The QuEChERS Method

Background Information
and Recent Developments

Michelangelo Anastassiades
CVUA Stuttgart

CRL-SRM
1
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Outlook
v Classical Multiresidue Methods (MRMs)
Evolution
Limitations and Expectations
v Original QuEChERS-Method
Strategy of Method Development (Background Info)
v Recent Developments in QuEChERS Methodology
pH-Adjustment (during extraction, in final extracts)
Improved Selectivity (extraction, cleanup)
Expanded Matrix Spectrum (dry food, fatty food)
v Experiences of its Implementation in the Lab
v Method Validation
EU-Proficiency Tests (incurred and fortified residues)
CRL-SRM
2
Inter-laboratory Ring Tests
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
Pesticide Residue Analysis:
Sample Processing

Sample Preparation
Multi- and Single-Residue Methods

Measurement

Data Processing
CRL-SRM
3
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
Multiresidue Methods (MRMs):
Aim of MRMs:
Cover as many pesticides as possible from a single sample
portion employing a single sample preparation procedure

But, still
more than one determinative analysis run
is required to cover all analytes of interest 80000

70000

w ith su fficie n t se le ctivity a n d se n sitivity

60000

50000

40000

30000

20000

10000

4 5 6 7 8 9 mi n

The broader the spectrum of analytes covered by the MRM,

The less additional methods are required to cover all analytes
The more efficient and economical the analysis
CRL-SRM
4 Less time, personnel, materials...
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
Early MRM Evolution...
Simple but of
narrow scope
(OCs)
Intermediate
Expanded scope (to cover polar OPs)
Very complex since determ. analysis instr.
of poor selectivity and specificity

Novel
Simplicity,
streamlining,
cost reduction
miniaturization,
automation
CRL-SRM
5
Community Reference Laboratory
Pesticide Residues
1960
1st 1970
Joint CRL-Workshop - Stuttgart, 06/12/2006 1980 1990 2000 2010
using Single Residue Methods
 MRM Evolution:

Technical Development
always follows the way from the Primitive
via the Complicated
to the Simple

Antoine de Saint-Exupry (1900-1944)

CRL-SRM
6
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
Factors that pushed the Developm. of New Approaches
Environmental, Health-Related and Economic Factors

Need to Assess Risks for Humans

and Environment

Need to Improve
Productivity and Sample Throughput
& to Reduce TAT and Costs

Need to Reduce Solvent Consumpt.

Advancements in Instrumentation (Electronics, Robotics, IT...)

GC/ITD, MSD, MS-MS, TOF, PTV

LC/MS, MS-MS, TOF
S elective d etecto rs E C D , N P D , F P D
CRL-SRM
7
Community Reference Laboratory
Pesticide Residues
1960
1st 1970
Joint CRL-Workshop - Stuttgart, 06/12/2006 1980 1990 2000 2010
using Single Residue Methods
 Typical Classical MRM
Weigh sample (e.g. 50 g)

Add acetone and blend

Filter by suction

Add non-polar solvent (and salts),

perform (multiple) partitioning

Dry and filter organic phase

Evaporate
Reconstitute, perform GPC cleanup
Evaporate
Perform fractionated cleanup on silica
Evaporate
Analysis
Transfer fractions in GC-Vials
CRL-SRM
8
by GC-ECD,
CommunityFPD, NPD
Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
Pesticides and Co-extractives...
Amino acids Flavonoids/Anthocyanes Fatty Acids Phytosterols
-5 - -1 Polarity
0-6
range covered - by
. trad.8.5-11.5
MRMs

6
8
5
(pH dependent)
Monoterpenes
Vit. E
Sugars 2.5-5.5

11
.

5
- -- Pyrethroids (~45)
5

3.8 - 8.3
Acidic Pesticides (~40)
OCs (~20) Carotenoids
pH dependent 11-18
3.5 - 7.0
Strepto- Ureas (~ 30)
mycin 1.6 - 5.9 Chlorophyll
-7.5 OPs (~95) 17.2
-0.9 - 5.7
Glyphosate Carbamates (~30)
- -0.4 - 5.5 TGs
4

PAHs PBDEs 20-24

Basic Pesticides 3.3 6.8 6.2 - 9.5
Quats PCBs
pH dependent Phthalates
- . -- . LogKow
4
5

2
8

2.5 - 6 5 8.5Community Reference

CRL-SRM
9
Laboratory
Pesticide Residues
-5 -4 -3 -2 -1 0
1st Joint CRL-Workshop - Stuttgart, 06/12/2006 1 2 3 4 5 6 7 8 9 10 11
using Single Residue Methods
Scope and performance of classical MRMs

CRL-SRM
10
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
Typical inefficiencies of classical MRMs
Main Drawbacks Consequences

Wasteful:
Large Sample-Sizes Solvent & Material
M a cro -A p p ro a ch
Critical for
Limited Scope
Environment
(p o la rs )
& Health
Analysts Exposure Time-Consuming
to Solvents Troublesome Expensive
Unpopular
Limited LC
Too Many Amenability Error-Prone
Complicated Too many
Steps Additional Methods
required
CRL-SRM
11
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Sample Processing

Sample Preparation
has traditionally been
Sample Preparation
the tleneBo
Multi- andtSingle-Residue ck
Methods

of Pesticide Residue Analysis

Measurement

Data Processing
CRL-SRM
12
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
Desirable Characteristics of MRMs

Fast (as Few Steps as Possible)

Easy to Perform
Inexpensive
Low Solvent Consumption
Safe for Personnel and Environment
Selective
Rugged and precise
Achieve Good Recoveries for a Broad Analyte Spectrum
Thus Reducing the need to run Single (-Group) Residue Methods

CRL-SRM
13
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
Some Novel Sample Preparation Techniques
Focusing on Automation
SFE
PLE
Focusing on Automation and/or Miniaturization
SPME/SBSE
MSPD
Focusing at Simplification of Classical Methods
SPE of water-diluted extracts
Partitioning on Macroporous Sorbents
QuEChERS

CRL-SRM
14
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 QuEChERS - Original-Method
Weigh 10 g of Sample (50 mL Teflon-Tube)

Add 10 mL Acetonitrile
Shake Vigorously 1 min
Add 4 g MgSO4 and 1 g NaCl
Shake Vigorously 1 min
Add ISTD-Solution
Shake 30 s and Centrifuge
Take Aliquot and Add MgSO4 and Sorbent(s)
Shake 30 s and Centrifuge
Anastassiades et al.
(A d d A n alyte P ro tectan ts , ad ju st p H )
JAOAC Int. 86 (2003) 412-431
GC-MS (and LC-MS)

