Hair decontamination procedure prior to

multi-class pesticide analysis

Radu-Corneliu Duca, Emilie Hardy, Guillaume Salqubre and
Brice M. R. Appenzeller*
Although increasing interest is being observed in hair analysis for the biomonitoring of human exposure to pesticides, some
limitations still have to be addressed for optimum use of this matrix in that specic context. One main possible issue concerns
the need to differentiate chemicals biologically incorporated into hair from those externally deposited on hair surface from
contaminated air or dust. The present study focuses on the development of a washing procedure for the decontamination
of hair before analysis of pesticides from different chemical classes. For this purpose, three different procedures of articial
contamination (with silica, cellulose, and aqueous solution) were used to simulate pesticides deposition on hair surface. Sev-
eral washing solvents (four organic: acetone, dichloromethane, methanol, acetonitrile; and four aqueous: water, phosphate
buffer, shampoo, sodium dodecylsulfate) were evaluated for their capacity to remove articially deposited pesticides from
hair surface. The most effective washing solvents were sodiumdodecylsulfate and methanol for aqueous and organic solvents,
respectively. Moreover, after a rst washing with sodium dodecylsulfate or methanol, the majority of externally deposited
pesticides was removed and a steady-state was reached since signicantly lower amounts were removed by additional second
and third washings. Finally, the effectiveness of a decontamination procedure comprising washing with sodium dodecyl-
sulfate and methanol was successively demonstrated. In parallel, it was determined that the nal procedure did not affect
the chemicals biologically incorporated, as hair strands naturally containing pesticides were used. Such a procedure appears
to remove in one-shot the fraction of chemicals located on hair surface and does not require repeated washing steps.
Copyright 2014 John Wiley & Sons, Ltd.
Keywords: hair; decontamination; washing procedure; pesticides; multi-class analysis
Introduction
Although increasing interest is being observed in hair analysis for
the biomonitoring of human exposure to pesticides, some limita-
tions still have to be addressed for optimum use of this matrix in
that specic context. One main possible issue concerns the need
to differentiate chemicals biologically incorporated into hair from
those externally deposited on hair surface due to contaminated
air or dust. Chemicals incorporated by means of biological mech-
anisms, likely to be located in the whole hair structure, can be
interpreted as representative of the internal dose people have
undergone during the period of hair growth. On the contrary,
chemicals externally deposited only represent recent contamina-
tion and are therefore less biologically relevant. Ideal decontam-
ination procedure therefore has to remove external chemicals
(deposited on the cuticle surface) without affecting internally
incorporated compounds (present in the bulk of the matrix).
In forensic sciences, decontamination of hair prior to drug
analysis is an important step, since false positive samples might
have legal implications. Thus, decontamination has been
extensively documented and several washing procedures with
different degrees of complexity have been proposed.
[1]
Hair
decontamination before drug testing is generally performed with
organic solvents (e.g. dichloromethane, acetone, methanol,
acetonitrile) and/or with aqueous solutions of detergents (e.g.
sodium dodecylsulfate) or buffers (e.g. phosphate buffer).
[16]
Unlike for drug testing, the decontamination with organic solvents
before environmental pollutants analysis was less used
[711]
and
more gentle decontamination solvents like water
[1214]
and water
with shampoo
[1520]
were preferred. Nevertheless, no standardized
washing procedure removing the totality of the external contami-
nation without affecting the drugs biologically incorporated into
hair is available yet.
[21]
Still, the importance of hair decontamination
was acknowledged by the Society of Hair Testing (SoHT) and rec-
ommendations for washing were made: both organic and aqueous
solvents should be used, the efciency of the washing procedure
should be tested and, if required, additional clean-up steps should
be taken.
[21]
An important step in the development of a decontamination
procedure is to produce articially contaminated specimens to
be used for testing the efciency of the washing to remove
chemicals deposited on hair surface. The most classic contamina-
tion procedure is the utilization of a soaking aqueous solution
containing the compounds of interest.
[4,6,22,23]
Even if this arti-
cial contamination is effective, it reects neither the external de-
position of compounds on hair surface by biological uids nor
the environmental contamination occurring mainly through air-
borne dust particles. Moreover, an important limitation associ-
ated with hair dipping in aqueous solutions is the possible
irreversible penetration of the compounds within the hair shaft.
[6]
* Correspondence to: Brice M. R. Appenzeller, Laboratory of Analytical Human
Biomonitoring, Centre de Recherche Public de la Sant (CRP-Sant), 162A,
Avenue de la Faencerie, L-1511 Luxembourg, Luxembourg. E-mail: brice.
appenzeller@crp-sante.lu
Laboratory of Analytical Human Biomonitoring, CRP-Sant, 162A avenue de la
Faencerie, L-1511 Luxembourg, Luxembourg
Drug Test. Analysis 2014, 6, 5566 Copyright 2014 John Wiley & Sons, Ltd.
Research article
Drug Testing
and Analysis
Received: 31 October 2013 Revised: 7 February 2014 Accepted: 24 February 2014 Published online in Wiley Online Library
(www.drugtestinganalysis.com) DOI 10.1002/dta.1649
5
5
In the last decade, more realistic articial contamination proce-
dures were also used: hair incubation with spiked blood to dem-
onstrate postmortem external contamination,
[24]
hair briey
dipped in authentic urine from a methamphetamine user to in-
vestigate the contamination/decontamination of pubic hair,
[25]
hair coating with drugs followed by sweat conditioning using
synthetic sweat to study drug contamination/decontamination
in the analysis of human hair,
[6]
and subjects hair own contamina-
tion using their hands powdered with a drug mixture.
