Certification of a reference material

for herbicides in water by way of

inter laboratory comparison
B. Lalrea, V. Le Diourona, M. Dsenfanta,
K. El Mrabeta,b V. Pichonb, J. Vialb, G. Hervoeta
a Laboratoire National de Mtrologie et dEssais (LNE), 1 rue Gaston Boissier,
75724 Paris cedex 15, France
b Laboratoire Environnement et Chimie Analytique (LECA) UMR-CNRS 7121,
Ecole Suprieure de Physique et Chimie Industrielles, 10 rue Vauquelin,
75231 Paris Cedex 05, France
ABSTRACT: The certified reference materials (CRMs) are among the most appropriate tools for the
traceability and validation of analytical methods. Although their number seems to be high 20,000, they
only cover a small part of the analytical needs for environmental monitoring. Until now, none were
available for analyzing pesticides in water.
After an intralaboratory feasibility study and an evaluation of the behavior of this material during
interlaboratory testing, the LNE has produced and certified a CRM for the triazine and phenylurea
analysis in water samples.

Introduction
In accordance with the European outline directive on water (DCE), the
department of ecology published memorandum service instruction DCE 2006/16
relating to the constitution and implementation of a monitoring program for the
different water categories. Because of this, quality of rivers and ground water is
regularly monitored. Pesticide content, a micropollutant considered as a priority by
the European Union in particular is verified.
In order to achieve this monitoring, several measures are taken daily by different
laboratories. It is important to be able to compare results. This objective can only be
reached by ensuring traceability of these analyses with the help of different tools
such as certified reference material (CRM), the link to national standards through
uninterrupted chains of comparison and a participation in interlaboratory testing.
CRMs have the advantage of also being able to evaluate accuracy during validation
of analytical methods implemented. They can be of two types: those destined to
calibrate measurement systems and matrices making it possible to consider all
steps from the preparation of the sample.

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Until now, there were none for the analysis of pesticides in water. That is why
LNE in cooperation with LECA decided to develop one. This project started four
years ago with a feasibility study of such a material. The behavior of products
developed was then evaluated during an interlaboratory comparison.
A group of CRMs was developed in March 2006 and certified in June 2006 for
the analysis of triazines and phenylureas in drinking water.
Previous studies: feasibility and behavior of RM
The following components which belong to two pesticide families were selected:
triazines: deisopropylatrazine (DIA), deethylatrazine (DEA), simazine, atrazine,
terbutylazine and terbutryne.
phenylureas: chlortoluron, diuron, isoproturon and linuron.
Completed in 2002, this choice, except for terbutylazine and DIA, is based on
the frequency of detection of these triazines and phenylureas in water [1]. In
addition, they are indexed in the list of 50 pesticides sold in larger quantities than
500 tons per year in Europe. Terbutylazine, following the ban on the use of atrazine,
is part of the mix used for its replacement in corn cultures and thus was added.
Despite its usage ban, atrazine can still be detected in water; because of this, two of
its metabolites, DEA and DIA were included in this study.
Feasibility study
In order to achieve the most complete kit possible, two types of reference
materials (RM) were retained:
sealed vials: targeted pesticides are stored:
-

in a solution of acetonitrile,

dried after evaporation of the solvent used for their preparation.

solid phase extraction (SPE) cartridges called cartridges in this document: a

water sample spiked with each compound is percolated on the support in order to
retain them with other molecules in the sample. Two polymeric supports were
evaluated: a divinylbenzene copolymer functionalized with N-vinylpyrrolidone
(Oasis HLB, Waters) groups and a polystyrene divinylbenzene copolymer
(ENVI Chrom P, Supelco). Analysis laboratories will then have to carry out the
elution of compounds.
These materials were prepared with two levels of concentration, targeted in
relation to regulation (0.15 g/l for drinking water and 0.50 g/l in surface water)

