystrobin

MS[1];1.538..1.552;-1.0*MS[1];1.571..1.577; / ESI+ / 20080307_SMPL3

50 100 150 200 250 300 350 400 450 500m/z

0

10

20

30

40

50

60

70

80

90

100

110

120

130

Intensity (136863)x103

137.14217 219.18489

205.20365

437.35451

203.18488

221.19540135.12325 237.19225

273.26460

438.35638153.13249 341.32947

195.14294

355.3087095.09818 169.15523371.30434

440.37286274.26800254.2166781.08691 123.12503

339.31240 457.37870435.34370391.31387

MS[1];1.538..1.552;-1.0*MS[1];1.571..1.577; / ESI+ / 20080307_SMPL3

292 294 296 298 300 302 304 306 308m/z

0

1000

2000

3000

4000

5000

6000

7000

8000

Intensity (8141)

297.06266

299.06012

300.26710

295.20289298.25271

305.25600

Imazalil in citrus fruit detected in 1 minuteImazalil in citrus fruit detected in 1 minute

Cl pattern

ElementElement compositioncomposition

direct screening

of peel

Concentration measured in crude extract by DART TOF

MS was 34 µg/kg.