Anal Bioanal Chem (2007) 388:1393–1414
DOI 10.1007/s00216-007-1289-9
REVIEW
Application of solid-phase microextraction
in analytical toxicology
Fritz Pragst
Received: 15 February 2007 / Revised: 28 March 2007 / Accepted: 29 March 2007 / Published online: 3 May 2007
# Springer-Verlag 2007
Abstract Solid-phase microextraction (SPME) is a mini-
aturized and solvent-free sample preparation technique for
chromatographic–spectrometric analysis by which the ana-
lytes are extracted from a gaseous or liquid sample by
absorption in, or adsorption on, a thin polymer coating
fixed to the solid surface of a fiber, inside an injection
needle or inside a capillary. In this paper, the present state
of practical performance and of applications of SPME to
the analysis of blood, urine, oral fluid and hair in clinical
and forensic toxicology is reviewed. The commercial
coatings for fibers or needles have not essentially changed
for many years, but there are interesting laboratory
developments, such as conductive polypyrrole coatings for
electrochemically controlled SPME of anions or cations and
coatings with restricted-access properties for direct extrac-
tion from whole blood or immunoaffinity SPME. In-tube
SPME uses segments of commercial gas chromatography
(GC) capillaries for highly efficient extraction by repeated
aspiration–ejection cycles of the liquid sample. It can be
easily automated in combination with liquid chromatogra-
phy but, as it is very sensitive to capillary plugging, it
requires completely homogeneous liquid samples. In
contrast, fiber-based SPME has not yet been performed
automatically in combination with high-performance liquid
chromatography. The headspace extractions on fibers or
needles (solid-phase dynamic extraction) combined with
GC methods are the most advantageous versions of SPME
F. Pragst (*)
Institute of Legal Medicine, University Hospital Charité,
Hittorfstr. 18,
14195 Berlin, Germany
e-mail: fritz.pragst@charite.de
because of very pure extracts and the availability of
automatic samplers. Surprisingly, substances with quite
high boiling points, such as tricyclic antidepressants or
phenothiazines, can be measured by headspace SPME from
aqueous samples. The applicability and sensitivity of SPME
was essentially extended by in-sample or on-fiber derivati-
zation. The different modes of SPME were applied to
analysis of solvents and inhalation narcotics, amphetamines,
cocaine and metabolites, cannabinoids, methadone and other
opioids, fatty acid ethyl esters as alcohol markers, γ-
hydroxybutyric acid, benzodiazepines, various other thera-
peutic drugs, pesticides, chemical warfare agents, cyanide,
sulfide and metal ions. In general, SPME is routinely used
in optimized methods for specific analytes. However, it was
shown that it also has some capacity for a general screening
by direct immersion into urine samples and for pesticides
and other semivolatile substance in the headspace mode.
Keywords Drug analysis by solid-phase microextraction .
Headspace extraction . In-tube solid-phase microextraction .
Sample preparation . Solid-phase microextraction .
Volatile poisons
Introduction
In forensic and clinical toxicology the extraction of blood
(serum, plasma), urine, gastric content, other body fluids or
tissue samples is an important step of almost every analysis
in order to separate the toxic compounds or their metabo-
lites from the complex biological matrix and to exclude a
disturbing response of matrix constituents in the subsequent
chromatographic–spectrometric analysis. The increasing
sensitivity and selectivity of the analytical techniques has
enabled a miniaturization and simplification of the sample-
1394 Anal Bioanal Chem (2007) 388:1393–1414

preparation techniques. One of the most challenging

developments in this area is solid-phase microextraction
(SPME), which was invented by Pawliszyn and coworkers
in 1989 [1] and which has found widespread analytical
applications, among them many in toxicological analysis.
In the original version, the analytes are adsorbed or
absorbed from the sample by a coated fiber and transferred
into the injector of the gas- or liquid-chromatographic
system where they are desorbed by evaporation at high
temperature or by dissolution in the mobile phase. In later
developments, the principle was extended to extraction in
coated capillaries (in-tube SPME) [2], to solid-phase
dynamic extraction by internally coated syringes (SPDE)
[3], to stir-bar sorbtive extraction (SBSE) methods [4] and
to membranes [5, 6]. The general advantages of SPME in
comparison with other extraction methods are that it works
without any organic solvent, including solvent-free injec-
tion, and that it can be performed fully automatically. In
addition, the headspace version of SPME has the advantage
of cleaner extracts and is superior to usual headspace gas
chromatography (GC) because of accumulation of the
analytes on the fiber which can be injected into the gas
chromatograph without an excess of air. Meanwhile, a huge
number of SPME papers were published showing the wide
versatility of the technique, and progress was summarized
in books [7–9] and reviews [10–18].
In the present paper, an overview of the experimental
possibilities and of the applications to forensic and clinical
toxicology shall be given. It will be limited to the use of
Fig. 1 Devices for solid-phase microextraction (SPME): a SPME
fibers, to in-tube SPME and to SPDE, whereas other
fiber, b solid-phase dynamic extraction syringe, c capillary for in-tube
approaches such as SBSE and the use of membranes will SPME. GC gas chromatograph
be excluded.

Practical performance of SPME
In the practical performance of SPME, a number of
experimental details must be considered and specified for
the problem under investigation. This optimal choice of the
analytical conditions depends on the nature and composi-
tion of the matrix, on the properties of the analytes and on
the required sensitivity and accuracy. In this section some
essential aspects shall be considered.
SPME coatings
The extraction and enrichment of the analytes in SPME
occurs by a thin layer of a suitable polymer at the surface of
a fused silica fiber, on the inner wall of a stainless steel
syringe or within a fused-silica capillary (Fig. 1). The
efficiency of the extraction depends decisively on the
properties of these coatings.
SPME fibers
The fibers are arranged in assemblies in which the coated
fiber is fixed on a steel rod which can be exposed or
withdrawn into a needle (Fig. 1a). The needle protects the
fiber and enables penetration of the septa of the sample
vials or of the chromatographic injector without damage.
The only manufacturer of commercially available SPME
fibers is Supelco (Bellefonte, PA, USA), which supplies
fibers consisting of fused silica (length 1 cm, diameter 110
μm) or StableFlex (a flexible fused-silica core, less
breakable, lengths 1 or 2 cm) coated with seven different
single-polymer or mixed-polymer materials (Table 1). The
coatings have different film thickness between 7 and 100
μm and can be nonbonded or bonded to the core. They are
either in a quasi-liquid state (polydimethylsiloxane—
PDMS, polyacrylate—PA, carbowax—CW) or porous solid
particles imbedded into a PDMS or a CW layer (poly-
divinylbenzene, Carboxen—CAR, templated resin–TPR).
General advice for proper selection of the fiber depending
Anal Bioanal Chem (2007) 388:1393–1414 1395

Table 1 Commercially avail-

SPME fiber Coating
able solid-phase microextraction
(SPME) fibers and solid-phase PDMS Polydimethylsiloxane, 100 μm
dynamic extraction (SPDE) PDMS Polydimethylsiloxane, 30 μm
syringes PDMS Polydimethylsiloxane, 7 μm
PA Polyacrylate, 85 μm
PDMS/DVB Polydimethylsiloxane/divinylbenzene, 65 μm
CW/DVB Carbowax/divinylbenzene, 65 μm
CAR/PDMS Carboxen/polydimethylsiloxane, 75 μm
CW/TPR Carbowax/templated resin, 50 μm
DVB/CAR/PDMS Divinylbenzene/Carboxen on polydimethylsiloxane coating, 50/30 μm

SPDE syringe
PDMS Polydimethylsiloxane, 50 μm
PDMS/AC Polydimethylsiloxane + 10% activated charcoal, 50 μm
CT-5 5% diphenyl/95% polydimethylsiloxane , 10 and 50 μm
PEG Poly(ethylene glycol), 10 and 50 μm
CT-1701 14% cyanopropyl/86 % polydimethylsiloxane , 50 μm
CT-225 50% cyanopropyl/50 % polydimethylsiloxane , 50 μm

on polarity, molecular size and volatility of the analytes was discharge (Fig. 2b). These coatings can be used for selective
given [7] and applications for a variety of analytes and EC-SPME of cations such as protonated dopamine.
matrices are accessible from the Supleco Web site. Most For direct extraction from blood, the principle of
applications of SPME in toxicological analysis use one of restricted-access materials (RAM) was adopted from solid-
these fibers. phase extraction (SPE) columns also for SPME [28].
Procedures for noncommercial fiber preparation and Commercial 35-μm LiChrospher RP-18 alkyldiol-silica
corresponding applications were reviewed by Dietz et al. RAM particles were glued to a cleaned stainless steel wire.
[19]. Sol–gel technology [20], electrochemical procedures, These fibers could be applied to blood samples without
e.g., anodic deposition of polypyrrole (PPY) on a platinum previous protein precipitation and were even suitable for in
wire [21, 22], and physical deposition (subsequent dipping vivo extraction directly from animal veins. Finally, also
of the support material into polymer solution) are applied SPME fibers with molecularly imprinted polymers were
techniques. In the sol–gel technique hydroxyl-terminated
siloxane polymers or mixed polymers with polyethylene or
polypropylene glycols are bonded to the Si–OH groups at
the fused-silica surface. Examples prepared in this way are
ultrathin phenyl-functionalized fibers (thickness 0.2–1 μm)
[23] or fibers with benzo-15-crown-5 modified coatings
with an improved selectivity toward aromatic compounds
[24]. Coatings prepared by sol–gel technology have
increased thermal and solvent stability. Another approach
is the fixation of activated carbon powder on the fiber with
silicone adhesive prepared by hydrolysis and polyconden-
sation of chloropropylsiloxane and chloropropylphenylsi-
loxane [25].
PPY coatings are intrinsic conducting polymers which
are positively charged. The properties can be varied by
inclusion of suitable counteranions such as dodecyl sulfate
[26]. If deposited on a metal wire, PPY can be electrochem-
ically discharged and recharged, simultaneously releasing or
Fig. 2 Electrochemically controlled SPME of anions or cations on
taking up counteranions. For this reason, PPY-coated fibers polypyrrole (PPY) coatings [26, 27]. The conductive polymer coating
can be used for electrochemically controlled SPME (EC- is deposited on a platinum wire. a At positive potential, anions A- are
SPME) of anions such as ferrocyanide or methylarsonate as absorbed as counterions to the positively charged polymer. On
potential reversal, A- are desorbed as the polymer is discharged. b
shown in Fig. 2a [27]. However, PPY coatings can be
Polystyrenesulfonate (PSS-) is imbedded as an immobilized polyanion
prepared with bulky and immobile anions such as polysty- in the PPY coating. In this case, cations C+ are adsorbed at negative
renesulfonate, which are not released by electrochemical potential and released at positive potential
1396
prepared [29, 30]. Fiber coatings with a high selectivity for
clenbuterol and bromobuterol from urine were obtained by
silylation of the silica fiber and subsequent immersion for
12 h under UV irradiation in a polymerization solution
composed of clenbuterol, methacrylic acid, ethylene glycol
dimethacrylate and azobis(isobutyronitrile) dissolved in
acetontrile [29]. Another novel strategy is the synthesis of
monolithic imprinted fibers using silica capillaries as a
mold which are etched away after polymerization [30].
With propazine as the template molecule, fibers with high
recognition for triazine herbicides were prepared. Experi-
ments were also performed concerning the use of immu-
noaffinity fibers for drug extraction [31, 32]. The drug
antiserum was covalently immobilized on the surface of a
fused-silica fiber, which was modified before with 3-
aminopropyltriethoxysilane and glutaraldehyde, and was
used as a selective and sensitive extraction medium for the
immunoaffinity SPME in serum samples.
SPDE syringes
Commercial syringes are available from Chromtech
(Idstein, Germany). The gastight syringes have a volume
of 2.5 ml and the stainless steel needles (length 56 or
76 mm, inner diameter 500 μm) are internally coated with a
10- or 50-μm layer of six different coatings (Fig. 1b, Table 1).
In addition, a variety of specific custom coatings have been
developed by this company. In clinical or forensic applica-
tions, they are preferentially used for headspace sampling
[3, 33, 34].
In-tube SPME capillaries
This version of SPME is generally limited to liquid
samples. No special capillaries are supplied by any
manufacturers for in-tube SPME. Instead, sections of
conventional commercial capillaries intended for GC are
applied [2]. Correspondingly, a wide variety of coatings are
available. Good experiences have been described for a
capillary length of 60 cm and an internal diameter of
0.25 mm (Fig. 1c). As the layers are relatively thin (e.g.,
0.25 μm), the extraction equilibrium is quickly attained.
Applications were published with the coatings Omegawax
250 (poly(ethylene glycol) [35]), Supel-Q-PLOT (porous
divinyl polymer [36]). In addition, also individually
prepared coatings on capillaries were described. An in-tube
PPY film was synthesized by first passing a pyrrol/2-
propanol mixture (1:1, v/v) and then 0.2 M ferric
perchlorate in 0.4 M HClO4 through the capillary column
[22]. This procedure was repeated several times in order to
obtain sufficient film thickness. The film could be coated
only on polar pretreated capillaries, e.g., poly(ethylene
glycol). In order to further increase the extraction efficiency
Anal Bioanal Chem (2007) 388:1393–1414
by decreasing the capillary volume or by increasing the
extracting surface, a steel wire was inserted into the
capillary (“wire-in-tube configuration” [37]) or a 10-cm-long,
0.25-mm inner diameter polyetheretherketone (PEEK) tube
was packed with 280 filaments of the same lengths and a
diameter of 11.5 μm of the polymer Zylon® (fiber-in-tube
configuration [38]). Similarly, an 8-cm PEEK tube (inner
diameter 0.76 mm) was filled with molecularly imprinted
polymer particles for the selective extraction of propranolol
[39].
Headspace SPME
The extraction of volatile and semivolatile analytes from
the vapor phase above a sample is the most frequent
application of fiber-based SPME. The steps of this analysis
are shown in Fig. 3 and can be performed manually or
automatically. It is a further advantage besides the efficient
separation from nonvolatile disturbing matrix components
and the enrichment of the substances on the fiber that also
fiber damage by aggressive or irreversibly adsorbed matrix
components is minimized. Therefore, as a rule, more than
100 samples can be analyzed by the same fiber. The
coupling with GC is technically analogous to the injection
of a liquid sample and is combined with the reconditioning
of the fiber. The coupling with liquid chromatography (LC)
will be dealt with in a later section.
In general, the analysis is performed from aqueous
samples (blood, urine, hair hydrolysate, etc.). The thermo-
dynamics and kinetics of the partition processes between
the three phases (sample, headspace and fiber coating) were
thoroughly investigated by Pawliszyn [7, 9, 38]. There
exists a linear relationship between the amount of the
analyte on the fiber extracted and its initial concentration
cs,0 in the sample if the extraction time is long enough for
equilibration, the dynamic range of the method is not
exceeded and all other experimental conditions (sample
composition, temperature, volumes of sample and head-
space) are held constant (Eq.(1)).
n¼ const  cs;0 ; ð1Þ
where
const ¼Kfh Khs Vf Vs =ðKfh Khs Vs þKhs Vh þVs Þ: ð2Þ
Kfh and Khs are equilibrium constants for the fiber and
the headspace and the headspace and the sample, respec-
tively, and Vf, Vh and Vs are the volumes of the fiber
coating, the headspace and the sample, respectively.
However, the time to reach the partition equilibrium
depends on the sample conditions and the nature of the
analyte and ranges from a few minutes to several hours.
Nevertheless, Eq. (1) proved to be valid from a theoretical
Anal Bioanal Chem (2007) 388:1393–1414 1397

