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
[email protected]@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.