[26]
Although such procedures might be relevant to clinical or forensic
contexts, they are not representative of hair surface contamina-
tion by organic pollutants present in air or dust; specic
approaches have to be developed for the latter purpose.
The present study is therefore focused on the development of
a washing procedure applied to hair samples articially contami-
nated with a mixture of different organic pollutants (pesticides)
from different chemical classes and thus displaying various
physicochemical properties. For the articial contamination, three
different media were used: two types of solid particles (silica and
cellulose) previously contaminated with the pesticide mixture to
simulate contamination from dust or airborne particles, and aque-
ous solution of the target compounds used for comparison. The
washing procedures applied to the contaminated hair were eval-
uated according to the criteria proposed within a review paper
previously published:
[27]
1. A signicant part of the chemicals has to be removed dur-
ing the rst washing.
2. No more pesticides or signicantly lower amounts are re-
moved by additional washings.
3. The part of chemicals extracted from hair after pulverization of
the matrix should not be affected by the washing procedure.
For this purpose, a rst step consisted in evaluating several
solvents for their capacity to remove the articially deposited
pesticides from the hair surface. Four organic solvents (acetone,
dichloromethane, methanol, acetonitrile) and four aqueous sol-
vents (water, phosphate buffer pH7, shampoo, and 5% sodium
dodecylsulfate) were tested. In a second step, the most effective
washing solvents (organic and aqueous, respectively) were evalu-
ated to determine whether a steady-state was reached after the
rst washing, by doing three successive washings. The third step
consisted of testing the effectiveness of the nal washing proce-
dure chosen based on the results obtained from the previous
steps. Finally, hair samples naturally containing pesticides were
submitted to the washing procedure to determine whether
pesticide concentration was affected.
Materials and methods
Chemicals and reagents
Pesticides analytical standards with purities higher than 97%,
except for permethrin and cypermethrin (94% purity), were
purchased from Dr Ehrenstorfer GmbH (Augsburg, Germany).
Individual stock solutions at 1 g L
-1
of all standards were prepared
by exact weighing and dissolution in acetonitrile. A working solu-
tion containing the targeted molecules at 10 mgL
-1
in acetonitrile
was prepared. Acetonitrile, methanol, dichloromethane, acetone
and hexane were purchased from Biosolve (Valkenswaard, The
Netherlands). A phosphate buffer solution (1 M, pH7.0) was pre-
pared using monosodium phosphate and disodium phosphate.
Sodium dodecylsulfate (SDS), monosodium phosphate, disodium
phosphate, formic acid, silica and cellulose were provided by
Sigma-Aldrich (Bornem, Belgium). The shampoo used for hair
washing tests was Baby shampoo (Mustela, Laboratoires
Expanscience, Courbevoie, France). Ultrapure water was produced
using a Milli-Q gradient water system (Millipore, Brussels, Belgium).
Hair articial contamination
Hair specimens were collected from the Laboratory staff mem-
bers (Caucasian, male, average age of 31 years). The collected
samples were initially analyzed for pesticides content. Two hair
specimens (brown) were nally selected: one containing low
levels of pesticides was used for the preliminary selection of de-
contamination solvents (after articial contamination); and one
naturally containing several pesticides was used to test the nal
washing procedure. Except for individuals self-washing with
shampoo, no cosmetic treatment was reported.
To simulate pesticide deposition on hair surface, three
different contamination procedures were used: two dry contam-
inations using silica or cellulose particles, and one wet con-
tamination using an aqueous solution. The articial dry
contamination procedures were used to simulate the natural
solid-solid transfer of pesticides from dust to the hair shaft. Silica
(particle size: 210300 m), a hydrophilic inorganic support, was
spiked with a mixture of pesticides to reach a nal concentration
of 0.5 g g
-1
. Then, 1 g hair was mixed with 10 g silica for 24 h at
25 rpm, using an end-over-end mixer (Intelli-Mixer RM-2 L, Elmi
Ltd, Riga, Latvia). Cellulose (particle size: 2560 m), a lipophilic
organic support, was spiked with a pesticide mixture to reach a
nal concentration of 1 g g
-1
. Five grams of cellulose were gently
mixed with 1 g hair sample for 24 h at 25 rpm, using an end-over-
end mixer. The articial wet contamination, classically used to test
the efciency of hair washing solvents,
[4,6,22,23]
was done using
aqueous solution containing the target pesticides at 0.2 g mL
-1
.
A hair sample (1 g) was dipped in 100 mL pesticides aqueous solu-
tion for 1 h. The amount of pesticides added to each media (silica,
cellulose, and water) was adjusted with regard to its density, in or-
der to ensure that for each assay, regardless of the contamination
path, 1 g of hair was in contact with 20 g of each pesticide.
Hair decontamination
The present paper is dealing with the development of a washing
procedure prior to multi-residue pesticides determination in hair.
The washing efciency was calculated as the ratio of the pesti-
cide concentration between the washed and non-washed hair
samples, expressed as percentage of decontamination (Table 2).
The nal procedure was as follow: hair (100 mg) was successively
washed for 5 min. with sodium dodecylsulfate, followed by a sec-
ond 5 min. washing with methanol using an orbital shaker at
350 rpm (Incubator Shaker G25, New Brunswick, NJ, USA). Then
the sample was allowed to dry at room temperature prior to pul-
verization for pesticides analysis.
Pesticides analysis in hair samples
Hair samples were extracted and analyzed according to
Salqubre et al.,
[11]
with minor modication and adaptation. Fifty
mg of pulverized hair were extracted overnight at 40 C, using
1 mL of acetonitrile:water (80:20). The extract was centrifuged
for 10 min at 5000xg. Four-hundred L supernatant were
R.-C. Duca et al.