Fig. 3 Steps in the analysis by

headspace SPME (HS-SPME).
The fiber is withdrawn into the
needle outside and for penetra-
tion of the septa of the vial and
the GC injector and is exposed
for absorption or desorption
of the analytes in the vial and
the injector

point of view as well as in practice if the extraction is
interrupted after a constant time before the equilibrium is
attained [41].
In practice, there are some severe limitations to the
validity of Eq. (1). It can be seen from Eq. (2), that the
proportionality factor contains the equilibrium constants Kfh
and Khs for the fiber and the headspace and the headspace
and the sample, respectively. But, both Kfh and Khs can be
strongly affected by variations in the sample composition.
The vapor pressure of an analyte above the aqueous sample
(that means Khs) can be decreased in the presence of small
amounts of organic solvents such alcohol or of surfactants
which increase the solubility in the aqueous phase. On the
The experimental optimization of HS-SPME methods
has been described in a large number of papers and is
always performed according to the same scheme: choice of
fiber, amount of sample, sample pH, addition of salt,
temperature and time of extraction, temperature and time of
desorption plus reconditioning in the GC injection port. An
important characteristic of the method is the absolute
25
Analyte / Int. standard

EDDP
other hand, an excess of volatile matrix constituents can 20
Peak area ratio

lead to a saturation of the fiber coating and can decrease Kfh

for the analyte. This is particularly severe for fibers with
solid porous coatings such as PDMS/divinylbenzene 15
(DVB), CW/TPR or CAR (Table 1) which extract primarily
Methadone
via adsorption. In this case, the amount extracted is
10
described by an adsorption isotherm with only a small
quasi-linear range and a saturation at higher concentrations.
In contrast to the absorption in liquid coatings, adsorption is 5
a competitive process which leads to mutual displacement
from the adsorption sites between different analytes or
0
analytes and matrix constituents; therefore, an internal
0 5 10 15 20 25 30 35
standard is always required for quantitative headspace
SPME (HS-SPME) measurements. Since the effects de- Concentration (ng/mg)
scribed above are quite different for different substances, Fig. 4 Calibration curve of the HS-SPME determination of metha-
the internal standard should have a structure very similar to done and its main metabolite 2-ethylene-1,5-dimethyl-3,3-diphenyl-
that of the analyte. Deuterated standards are most suitable, pyrrolidine (EDDP) with methadone-d9 and EDDP-d3 as internal
standards. The deviations from linearity are explained by different
but even in that case strong deviations of the calibration
affinity of the deuterated and nondeuterated compounds for the
curve from linearity are possible as was found for adsorption sites of the polydimethylsiloxane/divinylbenzene fiber
methadone/methadone-d9 [42] (Fig. 4). and discrimination effects [42]
1398
extraction yield which is determined by comparison of the
GC–mass spectrometry (MS) peak area obtained from the
same amount of sample after HS-SPME and after direct
injection. From a structural point of view, this yield
depends mainly on the volatility and the hydrophobicity of
the analyte. The values range from 0.1 to more than 60 %
[43]. They are much higher from pure aqueous buffer than
from blood or hair hydrolysate. Obviously, the organic
matrix constituents increase the analyte solubility in the
sample by complexation and tenside effects. Nevertheless,
it is surprising that drugs such as phenothiazines and
tricyclic or tetracyclic antidepressants with boiling points
well above 250 °C can be headspace-extracted from such
very low concentration aqueous solutions. A possible
explanation is that these “semivolatile” hydrophobic sub-
stances are accumulated at the surface of the aqueous
phase.
Since the fiber is completely arranged in the sample vial,
it usually has the same temperature as the sample. The HS-
SPME yield could be increased by cooling the fiber
(increasing Kfh in Eq. (2)). For this purpose, an internally
cooled coated fiber device was recently developed [44]. The
coating was deposited at the lower closed end of a 0.6-mm
inner diameter stainless steel tube which contained a
thermocouple for control of the temperature and an inner
stainless steel tube (inner diameter 0.15 mm, outer diameter
0.30 mm) for delivering liquid CO2 for cooling. The rate of
liquid CO2 inflow was adjusted by a solenoid valve in such a
way that the preset temperature was maintained. With use of
this device, the fiber temperature could be held at 0 °C in a
sample vial of 200 °C. The sensitivity could be significantly
improved in applications with this cooled fiber [45, 46].
In analogy to static multiple headspace extraction GC, a
multiple HS-SPME technique was developed [47, 48]. This
technique allows the direct analysis of solid and heteroge-
neous materials (e.g., tissues) by repeated measurement of
the same sample with an easy possibility to eliminate the
influence of the matrix. Until now no applications to human
material have been described.
Headspace SPDE
Although headspace SPDE (HS-SPDE) has obvious advan-
tages compared with HS-SPME (larger coating volume,
shorter extraction time, better mechanical stability), only
limited use has been described in the literature [3, 33, 34,
49–51]. The method is always performed automatically. In
a typical experiment, the sample is placed in a 10-ml
headspace vial in the heating station of the sampler, and
after preincubation the internally coated needle of the
heated 2.5-ml gastight syringe is injected into the headspace
through the septum. A 1-ml volume of the headspace is
aspirated and returned through the capillary 50 times with a
Anal Bioanal Chem (2007) 388:1393–1414
plunger speed of 50 μl/s. Then, the syringe is filled with 2.5
ml of high-purity gas (e.g., nitrogen) from a gas station and
the needle is placed into the injection port of the gas
chromatograph. There, the analytes are desorbed at high
temperature with a plunger speed of 10 μl/s and are
concentrated on the capillary column inlet at sufficiently
low oven temperature. For cleaning and reconditioning and
in order to avoid carryover, the syringe and the needle are
purged with inert gas in an optional fiber cleaning station at
a suitable temperature between 30 and 350 °C. Another
advantage in comparison with HS-SPME is that the needle,
a considerable part of which is outside the sample vial, can
be cooled more easily by a Peltier SPDE extraction cooler
supplied by the manufacturer. This is particularly important
for volatile substances such as solvents and HCN.
Direct SPME with fibers from liquid samples
The direct extraction of the analytes by immersion of the
SPME fiber into the solution (direct immersion SPME—
DI-SPME) has to be performed for analytes with no or very
low volatility. From a thermodynamic point of view, the
same principles have to be taken into account for a good
extraction recovery as in the headspace version. However,
since the sample is in direct contact with the fiber, strong
acidic or alkaline conditions must be avoided. Fibers with
polymers bonded to the silica surface are preferentially
suitable for DI-SPME (Table 1). In a typical experiment,
the fiber is exposed to the liquid sample at constant
temperature between 30 and 100 °C and with constant
agitation (mostly stirring) for 15–60 min and then directly
or after a very short washing step is introduced into the GC
or high-performance LC (HPLC) injection system.
The sample pretreatment is different for urine or blood,
serum and plasma. Urine samples are mostly diluted (e.g.,
1:10 v/v) with a suitable buffer and filtrated through a
Millipore filter in order to suppress matrix effects and to
prevent contamination of the fiber. For whole blood, plasma
or serum samples, a deproteination is recommended for two
reasons. Firstly, the proteins may be adsorbed at the fiber
and prevent the efficient extraction of the drug. In addition,
they increase the matrix viscosity and reduce the extraction
rate. Secondly, the extraction yield is strongly affected by
the drug–plasma protein binding, which may be different
from sample to sample. For instance, in the case of
lidocaine, by direct extraction from buffered plasma a yield
of only 0.4% was obtained after equilibration, whereas after
deproteination the yield was 1.0–1.3% [52]. Usually,
perchloric acid or trichloroacetic acid are used for depro-
teination. It has to be taken into account that a part of the
drug is removed together with the protein. By consideration
of the pK values, the pH has to be chosen in a way that the
analytes are in the electroneutral state. As a rule, the
Anal Bioanal Chem (2007) 388:1393–1414
recovery is strongly increased by addition of salts (out-
salting effect).
The extraction process may be affected by bubbles on
the lipophilic fiber surface, which can be removed by
repeated immersion and retracting the fiber from the
solution. The extraction rate is increased at elevated temper-
atures. For this reason, DI-SPME is preferentially performed
at 50–100 °C. Usually, the fiber is not washed between
extraction and injection into the chromatographic system;
however, particularly for combination with GC, a short
washing step (e.g., 5-s immersion in deionized water) had an
important role in preventing protein burden in the fiber when
extracting blood samples. Furthermore, salt accumulation on
the surface is avoided. In other cases, a washing sequence
proved to be useful: 20 s water, 20 s 50% methanol, 2-min
drying. If aggressive conditions are avoided, the fibers can
be quite stable also in direct contact with the samples. More
than 100 reproducible extractions were performed with the
same fiber in several DI-SPME studies [53–55].
Interfacing SPME with LC
Whereas the injection of the SPME fiber or of the SPDE
needle into the gas chromatograph is completely analogous
to the injection of liquid samples and is very easily
accomplished, there is not such an obvious possibility for
LC. Strategies for coupling SPME with LC were reviewed
by Zambonin [56] and Lord [57]. At present, there are only
two ways to solve the problem: the SPME fiber interface
and the in-tube SPME.
Fig. 5 a Manual SPME fiber
interface for high-performance
liquid chromatohgraphy (HPLC)
in the dynamic desorption posi-
tion. b Heated interface from
[59, 60]
1399
SPME fiber interface
No automated performance has been described to date. The
practical device for manual on-line desorption from the SPME
fiber which is commercially available from Supelco has not
essentially changed since it was first introduced by Pawliszyn
in 1995 [58]. The principle is shown in Fig. 5a. The T-shaped
desorption chamber is located in the position of the injection
loop of a usual Rheodyne six-port valve. After fiber
introduction and exposition into the 75-μl desorption
chamber under ambient pressure in the loading position, it
is sealed by a PEEK ferrule around the support rod of the
fiber in order to withstand the HPLC precolumn pressure.
Then, a solvent for dissolution of the analytes from the fiber
can be introduced by the syringe (static desorption),
followed by switching the valve to the injection position or
it is immediately switched to the injection position for
desorption by the mobile phase (dynamic desorption). The
main problems are the possible damage of the fiber and the
insufficient elasticity of the ferrule used to tightening and
release the fiber rod in repeated use.
Furthermore, peak broadening or tailing has been noted.
The reasons are slow dynamic desorption or too large a
volume of the desorption chamber as well as higher eluent
strength of the desorbtion solvent in the static mode. This
has been overcome by shortening the injection time (fast
switching back to the loading position) or by reducing the
flow rate during injection.
An improved heated interface was developed by
Rodrigues et al. [59, 60] (Fig. 5b). Heating to 50 °C of
1400
the 60-μl desorption chamber filled with the mobile phase
improved the detection sensitivity by almost a factor of 10.
In-tube SPME
In contrast to fiber SPME, in-tube SPME can be completely
automated using commercial autosamplers [2, 57]. The
principle and the extraction steps are shown in Fig. 6.
Generally, the capillary is placed either between the needle
of an autosampler and the injection valve or in place of the
injection loop of the valve. For extraction, the sample is
aspirated and dispensed to and from the capillary a number
of times until either an equilibrium is reached or sufficient
analyte is extracted for the desired sensitivity. For static
desorption, a solvent is drawn into the capillary and then
with the desorbed analytes is sent into the injection loop of
the valve. If desorption is efficient in the initial mobile
phase, the mobile phase is directly passed through the
capillary to the column for dynamic desorption. Owing to
the potential for plugging by heterogeneous impurities, the
technique is limited to homogeneous and fairly pure
samples. Similar to the other types of SPME, an in-tube
SPME method must be optimized for several parameters.
For example, for a capillary column (inner diameter
0.25 mm, length 60 cm) with a volume of 30 μl and a
capacity of the injection needle of 10 μl, 30–40-μl draw/
ejection volume, 50–100-μl/min draw/ejection flow rate and
ten to 15 draw/ejection cycles proved to be optimal [2]. For
avoidance of carryover, washing or rinsing can easily be
included into the automated procedure.
Derivatization in SPME
Generally, derivatization in combination with SPME is
important if the extract is analyzed by GC in order to
improve the extraction or the chromatographic performance.
The reaction may occur in the sample, on the fiber or in the
injection port. It will be shown in a later section for the
Fig. 6 In-tube SPME as imple-
mented with a commercial
HPLC autosampler. a Extraction
by repeated aspiration and dis-
pensing of the sample to and
from the capillary. b Dynamic
desorption by the mobile phase
Anal Bioanal Chem (2007) 388:1393–1414
example of amphetamines that all of these alternatives can
successfully be applied to the same substance.
In-sample derivatization
In this case the reagent is added directly to the sample and
the derivative is formed before SPME. There is no change
in the principle of the performance of SPME. Derivatization
increases the extraction yield because of higher volatility and
lipophilicity of the derivatives. The reagent must be sufficiently
stable to hydrolysis since, as a rule, SPME is performed from
aqueous samples. This is the case for alkylchloroformates
which are used to transform amino groups into the
corresponding alkyl carbamates [61–64] and, in combination
with pyridine, carboxylic acids into the corresponding esters
[65, 66]. For instance, benzoylecgonine [65] and valproic acid
[66] were determined in this way by SPME. Another example
is the transformation of busulfane [1,4-bis-(methylsulfonoxy)
butane] into 1,4-diiodobutane with sodium iodide [67].
However, most derivatization reagents are sensitive to
hydrolysis. They can be used if dry residues of extracts from
a biological sample are cleaned up and injected using the HS-
SPME technique. As an example, cannabinoids in a dry residue
of a hair extract were derivatized with N,O-bis(trimethylsilyl)
trifluoroacetamide (BSTFA) in combination with HS-SPME,
leading to a fivefold increase in sensitivity in comparison with
liquid injection of the derivatives [68]. In the same way, 24
drugs were detected from the residue of liquid/liquid extracts
by addition of 2 μl N-methyl-N-trimethylsilyltrifluoroaceta-
mide (MSTFA) and subsequent HS-SPME at 200 °C or by
exposition of the fiber which was doped with an acetanhy-
dride/pyridine agent into the headspace vial at 200 °C for in
situ on-fiber derivatization [69].
On-fiber derivatization
The on-fiber derivatization may occur simultaneously with
extraction, or subsequent to extraction. In the first case, a
Anal Bioanal Chem (2007) 388:1393–1414 1401