Chemistry Statistics /Gas Analysis

and they are stored under different temperature conditions: Tambiant, T=0.5C and
T=-18C for a year. Every month, the evolution of compound concentration was
studied by the analysis of vial and cartridge contents.
The detailed presentation of results for each compound, each type of material
and each storage condition was the subject of publication [2]. Between the
beginning and ending of the study, the evolution was quantified in percentage of the
quantity found in relation to the one initially introduced (rate of recovery). To
summarize, compounds are classified into three families of behaviors:
non-usable family (Figure 1): either the compounds were totally degraded or
their recovery level after nine months of study are lower than 10%;
family with trend (Figure 2): compound concentration evolves over time;
family without trend (Figure 3): pesticide concentration does not evolve
significantly over time.

Recovery rate (%)

Linuron
120
100
80
60
40
20
0
0

Time (months)

Figure 1. Non-usable family example: Linuron stored in vial after dry evaporation
and conservation at ambient temperature

Recovery rate (%)

DIA
120
100
80
60
40
20
0
0

Time (months)

Figure 2. Family with trend example DIA stored in vial after dry evaporation
and conservation at ambient temperature

Transverse Disciplines in Metrology

Recovery rate (%)

Atrazine
120
100
80
60
40
20
0
0

Time (months)

Figure 3. Family without trend example Atrazine stored in vial after dry evaporation
and conservation at T=-18C

The results have shown that compounds are stable when then are in a solution of
acetonitrile when temperature becomes ambient, whereas it is necessary to store
them at -20C when they are dried in a vial. When they are fixed on cartridges,
storage temperature must be lower than 0.5C. Observed behaviors are identical
regardless of the level of concentration.
The retained RM was then a kit with:
vials containing pesticides in acetonitrile;
cartridges (Oasis HLB, Waters) treated by percolation of drinking water
containing selected pesticides.
As a precaution, they will be stored at -20C.
RM behavior during an interlaboratory comparison
In order to test the conditions of RM usage (sending conditions, reception and
analysis with different methods), an interlaboratory test involving about 15
participants was organized [3].
The variation factors of both components of the material tested vary between 14
and 30% depending on compounds, which is very satisfactory for a circuit involving
laboratories using different methods compared to other campaigns [4,5].
In view of the results (feasibility and interlaboratory test), a certification
campaign was then completed.

Chemistry Statistics /Gas Analysis

Material production and certification

This reference material, in the form of a kit is made up of:
two vials containing approximately 1.2 ml of a herbicides solution (atrazine,
deethylatrazine, deisopropylatrazine, simazine, terbutryne, terbutylazine,
chlortoluron, linuron, diuron, and isoproturon) in acetonitrile (concentration for
each pesticide | 0.1 mg/l);
three cartridges on which 250 ml of a drinking water spiked with a solution of
herbicides in acetonitrile was percolated, in order to reach a concentration of
approximately 0.50 g/l of each compound in water.
This material is destined to calibrate the measurement devices and/or to validate
the analytical methods for the determination of herbicides in water.
Preparation of materials
The preparation of materials required the creation of multi compound solutions
by consecutive dilutions of a mother solution obtained with pure compounds.
After preparation, the solution is transferred into vials which are immediately
sealed under nitrogen.
For the cartridges, 180 l of tap water were taken the same day. Before
transferring to a cartridge according to the protocol described below, 5 l of water are
spiked with 5 ml of a pesticide solution.
Cartridges are first conditioned with 3 ml of acetonitrile, 3 ml of methanol, 3 ml
of water then 250 ml of spiked water are percolated. Water rinsing and nitrogen flow
drying steps are then realized before storing them in specific conditions (protected
from light and at a temperature lower than 20C). All these steps were conducted
with the ASPEC XL IV robot (GILSON).
There was a prior verification that vial seals would not alter the solution inside
and that the reliability of the robot was sufficient to guarantee good cartridge
homogenity.
Vial and cartridge preparations required three days and three weeks respectively.
400 sealed vials and 600 cartridges were produced.

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Homogenity
The homogenity was verified on two series of ten vials and ten cartridges (Table 1).