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collected and diluted in 7.6 mL phosphate buffer (1 M, pH7.0)
prior to solid phase micro extraction-gas chromatography-
tandem mass spectrometry (SPME-GC-MS/MS) analysis. The
remaining supernatant was dried under a gentle stream of ni-
trogen and recovered in a solution of water:acetonitrile:formic
acid (94.5/5/0.5, v/v/v) for ultra performance liquid chroma-
tography-tandem mass spectrometry (UPLC-MS/MS) analysis.
Stable-isotope-labelled derivatives were used as internal stan-
dards to compensate any possible extraction losses (Table 1).
SPME-GC-MS/MS analysis
A direct immersion SPME was carried out using a 65 m mixed
polydimethylsiloxane/divinylbenzene (PDMS/DVB) bre. The ex-
traction was performed at 60 C for 80 min. The bre desorption
was done at 260 C for 15 min. GC analyses for pesticides were
performed using an Agilent Technologies GC system (7890A),
and separations were carried out using an HP-5MS 5% Phenyl
Methyl Silox capillary column (30 m x 250 m, 0.25 m particle
size) from Agilent Technologies (Santa Clara, CA, USA). The
analytes were separated using helium as carrier gas at a ow rate
of 1.2 mL/min. The oven temperature program started at 70 C for
5 min, followed by three successive linear increases of 10 C/min
to 200 C, 2 C/min to 240 C, and 10 C/min to 300 C; tempera-
ture was held for 3 min. MS/MS analysis was carried out using
an Agilent GC-MS TQ 7000A tandem quadrupole mass spectrom-
eter (Agilent Technologies, Santa Clara, CA, USA). The mass spec-
trometer was operated in negative chemical ionization mode
using methane as the reagent gas. The specic MS/MS parame-
ters for the GC-amenable compounds are shown in Table 1. The
methods quantication limits (LOQ) for GC-amenable target
analytes ranged from 0.02 to 0.5 pg/mg hair, with most LOQs
lower than 0.2 pg/mg hair.
UPLC-MS/MS analysis
LC analyses were performed using an Acquity UPLC system, and
separations were carried out using an Acquity UPLC BEH C18
Table 1. MS/MS parameters for pesticides determination
Compound Name RTW
a
Quantier Transition CE
b
(V) Qualier Transition CE
b
(V)
GC amenable molecules
Triuraline 10.0 - 20.1 335 305 9 335 46 14
Triuralin-D14 10.0 - 20.1 349 319 9
-HCH 10.0 - 20.1 255 35 5 253 35 5
-HCH 10.0 - 20.1 255 35 5 253 35 5
-HCH D6 10.0 - 20.1 261 35 5
Diazinon 10.0 - 20.1 169 95 18 169 141 13
Chlorpyriphos methyl 20.1 - 23.2 285 95 8 287 95 8
Parathion methyl 20.1 - 23.2 263 154 5 263 141 17
Aldrin 20.1 - 23.2 237 35 32 328 35 22
Chlorpyriphos ethyl 20.1 - 23.2 313 189 12 313 95 23
Parathion ethyl 20.1 - 23.2 291 154 4 291 169 12
Fipronil 23.2 - 25.1 366 318 11 400 331 6
-Endosulfan 23.2 - 25.1 406 35 4 404 35 4
p,p-DDE 23.2 - 25.1 318 35 1 316 35 1
p,p-DDE D8 23.2 - 25.1 326 35 1
Dieldrin 25.1 - 29.0 380 35 3 378 35 3
-Endosulfan 25.1 - 29.0 406 35 6 404 35 6
-Endosulfan D4 25.1 - 29.0 410 35 6
p,p-DDT 29.0 - 35.0 281 35 15 71 35 8
p,p-DDT D8 29.0 - 35.0 71 35 8
Diufenican 29.0 - 35.0 394 231 13 394 161 12
Permethrin 35.0 - 47.0 207 35 8 209 37 8
Cypermethrin 35.0 - 47.0 207 35 8 209 37 8
LC amenable molecules
Thiamethoxam 1.97 - 2.37 292 181 21 292 211 12
Clothianidin 2.25 - 2.65 250 132 18 250 169 12
Clothianidin D3 2.25 - 2.65 253 172 12
Imidacloprid 2.40 - 2.80 256 175 20 256 209 14
Propoxur 3.64 - 4.04 211 111 14 211 168 7
Carbofuran 3.69 - 4.11 223 123 22 223 165 12
Tebuconazole 5.37 - 5.79 309 70 18 309 125 34
Tebuconazole D6 5.37 - 5.79 315 72 18
Propiconazole 5.69 - 6.13 343 159 28 345 161 28
a
Retention time window.
b
Collision energy.
Hair decontamination
Drug Testing
and Analysis
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column (100 2.1 mm, 1.7 m particle size) from Waters
(Manchester, UK). The C18 column was equilibrated at 40 C.
The analytes were separated using a mobile phase consisting of
water:acetonitrile:formic acid, 94.5/5/0.5, v/v/v (eluent A) and ace-
tonitrile:formic acid, 99.5/0.5, v/v (eluent B), at a ow rate of
0.6 mL/min. The chromatographic separation program started at
1% eluent B for 0.5 min, than increased linearly to 99% in
8.5 min; composition that was held for 0.4 min before returning
to the initial conditions and was followed by an equilibration
time of 0.5 min. The total run time was of 10 min. Tandem mass
analysis was carried out using a Waters Acquity TQD tandem
quadrupole mass spectrometer (Manchester, UK). The instrument
was operated using an electrospray source in positive mode. The
specic MS/MS parameters for the LC-amenable compounds are
shown in Table 1. The methods LOQs for LC-amenable target
analytes ranged from 1 to 5 pg/mg hair, values which are higher
than for GC-amenable compounds partially explained by SPME
advantage to use the integrality of sample extract.