derivatization reagent with low volatility is deposited on the
fiber prior to HS-SPME. An example is the derivative HS-
SPME of carboxylic acids with 1-pyrenyldiazomethane
(Fig. 7) [70, 71]. An advantage of this procedure is that
traces of volatile substances can be captured as a derivative
on the fiber and can be detected with very high sensitivity.
Further examples are aldehydes or acetone from blood
which are on-fiber derivatized with O-(2,3,4,5,6-
pentafluorobenzyl)hydroxylamine [72, 73]. For on-fiber
derivatization subsequent to extraction, the fiber is exposed
after SPME to the vapor phase above the derivatization
reagent in a separate vessel. Reagents used for this purpose
are MSTFA [74], N-methyl-N-(tert-butyldimethylsilyl)tri-
fluoroacetamide [75], hexamethyldisilazane [76], N-meth-
ylbis(trifluoroacetamide) [77], pentafluorobenzoyl chloride
[78] or trifluoroacetic anhydride [79]. Complications of the
on-fiber version can be loss of substance or derivative in
the derivatization vial and carryover.
Derivatization in the injection port of the gas
chromatograph
An example is the direct ion-pair extraction of alkylben-
zensulfonates in the presence of tetrabutylammonium ions
on a PDMS fiber [80]. The ion pairs are transformed into
the corresponding sulfonated butyl esters in the injection
port of the gas chromatograph at 300 °C. In the same way,
carboxylic acids are methylated by coextracted phenyl-
trimethylammonium ions in the injection port [81].
Applications of SPME in analytical toxicology
The detection and quantitative determination of illicit and
therapeutic drugs, pesticides, solvents and other poisons
from blood (serum, plasma), urine, hair or human tissues is
one of the most important applications of SPME in
combination with chromatographic–spectroscopic methods
and was reported in a large number of papers. In general,
the samples must be brought into a homogeneous aqueous
solution by a suitable pretreatment such as homogenization,
protein precipitation or centrifugation. Hair samples must
be first digested by NaOH or extracted with a suitable
solvent. Apart from these matrix-dependent differences in
sample pretreatment, the SPME conditions are determined
by the structure and the properties of the analyte (volatility,
lipophilicity, pKa) rather than by the sample material.
The use for toxicological screening procedures (general
unknown analysis) has not yet been systematically evalu-
ated although there have been some investigations in this
direction [53, 69, 82–84]. As an example, 40 stimulants or
narcotics were identified from urine with detection limits

Fig. 7 Practical performance of

on-fiber derivatization for deter-
mination of fluoroacetic acid
from blood [71]. Pyrenyldiazo-
methane (PDAM) is loaded onto
the fiber from the n-hexane
solution in the washing station
of the sampler. During the sub-
sequent headspace extraction,
the acid is on-fiber-transformed
into pyrenylmethyl fluoroace-
tate, which is measured by gas
chromatography–mass spec-
trometry (GC-MS) with high
sensitivity
1402
between 20 and 200 ng/ml by DI-SPME with a PDMS/
DVB fiber and GC-MS in full-scan mode [53]. More than
100 samples could be measured with one fiber.
However, most applications of SPME have been
described for single compounds or small groups of
compounds. Some toxicologically relevant substance
groups or single substances shall be described in this part
of this review. The SPME mode and the analytical
conditions are presented in Table 2.
Solvents and inhalation narcotics
HS-SPME or HS-SPDE in combination with GC-MS can
be an efficient alternative to static headspace GC-MS in the
analysis of volatile substances because of the accumulation
at the fiber and since the injection of excessive air is
avoided. In a screening procedure, detection limits between
5 and 120 pg/ml were described for 14 halogenated
alkanes, five halogenated alkenes and ten aromatic com-
pounds in blood [85]. Halothane was quantitatively
determined in blood of two homicide victims using
enflurane as an internal standard [86] and chloroform, a
toluene/xylene mixture and a brake cleaner consisting of
various alcohols and alkanes were detected in sniffing
fatalities with probable autoerotic background [87]. HS-
SPME was also used for elucidation of fatal poisoning
cases with dichloromethane [88] and a mixture of trichlo-
roethylene and tetrachloroethylene [89]. The method
appeared to be 10 times more sensitive in determination
of toluene in blood and urine of glue sniffers than the usual
headspace technique [90]. Dichlorobenzene isomers were
determined from blood of a patient who had drunk a
pesticide-like material [91].
Amphetamines and related substances
A general screening procedure for illegal drugs using
SPME has not been described yet but there have been
some reports about the simultaneous detection of several
drugs [92–95]. Most applications of SPME in illegal drug
analysis from human samples were performed for amphet-
amines and related designer drugs [35, 49, 62–64, 77–79,
93–116]. The technique was applied to blood [97, 102, 103,
105, 115], urine [35, 64, 66, 79, 96, 99–101, 107, 110, 111,
113, 115, 116], hair [49, 62, 63, 77, 95, 98, 106, 108, 114]
and oral fluid [93, 94, 108]. Amphetamine, methamphet-
amine, methylenedioxymethamphetamine and methylene-
dioxyamphetamine (MDA) were involved in almost all the
studies. In addition, methylenedioxyethamphetamine [62,
77, 93, 108, 110], N-methyl-1-(1,3-benzodioxol-5-yl)-2-
butanamine [77, 93, 108], 1-(1,3-benzodioxol-5-yl)-2-
butanamine [77], dimethamphetamine [99], fenfluramine
Anal Bioanal Chem (2007) 388:1393–1414
[103], selegiline and desmethylselegiline [115], phenter-
mine [104], phenethylamine [104], 4-bromo-2,5-dimetho-
xyphenethylamine [104] and norephedrine [111] were
included in some methods. A screening procedure for 21
amphetamine-related compounds in urine was described
[100].
As the substances are stable in an alkaline medium, hair
samples were generally digested with aqueous NaOH prior
to or in combination with SPME. The technique was
applied in the headspace mode and in combination with
GC-MS in almost all the studies. The method proved to be
very efficient and enabled the determination of amphet-
amine and methamphetamine from 1-cm segments of a
single hair (weight 0.08 mg) with detection limits of
0.25 ng/mg [114]. In one study in which other drugs were
included [95], 1 M HCl was used for hair extraction in
order to avoid alkaline hydrolysis of cocaine.
Although these substances can be detected in a non-
derivatized state [93, 95, 96, 98–100, 108, 110], the
performance was much improved by derivatization. All kinds
of combinations of SPME with derivatization were realized
(Fig. 8). For in-sample derivatization, ethylchloroformate
[104], propylchloroformate [101, 114], butylchloroformate
[62, 94], hexylchloroformate [116] and 9-fluorenylmethyl-
chloroformate [64] were used. The alkylcarbamates formed
were sufficiently volatile for headspace extraction. In case of
the 9-fluorenylmethyl derivative, DI-SPME and HPLC with
fluorescence detection were applied [64]. Other in-sample
derivatization procedures in blood samples were performed
with pentafluorobenzyl bromide in the presence of tri-n-
propylamine [105] and in hair hydrolysis solution with
heptafluorobutyryl chloride [106]. For enantioselective deter-
mination, the amphetamines were in-sample-derivatized with
o-phthaldialdehyde/N-acetyl-L-cysteine and the products
were extracted by DI-SPME and were analyzed by HPLC
with fluorescence detection [111]. As a disadvantage, this
derivatization is possible only for primary amino groups
(e.g., amphetamine, MDA, norephedrine). Amphetamines
with both primary and secondary amino groups were in-
sample-derivatized with (S)-(-)-N-(trifuoroacetylprolyl) chlo-
ride for HS-SPME in combination with GC-MS [112]. In this
way, the enantiomers of amphetamine and methamphetamine
were well separated with detection limits of 30 ng/ml from
urine.
On-fiber derivatization was in most cases subsequent to
the extraction performed by exposing the fiber to the vapor
of trifluoroacetic anhydride [79], heptafluorobutyric anhy-
dride [102], N-methylbis(trifluoroacetamide) [49] or penta-
fluorobenzoyl chloride [78]. As an example for
derivatization simultaneously with extraction, the fiber
coating was loaded with pentafluorobenzoyl chloride in
the vapor phase above the reagent and then directly
Anal Bioanal Chem (2007) 388:1393–1414 1403

Table 2 Applications of SPME for analysis of toxic compounds

Substance Matrix SPME mode Coating Extraction Detection LOD Remarks References
conditions (ng/ml or
ng/mga)