Table 1. Vial and cartridge homogenity

Stability
During the life span of the material, analyses are done each month. Until now
and in accordance with the feasibility study, no significant evolution of
concentrations was detected as is shown for example in Figures 4 and 5.

Chemistry Statistics /Gas Analysis

Vial - Chlortoluron
Phial

Conc entration (mg/l)

0.1200
0.1000
0.0800
0.0600
0.0400
0.0200
0.0000
3/24/2006

6/22/2006

9/20/2006

12/19/2006

Figure 4. Stability of chlortoluron in solution in the vials

DEA- Thimble
Cartridge
0.60

Concentration (g/l)

0.50
0.40
0.30
0.20
0.10
0.00

3/24/2006

6/22/2006

9/20/2006

12/19/2006

Figure 5. DEA stability in cartridge

Inter laboratory testing

16 laboratories with different techniques participated in this test (Table 2).
Samples (3 vials and 3 cartridges) were sent on April 4th 2006 and all results
were received on May 20th 2006.

10

Transverse Disciplines in Metrology

Laboratory

Bouisson Bertrand Laboratoires

MMA/APO
BRGM ORLEANS
CTCPA
Dpartement de l'conomie et de
la Sant (DES)
Service de Protection de la
Consommation (SPCo) - Geneva
IANESCO
Institut Dpartemental dAnalyse
et de Conseil
Ple Organique de l'INERIS
ISSeP (Institut de Scientifique de
Service Public) - Belgium
LECA / ESPCI
LDA 22
Laboratoire Dpartemental
DAnalyses de la Drme
Laboratoire national de
mtrologie et dessais (LNE)
Micropolluants Technologie
Nestl Waters MT
Laboratoire chimie - PTC Waters
Ple Analytique des Eaux
/LABO/CEE Plouzan
SGS Multilab Laboratoire de
lEssonne

Analytical
techniques

CPL SM2
(67% of
participants)
CPL UV
(28% of
participants)
CPG TSD
(5% of
participants)

CPG: gas chromatography,

CPL: liquid chromatography,
SM2: tandem mass spectrometry,
TSD: thermoionic detector,
UV: UV absorption.
Table 2. Laboratories participing in the certification testing and implemented techniques

Results
Raw results for cartridges and vials are grouped in Tables 3 and 4 respectively.

Chemistry Statistics /Gas Analysis

labo
A

Atrazine
0.093
0.100
0.095
0.106
0.110
0.110
0.076
0.088
0.096
0.093
0.095
0.102
0.111
0.111
0.107
0.094
0.093
0.096
0.102
0.100
0.104
0.110
0.110
0.105
0.109
0.110
0.113
0.108
0.108
0.110
0.114
0.117
0.133
0.086
0.085
0.087
0.108
0.085
0.107
0.106
0.105
0.107

DEA
0.082
0.092
0.083
0.099
0.101
0.101
0.054
0.068
0.077
0.094
0.103
0.096
0.097
0.101
0.093
0.084
0.082
0.082
0.097
0.094
0.099
0.130
0.135
0.125
0.098
0.102
0.106
0.098
0.098
0.098
0.115
0.117
0.140
0.090
0.083
0.084
0.118
0.095
0.103
0.097
0.097
0.099

DIA
0.079
0.079
0.084
0.100
0.102
0.100
0.076
0.084
0.087
0.079
0.079
0.067
0.094
0.100
0.095
0.096
0.093
0.100
0.096
0.091
0.097
0.115
0.110
0.100
0.070
0.070
0.076
0.084
0.084
0.084
0.103
0.102
0.113
0.082
0.081
0.089
0.154
0.129
0.163
0.078
0.079
0.081

Simazine
0.093
0.103
0.096
0.133
0.137
0.136
0.100
0.101
0.103
0.105
0.105
0.107
0.130
0.120
0.126
0.100
0.100
0.100
0.116
0.111
0.117
0.130
0.130
0.125
0.130
0.132
0.142
0.119
0.119
0.117
0.112
0.111
0.121
0.084
0.087
0.098
0.116
0.100
0.109
0.108
0.108
0.109