Results and discussion
Preliminary selection of decontamination solvents
In the rst step of the washing procedure development, eight
washing solvents water, phosphate buffer pH7, shampoo, so-
dium dodecylsulfate, acetone, dichloromethane, methanol, and
acetonitrile were evaluated for their ability to remove chemicals
from the surface of articially contaminated hair samples. For this
preliminary evaluation, 20 representative pesticides from several
chemical classes were targeted: organochlorines (p,p-DDE,
HCH-beta/gamma, dieldrin, endosulfan-alpha/beta), pyrethroids
(permethrin and cypermethrin), organophosphates (chlorpyri-
phos ethyl, parathion ethyl and diazinon), neonicotinoids
(clothianidin, imidacloprid, thiametoxam), azoles (tebuconazole
and propiconazole), carbamates (carbofuran and propoxur),
pronil and triuraline.
One hundred mg hair samples, articially contaminated with
silica, cellulose and aqueous solution containing the 20 target
pesticides, were washed with each solvent separately. All the
samples have been treated simultaneously and in ve replicates.
Removing deposit chemicals from hair contaminated with silica
The efciency of the washing to remove the externally deposited
chemicals from hair contaminated with silica was depending on
the solvent (Figure 1). Using water, the part of chemicals re-
moved from hair ranged from 4% (p,p-DDE) to 90%
(thiametoxam), and was above 50% for ve compounds. The de-
contamination efciency obtained with phosphate buffer ranged
from 2% (triuraline) to 88% (thiametoxam), and was above 50%
for nine compounds. Decontamination with shampoo removed
from 1% (permethrin) to 96% (thiametoxam) of deposited
chemicals, with eight compounds decreased by more than 50%.
Using sodium dodecylsulfate, the decontamination efciency
ranged from 12% (propiconazole) to 98% (thiametoxam and
clothianidin) removal, with a concentration decreased by more
than 50% for 15 compounds. Among the aqueous solvents, the
most efcient decontamination of hair samples articially con-
taminated with silica was achieved by washing with sodium
dodecylsulfate.
The decontamination efciency reached using acetone ranged
from 31% (permethrin and propoxur) to 85% (endosulfan-alpha
and triuraline), with 16 compounds whose concentration was
decreased by more than 50%. For dichloromethane, the part of
chemicals removed ranged from 7% (permethrin) to 92% (endo-
sulfan-alpha and triuraline), with 17 compounds decreased by
more than 50%. Using methanol, the decontamination removed
from 54% (propiconazole) to 96% (diazinon) of deposited
chemicals, and the concentration was decreased by more
than 50% for the totality of target compounds. The de-
contamination obtained with acetonitrile ranged from 10%
(carbofuran) to 83% (diazinon), and the concentration of 9
compounds was decreased by more than 50%. Methanol
was therefore found to be the most appropriate washing
organic solvent to remove external contamination from hair
contaminated with silica.
Removing deposit chemicals from hair contaminated with cellulose
The capacity of the washing to remove externally deposited
chemicals from hair contaminated with cellulose was also highly
depending on the solvent (Figure 2). The percentage of
chemicals removed using water ranged from 2% (permethrin)
to 100% (propiconazole), with 14 compounds out of the 20 target
pesticides having their concentration decreased by more than
50%. The decontamination percentages obtained using phos-
phate buffer ranged from 3% (permethrin) to 100% (propicona-
zole), and 13 compounds were decreased by more than 50%.
Decontamination with shampoo removed from 8% (endosulfan-
beta) to 100% (thiametoxam and propiconazole) of the deposited
chemicals, and removed more than 50% of 14 compounds. Using
sodium dodecylsulfate, the percentage of decontamination
ranged from 36% (permethrin) to 100% (clothianidin, thiameto-
xam and propiconazole), and the concentration of all the com-
pounds was decreased by more than 50%, except permethrin.
Among the aqueous solvents, the most appropriate decontami-
nation of hair samples articially contaminated with cellulose
containing the target pesticides was achieved using sodium
dodecylsulfate.
The percentage of chemicals removed using acetone ranged
from 25% (permethrin) to 100% (chlorpyriphos ethyl, thiameto-
xam, clothianidin, tebuconazole and propiconazole), 4% to
100% for dichloromethane and 32% to 100% for acetonitrile. All
chemicals, except permethrin, had their concentration decreased
by more than 50% upon washing with acetone, dichloromethane
and acetonitrile. All the chemicals, including permethrin, were re-
moved from the articially contaminated hair upon washing with
methanol. The percentage of chemicals removed ranged from
69% (permethrin) to 100% (chlorpyriphos ethyl, clothianidin,
tebuconazole and propiconazole). Methanol was found to be
the most efcient organic solvent to remove external contamina-
tion from hair contaminated with cellulose.
Removing deposit chemicals from hair contaminated with aqueous
solution
The capacity of washing externally deposited chemicals from hair
contaminated by dipping in an aqueous solution also depended
on the solvent nature, but was signicantly less effective than for
silica- and cellulose-contaminated hair (Figure 3). The part of
chemicals removed using water as washing solvent ranged from
0% (p,p-DDE, endosulfan-beta, parathion ethyl and diazinon) to
37% (pronil). Phosphate buffer decontamination was even less
effective than water and ranged from 0% (p,p-DDE, endosulfan-
beta, chlorpyriphos ethyl, diazinon, imidacloprid and tebuco-
nazole) to 23% (cyphermetrin). The decontamination using
R.-C. Duca et al.