Solvents, inhalation narcotics

31 volatile Blood HS- 75 μm NaF, Na2CO3, GC-MS 0.005–0.12 Spiked samples [85]
compounds SPME CAR/PDMS 40 °C, 6 min
Halothane Blood a. o. HS- 100 μm PDMS (NH4)2SO4,H2SO4 GC-MS 4 2 homicides [86]
SPME 100 °C, 15 min
Chloroform Blood a. o. HS- 100 μm PDMS (NH4)2SO4, GC-MS – Sniffing fatality [87]
SPME 100 °C, 15 min
Toluene Blood a. o HS- 100 μm PDMS (NH4)2SO4, GC-MS – Sniffing fatality [87]
SPME 100 °C,15 min
o-Xylene, m-xylene, Blood a. o HS- 100 μm PDMS (NH4)2SO4, GC-MS – Sniffing fatality [87]
p-xylene SPME 100 °C,15 min
Dichloromethane Urine HS- 75 μm NaCl, 22 °C, GC-MS 0.005 Accident [88]
SPME CAR/PDMS 30 min
Trichloroethylene Urine HS- 75 μm NaCl, 22 °C, GC-MS 0.005 Workplace occupation [88]
SPME CAR/PDMS 30 min
Perchloroethylene Urine HS- 75 μm NaCl, 22 °C, GC-MS 0.005 Workplace occupation [88]
SPME CAR/PDMS 30 min
Trichloroethylene Blood a. o. HS- 100 μm PDMS No additives, GC-ECD 5 Fatal poisoning [89]
SPME 60 °C,1 min
Tetrachloroethylene Blood a. o HS- 100 μm PDMS No additives, GC-ECD 3 Fatal poisoning [89]
SPME 60 °C,1 min
Toluene Blood, HS- 100 μm PDMS Dilution 1:3 H2O, GC-MS – Glue sniffer [90]
urine SPME 60 °C, 1 min
3 isomeric Blood HS- 100 μm PDMS No additives, GC-MS 20 Poisoning case [91]
dichlorobenzenes SPME 30 °C, 15 min

Amphetamines and related substances

A, MA Blood HS- 100 μm PDMS Borate buffer, GC-MS 0.5 Derivatized with PFBBr [105]
SPME 90 °C, 30 min
A, MA Urine HS- 100 μm PDMS KOH, NaCl, GC-MS 0.3 Derivatized with HFBA [107]
SPME 100 °C, 20 min
A, MA Oral fluid DI- 100 μm PDMS NaHCO3,K2CO3, GC-MS 0.5-5 Derivatized with BCF [94]
SPME RT, 20 min
A, MA Hair HS- 50 μm PDMS NaOH, 50 °C, GC-MS 0.04–0.05 Derivatized with MBTFA [49]
SPDE 50 cycles, 1 ml
MDMA, MDE, MDA Urine DI- 100 μm PDMS pH 10.8, RT, GC-MS 5–15 Derivatized with PCF [101]
SPME 16 min
MDMA, MDE, MDA Hair HS- 50 μm PDMS NaOH, 50 °C, GC-MS 0.03–0.19 Derivatized with MBTFA [49]
SPDE 50 cycles, 1 ml
MDMA, MDE, MDA Oral fluid DI- 30 μm PDMS NaOH, NaCl, RT, GC-MS 1–5 [93]
SPME 30 min
MBDB, BDB Hair HS- 50 μm PDMS NaOH, 50 °C, GC-MS 0.07–0.18 Derivatized with MBTFA [49]
SPDE 50 cycles, 1 ml
Fenfluramine Blood HS- 100 μm PDMS K2CO3, NaCl, GC-MS 5 Derivatized with ECF [104]
SPME 80 °C, 15 min
Selegiline/ Blood, HS- DVB/CAR/ NaOH, NaCl, GC-MS 0.01–0.05 Parkinson disease patient [115]
norselegiline urine SPME PDMS 90 °C, 25 min

Cocaine and metabolites

Cocaine, Plasma DI- 100 μm PDMS pH 9, NaCl, RT, GC-MS 11–19 Drug abusers [121]
cocaethylene SPME 25 min
Cocaine, Urine DI- 100 μm PDMS pH 9–10, RT, GC-MS 5 Patient in withdrawal [120]
cocaethylene SPME 20 min treatment
Cocaine, Hair DI- 100 μm PDMS pH 8.5, NaCl, RT, GC-MS 0.1 Digested with pronase [117]
cocaethylene SPME 25 min dithiothreitol
Cocaine, Sweat DI- 100 μm PDMS pH 5, RT, 20 min GC-MS 12.5 ng/patch PharmCheck™ sweat patch [118]
cocaethylene patches SPME
Benzoylecgonine Urine DI- 100 μm PDMS pH 7, 55 °C, GC-MS 0.03 Derivatized with HCF, [65]
SPME 10 min confirmed with Meth.
Benzoylecgonine Hair DI- 100 μm PDMS pH 9–10, RT, GC-MS 0.5 MeOH extractoin, derivatized [117]
SPME 20 min with BCF

Cannabinoids
THC, CBD, CBN Oral fluid DI- 30 μm PDMS No reagents, RT, GC-MS 1 [123]
SPME 30 min
THC, CBD, CBN Hair HS- 50 μm PDMS Na2CO3, 90 °C, GC-MS 0.09–0.14 Derivatized with MSTFA [50]
SPDE 20 min
THC, CBD, CBN Hair HS- 100 μm PDMS Extr. residue, GC-MS 0.01 Derivatized with BSTFA/TMCS [68]
SPME 125 °C, 20 min
1404 Anal Bioanal Chem (2007) 388:1393–1414

Table 2 (continued)
Substance Matrix SPME mode Coating Extraction Detection LOD Remarks References
conditions (ng/ml or
ng/mga)

Opioids and related substances

Methadone, EDDP Plasma DI- 100 μm PDMS pH 9, RT, GC-MS 5–9 Methadone patients [127]
SPME 30 min
Methadone, EDDP Urine DI- 100 μm PDMS pH 11, NaCl, RT, GC-NPD 1 Drug addicts [92]
SPME 30 min
Methadone, EDDP Oral fluid DI- 100 μm PDMS pH 9.2, NaCl, RT, GC-MS 8–40 Methadone patients [125]
SPME 30 min
Methadone, EDDP, Hair HS- 65 μm NaOH, HCl, GC-MS 0.03–0.05 Drug fatalities [42]
EMDP SPME PDMS/DVB 110 °C, 20 min
Methadone, EDDP Hair HS- 50 μm PDMS Na2CO3, 90 °C, GC-MS- 0.006–0.009 Derivatized with MBTFA [33]
SPDE 20 min MS
Tramadol Plasma HS- 65 μm NaOH, 100 °C, GC-MS 0.2 Healthy volunteers [128]
SPME PDMS/DVB 30 min
Tramadol Hair HS- 85 μm PA NaOH, 90 °C, GC-MS 0.1 Chronic treatment [43]
SPME 20 min
Fentanyl Plasma HS- PDMS, own pH 12, 85 °C, GC-MS 0.01 Fentanyl patch treatment [129]
SPME preparation 30 min
Sufentanil Plasma DI- PDMS/DVB pH 10.6, NaCl, RT, GC-MS 0.4 Intensive care patients [130]
SPME 30 min

Ethanol and alcohol markers

Ethanol Blood, HS- 85 μm PA (NH4)2SO4, GC-FID 1,000 Postmortem samples [131]
urine SPME 60 °C, 1 min
Fatty acid ethyl esters Hair HS- 65 μm pH 7.4,Na2SO4, GC-MS 0.01–0.04 Detection of alcohol abuse [132, 133]
SPME PDMS/DVB 90 °C, 30 min
Fatty acid ethyl esters Skin HS- 65 μm pH 7.4,Na2SO4, GC-MS 0.3–1 Wipe and patch test [135]
surface SPME PDMS/DVB 90 °C, 30 min
lipids

γ-Hydroxybutyric acid
GHB, GBL Plasma, HS- CW/TPR, H2SO4, Na2SO4, GC-MS 50–100 Conversion of GHB to GBL [136, 138]
urine SPME CAR/PDMS 65 °C, 15 min
GHB Water DI- 70 μm No additives, RT, GC-MS 1,500 Derivatized with BSTFA/TMCS [139]
SPME CW/DVB 15 min
GHB Urine DI- 100 μm PDMS pH 7, 40 °C, 20 min GC-MS 200 Derivatized with HCF/pyridine [137]
SPME

Benzodiazepines
Benzophenones Urine DI- 100 μm PDMS pH 9.4, RT, 30 min GC-ECD 80–160 Hydrolysis of 8 benzodiazepines [141]
SPME
6 benzodiazepines Urine, DI- 85 μm PA pH 6.0, NaCl, RT, GC-NPD 3–140 Volunteer [140]
serum SPME with octanol 15 min
7 benzodiazepines Urine, In-tube Supelco-Q-plot pH 8.5, 10 draw/ LC-ESI- 0.2–2 60 cm capsule, spiked samples [36]
serum SPME capillary eject cycles MS
4 benzodiazepines Whole DI- ADS-coated fiber No additives, RT, LC-ESI- 20–35 Volunteers [28]
blood SPME 30 min MS
Midazolam Plasma DI- 85 μm PA pH 6.5, 50 °C, GC-MS 1 [142]
SPME 10 min
Delorazepam Urine, DI- 65 μm pH 6.5 or 7.4, RT, HPLC- 5 Study of albumin interaction [145, 146]
serum SPME PDMS/DVP 30 min DAD

Various therapeutic drugs

8 barbiturates Urine DI- 65 μm NaCl, RT, 20 min GC-MS 1 Patient sample [54]
SPME CW/DVB
7 barbiturates Blood, DI- 85 μm PA pH 6-7, 60 °C, GC-MS 50–600 Patient sample [153]
urine SPME 60 min
Thiopental, Hair DI- 50 μm pH 5–6, 30 °C, GC-MS- 0.05–0.07 DFSA case [152]
pentobarbital SPME CW/TPR 20 min MS
Ketamine Hair HS- 100 μm PDMS K2CO, 90 °C, GC-MS 0.7 Hair extraction 1 M HCl [95]
SPME 5 min
Propofol Blood, HS- 100 μm PDMS 100 °C, 25 min GC-MS – Death case [155]
tissues SPME,
4 tricyclic Urine In-tube 20 cm, pH 12, 10 min, HPLC- 5 Wire in-tube configuration [35]
antidepressants SPME 0.25μm PDMS 80 μl/min UV
4 tricyclic Plasma DI- 65 μm pH 10, 65 °C, HPLC- 50–75 Spiked samples [159]
antidepressants SPME PDMS/DVB 40 min DAD
Anal Bioanal Chem (2007) 388:1393–1414 1405

Table 2 (continued)
Substance Matrix SPME mode Coating Extraction Detection LOD Remarks References
conditions (ng/ml or
ng/mga)

4 tricyclic Blood HS- 100 μm PDMS NaOH, 100 °C GC-FID 30–50 Animal experiments [156]
antidepressants SPME
3 tetracyclic Blood HS- 100 μm PDMS NaOH, 120 °C, GC-MS 5–25 Death case [157]
antidepressants SPME 45 min
7 tricylic or tetracyclic Hair HS- 100 μm PDMS NaHCO3, NaCl, GC-MS 0.03–0.10 Death cases, metabolized [43, 62]
antidepressants SPME 100 °C, 30 min with MCF
6 SSRI Urine DI- 65 μm NaHCO3 NaCl, GC-MS 0.4 Patient sample, derivatized [160]
antidepressants SPME PDMS/DVB 100 °C, 30 min with acetanhydride
Fluoxetine, Plasma DI- 65 μm pH 9.0, 50 °C, HPLC- 5–10 Spiked samples [60]
norfluoxetine SPME PDMS/DVB 30 min UV
11 phenothiazines Blood, DI- 85 μm PA pH 8, KClO4, LC-MS- 0.2–200 Volunteer samples [161]
urine SPME 40 °C,60 min MS
L-Mepromazine Plasma DI- 100 μm PDMS NaOH, NaCl, GC-NPD 2 Spiked samples [162]
SPME 30 °C, 30 min
Clozapine Plasma DI- 100 μm PDMS NaOH, NaCl, GC-NPD 30 Patient samples [163]
SPME 30 °C, 30 min
5 local anesthetics Blood HS- 100 μm PDMS NaOH, 120 °C, GC-MS 10–500 Death case [164]
SPME 45 min
Lidocaine Hair HS- 65 μm NaOH,Na2SO4, GC-MS 0.1 Drug fatalities [165]
SPME CW/DVB 70 °C,30 min
Lidocaine Plasma DI- 100 μm PDMS pH 9.5, NaCl, RT, GC-FID 5 Spiked samples [162]
SPME 45 min
6 antiepileptics Plasma DI- 65 μm pH 7.0, NaCl, GC-TSD 50–200 Therapeutic drug monitoring [167]
SPME CW/DVB 30 °C, 15 min
5 anticonvulsants Plasma DI- 50 μm pH 5.0, NaCl, HPLC- 2,000 Spiked samples [159]
SPME CW/TPR 30 °C, 30 min DAD
Valproic acid Plasma HS- 100 μm PDMS NaCl, 80 °C, GC-MS 300 In-sample-derivatized with [66]
SPME 20 min IBCF/ethanol
9 beta-blockers Serum, In-tube Omegawax pH 8.5, RT, 15 draw/ LC-MS 0.1–1.2 Spiked and patient samples [168]
urine SPME 250 capillary eject cycles
Verapamil, Plasma, In-tube PPY, 3 times pH 11, 40 draw/ LC-MS 2–6 Metabolism study [169]
gallopamil urine SPME coated eject cycles
Clenbuterol Water DI- 85 μm PA pH 12, RT, GC-MS 0.2 On-fiber-derivatized with HMDS [76]
SPME 60 min
Naproxen and Urine DI- 100 μm pH 3, NaCl, 20 °C, HPLC- 30 Patient samples [170]
metabolites SPME CW/TPR 30 min UV
Ibuprofen and Urine DI- PDMS/DVB pH 3.8, NaCl, RT, HPLC- 250 Patient samples [55]
glucuronide SPME 30 min UV
11 corticosteroids Urine DI- 65 μm NaCl, RT, 15 min LC-MS 4–30 Spiked samples [171]
SPME CW/TPR
Sterols Serum DI- 85 μm PA KCl, 90 °C, 90 min GC-FID 250–1,100 On-fiber-derivatized with [172]
SPME BSTFA