Terburtryne
0.106
0.116
0.109
0.112
0.116
0.116
0.066
0.086
0.107
0.121
0.123
0.122
0.128
0.130
0.125
0.102
0.093
0.098
0.134
0.130
0.136
0.160
0.150
0.150
0.138
0.142
0.145
0.121
0.123
0.121
0.109
0.107
0.138
0.083
0.082
0.087

0.128
0.129
0.133

Terbutylazine
0.094
0.100
0.099
0.097
0.102
0.102
0.042
0.056
0.068
0.089
0.095
0.091
0.105
0.109
0.106
0.086
0.085
0.086
0.105
0.101
0.106
0.110
0.110
0.100
0.100
0.102
0.107
0.103
0.104
0.102
0.091
0.092
0.106
0.075
0.072
0.073
0.165
0.139
0.154
0.104
0.104
0.105

Chlortoluron
0.118
0.121
0.128
0.102
0.109
0.106
0.105
0.111
0.112
0.095
0.111
0.098
0.107
0.111
0.109
0.125
0.115
0.132
0.119
0.114
0.121
0.130
0.130
0.125
0.117
0.124
0.134
0.107
0.107
0.106
0.129
0.122
0.152
0.085
0.087
0.090
0.135
0.112
0.130
0.129
0.133
0.134
0.120
0.096
0.115

Diuron
0.104
0.100
0.097
0.095
0.098
0.097
0.114
0.112
0.111
0.096
0.096
0.097
0.100
0.101
0.096
0.126
0.117
0.123
0.110
0.108
0.111
0.110
0.110
0.105
0.105
0.107
0.110
0.104
0.104
0.104
0.109
0.115
0.117
0.093
0.086
0.083
0.115
0.093
0.106
0.103
0.103
0.101
0.099
0.082
0.104

Isoproturon
0.114
0.123
0.120
0.109
0.111
0.110
0.107
0.117
0.119
0.132
0.106
0.114
0.121
0.126
0.117
0.161
0.145
0.155
0.130
0.126
0.132
0.145
0.145
0.135
0.130
0.132
0.139
0.123
0.124
0.122
0.133
0.136
0.147
0.104
0.099
0.107
0.155
0.120
0.140
0.125
0.127
0.128
0.162
0.125
0.148

Linuron
0.116
0.123
0.107
0.105
0.110
0.110
0.057
0.075
0.084
0.106
0.106
0.095
0.111
0.131
0.119
0.119
0.123
0.124
0.119
0.116
0.120
0.140
0.145
0.140
0.121
0.110
0.126
0.113
0.111
0.114
0.138
0.126
0.144
0.076
0.076
0.087
0.113
0.098
0.106
0.116
0.116
0.117
0.113
0.086
0.112

Table 3. Laboratory results for cartridges expressed in g/l of water

Lab
A
A
A
B
B
B
C
C
C
D
D
D
E
E
E
F
F
F
G
G
G
H
H
H
I
I
I
J
J
J
K
K
K
L
L
L
M
M
M
N
N
N
O
O
O
P
P
P

Atrazine
0.385
0.400
0.369
0.415
0.408
0.419
0.433
0.423
0.387
0.336
0.320
0.324
0.379
0.384
0.357
0.341
0.359
0.321
0.401
0.391
0.404
0.405
0.400
0.430
0.430
0.420
0.440
0.422
0.429
0.430
0.453
0.434
0.404
0.330
0.380
0.360
0.310
0.332
0.327
0.454
0.411
0.441
0.422
0.463
0.470

DEA
0.451
0.466
0.429
0.503
0.488
0.522
0.603
0.511
0.468
0.424
0.440
0.457
0.455
0.426
0.448
0.451
0.408
0.499
0.504
0.518
0.565
0.555
0.565
0.560
0.420
0.570
0.500
0.503
0.501
0.502
0.451
0.440
0.400
0.430
0.420
0.467
0.453
0.421
0.538
0.449
0.462
0.596
0.631
0.634