CRL-SRM
15
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Procedure in Pictures
1. Initial Extraction Step

Weigh 10 g Sample

Add 10 mL MeCN

Shake Intensively
for 1 min

CRL-SRM
16
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Procedure in Pictures
2. Extraction/Partitioning Step

(Pre-)Weigh
4 g MgSO4 + 1 g NaCl

Add to the Tube

Shake Intensively
for 1 min

CRL-SRM
17
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Procedure in Pictures
3. Addition of ISTD and Centrifugation

Add ISTD

Shake for 30 s

Centrifuge (ca. 5 min)

Separated Raw Extract

CRL-SRM
18
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Procedure in Pictures
4. Dispersive SPE Step
(Pre-) Weigh
MgSO4 and PSA

Add Extract to Tube

and Shake ca. 30 s

Centrifuge (ca. 2 min)

Cleaned up Extract

CRL-SRM
19
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Simplifications Introduced in the Method
Time Consuming, Complicated or Simplified Alternatives
Error Prone Steps of traditional MRMs
Use of Ultra-Turrax during Initial Extraction Shaking
Filtration Centrifugation
Multiple LL-Partitioning Steps S in g le P artitio n in g (O n -Line-A p p ro ach )
and Isolation of Entire Extract Take Aliquots (Use ISTD)
Use of a Lot of Glassware Extraction/Partitioning in Single Vessel
Evaporation/Reconstitution Large Volume Injection; Sensitive Instr.

Trad. Cleanup w. Columns (SPE, GPC) Dispersive SPE

Sample Processing/Homogenization No Way Around this!!

a Goal achieved: Simple and Streamlined MRM

Few working steps,
Convenient to perform
Low Material- and Solvent consumption
CRL-SRM
20
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Strategies in the Development
of the QuEChERS-Method

STREAMLINED AND
SIMPLE...

ECONOMICAL...

FIT FOR PURPOSE...

CRL-SRM
21
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Method Development - Aspects Considered

Initial Extraction & Extraction/Partitioning Step

Choice of Extraction Solvent and Sample/Solvent Ratio
Sample Amount
Blending Vs. Shaking (Incurred Residues)
Influence of Sample pH on Recov. (Ionization, Degradation)
Type and Amount of Salts Used to Induce Phase Separation
Selectivity (Gravimetric Anal. of Extracts, GC-Interferences)
Use of ISTD (Check that Recovery-Correction is minimal)

Cleanup (Dispersive SPE)

Type and Amount of Sorbent and MgSO4
Selectivity (Gravim. Anal. of Extracts, GC-Interferences)

Instrumental Analysis
Matrix Effects (Influence of Cleanup)
CRL-SRM
22 U se o f A n a lyte P ro te cta n ts
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Method Development
Choice of Acetonitrile as Solvent
Selective (Few Co-Extractives but still broad pesticide Spectrum covered)
Compatible with LC- and SPE-Applications
Not Chlorinated
Miscible with Water (Good for Initial Extraction)
Separ. from Water-Phase by Salt-Add. (No Non-Polar Solv. Needed)
Easier to Remove Water (with MgSO4) than from Acetone

Q Difficult to Evaporate
But PTV with Solvent Venting
Q High Expansion Volume could be used
Q Not Compatible With NPD
Q Not Compatible with GPC (But, Lipid-Co-Extraction is Low)
Q Low Lipid Solubility
Losses of non-polar pesticides (Recov. consistent at same Lipid/solvent ratio)
Accessibility problems of pesticides enclosed in Lipid particles (Ultra Turrax)
Q Rel. Toxic (But, Method Performed in a Closed Vessel, thus minimal exposure)
CRL-SRM
23
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Method Development
Acetonitrile vs. other Solvents
Residual co-extracted matrix components in mg/mL

3,50
Extracts of a
3,00
Mix of Fruits and vegetables
2,50 using QuEChERS
2,00

1,50
1,00
0,50
-
c
N

ne
SA
A

SA
SA
eC

Et

to
rP

rP
rP
M

ce
te

te
te

A
af

af
af

c
N

ne
A
eC

Et

to
M

ce
A

(see AOAC publ.)

CRL-SRM
For More Details and Comparison with EtAc , AcetoneCommunity
24
Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
Method Development
Sample Amount and Sample/Solvent Ratio
Sample Amount: 10 g
Miniaturization improves efficiency
Less material consumption
Reduced costs
Important:
Good homogenization is needed (e.g. use of dry ice)
Studies: Acceptable variations for 10 g subsamples using cryogenic milling

Sample:Solvent Ratio: 1:1 (w/vol) gives 1g/mL

Still good recoveries of incurred residues (polar and non-polar)
N o e va p o ra tio n o f fin a l e xtra ct n e ce ssa ry
1g/mL is enough when using modern instruments (PTV in GC is better)

CRL-SRM
25
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 5500000
Method Development -
MgSO4 / NaCl 4:1 for the Partitioning
5000000
Step
4500000
Many Salts tested
4000000 + 2 g NaCl
MgSO4 gave best salting-out of ACN
3500000 Methamidophos
and Best Overall Recoveries + 1 g NaCl Rec. 80-85 %
(especially for polar pesticides) 3000000

2500000 + 0.5 g NaCl

However: too much water in ACN-
phase and too many Polar Co- 2000000
HMF
extractives (e.g. Sugars) 1500000 GC-Degradant of Fructose
used as indicator
NaCl Addition increases Selectivity 1 0 0 0 0 0 0
Methamidophos
Less Water (and Sugar) in ACN-Phase 4g MgSO4 Rec. 95-100 %
500000
NaCl is used to Control Selectivity + 0 g NaCl
0
6 .0 0 8 .0 0 1 0 .0 01 2 .0 01 4 .0 01 6 .0 01 8 .0 02 0 .0 02 2 .0 02 4 .0 02 6 .0 02 8 .0 03 0 .0 03 2 .0 03 4 .
T im e -->
CRL-SRM
26
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Method Development -
MgSO4 / NaCl 4:1 for the Partitioning Step

Extraction of Honey (5g/10 mL MeCN)

mg co-extractives/mL extract

3,8
4,0
3,0
1,9 1,7
2,0

1,0 0,6

0,0
Partitioning with 4g MgSO 4 Partitioning with 4g MgSO 4 + 1g NaCl
(Original QuEChERS)