Pesticides and chemical

warfare agents
22 organophosphorous Blood HS- 100 μm PDMS (NH4)2SO4,H2SO4, GC-MS 10–300 Spiked samples [175]
pesticides SPME 120 °C, 15 min
5 organophosphorous Blood, HS- 85 μm PA NaCl, 70 °C, GC-NPD 2–55 Postmortem samples [177]
pesticides tissues SPME 20 min
Parathion-methyl/ Blood HS- 100 μm PDMS (NH4)2SO4,H2SO4, GC-MS 20–50 Spiked samples [174]
-ethyl SPME 90 °C, 15 min
Parathion, paraoxon Blood, DI- 65 μm Dilution 1:10 H2O, GC-MS 3–25 25 poisoning cases [178]
urine SPME CW/DVB 60 °C, 60 min
Parathion-methyl Blood, HS- 85 μm PA NaCl, 70 °C, GC-MS 1 Death case [176]
tissues SPME 20 min
Malthion Blood HS- 100 μm PDMS (NH4)2SO4,H2SO4, GC-MS 1,000 Death case [173]
SPME 90 °C, 15 min
Quinalphos Blood, DI- 65 μm Dilution 1:10 H2O, GC-MS 2-10 36 authentic samples [179]
urine SPME CW/DVB 60 °C, 60 min
Fluoroacetic acid Serum, HS- DVB/CAR/ H2SO4, Na2SO4, GC-MS 20 Derivatized with [71]
urine SPME PDMS 90 °C, 30 min pyrenyldiazomethane
Fluoroacetamide Blood, DI- 100 μm PDMS pH 7.0, TEABr, GC-MS 1,000 Fatal poisoning [184]
urine SPME 70 °C, 25 min
Chlorovinylarsounous Urine HS- 100 μm PDMS NH4Ac, 70 °C, GC-MS 0.007 Derivatized with [190]
acid SPME 10 min 1,3-propanedithiol
1406 Anal Bioanal Chem (2007) 388:1393–1414

Table 2 (continued)
Substance Matrix SPME mode Coating Extraction Detection LOD Remarks References
conditions (ng/ml or
ng/mga)

Other poisons
Cyanide Whole HS- 75 μm H3PO4, Na2SO4, GC-MS 10 Fire victim [192]
blood SPME CAR/PDMS 30 °C, 10 min
Sulfide Blood DI- 100 μm PDMS RT, 20 min GC-MS 10 In-sample-derivatized [193]
SPME with PFBBr
Strychnine Blood DI- 65 μm Dilution 1:10 H2O, GC-MS 7 3 poisonings [194]
SPME CW/DVB RT, 20 min
Lead Whole HS- 70 μm pH 4, RT, 10 min GC-FID 4 In-sample-derivatized [195]
blood SPME PDMS-DVB with STEB

a
The limit of detection (LOD) is given in nanograms per milliliter for blood, serum, plasma, urine and oral fluid and in nanograms per milligram
for hair.
A amphetamine, ADS alkyldiol–silica particles, a.o. and others, BCF butylchloroformate, BDB 1-(1,3-benzodioxol-5-yl)-2-butanamine, BSTFA N,
O-bis(trimethylsilyl)trifluoroacetamide, CAR Carboxen, CBD cannabidiol, CBN cannabinol, CW carbowax, DAD photodiode array detection,
DFSA drug-facilitated sexual assault, DI direct immersion, DVB divinylbenzene, ECD electron capture detection, ECF ethylchloroformate,
EDDP 2-ethylene-1,5-dimethyl-3,3-diphenylpyrrolidine, EMDP 2-ethyl-5-methyl-3,3-diphenyl-1-pyrroline, ESI electrospray ionization, FID
flame ionization detection, GC gas chromatography, GHB γ-hydroxybutyric acid, GBL γ-hydroxybutyrolactone, HCF hexylchloroformate,
HFBA heptafluorobutyric anhydride, HMDS hexamethyldisilazane, HPLC high-performance liquid chromatography, HS headspace, IBCF
isobutylchloroformate, LC liquid chromatography, MA methamphetamine, MBDB N-methyl-1-(1,3-benzodioxol-5-yl)-2-butanamine, MBTFA N-
methylbis(trifluoroacetamide), MCF methylchloroformate, MDA methylenedioxyamphetamine, MDE methylenedioxyethamphetamine, MDMA
methylenedioxymethamphetamine, MS mass spectrometry, NPD nitrogen-phosphorous detection, PA polyacrylate, PCF propylchloroformate,
PFBBr pentafluorobenzyl bromide, PDMS polydimethylsiloxane, PPY polypyrrole coating, RT room temperature, SSRI selective serotonin
reuptake inhibitor, STEB sodium tetraethylborate, TEABr tetraethylammonium bromide, THC Δ9 -tetrahydrocannabinol, TMCS trimethylchlor-
osilane, TPR templated resin, TSD thermionic-specific detection, UV variable-wavelength UV detection

immersed into the alkalized urine sample [78]. Another way
of simultaneous derivatization and HS-SPME was to insert
a small vessel with a mixture of heptafluorobutyric
anhydride and heptafluorobutyric chloride into the head-
space vial with the sample [107]. Then, the vapor of the
reagent reacted with the amphetamines at the sample
surface, in the vapor phase or on the fiber. For in-injector
derivatization, heptafluorobutyric anhydride was injected
into the gas chromatograph immediately prior to fiber
injection [97].
HS-SPME has been mostly preferred for amphetamines,
but methods based on DI-SPME for urine [64, 101, 111] or
oral fluid [93] have also been developed. In-tube SPME and
LC–electrospray ionization (ESI)–MS or HPLC–UV detec-
tion for determination of these compounds has been less
frequently described [35, 103, 113].
Cocaine and metabolites
The detection of cocaine and cocaethylene from different
human sample materials by SPME was described in a series
of papers [65, 93, 94, 95, 117–120]. Benzoylecgonine was
included only in two studies after in-sample derivatization
with hexylchloroformate or butylchloroformate [65, 117].
Although cocaine is sufficiently volatile for HS-SPME
[105], direct immersion of the fiber was generally preferred.
Cocaine and cocaethylene were determined from plasma
[121], urine [65, 120] and sweat patches [118]. For
detection in hair, the sample matrix had to be extracted
first by 1 M HCl [95] or by methanol [117] or to be
enzymatically digested by pronase and dithiotreitol [119].
Cannabinoids
Δ9-Tetrahydrocannabinol, cannabidiol and cannabinol
were analyzed only from oral fluid [93, 94, 122, 123]
and hair [50, 68, 74, 124] by use of SPME in combination
with GC-MS. Generally PDMS fibers appeared to be most
suitable for these highly lipophilic substances. The
detection in oral fluid was performed by direct immersion
of the fiber and without derivatization. An improvement
of the sensitivity was achieved by acidifying the oral fluid
with acetic acid [122]. It is assumed that the cannabinoids
were sequestered in the buccal cavity rather than excreted
with the saliva. Different approaches were used for
determination in hair. DI-SPME after hydrolysis of the
hair sample with NaOH and subsequent neutralization
provided detection limits of 0.1–0.2 ng/mg [124]. The
fully automated analysis of these three compounds by HS-
SPME or HS-SPDE directly from the hair hydrolysate
(prepared with 1 M NaOH + K2CO3) and subsequent
headspace derivatization with MSTFA enabled lower
detection limits (0.05–0.14 ng/mg) [50, 74] to be
acheived. A further improvement of sensitivity (limit of
detection 0.01–0.02 ng/mg) was possible if HS-SPME
was performed from the residue of the evaporated liquid/
liquid extract in presence of BSTFA as the derivatization
reagent [68].
Anal Bioanal Chem (2007) 388:1393–1414 1407

HN O HN O
R HN C6F5
O Alk O
in ylch
sa
R mp lorof ro-
C = l
CH e, H rma
o
9-Fluorenymethyl- f luo mide
4H S- tes nta ro MS
9, C 3 , C SP ,
ME
chloroformate Pe nzylb e, C-
6H 2H e pl G
13 , 5 , C In-sample b m ,
GC 3 H sa ME
HN C6F5 -M 7 , DI-SPME In- -SP
C6 F S HPLC-FL HS
-
5 CO
Cl, o
O DI-S n fib
PME er H7F3-COCl
, GC
-MS In sample

er NH2 HS-SPME HN C3F7

n fib GC-MS
BA, O S
H F C-M
P M E, G o-P O
HS- S N- hth
HN C3F7 N-trifluoro- ac ald
In ety ia
FA acetylprolyl l
DI am l-L-c dehy
s
O BT S chloride -S pl y ste de/
,M
A -M In sample
PM e
ine
FA er GC E,
HP
b HS-SPME
T Fi M E LC
On -SP GC-MS -F
L
HS
COOH
N S
HN CF3
HN *N * NH

O CF3 O
O
Fig. 8 Methods for derivatization of amphetamines in combination with SPME. DI direct immersion FL fluorescence, HFBA heptafluorobutyric
anhydride, HS headspace, MBTFA N-methylbis(trifluoroacetyl)amide, TFAA trifuoroacetanhydride

