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Pesticides and PCB in Food and Feed According to Method L 00.00-34

(§ 64 LFGB)

What LCTech does

supplies innovative products and methods for the preparation and analysis of food, feed and environmental samples.

The Extended and Revised DFG S19 Method L 00.00-34 § 64 LFGB (former § 35 LMBG)

Official Manual of Test Methods According to § 64 LMBG

Modular Multiple Analytical Method for the Determination of Pesticide Residues in

Foodstuffs(Modified Version of the DFG Method S 19)

Analysis of FoodstuffsL 00.00

34

"Beuth" publishing house www.beuth.de

Content

General Questions

The Fields of Application

The Method in Detail

General Questions

Why to analyse e.g. pesticides?

Why to use a MMM (Modular Multi-Residue Method)?

Why to analyse e.g. pesticides?

165149

124121105100

6884

Vegetable

Fruit

Number of different pesticides found

736949

158144

121100

8368

4046444037

1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2005

?

Why to analyse e.g. pesticides?

• Improved detection technologies

• Resistant pests on crops (e.g. grape 10 20 pesticdes)

• Old or forbidden pesticides

?

Occurrence of pesticides (40.000 samples)

Pesticide Occurrence in samples

Carbendazim 1931Chlorpyrifos 1735Iprodion 1447Bromide (inorg.) 1383Azoxystrobin 1245Imazalil 921Cyprodinil 889Thiabendazol 748Cypermethrin 703Captan 673Metalaxyl/Metalaxyl M 670Fludioxonil 623Pyrimethanil 609Bromopropylat 603Malathion 582

Example Turkish Grapes

Compound Application Conc. mg/kg

MRL mg/kg % MRL

Quinalphos Insectizide 0,22 0,05 440,00Imazalil Fungicide 0,14 0,02 700,00

Unfortunately very often 10 to 15 different pesticides can be found on grapes!

Why to use a MMM (Modular Multi-Residue Method)?

Sample Reception

Why to use a MMM (Modular Multi-Residue Method)?

• Pesticides can be found in nearly any food or feed

• Matrices differ strongly in their composition (e.g. fat, water, acid, carbohydrate, …)

• In general the analytes are not known

• Single or group methods are not practical in routine labs for a general use

• > 1000 pesticides are sold world-wide• In Germany, residues of approx. 550 pesticides in > 25

crop types and 5 animal materials are regulated• Each year 10 new pesticides are registered in Germany• Additionally, residue chemists have to determine

forbidden pesticides and contaminants (e.g. DDT, PCB, Nitro musk, TBT)

Conclusion: Multi-residue methods are needed!

Why to use a MMM (Modular Multi-Residue Method)?

• DFG S19

• Pesticides (X-, P-, N-containing) Pyrethroids

• Polyhalogenated Biphenyls

• Non-polar to semi-polar

The Fields of Application

Gel Permeation Chromatography GPC

Eluate 1 65% Hexane 35% Toluene

Mini Silica Column Chromatography MSC

Eluate 2

100% Toluene

Eluate 3 5% Acetone 95% Toluene

Eluate 4 20% Acetone 80% Toluene

Eluate 5 100% Acetone

Extraction modules

E1 E2 E3 E4 E5 E6 E7 E8 E9

The Method in Detail

MSDPNDFPD

ECD ECD ECD FPD PND

LC-MS

Extraction Modules

Since 1995 ‚NEW‘

Till 1995 ‚OLD‘

Ethyl acetate Cyclohexane

Dichloro methane

Extraction Modules

• Extraction modules are choosen according to matrix composition

• E1/E4: water > 70 %, fat < 2,5 %

• E2/E5: water < 70 %, fat < 2,5 %

• E3: see E1, but high acid content (buffered to pH 7)

•Type: Extraction with liquid/liquid partitioning

• Typically for vegetables, fruit, juice

Extraction Modules

• Why is the water content so important?

• Adjusting to 100 mL total water content (E1 – E5)

• Soaking time (30 min) is essential, if water < 70 %!