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shampoo as washing solvent ranged from 0% (endosulfan -alpha/-
beta and propiconazole) to 39% (pronil). Sodium dodecylsulfate
was found to have the highest decontamination capacity on
dipped hair, ranging from 0% (endosulfan-beta, parathion ethyl
and propiconazole) to 95% (cypermethrin), with 10 compounds
having their concentration decreased by more than 50%.
Figure 1. Solvents capacity of washing externally deposit chemicals from hair articially contaminated with silica. The percentage of externally de-
posited pesticides that was not removed upon washing is presented for: A - Organochlorides; B - Pyrethroids and Organophosphates; C - Neonicotinoids;
D - Azoles, Carbamates, Fipronil and Triuraline. The level of pesticides determined in hair articially contaminated with silica without washing was ar-
bitrary set at 100% of hair external contamination and was presented as No Wash (red line).
Hair decontamination
Drug Testing
and Analysis
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Using acetone, decontamination ranged from 0% (endosulfan -
alpha/-beta, parathion ethyl, tebuconazole, carbofuran and
propoxur) to 56% (clothianidin), and only two compounds were
decreased by more than 50%. Using dichloromethane, the de-
contamination ranged from 0% (dieldrin and endosulfan-alpha)
to 57% (cypermethrin), with again only two compounds having
their concentration decreased by more than 50%. The decontam-
ination using methanol as washing solvent was the most
effective among all the organic solvents and ranged from 0%
(endosulfan-alpha) to 94% (pronil), with nine compounds
Figure 2. Solvents capacity of washing externally deposited chemicals from hair articially contaminated with cellulose. The percentage of externally
deposited pesticides that was not removed upon washing is presented for: A - Organochlorides; B - Pyrethroids and Organophosphates; C -
Neonicotinoids; D - Azoles, Carbamates, Fipronil and Triuraline. The level of pesticides determined in hair articially contaminated with cellulose with-
out washing was set to100% of hair external contamination and was presented as No Wash (red line).
R.-C. Duca et al.
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and Analysis
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Figure 3. Solvents capacity of washing externally deposited chemicals from hair articially contaminated by dipping. The percentage of externally de-
posit pesticides that was not removed upon washing is presented for: A - Organochlorides; B - Pyrethroids and Organophosphates; C - Neonicotinoids; D
- Azoles, Carbamates, Fipronil and Triuraline. The level of pesticides determined in hair articially contaminated by dipping without washing was ar-
bitrary set to 100% of hair external contamination and was presented as No Wash (red line).
Hair decontamination
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and Analysis
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whose concentration was decreased by more than 50%. Deco-
ntamination with acetonitrile was similar to acetone and
dichloromethane, and ranged from 0% (dieldrin, HCH-gamma,
endosulfan-alpha/-beta, diazinon, imidacloprid, propiconazole
and carbofuran) to 58% (cypermethrin), with two compounds de-
creased by more than 50%. Even if the percentage of chemicals
removed was lower than for hair samples contaminated with sil-
ica and cellulose, sodium dodecylsulfate and methanol were still
the most efcient solvents.
The hair contamination model based on hair dipping could be
considered inappropriate since not reecting natural external
contamination, which is mainly due to dry contamination from
dust present in the individuals surroundings. Moreover, the hair
dipping in aqueous solution has two main limitations that can ex-
plain the low percentages of chemical removal upon washing:
rst, the irreversible penetration of compounds within the hair
shaft,
[6]
and secondly, the important transfer of compounds to
the hair resulting in unrealistic levels of pesticides deposited
on hair shaft (about ten times higher than for the dry contamina-
tions). Nevertheless, the issue associated with wet contamination
should not be disregarded, since the dry contamination might
become aqueous in high-humidity environment or in presence
of perspiration. Further assays involving aqueous contamination
with lower nal levels of pesticides deposited on (or incorporated
in) hair might therefore provide relevant information and allow
assessing the decontamination in more realistic conditions.
For environmental exposure studies the complete certainty of
the source of pesticide (external vs. incorporated) may not be
as critical as in some other applications of hair analysis (e.g. foren-
sic determination of use of illicit drugs). As stated in a previous re-
view paper, the part of externally deposited compound that
might become unremovable (due to wet conditions for in-
stance) can eventually (and reasonably) be considered as repre-
sentative of the history of the exposure.
[27]
The possible risk of
misinterpretation is actually associated with the part of externally
deposited compound that remains easily removable (ERC for
easily removable compounds) and can thus be removed by
peoples self-hair washing with shampoo. Since samples from
different subjects have to be treated the same for reliable com-
parison, the time between hair sampling and individuals last
bath/shower can be a source of variability, since ERC might have
been removed or not.
[27]
A washing procedure prior to hair anal-
ysis is therefore a necessary precaution, in order to have samples
similarly treated.
Successive washing with sodium dodecylsulfate or methanol
As presented here, sodium dodecylsulfate and methanol were
found to be the most efcient solvents to remove pesticides from
hair, whatever the articial contamination (with silica, cellulose,
or aqueous solution). The key question at this stage of the devel-
opment was whether to use sodium dodecylsulfate or methanol,
or both of them. Sodium dodecylsulfate and methanol were
equivalent regarding the number of pesticides whose concentra-
tion was decreased by more than 50% as well as the intensity of
average decontamination. Nevertheless, differences in their
washing properties are to be noticed mainly concerning organo-
phosphates and neonicotinoids (Figures 1 and 2). Sodium
dodecylsulfate was more effective than methanol to remove
the neonicotinoids externally deposit on hair. On the contrary,
methanol was more effective than sodium dodecylsulfate to re-
move the organophosphates deposited on hair. These results
are in concordance with the physicochemical properties of
these pesticides. Indeed, imidacloprid (neonicotenoid) is
water soluble (610 mg/L) and has an octanol/water partition
coefcient (Log P) of 0.57. At the opposite, diazinon
(organophosphate) has lower water solubility (60 mg/L)
and a Log P of 3.69.