Raw Extract After PSA

NaCl reduces the amount of co-extracted matrix

CRL-SRM
27
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
Method Development
Shaking vs. Blending
Advantages
No Exposure to Metal Surfaces
Can Be Done by Hand and in Parallel
No Cleaning of Jar and Blender Between Samples
No Carry Over Between Samples
Only One Container necessary
Safer (Closed Vessel)
Less Noisy than Blending
No Frictional Heat
- May be less reproducible that blending
Pesticides from Fruits + Veg.: Ultra-Turrax usually not necessary
CRL-SRM

Community
28Checked with Incurred Residues (Cryogenic milling)
Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
Method Development -
Dispersive SPE for Cleanup
Advantages over classical SPE with Cartridges
No SPE Manifold, Vacuum/Pressure,
No Conditioning,
No problems w. Channeling, Flow Control, Drying-Out,
No Elution Step Needed,
No Add. Vessels for Eluent Collection,
No Dilution of Extract
No Evaporation,
Less Sorbent Needed,
Faster and Cheaper,
No Experience Needed.
W h e n Chemical Filtration is n e e d e d
CRL-SRM
Dispersive SPE is a se rio u s o p tio n
29
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
Dispersive SPE Removal of Co-extractives
PSA Cleanup and effect on pH
3,5 10
2,9 9
3,0
pH 8.9 8
pH 8.4
2,5 7
6

pH value
2,0
mg/mL

5
1,5 pH 5.4 1,3 4
1,0
1,0 3
2
0,5
1
0,0 0
No PSA PSA 25 mg/mL PSA 50 mg/mL

Amount of Co-extractives in the extract (mg/kg)

pH of Acetonitril Extract

Drawbacks: Solutions:
pH goes up (degradation risk) Addition of Acids (see later)
Matrix-Induced Analyte
CRL-SRM
30 Protection in GC reduced
Community Reference Laboratory
Pesticide Residues
Addition of Analyte Protectants
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Impact of Matrix-Effects
M atrix-In d u ced P eak E n h an cem en t

CRL-SRM
31
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
M atrix-In d u ced P eak E n h an cem en t E ffect
GC-Liner

GC-Capillary

Active Sites (on Surface of GC-Liner & Column )

(Siloxanes & deposited non-volatile matrix-co-extractives)
Analytes (in teract w ith A ctive S ites w h ich cau ses )
Unwanted Retention/Tailing
Quasi-catalysed degradation (susceptible compounds)
Matrix-Components (in Excess)
CRL-SRM Bloc- Stuttgart,
active
Community Reference Laboratory
Pesticide Residues
sites and protect analytes using Single Residue Methods
1st Joint
32
CRL-Workshop 06/12/2006
Analyte: Atrazine ; Matrix: Strawberry
RT= 8,80 min

30000
WITH Matrix co-extractives
25000
(Strawberry-Extract)

20000 WITHOUT Matrix co-extractives

(in pure solvent)
15000
e.g. Calibration standard
10000 RT= 8,92 min
Stronger Tailing
5000
Apex-Shift towards longer RTs!
0
8.70 8.75 8.80 8.85 8.90 8.95 9.00 9.05 9.10 9.15 9.20 9.25 9.30 9.35 9.40 9.45

Ratios: Matrix-Induced Peak Enhancement

Peak-Areas: ~ 1,5:1 a OVERESTIMATION OF RESULTS!!
Peak-Heights: ~ 4:1
CRL-SRM
33 Peak-Width (at half height): ~ 1:3
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Analyte Protectants
Principle
Protection Cleaned-up Extract + AP
Standard
+ AP

Raw Extract Addition of

A n alyte
PSA P rotectan ts
cleanup (AP)
Cleaned up
Extract
Standard in Pure Solvent

Analyte Protectants Reduce:

CRL-SRM
34
Analyte Interactions with Active
Community Reference Laboratory
Sites Pesticide Residues
and thus
1st Joint CRL-Workshop Errors
- Stuttgart, Related to Matrix-Induced Peak Enhancement
06/12/2006 in GC
using Single Residue Methods
Analyte Protectants-
Reduction of Matrix Induced Enhancement Errors
Errors eliminated if: Response in Matrix/Response in Solvent ~ 1
coumaphos

azinphos-methyl

imazalil

endosulfan I

cyprodinil
Response in Matrix
fenthion
Response in Solvent
dichlofluanid

carbaryl

chlorpyriphos
Overestimations
metalaxyl 1,0 when using
vinclozolin
Standards in Solvent
omethoate

acephate

mevinphos

methamidophos

0,00 0,50 1,00 1,50 2,00 2,50 3,00

no AP AP
Without withAP
With AP
AP was added to both :
CRL-SRM
Sample
35
Extract and Calibration Standard (in pure
Community Reference Laboratory
Pesticide Residues
Solvent)
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
Analyte Protectants Examples
V a rio u s C o m p o u n d s T e ste d fo r P ro te ctive P o te n tia l.
Best Protection : Polyhydroxy-Compounds (sugars, ~derivatives)

Examples: H
O O
HO
OH

OH OH
Ethylglycerol O
HO O
HO
OH
OH OH HO OH

Sorbitol -Gulonolactone

Give broadly eluting peaks protection over a wide volatility range

CRL-SRM
36
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
Analyte Protectants-
Desirable properties
v Strong interactions with active sites (H-Bond activity)
v Simmilar volatility to analytes to be protected (so that
protection extents during entire run)
v Soluble in sample extract
v Not accumulating in GC-system
v Not reactive with analytes (not inducing their degradation)
v Minimal interference with analyte detection (small m/z)
v Not deteriorating GC-column separation performance
v Cheap and not hazardous

CRL-SRM
37
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
CRL-SRM
38
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
CRL-SRM
39
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 QuEChERS
New Developments

CRL-SRM
40
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 QuEChERS Further Improvements
Some Issues Addressed
pH-issue
Stability of pH-labile Compounds
Recoveries of Ionizable Compounds

Selectivity Issue
Of Extraction/partitioning
Of Cleanup
Lipids, Sugars
Chlorophyll, Carotenoids

Expanding Matrix Spectrum OIL

Fatty Commodities
CRL-SRM
41 Dry Commodities
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 The pH Issue
Recoveries of Ionizable Compounds
Stability of pH-labile Compounds
Selectivity of Extraction (see later)

CRL-SRM
42
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 pH-issue Ionization of Pesticides
Some pesticides get ionized at low or high pH-values
Acids: HX H+ + X-
Bases: B + H+ BH+
Ionic form prefers to stay in the water phase

pH-Range of agricultural
samples: ~2.5 7

CRL-SRM
43
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
Pka-Values of Acidic and Basic Pesticides