• Ratio of water / extraction solvent is the same

Extraction Modules

• Extraction modules are choosen according to matrix composition

• E6: Water 0 %, fat 100 % Oil, Tallow

• E7: Water low, fat > 2,5 % Nuts, Oil Seeds

• E8: Water high, fat > 2,5 % Meat, Fish, Cheese

• E6: Dissolving fat in GPC eluent

• E7: Extraction in the presence of large amounts of fat

• E8: Cold extraction with hexane / acetone

Extraction Modules

• Automated extraction with the ASE system from Dionex

• Typically used for dry matrices (Tea, Cereals, Spices)

• Extraction is performed with elevated temperature (120 °C) and pressure (100 bar)

• ‚Pressure Cooking Pot‘-Principle

Extraction Modules

Gel Permeation Chromatography Module (GPC)

• See previous presentation:

• ‘Introduction into Gel Permeation Chromatography’

Gel Permeation Chromatography GPC

Eluate 1 65% Hexane 35% Toluene

Mini Silica Column Chromatography MSC

Eluate 2

100% Toluene

Eluate 3 5% Acetone 95% Toluene

Eluate 4 20% Acetone 80% Toluene

Eluate 5 100% Acetone

Extraction modules

E1 E2 E3 E4 E5 E6 E7 E8 E9

MSDPNDFPD

ECD ECD ECD FPD PND

LC-MS

OCP Pesticides < 3 ppb

OPP Pesticides < 3 ppb

MS-NCI

Module C1 – C2 Mini Silica Column Cleanup

GPC Concentrate 5,0 mL

Isooctane Concentrate

Solvent Exchange

Mini Silica Column

Ethyl acetate musn‘t be present!

Eluate 1 65% Hexane 35% Toluene

Eluate 2

100% Toluene

Eluate 3 5% Acetone 95% Toluene

Eluate 4 20% Acetone 80% Toluene

Eluate 5 100% Acetone

Combined Combined

Eluate 0

100% Hexane

PCB expected

Module C1 – C2 Mini Silica Column Cleanup

Pesticide GPC E 1 E 2 E 3 E 4 E 5 ng / µl notes

Aldrine 120-150 5 0 0 0 0 0,9 Eluate 1 (ECD) + Rt check

HCB 140-165 5 0 0 0 0 0,5 "evaporate" - check

Trifluraline 100-130 5 0 0 0 0 0,9 Eluate 1 (ECD)

Bromophos 120-150 4 2 0 0 0 1,8 Eluate 1 (NPD + FPD)

Bifenthrine 60-120 2 3 0 0 0 4,5 GPC cut - large molecules

Endosulfansulfate 100-140 0 5 0 0 0 0,5 Eluate 2 (ECD)

Parathion 110-140 0 4 1 0 0 2,7 Eluate 2 (NPD + FPD)

Chinomethionate 170-200 0 1 4 0 0 27,0 GPC cut end

Bromopropylate 95-135 0 0 4 2 1 2,7 Eluate 3 + 4 (ECD)

Captane 120-150 0 0 5 0 0 45,0 Eluate 3 (NPD) // + Insert check

Metalaxyl 115-150 0 0 0 5 1 45,0 Eluate 4 (NPD)

Triamiphos 125-160 0 0 0 0 4 90,0 Eluate 5 (NPD + FPD)

GPC and Mini-Silica Testmix

Official Manual of Test Methods According to § 64 LFGB

L Analysis of FoodstuffsResidue analysis of plant protective substances

in Food (Table of relative retention time of plant

protective substances with gas chromatography

00.00

38

Analysis of Foodstuffs

Pesticide CAS-Nr.RRT

HP5MS

(Parathion)

RRT DB35MS

(Parathion)

Ion 1 Ion 2 Ion 3 Ion 4 MS

D

EC

D

NP

D

Dichloranilin, 3,5- 626-43-7 0,47 0,54 161 163 90 99 xxx xx x

Acephate 30560-19-1 0,47 0,58 136 94 95 96 xx x xx

Aclonifen 74070-46-5 1,40 1,34 264 266 212 77 xxx x x

Acrinathrin, 1 101007-06-1 1,57 1,41 207 181 281 208 xx xxx xx

Acrinathrin, 2 101007-06-1 1,58 1,46 181 208 207 93 xxx xxx xx

Alachlor 15972-60-8 0,89 0,88 160 188 146 237 xxx xxx x

Aldrin 309-00-2 0,97 0,93 263 66 265 261 xxx xxx

Allidochlor 93-71-0 0,40 0,47 56 138 132 70 xxx xx xx

Alodan 2550-75-6 1,09 1,04 229 272 237 273 xx xxx

Ametryn 834-12-8 0,89 0,93 227 212 170 185 xxx xx

Amidithion 919-76-6 0,96 1,03 125 131 93 143 x x x

Aminocarb 2032-59-9 0,73 0,79 151 150 136 208 xx xx

Amitraz 33089-61-1 1,56 1,46 121 162 132 293 xx x

696 peaks with Rt times

Thanks for your attention!

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