Opioids was achieved for sufentanil using DI-SPME with fentanyl as

Opiates in a narrower sense (morphine, codeine, 6-acetylmor-
phine, buprenorphine, etc.) have not yet been analyzed from
human materials by use of SMPE. However, there are reports
about methadone [31, 33, 92, 95, 125–127], tramadol [43,
128], pethidine [92], fentanyl [129] and sufentanil [130].
Methadone and its main metabolite 2-ethylene-1,5-dimethyl-
3,3-diphenylpyrrolidine (EDDP) were analyzed by DI-SPME
from plasma [127], urine [92] and oral fluid [125] at pH 9–
11. Because of the stability to strong bases, methadone,
EDDP and 2-ethyl-5-methyl-3,3-diphenyl-1-pyrroline could
be determined with high sensitivity by HS-SPME [42] or
HS-SPDE [33] directly from the hair hydrolysate with 1 M
NaOH. DI-SPME after pronase E hydrolysis of hair led only
to higher detection limits [126].
Tramadol was analyzed by HS-SPME from strong alkal-
ized plasma [118] as well as directly from the alkaline hair
hydrolysate [43]. Fentanyl was determined by HS-SPME
from plasma of patients treated with Duragesic patches with
a detection limit of 0.03 ng/ml after protein precipitation and
addition of excess K2CO3 [129]. A sensitivity of 0.4 ng/ml
an internal standard [130].
Alcohol and minor metabolites of ethanol
Although ethanol was determined from blood, urine or
vitreous humor by HS-SPME and GC with high sensitivity
[131], this technique is not a real alternative to the
established static headspace methods. However, HS-SPME
in combination with GC-MS proved to be a very
advantageous way for analysis of fatty acid ethyl esters
(FAEE) in hair [132–134] and skin-surface lipids [135].
FAEE are side metabolites of ethanol which accumulate in
hair and, for this reason, can be used as direct markers for
diagnosis of chronic excessive alcohol consumption. It is a
group of about 20 substances, from which the most
abundant substances, i.e., ethyl myristate, ethyl palmitate,
ethyl oleate and ethyl stearate, were chosen for routine
analysis. In order to avoid ester hydrolysis, the hair is first
extracted by a dimethyl sulfoxide/n-heptane mixture. The
n-heptane phase is evaporated and the residue is subjected
to HS-SPME from aqueous phosphate buffer pH 7.4 and
1408
saturated NaCl using a PDMS/DVB fiber. Under these
conditions, the FAEE are well separated from the excess of
free fatty acids and less volatile lipids in the hair extracts.
Using deuterated internal standards, the automated method
proved to be very robust in practical routines with detection
limits between 0.015 and 0.04 ng/mg.
γ-Hydroxybutyric acid
SPME proved to be a suitable technique for the fast
detection of γ-hydroxybutyric acid (GHB) in blood or urine
[116, 136–139]. This substance is to an increasing extent
abused for recreational purposes and in drug-facilitated
crimes. Since the substance is very hydrophilic, it is either
transformed into γ-butyrolactone (GBL) for HS-SPME by
addition of an acid to the sample [136, 138] or derivatized
with hexylchloroformate in the presence of pyridine to
hexyl-β-carboxypropylcarbonate [116, 137] for DI-SPME.
In another approach, GHB is adsorbed from the solution by
DI-SPME and is subsequently on-fiber-derivatized in the
vapor of BSTFA/trimethylchlorosilane [139]. The derivati-
zation reactions used for determination of GHB by use of
SPME are shown in Fig. 9. Whereas both DI-SPME
methods were able to differentiate between GHB and
GBL [137, 139], a second analysis without addition of an
acid is necessary for independent determination of GBL in
the HS-SPME methods [136, 138].
Benzodiazepines
Benzodiazepines have been generally extracted by DI-
SPME from urine, serum or plasma [28, 36, 140, 148].
Benzophenones formed by acidic hydrolysis of eight
benzodizaepines in urine were measured by GC–electron
capture detection [141]; however, the benzodiazepines
themselves were preferentially analyzed in combination
Fig. 9 Methods for derivatiza-
tion of γ-hydroxybutyric acid O
HO
(GHB) for determination by
SPME and GC-MS. BSTFA N,
OH
O-bis(trimethylsilyl)trifuoraceta-
mide GBL γ-butyrolactone, GHB
TMCS trimethylchlorosilane
BSTFA, TMCS
in sample
DI-SPME, GC-MS
(CH3)3Si
O O
O OH
Anal Bioanal Chem (2007) 388:1393–1414
with LC. For fiber-based SPME, the immobilization of
octanol on a PA fiber prior to extraction was shown to
increase significantly the recovery [140]. As a rule, in
serum or plasma samples protein had to be precipitated
before extraction [36, 140, 142]. On the other hand, DI-
SPME appeared also to be suitable to study protein binding
[146, 147]. In order to avoid disturbances by adsorption or
precipitation of proteins from the sample on the fiber,
biocompatible porous alkyldiol–silica particles (ADS) were
used both as a fiber coating [28, 144] and for in-tube SPME
[143] and were evaluated in the determination of benzo-
diazepines. ADS/SPME fibers can be applied to whole
blood without protein precipitation [28]. In the same way,
hydrophilic PPY and poly(ethylene glycol) coatings were
used in a specific design for direct extraction of diazepam,
nordazepam and oxazepam from the veins of beagles in
order to determine the pharmacokinetics of the drugs [142].
In this case, the probes were single use with external
calibration of the equilibrium state. As an alternative
method of calibration, a standard on the fiber technique
was tested [149–151]. In this method, a certain amount of
the standard diazepam-d5 is preloaded on the fiber. During
exposition to the bloodstream, diazepam is extracted, while
the standard diffuses into the blood. With a relatively short
exposition time, both adsorption of diazepam (analyte) and
desorption of diazepam-d5 (standard) are kinetically con-
trolled according to Eqs. (3) and (4), respectively:
nA =nA0 ¼ 1  expðαA tÞ; ð3Þ
nS =nS0 ¼ expðαS tÞ; ð4Þ
where nA is the amount of analyte extracted at time t, nA0 is
amount extracted at equilibrium, nS is the amount of
standard remaining on the fiber at time t, nS0 is the
preloaded amount of standard on the fiber and aA and aS
25% H2SO4- H2O
HS-SPME, GC-MS O
O
GBL
Hexylchloroformate/
Pyridine, in sample
DI-SPME, GC-MS
O
O O
O
Si(CH3)3
Anal Bioanal Chem (2007) 388:1393–1414
are time constants of mass transport, a function of diffusion
coefficients, distribution coefficients, geometry of the fiber
and agitation conditions.
aAt and aSt are almost identical if a deuterated standard
is used since both processes occur in the same time period
on the same fiber and depend in the same way on the
agitation conditions (isotropy of adsorption and desorp-
tion). Therefore, the amount of analyte nA0 can be
estimated from the preloaded amount of the standard nS0
and the simultaneously measured nA and nS (Eq. 5):
nA0  nA =ð1nS =nS0 Þ: ð5Þ
For exact calculation, the result can be corrected for the
slightly different diffusion coefficients of the analyte and
the standard. This is particularly important if standards
other than deuterated ones are used. The analyte concen-
tration in the sample CA can be calculated from nA0
according to Eqs. (1) and (2). This standard on the fiber
approach of SPME quantification is believed to have great
promise particularly in on-site or in vivo investigations
where an internal standard is difficult to apply and the
agitation conditions are difficult to control.
Various therapeutic drugs
SPME techniques were used for analysis of barbiturates [54,
69, 152, 153], chloral hydrate (trichloroethanol) [154],
propofol [155], ketamine [43, 95, 108, 116], tricyclic and
tetracyclic antidepressants [35, 43, 62, 82, 156–159],
serotonine reuptake inhibitors [160], neuroleptics [43, 161–
163], local anesthetics [52, 164–166], antiepileptics [66, 159,
167], beta-blockers [168], clenbuterol [76], verapamil [169],
anti-inflammatory drugs [55, 170] and steroid hormones
[171, 172]. Some examples are given in Table 2. A complete
survey is beyond the scope of this review.
Barbiturates were only accessible by DI-SPME. Except for
the addition of Na2SO4, urine could be extracted without any
pretreatment, whereas blood samples were first deproteinated
with HClO4 and then adjusted to pH 6–7 [54, 153]. For
sensitive determination of thiopental and pentobarbital, the
hair sample of the victim of a drug-facilitated sexual assault
was pulverized, extracted overnight with aqueous NaHCO3
and adjusted to pH 5.5 prior to DI-SPME [152].
Tricyclic antidepressants (amitriptyline, clomipramine,
imipramine and doxepine) and tetracyclic antidepressants
(maprotiline, mianserin, and setiptiline) were analyzed
from blood or plasma by DI-SPME [159] as well as by
HS-SPME under strong alkaline conditions [156, 157].
The desmethyl metabolites were not sufficiently sensitive-
ly detected in the HS-SPME approach, probably because
of too low volatility. It was shown for NaOH hydrolysis
solutions of hair samples that the desmethyl metabolites
can be included in the analysis by buffering with NaHCO3
1409
and in-sample derivatization with methylchloroformate
[62]. Tricyclic antidepressants were analyzed in urine
samples also by in-tube SPME [35]. DI-SPME and GC-
MS after in-sample acetylation of primary or secondary
amino groups (acetanhydride plus K2CO3) was used for
analysis of selective serotonin reuptake inhibitors (ven-
lafaxine, fluvoxamine, mirtazapine, fluoxetine, citalopram,
and sertraline) [160]. Fluoxetine and norfluoxetine were
also determined by DI-SPME and HPLC-UV detection in
plasma [60] as well as by HS-SPME and GC-MS in hair
[62]. In the latter case, fluoxetine was in-sample-derivat-
ized by methylchloroformate.
Phenothiazines with heavy side chains such as perazine,
thioridazine and flupenthixol were separated from whole
blood and urine by DI-SPME with extraction efficiencies of
0.0002–12 and 2.6–39.8%, respectively [161]. Nevertheless,
the detection limits (0.2–200 ng/ml in blood and 4–22 pg/ml
in urine) and were sufficiently low for practical use.
Levomepromazine was extracted from plasma in a similar
method with a recovery of 6.6% [162]. A DI-SPME method
for the neuroleptic clozapine in combination with GC–
nitrogen/phosphorous detection from plasma appeared to be
suitable for therapeutic drug monitoring [163]. The des-
methyl metabolite was not included. Despite the strong
alkaline medium (0.2 M NaOH), the 100-μm PDMS fiber
could be used for 75 samples.
Local anesthetics such as lidocaine, prilocaine, dibucaine,
mepivacaine, propipocaine or bupivacaine were sufficiently
volatile for HS-SPME [43, 164, 165]. Since the drugs are
stable to highly concentrated bases, the extraction can be
carried out in a one-step manner from the blood sample
diluted with 5 M NaOH [164] or in combination with the
alkaline hair digestion [43, 165]. The presence of lidocaine
in hair of 32 from 49 drug fatalities showed the wide
occurrence of this drug as an adulterant of illicit cocaine and
heroin preparations [165]. The DI-SPME of lidocaine from
plasma was chosen as a model system for theoretical
considerations for the determination of drug–plasma protein
binding by this method [162]. It was shown that SPME can
be used to obtain a good approximation of the plasma
protein binding, which was 74% for lidocaine.
A DI-SPME method for antiepileptics (lamotrigine,
carbamazepine, carbamazepine epoxide, phenytoin, pheno-
barbital, and primidone) was described for application in
pharmacokinetic studies and patient routine therapeutic drug
monitoring [167]. Valproic acid was in-sample-derivatized to
its ethyl ester with a mixture isobutylchloroformate and
pyridine in the presence of 25 vol% ethanol before
determination by HS-SPME and GC-MS [66]. However,
this drug can also be determined by HS-SPME without
derivatization from the acidified sample [63].
The analysis of beta-blockers is important for clinical
control, doping control and forensic analysis. In an automated
1410
in-tube SPME method in combination with LC-MS, nine