OPP 9.9

2,4-DB
Acids
4.8
Ioxynil 4
MCPP 3.8 pKa = pH above which
MCPA 3.1
compounds lay in
2,4-D 2.7
Clopyralid 2.3 deprotonized form
Dicamba 1.9

Nitenpyram 3.1

Prochloraz 3.8 Bases

Carbendazim 4.2 pKa = pH below
Thiabendazole 4.7
which compounds lay
Cyromazine 5.2
Imazalil 6.5 in protonized form
Fenpropimorph 7
Propamocarb 9.5

In traditional methods, using non-polar solvents, Community

pH-adjustment
CRL-SRM
1-2
Reference Laboratory
44 units > or < PKa is recommended for quantitative recoveries
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Basic Pesticides Not affected!
pKa = pH below which the compound
lays primarily in its protonized form

Propamocarb pKa of corresp. acid 9.5

Imazalil pKa of corresp. acid 6.5

Prochloraz pKa of corresp. acid 3.8

Thiabendazole pKa of corresp. acid 4.7 pH 2.9

Carbendazim pKa of corresp. acid 4.2

Fenpropimorph pKa of corresp. acid 6.9

Spiroxamine pKa of corresp. acid 7.0

0 20 40 60 80 100 120
Commodity: Apple; Recovery %
CRL-SRM
45
Fortif. Level: 0.1 mg/kg; Community Reference Laboratory
Pesticide Residues
Analysis:
1st Joint CRL-Workshop -LC-MS/MS;
Stuttgart, 06/12/2006 ESI (+) using Single Residue Methods
 pKa
Basic Pesticides and pH pH below which the compound
lays predominantly in its
Effect of pH on Recoveries (%) protonized form

Apple juice, Thiabendazole Imazalil

pH adjusted pKa = 4.7 pKa = 6.3
with H2SO4 EtAc QuEChERS EtAc QuEChERS
pH 3 54 90 51 92
pH 4 85 90 73 94
pH 5 96 84 84 86
pH 6 104 90 94 90

Despite theoretically unfavourable pH, the basic pesticides still

prefer to partition into the MeCN phase.
Possible Reason:
CRL-SRM
After
46
Community Reference Laboratory
partitioning ACN still contains a considerable amount of water
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
Acidic Pesticides Recovery-Drop at pH 6
LC-MS/MS, ESI (-), No PSA Cleanup 120

100
Recovery
%
80

60
max. pH 5.5
40

20
Clopyralid
Imazapyr

Picloram
Benazolin

Imazethapyr
4-CPA

Naphthylacetic acid
Dicamba

Imazaquin
MCPA
2,4-D

Fluoxypyr
2,4,5-T
Mecoprop
Triclopyr
2,4,5-TP

Propyzamid
2,4-DP

Bentazon

Bromoxynil
Ioxynil
0

Fluazifop

Bromacil

2,4-DB

MCPB
Lower pKa-Values General Trend Higher pKa Values
CRL-SRM
47
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 pH-Issue - Labile Compounds
Some Pesticides degrade at high or low pH-values!
In the sample (processing, storage)
Keep low temperature

During sample preparation

Work fast, adjust pH
Use frozen samples for analysis

MgSO4 + Water Heat

In the extract during storage (1 week common)
Keep low temperatures, adjust pH

SPE with PSA Extract pH > 8

CRL-SRM
48
Community Reference Laboratory
Pesticide Residues
1st Joint CRL-Workshop - Stuttgart, 06/12/2006 using Single Residue Methods
 Optimal pH for QuEChERS ?
Goals: Relevance:
Still good recoveries for the Strongest Acids Extraction
dicamba, 4-C P A , clo p yra lid (p H < 5 .5 ) Step

Still good protection for Base-Sensitives

tolylfluanid, dichlofluanid, captan, folpet, Extraction Step
d ico fo l, p yrid a te +
Extract Storage
Still good protection for Acid-Sensitives
sulfonylureas, pymetrozine, carbosulfan,
dioxacarb...

CRL-SRM
49
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 pH Adjustment in Extraction Step
Various Buffers tested
Compromise: Citrate Buffer at pH 5 to 5.5
4 g Magnesium sulphate anhydrous,
1 g Sodium chloride (still kept for better selectivity),
1 g Trisodium citrate dihydrate and
0.5 g Disodium hydrogencitrate sesquihydrate

Good recoveries even for most acidic pesticides (dicamba ....)

Acceptable recoveries for base- and acid-sensitive pesticides
Impoved Selectivity (less co-extractives from acidic samples)
No negative effect on PSA cleanup (unlike Acetate
CRL-SRM Community Buffer)
Reference Laboratory
Pesticide Residues
50
1st Joint CRL-Workshop - Stuttgart, 06/12/2006 using Single Residue Methods
Problem: T ed io u s W eig h in g o f S alts
Solution: R ap id & E asy P o rtio n in g b y S am p le D ivid ers

Also suitable to
portion the
sorbents for
d isp ersive S P E

Some companies offer ready-to-use mixturesCommunity forPesticide

QuEChERS
CRL-SRM Reference Laboratory
Residues
51
Partitioning
1st Joint Salts and Dispersive SPE Mixtures
CRL-Workshop - Stuttgart, 06/12/2006 using Single Residue Methods
Need to Adjust pH in Final Extracts
Goal: Avoid degradation of Base-labile
compounds in final extract.

Rec. % Tolylfluanid stability in extract

100 Also the case for:
90 7 days dichlofluanid,
80 13 days
70 captan,
60
50
pH folpet,
40 after PSA dicofol,
30
20 pyridate
10
0
pH 4 pH 5 pH 6 pH 7 pH 8 pH 9 MeCN
Measured pH in extract

Addition of formic acid (5% in ACN):

CRL-SRM
10 L p er m L extract b rin g s p H to ~ 5
52
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Rec. in % Primisulfuron-Methyl
Sulfonylureas, 120
7 Days Storage
Carbosulfan 100

80
13 Days Storage
acid labile... 60

40

20

0
pH 4 pH 5 pH 6 pH 7 pH 8 pH 9 MeCN
Pure
Measured pH of Extract

Rec. in % Carbosulfan
120
7 Days Storage
If these 100
13 Days Storage
compounds are 80

included in the 60

target spectrum 40

use an aliquot of 20

the final extract 0

pH 4 pH 5 pH 6 pH 7Community
pH 8 Reference MeCN
pH 9 Laboratory
CRL-SRM
before
53
acidifying Pure
Pesticide Residues
Measured pH of Extract
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Improving Selectivity

