©2008 Waters Corporation 1

Productos DisQuETM

para la preparación de muestra porQUECHERS

©2008 Waters Corporation 2

ReminderDispersive Sample Preparation“QuEChERS” Method

“Quick, Easy, Cheap, Effective, Rugged, Safe”

Where— Food laboratories

Application— High throughput sample preparation for pesticide screening analysis

Technique— Combination of salting-out LLE and matrix dispersion extraction

©2008 Waters Corporation 3

Dispersive Sample PreparationProduct offerings

PN 186004571

PN 186004572

PN 186004837

PN 186004832

Product Qty P/N

DisQuE Kits

DisQuE AOAC Dispersive Sample Preparation Kit 100 176001676

DisQuE CEN Dispersive Sample Preparation Kit 100 176001903

Individual Extraction tubes (Tube 1)

DisQuE 50 ml tube/AOAC - Acetate (1,5 g Sodium Acetate/6g MgSO4) 100 186004571

DisQuE 50 ml tube/CEN - Citrate (1,5 g Sodium Citrate/1g NaCl/4g MgSO4) 100 186004837

In dividual Clean-up tubes (tube 2)

DisQuE 2 ml tube - AOAC (150 mg MgSO4/50 mg PSA) 100 186004572

DisQuE 2 ml tube - AOAC/C18 (150 mg MgSO4/50 mg PSA/50mg C18) 100 186004830

DisQuE 2 ml tube - CEN (150 mg MgSO4/25 mg PSA) 100 186004831

DisQuE 2 ml tube - CEN/C18 (150 mg MgSO4/25 mg PSA/25mg C18) 100 186004832

DisQuE 15 ml tube / MgSO4/PSA (900 mg MgSO4/150 mg PSA) 50 186004833

DisQuE 15 ml tube / MgSO4/PSA/C18 (900 mg MgSO4/150 mg PSA/150 mg C18) 50 186004834

Bulk sorbents

Graphitized Carbon Black 25 g bottle 1 186004835

C18 100g bottle 1 WAT035672

©2008 Waters Corporation 4

DisQuE Kits

AOAC (Association of Analytical Communities )— worldwide provider and facilitator in the development, use, and

harmonization of validated analytical methods and laboratory quality assurance programs and services (primarily used in the US but European laboratories are considering their recommendations as well).

PN 176001676: DisQuE AOAC Dispersive Sample Preparation Kit

Description— DisQuE Tube 1 (Extraction):

o 50 mL centrifuge tubeo 1.5 g anhydrous sodium acetateo 6 g of anhydrous magnesium sulphate

— DisQuE Tube 2 (Clean-up):o 2 mL centrifuge tubeo 150 mg anhydrous magnesium sulphateo 50 mg of PSA

©2008 Waters Corporation 5

DisQuE ProcedureAOAC Method

Prepare Sample

Homogenize15 g sample

15 mL 1% Acetic Acid in ACN

Liquid FractionationShake for 1 minute

Centrifuge >1500 x g

CollectionRemove acetonitrile for clean-up

DisQuE ExtractionTube 1

Transfer1 mL Extract

Tube 1 to Tube 2

Shake vigously for 1 minuteCentrifuge >1500 x g

TransferCollect to autosampler vial

Dilute if necessary

DisQuE Clean-UpTube 2

©2008 Waters Corporation 6

DisQuE™ AOAC Application note

DisQuE extraction tested against 400 pesticide residue method

Matrix — Avocado, Baby food & Mango

Excellent recovery

Consistent results

©2008 Waters Corporation 7

DisQuE Kits

CEN (European Committee for Standardization )— business facilitator in Europe, removing trade barriers for European

industry and consumers. It provides a platform for the development of European Standards and other technical specifications.

PN 176001903: DisQuE CEN Dispersive Sample Preparation Kit

The European Norm has the number EN 15662:2008

Title:— Foods of plant origin - Determination of pesticide residues using GC-MS

and/or LC-MS/MS following acetonitrile extraction/partitioning and clean-up by dispersive SPE - QuEChERS-method

©2008 Waters Corporation 8

European harmonization of EN 15662:2008

The European Norm is accepted and adapted by the national “authorities” and a national document is produced.

Below you will find the national organizations and the reference for the national documents corresponding to the EN 15662:2008 norm

Country National Organization National Document ReferenceNetherlands NEN NEN-EN 15662:2008Germany DIN DIN EN 15662Switzerland SNV SN EN 15662:2009France AFNOR NF EN 15662Italy UNI UNI EN 15662:2009Sweden SIS SS-EN 15662:2008Norway SN NS-EN 15662:2008Finland SFS SFS-EN 15662:enIceland IST ÍST EN 15662:2008Estonia EVS EVS-EN 15662:2008Ireland NSAI I.S. EN 15662:2008

Country National Organization National Document ReferenceAustria ON OENORM EN 15662Hungary MSZT MSZ EN 15662:2009Poland PKN PN-EN 15662:2008 (U)Slovenia SIST SIST EN 15662:2009Croatia HZN HRN EN 15662:2008Lithuania LST LST EN 15662:2009Romania ASRO SR EN 15662:2009Malta MSA MSA EN 15662:2005Latvia LVS LVS EN 15662:2009

©2008 Waters Corporation 9

DisQuE ProcedureCEN Method

PN 176001903: DisQuE CEN Dispersive Sample Preparation Kit

Description— DisQuE Tube 1 (Extraction):

o 50 mL centrifuge tubeo 4 g magnesium sulfate anhydrouso 1 g sodium chlorideo 1 g trisodium citrate dihydrateo 0.5 g disodium hydrogencitrate

— DisQuE Tube 2 (Clean-up):o 2 mL centrifuge tubeo 150 mg anhydrous magnesium sulphateo 25 mg of PSAo 25 mg C18

©2008 Waters Corporation 10

DisQuE ProcedureCEN Method

Prepare Sample

Homogenize10 g sample* 10 mL ACN**

Liquid FractionationShake for 1 minute

Centrifuge 5 min>3000 g

CollectionRemove acetonitrile for clean-up

DisQuE ExtractionTube 1

Transfer1 mL Extract

Tube 1 to Tube 2

Shake vigously for 1 minuteCentrifuge >1500 x g

TransferCollect to autosampler vial

Dilute if necessary

DisQuE Clean-UpTube 2

*Depending on the matrices

**If highly acidic compounds add NaOH

©2008 Waters Corporation 11

DisQuE ProcedureCEN Method

The reason why the CEN norm came out was that the ACN extract made with the AOAC method was not clean enough. A cleaner extract is obtained with the citrate salt combination.

In our CEN kit tube 2 includes C18 (option with only PSA is available)

— C18 will remove co-extracted fat and waxes often present in these matrices. Their present affects the ruggedness of the further analysis.

— Graphitized Carbon Black is used if the samples has a high content of carotinoides (e.g. red pepper, corrots) or chlorophyll (e.g. spinach, rucola salad, vine leaves)

©2008 Waters Corporation 12

DisQuE Overview

The DisQuE 15 ml tube 2 is available for customers who wishes to use 6 ml of the ACN phase from tube 1. two version exist:— CEN PN 186004833 DisQuE 15 ml tube/MgSO4/PSA/C18— AOAC PN 186004834 DisQuE 15 ml tube/MgSO4/PSA