beta-blockers were determined from urine and serum samples
with detection limits between 0.1 and 1.2 ng/ml [168]. The
extraction recoveries of the 60-cm Omegawax capillary were
above 71% for serum and above 84% for urine. The β-2-
agonist clenbuterol, which promotes growth of muscle tissue
and is abused as a doping agent, can be detected in an
automated procedure by HS-SPME after in-sample derivati-
zation with hexamethyldisilazane [76].
Both nonsteroidal anti-inflammatory drugs naproxen
[170] and ibuprofen [55] are carboxylic acids and,
therefore, were analyzed in acidified urine (pH≈3) by DI-
SPME with HPLC-UV detection. In the case of ibuprofen,
chiral HPLC was performed. A screening procedure for 11
corticosteroids by DI-SPME and LC-MS-MS in urine
enables a sensitive detection in clinical samples [171].
The practical use of SPME for steroid hormones in doping
control was not found in the literature. A special derivati-
zation technique was applied for determination of sterols in
serum [172]. The PA fiber was first saturated in the
headspace with BSTFA and then immersed into the
saponified serum sample at 90 °C for 90 min.
Pesticides
Pesticides often have a relatively hydrophobic structure
and, therefore, are particularly suitable for analysis by
SPME. The method is therefore extensively used for
detection of almost all kinds of pesticides in air, soil, water,
wine and other beverages, seafood, grain, fruit or other
food. However, the application to human samples is
essentially limited to organophosporous compounds [173–
179]. Besides a screening procedure for 22 organophos-
phorus pesticides [175] special applications were described
for parathion [174, 178], methylparathion [176], malathion
[173], diazinon [177] and quinalphos [179]. Chlorinated
organics such as hexachlorobenzene, α-hexachlorocyclo-
hexane, β-hexachlorocyclohexane, γ-hexachlorocyclohex-
ane, dichlorodiphenyltrichloroethane (DDT) and its
derivatives, and some important congeners of the poly-
chlorinated biphenyls were determined by SPME from
breast milk [180]. p,p′-dichlorodiphenyldichloroethylene as
the main metabolite of DDT was analyzed from blood [181,
182]. Furthermore, a method for analysis of dinitroaniline
herbicides in human blood was developed [183].
The tasteless and insidious poisons fluoroacetic acid
and fluoroacetamide are generally difficult to determine
from biological material. Besides the HS-SPME method
with on-fiber derivatization for fluoroacetic acid (Fig. 7)
[71] also a DI-SPME method for fluoroacetamide was
published which uses tetraethylammoninum ions for
improvement of the extraction yield at the PDMS/DVB
fiber [184].
Anal Bioanal Chem (2007) 388:1393–1414
Chemical warfare
After a terrorist attack, rapid identification of the poison is
necessary. The fast analysis of samples from the scene, e.g.,
collected from air or with swabs from the floor or furniture,
may be requested. SPME offers a relatively safe possibility
of handling of samples of extremely dangerous chemical
warfare agents such as sarin, tabun or the nerve agent VX
expected in such incidents. Desorption ESI MS (LC-ESI-
MS-MS) was shown to be a sensitive method to detect
these organophosphorous poisons as well as bis(2-chlor-
oethyl) sulfide (sulfur mustard) [185, 186]. But, also usual
HS-SPME in combination with GC–electron impact–MS
offer sufficient sensitivity and specificity to identify sarin
[187], bis(diisopropylaminoethyl)disulfide (a degradation
product of the nerve agent VX) [188] and sulfur mustard
[189]. 2-Chlorovinylarsonous acid, a degradation product
of lewisite [dichloro(2-chlorovinyl)arsine], was determined
by HS-SPME from urine after in-sample derivatization with
1,3-propanedithiol [190]. A fiber with a phenol-based
polymer coating for selective sampling of nerve agents
from air was developed [191]. Analysis of chemical warfare
agents from human samples by SPME was not found in the
literature.
Cyanide, sulfide, strychnine, metal ions
Although cyanide is a typical poison with wide forensic
relevance, its determination from blood or other human
samples is still limited to special techniques or requires
tedious sample preparation such as microdistillation. A HS-
SPME/GC-MS–selected ion monitoring method for cyanide
directly from blood with acetonitrile-d3 as an internal
standard resulted in a total analysis time below 20 min
and a detection limit of 0.01 μg/ml with good integration
into the usual equipment of a toxicology laboratory [192].
The CAR/PDMS fiber appeared to be clearly superior to all
other fibers.
Sulfide was analyzed from blood with a detection limit
of 0.01 μg/ml using DI-SPME and GC-MS after in-sample
derivatization with pentafluorobenzyl bromide [193]. A
method for determination of strychnine from blood by DI-
SPME with a detection limit of 7 ng/ml was developed and
applied to three authentic poisonings [194].
Lead can be determined from whole blood by deproteina-
tion with trichloroacetic acid, neutralization to pH 4,
derivatization with sodium tetraethylborate to tertraethyllead
and subsequent HS-SPME on a PDMS/DVB fiber in
combination with GC– flame ionization detection [195].
The detection limit was 4 ng/ml. A differentiation between
inorganic lead and alkyllead is possible using deuterium-
labeled tetraethylborate and GC-MS [196]. Tin, mercury,
arsenic, antimony, chromium and selenium were analyzed in
Anal Bioanal Chem (2007) 388:1393–1414
a similar way from aqueous media [197–199], but there are
no applications for human samples.
Conclusions and outlook
In recent years, analytical methods have become more and
more sensitive and the trend of miniaturization is obvious.
Sample preparation must keep pace with this development.
SPME is an efficient and economic step in this direction. It
has found its way from an exotic technique to routine
application also in clinical and forensic toxicology laborato-
ries. From the extensive research work published on technical
progress and applications of the different modes of SPME it
follows that it has exceptional advantages in automated
routine techniques such as determination of cannabinoids or
amphetamines from hair, GHB from serum and urine or
organophosphorous pesticides from blood which were opti-
mized for all conditions. There are many procedures where
SPME is the more sensitive, the more selective or even the
much more convenient alternative. The advantages in
determination of cyanide as a classic poison by HS-SPME
are quite obvious. The combination with a large variety of
derivatization processes has essentially extended the field of
application. As shown for the example of amphetamines, the
capacity of this combination is by far not yet exhausted.
On the other hand, the practicability for general screening
procedures has been limited until now. HS-SPME is strongly
affected by matrix composition and the physicochemical
properties of the analytes and includes only a selection of
substances. Different from of the parent drugs, nor- and
hydroxymetabolites are frequently not detected. The pros-
pects for DI-SPME for this purpose are, similar to SPE, much
better but it will not replace the universality of liquid/liquid
extraction in general unknown analysis. Discrimination
effects by matrix constituents and deviations from linearity
in quantitative applications are another drawback.
In summary, SPME has proved to be a very useful
complement to the analytical equipment in a toxicology
laboratory and should always be taken into consideration
when a method for an analyte has to be updated or introduced.
In the same way, newly purchased GC instruments should
include the possibility of automated SPME performance.
References
1. Arthur CL, Pawliszyn J (1990) Anal Chem 62:2145–2148
2. Kataoka H (2002) Anal Bioanal Chem 373:31–45
3. Lipinski J (2001) Fresenius J Anal Chem 369:57–62
4. Kawaguchi M, Ito R, Saito K, Nakazawa H (2006) J Pharm
Biomed Anal 40:500–508
5. Bruheim I, Liu X, Pawliszyn J (2003) Anal Chem 75:1002–1010
6. Yang R, Xie W (2004) Forensic Sci Int 139:177–181
1411
7. Pawliszyn J (1997) Solid phase microextraction—theory and
practice. Wiley-VCH, New York
8. Pawliszyn J (1999) Applications of solid phase microextraction.
Royal Society of Chemistry, Cambridge
9. Sheppers Wercinski SA (1999) Solid phase microextraction: a
practical guide. Dekker, New York
10. Junting L, Peng C, Suzuki O (1998) Forensic Sci Int 97:93–100
11. Snow NH (2000) J Chromatogr A 885:445–455
12. Ulrich S (2000) J Chromatogr A 902:167–194
13. Mills GA, Walker V (2000) J Chromatogr A 902:267–287
14. Furton KG, Wang J, Hsu YL, Walton J, Almirall JR (2000) J
Chromatogr Sci 38:297–306
15. Theodoridis G, Koster EH, de Jong GJ (2000) J Chromatogr B
745:49–82
16. Pawliszyn J (2001) Adv Exp Med Biol 488:73–87
17. Vas G, Vekey K (2004) J Mass Spectrom 39:233–254
18. Theodoridis G, de Jong GJ (2005) Adv Chromatogr 43:231–271
19. Dietz C, Sanz J, Camara C (2006) J Chromatogr A 1103:183–192
20. Chong SL, Wang D, Hayes JD, Wilhite BW, Malik A (1997)
Anal Chem 69:3889–3898
21. Bagheri H, Es-Haghi A, Rouini MR (2005) J Chromatogr B
818:147–157
22. Wu J, Pawliszyn J (2001) J Chromatogr A 909:37–52
23. Azenha M, Malheiro C, Silva AF (2005) J Chromatogr A
1069:163–172
24. Wang D, Xing J, Peng J, Wu C (2003) J Chromatogr A 1005:1–12
25. Wang S, Wang Y, You H, Yao J (2006) Chromatographia
63:365–371
26. Mohammadi A, Yamini Y, Alizadeh N (2005) J Chromatogr A
1063:1–8
27. Wu J, Mullett WM, Pawliszyn J (2002) Anal Chem 74:4855–
4859
28. Walles M, Mullett WM, Pawliszyn J (2004) J Chromatogr A
1025:85–92
29. Koster EH, Crescenzi C, den Hoedt W, Ensing K, de Jong GJ
(2001) Anal Chem 73:3140–3145
30. Tamayo FG, Turiel E, Martin-Esteban A (2007) J Chromatogr A
(in press)
31. Yuan H, Mullett WM, Pawliszyn J (2001) Analyst 126:1456–
1461
32. Lord HL, Rajabi M, Safari S, Pawliszyn J (2006) J Pharm
Biomed Anal 40:769–780
33. Lachenmeier DW, Kroener L, Musshoff F, Madea B (2003)
Rapid Commun Mass Spectrom 17:472–478
34. Bicchi C, Cordero C, Liberto E, Rubiolo P, Sgorbini B (2004) J
Chromatogr A 1024:217–226
35. Kataoka H, Lord HL, Pawliszyn J (2000) J Anal Toxicol
24:257–265
36. Yuan H, Mester Z, Lord H, Pawliszyn J (2000) J Anal Toxicol
24:718–725
37. Saito Y, Kawazoe M, Hayashida M, Jinno K (2000) Analyst
125:807–809
38. Saito Y, Nakao Y, Imaizumi M, Takeichi T, Kiso Y, Kiyokatsu
(2000) J Fresenius J Anal Chem 368:641–643
39. Mullett WM, Martin P, Pawliszyn J (2001) Anal Chem 73:2383–
2389
40. Lord H, Pawliszyn J (2000) J Chromatogr A 885:153–193
41. Jiu A (1999) In: Pawliszyn J (ed) Applications of solid phase
microextraction. Royal Society of Chemistry, Cambridge, pp 22–37
42. Sporkert F, Pragst F (2000) J Chromatogr B 746:255–264
43. Sporkert F, Pragst F (2000) Forensic Sci Int 107:129–148
44. Chen Y, Pawliszyn J (2006) Anal Chem 78:5222–5226
45. Carasek E, Pawliszyn J (2006) J Agric Food Chem 54:8688–8696
46. Carasek E, Cudjoe E, Pawliszyn J (2007) J Chromatogr A
1138:10–77
47. Ezquerro O, Pons B, Tena MT (2003) J Chromatogr A 999:155–164
1412
48. Hakkarainen M (2007) J Biochem Biophys Methods 70:229–233
49. Musshoff F, Lachenmeier DW, Kroener L, Madea B (2002) J
Chromatogr A 958:231–238
50. Musshoff F, Lachenmeier DW, Kroener L, Madea B (2003)
Forensic Sci Int 133:32–38
51. Ridgway K, Lalljie SP, Smith RM (2006) J Chromatogr A
1124:181–186
52. Koster EH, Wemes C, Morsink JB, de Jong GJ (2000) J
Chromatogr B 739:175–182
53. Strano-Rossi S, Molaioni F, Botre F (2005) J Anal Toxicol
29:217–222
54. Hall BJ, Brodbelt JS (1997) J Chromatogr A 777:275–282
55. de Oliveira AR, Cesarino EJ, Bonato PS (2005) J Chromatogr B
818:285–291
56. Zambonin CG (2003) Anal Bioanal Chem 375:73–80
57. Lord HL (2007) J Chromatogr A (in press)
58. Chen J, Pawliszyn J (1995) Anal Chem 67:2530–2533
59. Rodrigues JC, Neto AJ, Fernandes C, Alves C, Contadori AS,
Lancas FM (2006) J Chromatogr A 1105:208–212
60. Fernandes C, Neto AJ, Rodrigues JC, Alves C, Lancas FM