At Extraction/partitioning Step
v pH
v Salts

At Cleanup Step
v Lipids, Sugars
v Chlorophyll, Carotenoids

CRL-SRM
54
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
Role of pH in the Selectivity of Extraction/Partitioning
Influence of pH in the Amount of Co-Extractives
Red Currant (pH adjusted with NaOH)
9
8,1
8
Raw Extracts
7
before Cleanup
Co-extracted Matrix

6
[mg/ml]

5
4,2
4

3 2,5 2,2
2

0
Natural pH pH 4 pH 5 pH 6
pH-Value

CRL-SRM
The higher the pH the less co-extractives
55
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Role of pH in the Selectivity of Extraction/Partitioning Step
Comparison of QuEChERS-Modifications
O r ig in a l Q u E C h E R S C it r a t e - B u f f e r e d A c e ta te -B u ffe re d
12
S a m p le p H 2 . 8 9 12
S a m p le p H 5 . 1 9 12
S a m p le p H 5 . 1 9
p H = 8 .3
m g /m L pH m g /m L p H = 7 .8 pH m g /m L pH
8 8 8
10 1 1 ,2 10 10
7 7 7

8 p H = 5 .4 6 8 6 8 6
p H = 5 .0 p H = 5 .1 p H = 5 .2
5 5 5
6 6 6
4 4 4
p H = 3 .2 p H = 3 .5
4
3 ,7 3
4 3 4 3
2 ,9
2 ,4 2 ,5
3 ,6 2 2 2
2 2 2
1 ,0
p H = 1 .6 0 ,8 1
Red Currant
0 ,5 1 1

0 0 0 0 0 0
R a w E x tra c t P S A 2 5
m g /m L
P S A 50
m g /m L
(aR adifferent
w E x tra c t P S A 2 5
m g /m L
one)
P S A 50
m g /m L
R a w E x tra c t P S A 2 5
m g /m L
P S A 50
m g /m L
m g c o -e x tr a c tiv e s/m L E x tr a c t m g c o -e x tr a c tiv e s/m L E x tr a c t m g c o -e x tr a c tiv e s/m L E x tr a c t
pH pH pH

Buffering to pH ~5 reduces amount of acidic co-extractives

CRL-SRM Community Reference Laboratory
Pesticide Residues
Acetate buffer negatively affects PSA cleanup efficiency
56
1st Joint CRL-Workshop - Stuttgart, 06/12/2006 using Single Residue Methods
 Selectivity of Cleanup
More than 50 SPE Sorbents and freezing-out tested!
Mainly removed:
v Amino-Sorbents, Alumina:
Acids (including fatty acids) Losses of
Sugars acidic pesticides
Pigments (Anthocyanes, some Chlorophyll)
v Carbon-based Sorbents: Losses of
Carotinoids, Chlorophyll, Sterols planar pesticides
v Reversed-Phase Sorbents: No losses
Lipids and Waxes observed
v Freeze-out:
No losses
Lipids and Waxes
CRL-SRM Community Reference Laboratory
observed
Pesticide Residues
1st
57
Sugars
Joint CRL-Workshop - Stuttgart, 06/12/2006 using Single Residue Methods
Use of Carbon Sorbents PSA
PSA not satisfying when
high contents of
carotinoids or chlorophyll

C Carbon Sorbents more Effective

Many tested, GCB (Graphitized Carbon Black) was best in handling
- Used in combination with PSA at small amounts
- Cleanup time (shaking) extended from 30 s to 2 min

Small GCB amounts are d ifficu lt to h a n d le

C
CRL-SRM
Pre-mixtures GCB/MgSO 4 (powder) facilitate weighing
Community Reference Laboratory
Pesticide Residues
58
1st Joint CRL-Workshop - Stuttgart, 06/12/2006 using Single Residue Methods
Problems with GCB:
Planar pesticides have a high
affinity towards GCB
e.g. hexachlorobenzene,
chlorothalonil, thiabendazole

But chlorophyll has higher affinity than all pesticides

C Final extract should remain slightly coloured!!

Anthracene may be used as surrogate QC standard.

Recoveries > 70% will indicate that no unacceptable
CRL-SRM
losses of pesticides have occurred.
59
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Removal of co-extracted lipids
by C18 or freezing out
5
4,20 OIL EXTRACTS
4,03
4
Extractives [mg/ml]

2
1,48
1,33 1,25 1,23 1,15 1,10
1

CRL-SRM
60
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Removal of co-extractives from
Whole-Wheat flour

CleanupofW hole-WheatFlour
Q uEChERS-Extracts

7
6
5
4
Co-extractives/mLextract

3
2
1
0
RawExtract C18 Freeze-out C18+PSA Freeze-out +PSA

CRL-SRM
61
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
Scope and Pereformance of QuEChERS

CRL-SRM
62
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
QuEChERS- Multiresidue-Method
Weigh 10 g of Frozen Sample
Changes introduced
Add 10 mL Acetonitrile to the method.

Add ISTD-Solution The method

Shake will become
Add 4 g MgSO4 / 1 g NaCl / Citrate Buffer official CEN method
(pH 5-5.5)
Shake & Centrifuge Optionally:
Acidic Pest. by LC-MS/MS
Mix an Aliquot w. MgSO4 & Sorbents, freeze-out
Shake & Centrifuge Optionally:
SUs by LC-MS/MS
Acidify extract to pH ~5
to protect base-sensitive pesticides

Multiresidue Analysis
CRL-SRM
63
O p tio n ally: A d d o th er A n alyte P ro tectan ts Community Reference
by GC-MS,
Laboratory
Pesticide Residues
LC-MS ...
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Broaden matrix spectrum
Dry commodities (cereals, dried fruits) Fatty Commodities

CRL-SRM
64
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Broaden matrix spectrum
Dry Commodities
E.g. cereals, dried fruits

Water-Addition prior to extraction

to weaken interactions of pesticides with
matrix and to ensure adequate partitioning.

Sample amount is reduced and water is brought to 10 mL

Co-extracted fat removed by freezing out or C18,
if necessary....
CRL-SRM
65
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Dry Commodities
Sample type Weigh Water Annotation
Fruit/Vegetables 10 g -
(water >80 %)
Fruit/Vegetables 10 g Xg X = 10 g water
(water 30-80 %) amount in 10 g sample
Cereals 5g 10 g
Dried fruits 5g 8.5 g Add water to
comminute, weigh
13.5 g of homogenate
Honey 5g 10 g
Spices 2g 10 g

CRL-SRM
66
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Broaden matrix spectrum
Fatty commodities
Commodities with a high lipid load, such as
avocados or plant oils can be employed.