[28]
Since the objective was to develop a general decontamination
procedure for multi-class pesticides analysis in hair, both sodium
dodecylsulfate and methanol were therefore necessary.
In the second step of the washing procedure development, it
was determined whether a steady-state was reached after a sin-
gle washing. Three successive washings were tested with either
sodium dodecylsulfate or methanol. All the samples were treated
simultaneously and in three replicates. Figure 4 presents the
removal of several representative pesticides from hair samples
contaminated with silica, upon repeated washings with sodium
dodecylsulfate or methanol.
The rst washing with SDS removed from 12% (propiconazole)
to 100% (clothianidin) of the pesticides and the average decon-
tamination percentage was 63%. The second wash with SDS re-
moved from 0% to 25% of the pesticides and the average
decontamination was of 8%. The third wash with SDS removed
from 0% to 5% and the average decontamination was 0.5%.
The rst washing with methanol removed from 54%
(propiconazole) to 95%(triuraline) of the pesticides and the average
decontamination percentage was 80%. The second wash with meth-
anol removed from 0% to 17% of the pesticides and the average de-
contamination was 4%. The third wash with methanol removed 0%
to 6% and the average decontamination percentage was 1.3%.
These results are in concordance with the few studies
previously published which highlighted the relevance of hair
washing prior to pollutants analysis.
[27]
Those studies compared
unwashed hair with hair washed with common surfactants just
before analysis, and reported that the levels of dioxins,
[15,29]
PCBs,
[16]
or some pesticides (e.g. organochlorines
[16]
and
carbamates
[30]
) were decreased in hair samples after washing.
Moreover, most of the decrease occurs after the rst washing
and additional washing has no further effect on the elimination
of chemicals.
[15,16]
In the same sense, Toriba et al.
[8]
studied the
effect of washing hair with organic solvents (i.e. methanol,
hexane and dichloromethane) prior to polycyclic aromatic hydro-
carbons (PAHs) analysis in hair. Although different results were
obtained depending on the compounds analyzed and the
solvents, they were in line with the results of the present study,
in that a part of the target molecules was removed by washing
and the most signicant part of the chemicals was removed after
the rst washing, even if some chemicals were also removed dur-
ing the second and the third washing. The latter difference might
be explained by the fact that washing was carried out without
agitation, contrary to the present study.
A single washing with sodium dodecylsulfate or methanol
therefore appeared to be sufcient to remove the majority of
the deposited pesticides and to reach a steady-state since no
more chemicals or signicantly lower amounts were removed
by additional washings.
Final washing procedure
In the third and nal step of the development, the efciency of a
washing procedure using both SDS and methanol was tested
R.-C. Duca et al.
Drug Testing
and Analysis
wileyonlinelibrary.com/journal/dta Copyright 2014 John Wiley & Sons, Ltd. Drug Test. Analysis 2014, 6, 5566
6
2
(Table 2). Hair naturally containing pesticides of a non-occupationally
exposed person was used. For this nal step, the list of target com-
pounds was enlarged with ve more pesticides: two organochlorines
(p,p-DDT and aldrin), two organophosphates (chlorpyriphos methyl
and parathion methyl), and one carboxamide (diufenican). Hair
strands were articially contaminated with silica, cellulose or aque-
ous solution containing pesticides. Hair samples (100mg) were rst
washed with sodium dodecylsulfate for 5min and then SDS was re-
moved. In a second step, the samples were washed with methanol
for another 5min. Hair bres were then gathered and allowed to
dry at roomtemperature prior to pulverization and extraction before
pesticides analysis. Washing hair samples with methanol after SDS
(and not before) allowed the entire removal of the latter solvent from
hair. This precaution is necessary as SDS is known to interfere
with the chromatographic separation and mass-spectrometric
determination.
[31,32]
The amount of pesticides removed from the surface of hair
contaminated with silica, using SDS-methanol washing, ranged
from 61% to 100%. For hair contaminated with cellulose, the to-
tality of externally deposited pesticides was removed upon
SDS-methanol washing. Even for dipped hair, the amount of
pesticides removed from the hair surface was above 50% for all
the pesticides except for p,p-DDT, dieldrin, HCH-gamma. The lat-
ter differences might be explained by the inuence the articial
contamination procedure might have on hair structure, com-
pounds deposition on hair surface and/or pesticides irreversible
incorporation. For the articial contamination of hair by dipping
in aqueous solution, the possible irreversible penetration of some
compounds within the hair shaft has been previously described
[6]
and is in concordance with the present study results. In the case
of the dry contamination procedures, the differences between sil-
ica and cellulose observed here could suggest that silica, as
tough mineral particle, could abrade hair surface during the con-
tamination process and hence allow pesticides to reach internal
areas of the hair bers and become less removable by washing.
At the opposite, cellulose, as soft organic particles, would limit
pesticide deposition on hair surface. A good illustration is provided
with -endosulfan, naturally present in hair at low levels of concen-
tration (0.410.10 pg/mg), for which upon SDS-methanol washing
of the dry contaminated hair samples, only for cellulose were
reached comparable concentration levels (1.00 0.10pg/mg) and
a decontamination percentage close to 100%, whereas the articial
Figure 4. Articially contaminated hair (by pesticides in silica particles), submitted to successive washings with sodium dodecylsulfate (right) or
methanol (left). The percentage of externally deposited pesticides that was not removed upon successive washing is presented for: (A) Organochlorides;
(B) Pyrethroids, Organophosphates and Neonicotinoids; (C) Azoles, Carbamates and Miscellaneous.