DIA
0.384
0.410
0.375
0.457
0.453
0.473
0.553
0.502
0.467
0.320
0.324
0.368
0.370
0.414
0.380
0.469
0.464
0.427
0.474
0.485
0.497
0.395
0.390
0.390
0.310
0.258
0.303
0.400
0.406
0.392
0.424
0.440
0.491
0.390
0.440
0.470
0.325
0.351
0.347
0.485
0.451
0.401
0.411
0.428
0.439

Simazine
0.305
0.328
0.314
0.442
0.432
0.447
0.345
0.334
0.303
0.304
0.304
0.308
0.353
0.366
0.324
0.300
0.326
0.300
0.378
0.374
0.382
0.360
0.355
0.370
0.440
0.420
0.460
0.389
0.390
0.394
0.350
0.339
0.346
0.340
0.400
0.350
0.247
0.261
0.269
0.451
0.361
0.390
0.364
0.388
0.392

Terburtryne
0.342
0.395
0.369
0.393
0.371
0.379
0.391
0.383
0.386
0.392
0.416
0.420
0.406
0.417
0.373
0.368
0.357
0.326
0.477
0.471
0.490
0.480
0.460
0.490
0.470
0.380
0.460
0.422
0.430
0.442
0.399
0.402
0.382
0.380
0.450
0.420
0.325
0.338
0.335

0.420
0.449
0.452

Terbutylazine
0.370
0.382
0.378
0.421
0.414
0.417
0.364
0.350
0.346
0.352
0.340
0.336
0.389
0.399
0.353
0.312
0.334
0.307
0.449
0.430
0.448
0.450
0.420
0.435
0.460
0.460
0.460
0.434
0.442
0.447
0.371
0.348
0.336
0.360
0.410
0.410
0.293
0.308
0.310
0.435
0.384
0.396
0.397
0.443
0.450

Chlortoluron
0.417
0.423
0.425
0.393
0.376
0.388
0.439
0.425
0.375
0.312
0.364
0.360
0.419
0.433
0.404
0.352
0.408
0.416
0.420
0.427
0.422
0.405
0.400
0.400
0.500
0.450
0.500
0.368
0.376
0.377
0.424
0.432
0.429
0.380
0.400
0.380
0.317
0.331
0.340
0.426
0.351
0.373
0.262
0.270
0.259
0.378
0.328
0.288

Diuron
0.250
0.226
0.328
0.400
0.378
0.387
0.454
0.441
0.388
0.372
0.360
0.404
0.404
0.398
0.396
0.436
0.460
0.500
0.443
0.452
0.452
0.415
0.380
0.405
0.430
0.390
0.440
0.404
0.415
0.425
0.386
0.382
0.408
0.300
0.340
0.300
0.415
0.438
0.448
0.439
0.356
0.388
0.435
0.446
0.449
0.321
0.255
0.259

Isoproturon
0.370
0.429
0.449
0.454
0.426
0.438
0.540
0.530
0.468
0.388
0.384
0.428
0.479
0.492
0.450
0.556
0.620
0.624
0.514
0.530
0.530
0.525
0.520
0.520
0.580
0.450
0.590
0.481
0.486
0.491
0.493
0.502
0.507
0.460
0.520
0.500
0.461
0.484
0.482
0.543
0.484
0.501
0.570
0.599
0.591
0.508
0.461
0.391

Table 4. Laboratory results for vials expressed in mg/l

Linuron
0.369
0.416
0.396
0.429
0.412
0.428
0.434
0.418
0.380
0.368
0.400
0.384
0.442
0.464
0.416
0.388
0.468
0.432
0.472
0.468
0.475
0.455
0.460
0.470
0.490
0.500
0.520
0.429
0.443
0.449
0.464
0.466
0.406
0.280
0.300
0.340
0.364
0.354
0.351
0.424
0.319
0.361
0.455
0.485
0.490
0.328
0.290
0.277

11