Problems:
Co-extracted lipids should be removed
prior to GC-analysis

Highly non-polar pesticides

may give recoveries < 70% (e.g. HCB and DDT)

Accessibility of residues may be limited (Ultra Turrax)

CRL-SRM
67
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
Recoveries of pesticides in high fat samples
Recoveries
100
%
75

50

25 1g
2g
0 3g
Hexachlorobenzene

Chlorpyriphos-methyl
HCH, gamma-

Pirimiphos-methyl
Endosulfan

Deltamethrin
Chlorpyriphos

Cypermethrin
p,p' -DDE

Fenthion

Diazinon

Malathion
BS138

Carbaryl
Dieldrin

Trifluralin
TPP
g Oil / 10 mL ACN
Values in
absense of
water

PCB 138 or 153 may be used as surrogate QC standards

Rec. > 70% will indicate that no unacceptable pesticide losses occurred

The tolerable lipid-amount depends on the selection of pesticides to be covered

e.g. for HCB 0.4 g lipids are still OK (>70% rec.), for DDE 1 g, for Endosulfane 5 g
(NOTE: In presence of water (ternary system) values are different, less lipid is tolerable)

Compromise for Oil samples: 2 g oil + 10 mL ACN

H C B a n d D D E g ive re co ve rie s < 7 0 %
CRL-SRM
1stbut
68 equilibrium is defined and recovery-correction is justified
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
Joint CRL-Workshop - Stuttgart, 06/12/2006
 Cleanup of Fermented Tea
extracts
Removal of Black-Tea Co-extractives Recoveries of
200 mg Tea/ml solvent
Methamidophos
100
35
31,0 88 CaCl2 removes more water
30 27,9 from the extract than MgSO4.
Thus interctions with the 104
Co-extractives [mg/ml]

25
21,2
20 58 sorbent (H-binding, ionic)
14,0
become stronger
15
Better Cleanup results
8,9
10 24 6,7 91
5 3,8
2,7 2,2
1,5
0,5
0
Cl2

aCl2

aCl2

PSA
xt

PSA

PSA

PSA

PSA

PSA

PSA
50 Ca
ectra

100 C

200 C

25

+ 25

+ 25

+ 25

+ 50

+ 75

+ 100
aw

Cl2 +

aCl2

aCl2

gSO4

gSO4

gSO4
Tea r

gSO4
50 Ca

100 C

200 C

150 M

150 M

150 M

150 M
CRL-SRM
Problem with CaCl2: recoveries of polar pesticides drop
69
Jointif CRL-Workshop
polar pesticides
Community Reference Laboratory

are not of interest CaCl2 / PSA is a serious cleanup

Pesticide Residues
option
using Single Residue Methods
1st - Stuttgart, 06/12/2006
 Release of covalently bound phenoxy-
acids by alkaline cleavage
Alkaline Hydrolysis for the release of phenoxy-acid pesticides
wheat sample

1400 At 70 C At Room Temperature

1200
pH 12 pH 10 pH 10 pH 9
Relativ concentration

1000 30 min 60 min 30 min 30 min

800 715
606 619
600
336
400 275
200 135 100
0

in
in
n

0m
m
mi

mi

mi

mi

l.
/30

ro
/30

/30

/60

/30

9/3

yd
12
12

10

10

10

oh
pH
pH
pH

pH

pH

pH

w/
RS
hE
EC
CRL-SRM Community Reference Laboratory

Qu
Pesticide Residues
70
1st Joint CRL-Workshop - Stuttgart, 06/12/2006 using Single Residue Methods
 Impact of QuEChERS-Implementation

More time for instrumental analysis

More time for QA/QC (incl. validation)
Broader analyte spectrum
Higher sample throughput and turnaround time
Less solvent consumption
Less lab space needed (hoods are empty)
Sample preparation more pleasant

CRL-SRM
71
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Impact of QuEChERS-Implementation

CRL-SRM
72
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Reduction of Solvent Consumption
15.000 savings in 1 year
just for solvent !!
mL Solvent/Sample (for ca. 2000 samples)
more pesticides than Becker
655

535 including basic & acidic pesticides,

but low recoveries for very polar ones

485
265 Many more pesticides covered
325 215

330 215 10
200 Sum [ml]
10
Other solvents
65 50 0 0
Organochlorine solvents
Becker, S-8 Becker, Mini Specht, S-19 CVUA-Method QuEChERS
up to 1990 1990-96 1993-96 1996-2002 since 2002
CRL-SRM
73
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 M ass M ig ratio n o f P erso n n el

CRL-SRM
74
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 YES!!
More finding s...
In 2004:
200 Pesticides
in Total
149

(CVUA Stuttgart) 121 124

105
100
84 144

69 73 68 121

49 100
83
68

46
40 44 40
Fruits 37
Vegetables

1995 1996 1997 1998 1999 2000 2001 2002 2003 2004
Nr. of different pesticides detected in fruits and vegetables

CRL-SRM
75
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Participation in
EU-Proficiency Tests
using the QuEChERS-Method

CRL-SRM
76
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
Using QuEChERS in EU - Proficiency Tests
180
Original Original Citrate- Citrate-
160 QuEChERS QuEChERS Buffered Buffered
140 Median of all (100-130) Labs
QuEChERS QuEChERS

120
100
80
60
2002 2003 2004 2005
40
20
0

CRL-SRM
77
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
Using QuEChERS in EU - Proficiency Tests
Unknown Pesticides and Unknown concentrations
Participants: 100-130 EU-Official Labs
Results:
All 57 identified (100%)
95% (54/57): within +/-30% from median concentration
82% (47/57): within +/-20%
53% (30/57): within +/-10%
On average +8% above the median
2002 2003 2004 2005

CRL-SRM
78
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 QuEChERS
Inter-Laboratory Validation Studies

CRL-SRM
79
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
GC-MS and LC-MS/MS Inter-Laboratory Validation Study (GDCh)

GC
Mean Orange Orange Apple Apple Lettuce Lettuce
0.25 0.025 0.25 0.025 0.25 0.025

Rec.: 101% 100% 105% 102% 100% 100%

RSD: 3% 4% 8% 9% 5% 3%
LC (+)

CRL-SRM
LC (-)
80
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 LC-MS/MS Inter-Laboratory Validation Study (BLAPS-Working Group)