Hair decontamination
Drug Testing
and Analysis
Drug Test. Analysis 2014, 6, 5566 Copyright 2014 John Wiley & Sons, Ltd. wileyonlinelibrary.com/journal/dta
6
3
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R.-C. Duca et al.
Drug Testing
and Analysis
wileyonlinelibrary.com/journal/dta Copyright 2014 John Wiley & Sons, Ltd. Drug Test. Analysis 2014, 6, 5566
6
4
contamination was higher in hair contaminated with cellulose
(2577 pg/mg) than with silica (125 3 pg/mg). Nevertheless,
these results conrmed that a washing procedure using rst so-
dium dodecylsulfate and secondly methanol, efciently removes
the pesticides deposited on hair.
Native hair samples (not submitted to articial contamination)
were also washed with SDS and methanol successively. The per-
centage of pesticides removed from hair then ranged from 5.4%
(HCH-gamma) to 100% (tebuconazole) of the total amount of
pesticides (determined in hair samples without washing). The
part of pesticides removed in the latter experiment is therefore
likely to correspond to externally deposited compounds whereas
the amount of pesticides still detected after washing would
correspond to biological incorporation.
Taking all together, the results obtained on SDS-methanol wash-
ing of both articially contaminated and native hair demonstrated
that the pesticides biologically incorporated into hair are not af-
fected by the decontamination procedure. A good illustration is
provided with propiconazole, for which the concentrations deter-
mined upon hair wash with SDS-methanol were 4.6 0.3pg/mg
in native hair, 4.00.5pg/mg in hair contaminated with silica and
3.40.7pg/mg in hair contaminated with cellulose. Although the
present study shall represent an important step in the development
of a standardized decontamination procedure before pesticide
analysis in hair, the results presented here were obtained from
Caucasian subjects who did not use cosmetics. As hair structure
might inuence incorporation, further studies should be conducted
on hair samples collected fromsubjects of different origins (African,
Asian) as well as on samples collected from people applying cos-
metic treatment that might have altered structure.
Conclusions
Evaluating the efciency of a washing procedure of hair before
analysis is a complex task which may be signicantly inuenced
by the protocol of articial contamination of the samples, as
demonstrated in the present work. Therefore, articial contami-
nation of samples has to be as close as possible to reality. Remov-
ing from hair surface compounds from different chemical classes
presents an additional challenge, as the efciency of a washing
solvent varies for different chemicals. It was demonstrated here
that washing hair with sodium dodecylsulfate and with methanol
successively appears to be the best procedure to remove
chemicals from different classes deposited on hair surface. Such
procedure appears to remove in a one-shot the fraction of
chemicals supposed to be located on hair surface and does not re-
quire repeated washing steps. Moreover, this decontamination
procedure does not seem to inuence the concentration of
chemicals located inside hair, which are considered being incor-
porated frombiological pathways and representative of exposure.
Acknowledgements
This study was carried out in the framework of the call for re-
search project 2010 of the national program Environmental
and Occupational Health (PNR EST) of the French Agency for
Food, Environmental and Occupational Health Safety (ANSES), with
the nancial support of the Ofce National de lEau et des Milieux
Aquatiques (ONEMA) supporting the implementation of the Plan
Ecophyto 2018, France. R.-C. Duca beneted from a postdoctoral
grant from the Fonds National de la Recherche (FNR) (AFR
1069412), Luxembourg.
References
[1] F. Pragst, M.A. Balikova. State of the art in hair analysis for detection
of drug and alcohol abuse. Clin. Chim. Acta 2006, 370, 17.
[2] P. Kintz, P. Mangin. Opiate concentrations in human head, axillary,
and pubic hair. J. Forensic Sci. 1993, 38, 657.
[3] M. Montagna, C. Stramesi, C. Vignali, A. Groppi, A. Polettini. Simulta-
neous hair testing for opiates, cocaine, and metabolites by GCMS: A
survey of applicants for driving licenses with a history of drug use.
Forensic Sci. Int. 2000, 107, 157.
[4] M.I. Schaffer, W.L. Wang, J. Irving. An evaluation of two wash proce-
dures for the differentiation of external contamination versus inges-
tion in the analysis of human hair samples for cocaine. J. Anal.
Toxicol. 2002, 26, 485.
[5] M.A. Balikova, V. Habrdova. Hair analysis for opiates: Evaluation of
washing and incubation procedures. J. Chromatogr. B 2003, 789, 93.
[6] T. Cairns, V. Hill, M. Schaffer, W. Thistle. Removing and identifying
drug contamination in the analysis of human hair. Forensic Sci. Int.
2004, 145, 97.
[7] V. Cirimele, P. Kintz, B. Ludes. Assessment of pesticide exposure by
hair analysis. Acta Clin. Belg. 1999, S1, 59.
[8] A. Toriba, Y. Kuramae, T. Chetiyanukornkul, R. Kizu, T. Makino, H.
Nakazawa, K. Hayakawa. Quantication of polycyclic aromatic hydro-
carbons (PAHs) in human hair by HPLC with uorescence detection:
A biological monitoring method to evaluate the exposure to PAHs.
Biomed. Chromatogr. 2003, 17, 126.
[9] H. Zhang, Z. Chai, H. Sun. Human hair as a potential biomonitor for
assessing persistent organic pollutants. Environ. Int. 2007, 33, 685.
[10] M.G. Margariti, A.M. Tsatsakis. Analysis of dialkyl phosphate metabo-
lites in hair using gas chromatography-mass spectrometry: A bio-
marker of chronic exposure to organophosphate pesticides.
Biomarkers 2009, 14, 137.
[11] G. Salqubre, C. Schummer, M. Millet, O. Briand, B.M.R. Appenzeller.