(2007) J Chromatogr B 847:217–223
61. Jonsson J, Kronstrand R, Hatanpaa M (1996) J Forensic Sci
41:148–151
62. Sporkert F, Pragst F (2001) In: Rasanan I (ed) Proceedings of
TIAFT 2000. Tallinna Raamatutrükikoja, Tallin, pp 429–439
63. Sporkert F (2002) Anwendungen der Headspace-Festphasenmik-
roextraktion in der forensischen Analytik unter besonderer
Berücksichtigung der Haaranalyse. Thesis, Humboldt University,
Berlin
64. Chafer-Pericas C, Campins-Falco P, Herraez-Hernandez R
(2004) Anal Biochem 333:328–335
65. Hall BJ, Parikh AR, Brodbelt JS (1999) J Forensic Sci 44:527–534
66. Deng C, Li N, Ji J, Yang B, Duan G, Zhang X (2006) Rapid
Commun Mass Spectrom 20:1281–1287
67. Abdel-Rehim M, Hassan Z, Blomberg L, Hassan M (2003) Ther
Drug Monit 25:400–406
68. Nadulski T, Pragst F (2007) J Chromatogr B 846:78–85
69. Staerk U, Külpmann WR (2000) J Chromatogr B 745:399–411
70. Mills GA, Walker V, Mughal H (1999) J Chromatogr B 730:113–122
71. Sporkert F, Pragst F, Hübner S, Mills G (2002) J Chromatogr B
772:45–51
72. Deng C, Zhang W, Zhang J, Zhang X (2004) J Chromatogr B
805:235–240
73. Deng C, Zhang X (2004) Rapid Commun Mass Spectrom
8:1715–1720
74. Musshoff F, Junker HP, Lachenmeier DW, Kroener L, Madea B
(2002) J Anal Toxicol 26:554–560
75. Rodriguez I, Carpinteiro J, Quintana JB, Carro AM, Lorenzo
RA, Cela R (2004) J Chromatogr A 1024:1–8
76. Engelmann MD, Hinz D, Wenclawiak BW (2003) Anal Bioanal
Chem 375:460–464
77. Musshoff F, Junker HP, Lachenmeier DW, Kroener L, Madea B
(2002) J Chromatogr Sci 40:359–364
78. Koster EH, Bruins CH, de Jong GJ (2002) Analyst 127:598–
602
79. Jurado C, Gimenez MP, Soriano T, Menendez M, Repetto M
(2000) J Anal Toxicol 24:11–16
80. Alzaga R, Pena A, Ortiz L, Bayona JM (2003) J Chromatogr A
999:51–60
81. Liu Y, Cho SR, Danielson ND (2002) Anal Bioanal Chem 373:64–69
82. Mosaddegh MH, Richardson T, Stoddart RW, McClure J (2001)
Ann Clin Biochem 38:541–547
83. Tranthim-Fryer DJ, Hansson RC, Norman KW (2001) J Forensic
Sci 46:934–946
84. Liu J, Hara K, Kashimura S, Kashiwagi M, Hamanaka T,
Miyoshi A, Kageura M (2000) J Chromatogr B 748:401–406
Anal Bioanal Chem (2007) 388:1393–1414
85. Blount BC, Kobelski RJ, McElprang DO, Ashley DL, Morrow
JC, Chambers DM, Cardinali FL (2006) J Chromatogr B
832:292–301
86. Musshoff F, Junker H, Madea B (2000) J Anal Toxicol 24:372–376
87. Musshoff F, Padosch SA, Kroener LA, Madea B (2006) Am J
Forensic Med Pathol 27:188–192
88. Poli D, Manini P, Andreoli R, Franchini I, Mutti A (2005) J
Chromatogr B 820:95–102
89. Dehon B, Humbert L, Devisme L, Stievenart M, Mathieu D,
Houdret N, Lhermitte M (2000) J Anal Toxicol 24:22–26
90. Kim NY, Park SW (2000) J Forensic Sci 45:702–707
91. Liu J, Hara K, Kashimura S, Hamanaka T, Tomojiri S, Tanaka K
(1999) J Chromatogr B 731:217–221
92. Myung SW, Kim S, Park JH, Kim M, Lee JC, Kim TJ (1999)
Analyst 124:1283–1286
93. Fucci N, De Giovanni N, Chiarotti M (2003) Forensic Sci Int
134:40–45
94. Yonamine M, Tawil N, Moreau RL, Silva OA (2003) J
Chromatogr B
95. Gentili S, Cornetta M, Macchia T (2004) J Chromatogr B
801:289–296
96. Centini F, Masti A, Barni Comparini I (1996) Forensic Sci Int
83:161–166
97. Nagasawa N, Yashiki M, Iwasaki Y, Hara K, Kojima T (1996)
Forensic Sci Int 78:95–102
98. Koide I, Noguchi O, Okada K, Yokoyama A, Oda H, Yamamoto
S, Kataoka H (1998) J Chromatogr B 707:99–104
99. Myung SW, Min HK, Kim S, Kim M, Cho JB, Kim TJ (1998) J
Chromatogr B 716:359–365
100. Battu C, Marquet P, Fauconnet AL, Lacassie E, Lachatre G
(1998) J Chromatogr Sci 36:1–7
101. Ugland HG, Krogh M, Rasmussen KE (1999) J Pharm Biomed
Anal 19:463–475
102. Lee MR, Song YS, Hwang BH, Chou CC (2000) J Chromatogr
A 896:265–273
103. Namera A, Yashiki M, Liu J, Okajima K, Hara K, Imamura T,
Kojima T (2000) Forensic Sci Int 109:215–223
104. Namera A, Yashiki M, Kojima T, Ueki M (2002) J Chromatogr
Sci 40:19–25
105. Okajima K, Namera A, Yashiki M, Tsukue I, Kojima T (2001)
Forensic Sci Int 116:15–22
106. Liu J, Hara K, Kashimura S, Kashiwagi M, Kageura M
(2001) J Chromatogr B 758:95–101
107. Huang MK, Liu C, Huang SD (2002) Analyst 127:1203–1206
108. Gentili S, Torresi A, Marsili R, Chiarotti M, Macchia T
(2002) J Chromatogr B 780:183–192
109. Brown H, Kirkbride KP, Pigou PE, Walker GS (2003) J
Forensic Sci 48:1231–1238
110. Raikos N, Christopoulou K, Theodoridis G, Tsoukali H,
Psaroulis D (2003) J Chromatogr B 789:59–63
111. Chafer-Pericas C, Campins-Falco P, Herraez-Hernandez R
(2006) J Pharm Biomed Anal 40:1209–1217
112. Wang SM (2005) J Chromatogr B 825:79–87
113. Fan Y, Feng YQ, Zhang JT, Da SL, Zhang M (2005) J
Chromatogr A 1074:9–16
114. Nishida M, Yashiki M, Namera A, Kimura K (2006) J
Chromatogr B 842:106–110
115. Kuriki A, Kumazawa T, Lee XP, Hasegawa C, Kawamura M,
Suzuki O, Sato K (2006) J Chromatogr B 844:283–291
116. Brown SD, Rhodes DJ, Pritchard BJ (2007) Forensic Sci Int (in press)
117. de Toledo FC, Yonamine M, de Moraes Moreau RL, Silva OA
(2003) J Chromatogr B 798:361–365
118. Follador MJ, Yonamine M, de Moraes Moreau RL, Silva OA
(2004) J Chromatogr B 811:37–40
119. Bermejo AM, Lopez P, Alvarez I, Tabernero MJ, Fernandez P
(2006) Forensic Sci Int 156:2–8
Anal Bioanal Chem (2007) 388:1393–1414
120. Yonamine M, Saviano AM (2006) Biomed Chromatogr
20:1071–1075
121. Alvarez I, Bermejo AM, Tabernero MJ, Fernandez P, Lopez P
(2007) J Chromatogr B 845:90–94
122. Hall BJ, Satterfield-Doerr M, Parikh AR, Brodbelt JS (1998)
Anal Chem 70:1788–1796
123. Fucci N, De Giovanni N, Chiarotti M, Scarlata S (2001) Forensic
Sci 119:318–321
124. Strano-Rossi S, Chiarotti M (1999) J Anal Toxicol 23:7–10
125. dos Santos Lucas AC, Bermejo A, Fernandez P, Tabernero MJ
(2000) J Anal Toxicol 24:93–96
126. Lucas AC, Bermejo AM, Tabernero MJ, Fernandez P, Strano-
Rossi S (2000) Forensic Sci Int 107:225–232
127. Bermejo AM, Seara R, dos Santos Lucas AC, Tabernero MJ,
Fernandez P, Marsili R (2000) J Anal Toxicol 24:66–69
128. Sha YF, Shen S, Duan GL (2005) J Pharm Biomed Anal 37:143–
147
129. Bagheri H, Es-Haghi A, Khalilian F, Rouini MR (2007) J Pharm
Biomed Anal 43:1763–1768
130. Paradis C, Dufresne C, Bolon M, Boulieu R (2002) Ther Drug
Monit 24:768–774
131. De Martinis BS, Martin CC (2002) Forensic Sci Int 128:115–119
132. Pragst F, Auwaerter V, Sporkert F, Spiegel K (2001) Forensic Sci
Int 121:76–88
133. Auwarter V, Sporkert F, Hartwig S, Pragst F, Vater H,
Diefenbacher A (2001) Clin Chem 47:2114–2123
134. Auwarter V, Kiessling B, Pragst F (2004) Forensic Sci Int
145:149–159
135. Pragst F, Auwarter V, Kiessling B, Dyes C (2004) Forensic Sci
Int 143:77–86
136. Frison G, Tedeschi L, Maietti S, Ferrara SD (2000) Rapid
Commun Mass Spectrom 14:2401–2407
137. Blair S, Song M, Hall B, Brodbelt J (2001) J Forensic Sci 46:688–693
138. Merckel C, Auwärter V, Simmert D, Pragst F (2003)
Toxichem Krimtech 70:93–98
139. Meyers JE, Almirall JR (2005) J Forensic Sci 50:31–36
140. Reubsaet KJ, Ragnar Norli H, Hemmersbach P, Rasmussen
KE (1998) J Pharm Biomed Anal 18:667–680
141. Guan F, Seno H, Ishii A, Watanabe K, Kumazawa T, Hattori
H, Suzuki O (1999) J Anal Toxicol 23:54–61
142. Frison G, Tedeschi L, Maietti S, Ferrara SD (2001) Rapid
Commun Mass Spectrom 15:2497–2501
143. Mullett WM, Levsen K, Lubda D, Pawliszyn J (2002) J
Chromatogr A 963 :325–334
144. Mullett WM, Pawliszyn J (2002) Anal Chem 74:1081–1087
145. Aresta A, Monaci L, Zambonin CG (2002) J Pharm Biomed
Anal 28:965–972
146. Zambonin CG, Aresta A (2002) J Pharm Biomed Anal 29:895–900
147. Yuan H, Pawliszyn J (2001) Anal Chem 73:4410–4416
148. Musteata FM, Musteata ML, Pawliszyn J (2006) Clin Chem
52:708–715
149. Chen Y, O’Reilly J, Wang Y, Pawliszyn J (2004) Analyst
129:702–703
150. Chen Y, Pawliszyn J (2004) Anal Chem 76:5807–5815
151. Zhou SN, Zhang X, Ouyang G, Es-Haghi A, Pawliszyn J (2007)
Anal Chem 79:1221–1230
152. Frison G, Favretto D, Tedeschi L, Ferrara SD (2003) Forensic
Sci Int 133:171–174
153. Iwai M, Hattori H, Arinobu T, Ishii A, Kumazawa T, Noguchi H,
Noguchi H, Suzuki O, Seno H (2004) J Chromatogr B 806:65–73
154. Thurau K, Goerke R, Vogt S, Perdekamp MG, Weinmann W
(2003) Arch Kriminol 211:90–97
155. Strehler M, Preuss J, Wollersen H, Madea B (2006) Arch
Kriminol 217:153–160
156. Lee XP, Kumazawa T, Sato K, Suzuki O (1997) J Chromatogr
Sci 35:302–308
1413
157. Namera A, Watanabe T, Yashiki M, Iwasaki Y, Kojima T (1998)
J Anal Toxicol 22:396–400
158. Alves C, Fernandes C, Dos Santos Neto AJ, Rodrigues JC,
Costa Queiroz ME, Lancas FM (2006) J Chromatogr Sci
44:340–346
159. Cantu MD, Toso DR, Lacerda CA, Lancas FM, Carrilho E,
Queiroz ME (2006) Anal Bioanal Chem 386:256–263
160. Salgado-Petinal C, Lamas JP, Garcia-Jares C, Llompart M, Cela
R (2005) Anal Bioanal Chem 382:1351–1359
161. Kumazawa T, Seno H, Watanabe-Suzuki K, Hattori H, Ishii A,
Sato K, Suzuki O (2000) J Mass Spectrom 35:1091–1099
162. Kruggel S, Ulrich S (2000) Ther Drug Monit 22:723–728
163. Ulrich S, Kruggel S, Weigmann H, Hiemke C (1999) J
Chromatogr B 731:231–240
164. Watanabe T, Namera A, Yashiki M, Iwasaki Y, Kojima T (1998)
J Chromatogr B 709:225–232
165. Sporkert F, Pragst F (2000) J Anal Toxicol 24:316–322
166. van Hout MW, Jas V, Niederlander HA, de Zeeuw RA, de Jong
GJ (2002) Analyst 127:355–359
167. Queiroz ME, Silva SM, Carvalho D, Lancas FM (2002) J
Chromatogr Sci 40:219–223
168. Kataoka H, Narimatsu S, Lord HL, Pawliszyn J (1999) Anal
Chem 71:4237–4244
169. Walles M, Mullett WM, Levsen K, Borlak J, Wunsch G,
Pawliszyn J (2002) J Pharm Biomed Anal 30:307–319
170. Aresta A, Palmisano F, Zambonin CG (2005) J Pharm Biomed
Anal 39:643–647
171. Volmer DA, Hui JP (1997) Rapid Commun Mass Spectrom
11:1926–1933
172. Domeno C, Ruiz B, Nerin C (2005) Anal Bioanal Chem
381:1576–1583
173. Namera A, Yashiki M, Nagasawa N, Iwasaki Y, Kojima T (1997)
Forensic Sci Int 88:125–131
174. Musshoff F, Junker H, Madea B (1999) Clin Chem Lab Med
37:639–642
175. Musshoff F, Junker H, Madea B (2002) J Chromatogr Sci
40:29–34
176. Tsoukali H, Raikos N, Theodoridis G, Psaroulis D (2004)
Forensic Sci Int 143:127–132
177. Tsoukali H, Theodoridis G, Raikos N, Grigoratou I (2005) J
Chromatogr B 822:194–200
178. Gallardo E, Barroso M, Margalho C, Cruz A, Vieira DN, Lopez-
Rivadulla M (2006) Anal Bioanal Chem 386:1717–1726
179. Gallardo E, Barroso M, Margalho C, Cruz A, Vieira DN, Lopez-
Rivadulla M (2006) J Chromatogr B 832:162–168
180. Rohrig L, Meisch HU (2000) Fresenius J Anal Chem
366:106–111
181. Kusakabe T, Saito T, Takeichi S (2001) J Chromatogr B 761:93–98
182. Kusakabe T, Saito T, Takeichi S (2003) Tokai J Exp Clin Med
28:131–138
183. Guan F, Watanabe K, Ishii A, Seno H, Kumazawa T, Hattori H,
Suzuki O (1998) J Chromatogr B 714:205–213
184. Cai X, Zhang D, Ju H, Wu G, Liu X (2004) J Chromatogr B
802:239–245
185. D’Agostino PA, Hancock JR, Chenier CL, Lepage CR (2006) J
Chromatogr A 1110:86–94
186. D’Agostino PA, Chenier CL, Hancock JR, Lepage CR (2007)
Rapid Commun Mass Spectrom 21:543–549
187. Schneider JF, Boparai AS, Reed LL (2001) J Chromatogr Sci
39:420–424
188. Hook GL, Kimm G, Koch D, Savage PB, Ding B, Smith PA
(2003) J Chromatogr A 992:1–9
189. Kimm GL, Hook GL, Smith PA (2002) J Chromatogr A
971:185–191
190. Wooten JV, Ashley DL, Calafat AM (2002) J Chromatogr B
772:147–153
1414
191. Harvey SD, Nelson DA, Wright BW, Grate JW (2002) J
Chromatogr A 954:217–225
192. Frison G, Zancanaro F, Favretto D, Ferrara SD (2006) Rapid
Commun Mass Spectrom 20:2932–2938
193. Liu J, Tan F, Feng X, Wang Y (2003) Wei Sheng Yan Jiu 32:620–621
194. Barroso M, Gallardo E, Margalho C, Avila S, Marques EP, Vieira
DN, Lopez-Rivadulla M (2005) J Chromatogr B 816:29–34
Anal Bioanal Chem (2007) 388:1393–1414
195. Yu X, Yuan H, Gorecki T, Pawliszyn J (1999) Anal Chem
71:2998–3002
196. Yu X, Pawliszyn J (2000) Anal Chem 72:1788–1792
197. Centineo G, Gonzalez EB, Sanz-Medel A (2004) J Chroma-
togr A 034:191–197
198. Diez S, Bayona JM (2006) Chromatogr Sci 44:458–471
199. Kaur V, Malik AK, Verma N (2006) J Sep Sci 29:333–345