Mean Cucumber Cucumber Lemon Lemon Wheat Wheat Raisins Raisins

0.1 0.01 0.1 0.01 0.1 0.01 0.1 0.01
Rec.: 97% 97% 98% 98% 96% 98% 95% 97%

RSD 7% 8% 7% 7% 5% 8% 6% 7%

Ethiofencarb
was oxidized
in cucumber

SUs degraded
CRL-SRM
81 in acidified extract
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
LC-MS/MS Inter-Laboratory Validation Study (BLAPS-Working Group II)

Mean Cucumber Cucumber Lemon Lemon Wheat Wheat Raisins Raisins

0.1 0.01 0.1 0.01 0.1 0.01 0.1 0.01
Rec.: 101% 98% 99% 94% 100% 101% 100% 97%

RSD 3% 8% 7% 8% 4% 9% 6% 11%

Oxidation

Acids, lost in
PSA cleanup
Degraded in the standard
solution provided

CRL-SRM
82
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
QuEChERS- Multiresidue-Method

Advantages
Rapid (8 Samples in Less Than 30 min)
Simple (No Laborious Steps, Minimal Sources of Errors)
Cheap (~1 Sample Prep. Materials for 1 mL Extract)
Low Solvent Consumption (10 mL Acetonitrile)
Practically no Glassware Needed
Wide Pesticide Range (Polar, pH-Dependent Compounds)
Extract in Acetonitrile (GC- and LC-Amenable)

CRL-SRM
83
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 www.QuEChERS.com

....the modified QuEChERS method

including all presented modifications
and a lot of background information is
available via the internet, as well as the
validation data.

www.quechers.com

CRL-SRM
84
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Thank you very much
for your Attention !

CRL-SRM
85
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Pesticides Online
Internet platform for
Pesticide Residue Analysts

www.pesticides-online.com

FEEL FREE TO REGISTER & TEST !!

CRL-SRM
86
Community Reference Laboratory
Pesticide Residues
using Single Residue Methods
1st Joint CRL-Workshop - Stuttgart, 06/12/2006
 Laboratory Researches Regarding Wear of the
Composite Materials for Making Brake Shoes

ERIKA ARDELEAN1*, ANA SOCALICI1, LIVIU PASCU2, VASILE PUTAN1, TEODOR HEPUT1
1
Univesity Politehnica Timisoara, Faculty Engineering of Hunedoara, 5 Revolutiei Str., 331128, Hunedoara, Romania
2
Technical College of Railway Transportation Anghel Saligny, 137 National Road, 335900, Simeria, Romania

The paper presents the results of the experiments on obtaining a composite material to be used in
manufacturing brake shoes for the rolling stock. The aim was to replace the classical material used at
present for the manufacturing of brake shoes: cast iron with a specially designed composite material. Six
recipes have been tested and compared to a standard sample made of phosphorous cast iron. The wear
trials carried out in laboratory conditions on the experimental test rods have led to results that entitle further
experiments that implies the manufacturing of brake shoes made of the composite material in question.
Keywords: organic materials, composite, noise, wear, brake shoes

The European Commission has taken a series of steps
and elaborated technical specifications for interoperability
related to railroad noise [1, 2], introducing limitations for
the rolling stock used in the European Union. These
limitations apply to the new or refurbished rolling stock,
which has to be equipped with silent breaking pads, meant
to cut down noise. The most economical means of
achieving that is by replacing the breaking system, as
replacing the whole vehicle fleet is extremely expensive.
It is recommended to replace the cast iron breaking pads
by pads made of composite materials.
They are efficient in noise reduction (by 10dB, which
means 50% of the noise produced by the cast iron brake
shoes) [2]. This post-refurbishing should be done for all the
European freight carriages which run over 10 000 km
annually and whose remaining life span is at least 5 years.
The profile industry developed several types of brake
shoes made of composite materials in order to replace the
conventional, cast iron pads, which represent the main
cause of rail and wheel roughness [2-4]. K brake shoes are
made of organic composite materials and have braking
characteristics that differ from those of the traditional pads.
They are highly efficient in terms of noise reduction, the
technology has been tested on new carriages, but it
supposes high refurbishing expenses. The LL brake shoes
need just minor adjustments of the breaking system, they
have breaking characteristics almost similar to the cast
iron pads and are made of either organic or sintered metal
composite materials. They offer noise reduction similar to
the K pads and are particularly meant for refurbishing.
Refurbishing leads to investment costs ranging from 200
to 700 million euro (LL pads) or 1,01,8 billion euro (K pads)
plus extra maintenance costs worth of 200-400 million
euro (added up until 2025, for both technologies) [4-11].
The European Commission [4] recommends the obtaining
of high quality composite materials for brake shoes in order
to significantly cut down both costs and noise.
Experimental part
The paper presents the laboratory experiments made
to obtain composite materials meant for the brake shoes
to be used by train engines or carriages, characterization
of the samples obtained and interpretation of the results.
For the laboratory trials of the composite materials
meant to be used to manufacturing brake shoes, the
following test rods have been established:
- a standard sample test rod made of P 10 phosphorous
cast iron (Ss);
- 6 test rods made of composite material (R1-R6).
The standard sample test rod was made of P 10
phosphorous cast iron [12, 13], a material that is used in
practice for the conventional brake shoes. The
characteristics of this standard sample have been taken
as reference for the composite material. Six experimental
tests rods were made for the composite material, using
the following components: Novolac, Hexamethylene-
tetramine, sulphur, Carbon fibre, graphite, aluminium, brass
and rubber. Figure 1 presents the component recipes.