Multi-class pesticide analysis in human hair by gas chromatography
tandem (triple quadrupole) mass spectrometry with solid phase
microextraction and liquid injection. Anal. Chim. Acta 2012, 710, 65.
[12] W.J. Luksemburg, R.S. Mitzel, R.G. Peterson, J.M. Hedin, M.M. Maier, M.
Schuld, H.D. Zhou, A.S. Wong. Polychlorinated dibenzodioxins and di-
benzofurans (PCDDs/PCDFs) levels in environmental and human hair
samples around an electronic waste processing site in Guiyu,
Guangdong province, China. Organohalogen Compd. 2002, 55, 347.
[13] A. Covaci, C. Hura, A. Gheorghe, H. Neels, A.C. Dirtu. Organochlorine
contaminants in hair of adolescents from Iassy, Romania.
Chemosphere 2008, 72, 16.
[14] C. Schummer, B.M.R. Appenzeller, M. Millet, R. Wennig. Determina-
tion of hydroxylated metabolites of polycyclic aromatic hydrocar-
bons in human hair by gas chromatographynegative chemical
ionization mass spectrometry. J. Chromatogr. A 2009, 1216, 6012.
[15] T. Nakao, O. Aozasa, S. Ohta, H. Miyata. Assessment of human expo-
sure to PCDDs, PCDFs and co. PCBs using hair as human pollution in-
dicator sample I: Development of analytical method for human hair
and evaluation for exposure assessment. Chemosphere 2002, 48, 885.
[16] L. Altshul, A. Covaci, R. Hauser. The relationship between levels of
PCB and pesticides in human hair and blood: Preliminary results.
Environ. Health Persp. 2004, 112, 1193.
[17] W. Tirler, G. Voto, M. Donega. PCDD/F and PCB levels in human hair.
Organohalogen Compd. 2006, 68, 1659.
[18] G. Zhao, Z. Wang, M.H. Dong, K. Rao, J. Luo, D. Wang, J. Zha, S.
Huang, Y. Xu, M. Ma. PBBs, PBDEs, and PCBs levels in hair of residents
around e-waste disassembly sites in Zhejiang Province, China, and
their potential sources. Sci. Total Environ. 2008, 397, 46.
[19] J.L. Tadeo, C. Snchez-Brunete, E. Miguel. Determination of
polybrominated diphenyl ethers in human hair by gas chromatogra-
phymass spectrometry. Talanta 2009, 78, 138.
[20] Y. Kang, H.S. Wang, K.C. Cheung, M.H. Wong, Polybrominated
diphenyl ethers (PBDEs) in indoor dust and human hair. Atmos.
Environ. 2011, 45, 2386.
[21] G.A. Cooper, R. Kronstrand, P. Kintz. Society of Hair Testing guide-
lines for drug testing in hair. Forensic Sci. Int. 2012, 218, 20.
[22] D.L. Blank, D.A. Kidwell. Decontamination procedures for drugs of
abuse in hair: Are they sufcient? Forensic Sci. Int. 1995, 70, 13.
[23] M. Schaffer, V. Hill, T. Cairns. Hair analysis for cocaine: The require-
ment for effective wash procedures and effects of drug concentra-
tion and hair porosity in contamination and decontamination.
J. Anal. Toxicol. 2005, 29, 319.
Hair decontamination
Drug Testing
and Analysis
Drug Test. Analysis 2014, 6, 5566 Copyright 2014 John Wiley & Sons, Ltd. wileyonlinelibrary.com/journal/dta
6
5
[24] P. Kintz. Segmental hair analysis can demonstrate external
contamination in postmortem cases. Forensic Sci. Int. 2012, 215, 73.
[25] S. Lee, E. Han, S. In, H. Choi, H. Chung, K.H. Chung. Analysis of pubic
hair as an alternative specimen to scalp hair: A contamination issue.
Forensic Sci. Int. 2011, 206, 19.
[26] G. Romano, N. Barbera, G. Spadaro, V. Valenti. Determination of
drugs of abuse in hair: Evaluation of external heroin contamination
and risk of false positives. Forensic Sci. Int. 2003, 131, 98.
[27] B.M.R. Appenzeller, A.M. Tsatsakis. Hair analysis for biomonitoring of
environmental and occupational exposure to organic pollutants:
State of the art, critical review and future needs. Toxicol. Lett. 2012,
210, 119.
[28] U.S. National Library of Medicine: http://www.chem.sis.nlm.nih.gov/
chemidplus/chemidheavy.jsp [14 April 2014].
[29] K.W. Schramm, T. Kuettner, S. Weber, K. Lutzke. Dioxin hair analysis
as monitoring pool. Chemosphere 1992, 24, 351.
[30] E.M. Ostrea, E. Villanueva-Uy, D.M. Bielawski, N.C.J. Posecion, M.L.
Corrion, Y. Jin, J.J. Janisse, J.W. Ager. Maternal hair an appropriate
matrix for detecting maternal exposure to pesticides during preg-
nancy. Environ. Res. 2006, 101, 312.
[31] M. Puchades, A. Westman, K. Blennow, P. Davidsson. Removal of so-
dium dodecyl sulfate from protein samples prior to matrix-assisted
laser desorption/ionization mass spectrometry. Rapid Commun. Mass
Spectrom. 1999, 13, 344.
[32] G.S. Winkler, L. Lacomis, J. Philip, H. Erdjument-Bromage, J.Q.
Svejstrup, P. Tempst. Isolation and mass spectrometry of transcrip-
tion factor complexes. Methods 2002, 26, 260.
R.-C. Duca et al.
Drug Testing
and Analysis
wileyonlinelibrary.com/journal/dta Copyright 2014 John Wiley & Sons, Ltd. Drug Test. Analysis 2014, 6, 5566
6
6