Fig.1. The component recipes [%] used in laboratory

experiments

* email: erika.ardelean@fih.upt.ro
MATERIALE PLASTICE 54 No.2 2017 http://www.revmaterialeplastice.ro 203
 The (cylinder - type) test rods having a diameter of
14mm and a height of about 30mm, were cast in metallic
moulds. The quantities of elements in each recipe were
determined by calculation and by preliminary trials. Part of
the phenolic resin (about 70%) went through a heating
process to 70oC, then the other components, including the
rest of solid phenolic resin (30%), was incorporated in it,
one at a time. The mould was filled with this mixture and
it was periodically pressed [7]. A homogenous distribution
of the components is necessary for obtaining a high quality
composite material.
The composite material in the mould was cold pressed
in a 2000MPa hydraulic press. The sample rod was sintered
before being removed from the mould. The optimal
sintered temperature, established by trials, was 200oC. From
each recipe three test rods had been produced (fig.2).
DIMENSIONAL CHARACTE-RISTICS OF THE SAMPLES
Fig.2. Samples
THE RESULTING DATA FOR THE EXPERIMENTAL SAMPLES
Fig. 3. Installation of testing abrasive
wear with abrasive disc
The resulting test rods were subjective to a wear test on
an installation of testing abrasive wear with abrasive disc
(fig.3).
The trials were performed according to STAS 963981
and consists in pressing the test rod made of the material
under examination against a rotating disc, in order to
determine certain abrasive wear characteristics [10]. HE
16 polishing silicon carbide paper was used as abrasive,
which is supplied as 310mm diameter discs. The length
of the wear spiral trajectory results from the combination
of the two movements (rotation and advance) and has the
value of 70m during 7.08 min. The assessment of the
behaviour of the test rods has been done by the gravimetric
method (mass loss). Table 1 gives the dimensional
characteristics of the test rods.
For each test rod, the mass wear, trajectory mass wear
rate and the relative wear were determined.
Relation (1) determines mass wear u:
204 http://www.revmaterialeplastice.ro
(1)
where:
m0 is the mass of the test rod before the trial, [g];
mf - the mass of the test rod after the trial, [g].
Relation (2) determines the trajectory mass wear rate:
(2)
where Lu is the wear trajectory, [m].
Comparing the test rod mass wear of the composite
material (uc) to the mass wear of the sample cast iron test
rod (uf), the calculated relative wear is, according to relation
(3):
(3)
Table 1
Table 2
Table 2 shows the resulting data for the experimental
samples.
Results and discussions
The analysis of the data shows that the lowest mass
wear of the composite material is that of sample rod R4.
As compared to the cast iron standard sample rod, the
sample rods made of composite material have a higher
mass wear, but their behaviour is similar to that of the
composite materials used at present in manufacturing
brake shoes [7, 8]. Experimental sample rod R6 showed
the highest wear.
Mass wear has also been analysed in terms of the
composition of the recipes (respectively the main
components). The aim was to find the optimal composition
of the chosen composite, so that it would show the slightest
mass wear. Thus, figure 4 shows the variation of the mass
wear according to the content of Novolac; the composite
MATERIALE PLASTICE 54 No.2 2017
having a content of Novolac between 35-40% shows the
highest wear resistance.
The analysis of the influence of the content of
Hexamethylenetetramine (used to turn the thermoplastic
resin into a thermo-rigid one), has led to the conclusion
that a content of 10% has a positive influence upon the
decrease of wear resistance (fig. 5).
It was used Carbon fibre in order to reinforce the
composite, and graphite to reduce the friction coefficient
powder. Mass wear was cut down to a minimum for an
interval within 17-19.5% Carbon fibre and graphite (fig. 6).
Figure 7 shows the variation of the relative height
reduction according to the content of aluminium powder
and brass powder.
Conclusions
The paper presents the results of the experiments on
obtaining composite materials meant for the
manufacturing of rolling stock brake shoes.

Fig. 4. Variation of relative

mass wear (a) and relative
height reduction (b)
according to the Novalac
content

Fig. 5. Variation of relative

mass wear (a) and relative
height reduction (b)
according to the
Hexametyltetramine
content

Fig. 6. Variation of relative

mass wear (a) and relative
height reduction (b)
according to the Carbon
fibre and graphite content

Fig. 7. Variation of
relative height reduction
and
according to the
aluminium powder (a)
and brass powder
content (b)

MATERIALE PLASTICE 54 No.2 2017 http://www.revmaterialeplastice.ro 205

 The analysis of the experiments and their results leads
to the following conclusions:
- the test rod with the highest resistance to abrasive
wear is R4; this could be explained by the proportion of
components and of the manufacturing technology (the
proportion of the components Novolac, Hexamethylene-
tetramine, carbon fibre and graphite ranged towards the
maximal limits);
- the resistance to abrasive wear is satisfactory in case
of test rods R1, R2 and R3, due to the fact that the
manufacturing technology is the same and the structure
of the components varies within narrow limits;
- test rod R6 has the highest resistance to abrasive wear,
and for that reason it has further been studied, in an attempt
to improve its behaviour under abrasive wear.
Considering the results, the best behaviour to wear of
the experimental composite materials, was the one of test
rods produced according to recipe R4. Experiments will
continue in order to finalize the recipe and the
manufacturing technology, in order to obtain the best values
for the indices under consideration.
References
1. *** EU Commission Decision 2006/66/EC concerning the technical
specification for interoperability relating to the material stock noise
of the conventional rail system.
2. Directive 2002/49/CE, JO L 189, 18.7.2002, p. 12.
3. ENACHESCU, G., STEFANESCU, M., Mat. Plast, 53, no.2, 2016, p.198
206 http://www.revmaterialeplastice.ro
4. Commission of European Communities, Measures to reduce rail
noise from the existing fleet, COM 432, Bruxelles, 2008.
5. NEHARKAR, P., PATIL, R.J.,SONAWANE, P.R., Study of friction and
wear for optimization of disc break material for reduction of brake
sound, International Journal of Research in Aeronautical and Mechanical
Engineering, 2 no. 6, 2014, p.137.
6. XEPAPADAKI, A., PAPANICOLAOU, G., KERAMIDAS, P., JIGA, G., Mat.
Plast, 47, no.2, 2010, p.153
7. PASCU, L., PhD thesis, Researches on Improving the Quality of
Brake Shoes Meant for Use with the Rolling Stock, Politehnica
Timisoara, Romania, 2015
8. OERTLI, J., Railway Noise Control In Europe, Current Status,
International workshop on railway Noise, Uddevalla Sweden, 2013,
p.1.
9. TUDOR, D., PETRESCU, H., HADAR, A., ROSU, D., Mat. Plast, 49,
no.2, 2012, p.123
10. STAS 9639 81, Testing of metals. Trying to wear on rotating disc
sanding paper
11. COCARD, M., MARSAVINA, L., Mat. Plast, 51, no.2, 2014, p.190
12. SUDARSHAN, T., SRIVATSAN, T. S., Role of phosphorus content on
porosity of cast irons, Journal of Materials Science, 27 no.4, 1992,
p.869.
13. ABBASI, H.R., BAZDAR, M., HALVAEE, A., Effect of phosphorus as
an alloying element on microstructure and mechanical properties of
pearlitic gray cast iron, Materials Science and Engineering, 444 no.1-
2, 2007, p.314.
Manuscript received: 7.07.2016
MATERIALE PLASTICE 54 No.2 2017