Forensic Chemistry 3 (2017) 8189

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Forensic Chemistry
journal homepage: www.elsevier.com/locate/forc

Cryofocusing capillary microextraction of volatiles (Cryo-CMV) as a

novel headspace extraction device for the analysis of volatile organic
compounds and smokeless powders
Anamary Tarifa, Natasha M. Kreitals, Jerome Mulloor, Sigalit Gura, Jos R. Almirall
Department of Chemistry and Biochemistry and International Forensic Research Institute, Florida International University, 11200SW 8th Street, Miami, Florida 33126, United States

a r t i c l e i n f o a b s t r a c t

Article history: A novel field sampling device for fast, sensitive, dynamic headspace sampling of volatile organic com-
Received 16 December 2016 pounds (VOCs) and smokeless powders from air is described. The capillary microextraction of volatiles
Received in revised form 10 February 2017 (CMV) is an inexpensive, preconcentration technique that results in fast sampling times (as low as
Accepted 16 February 2017
2 min) with high efficiency and sensitivity. Analysis of the CMV with GCMS is achieved with a thermal
Available online 20 February 2017
desorption probe to introduce the CMV directly into the injection port of a GC. A cryofocusing device con-
sisting of a Peltier-cooled module that operates down to 10 C was employed to improve the extraction
Keywords:
efficiency of the CMV for sampling both VOCs and smokeless powders.
VOC
Smokeless powders
The method performance using the CMV for the analysis of 17 VOCs, including the BTEX compounds
Capillary microextraction of volatiles (CMV) (Benzene, Toluene, Ethylbenzene, and Xylenes) is demonstrated using the criteria established by the
Cryo-CMV Environmental Protection Agency (EPA) for sorbent tube sampling of toxic organic compounds in ambient
Thermal separation probe with gas air (EPA TO-17). Experimental results with known standards yield absolute mass detection limits ranging
chromatography-mass spectrometry from 2.0 to 4.6 ng for the target compounds, in a 2 L air sample. A 35100% improvement in the analytical
performance was achieved by sampling at 10 C, in comparison to 20 C, for most compounds. Using
cryofocusing conditions, precision of 853 %RSD was achieved for headspace extraction for the majority
of the compounds with recoveries of as much as 37% and low breakthrough. Furthermore, the headspace
calibration curves show a linearity of 0.951 or better for the target compounds, suggesting the quantita-
tion capabilities of the CMV device.
The CMV method performance was further demonstrated for the detection of markers above the head-
space of smokeless powder. Experimental results with known standards, yield method detection limits of
2.0 ng and 1.3 ng for nitroglycerine (NG) and diphenylamine (DPA), respectively, in a 2 L air sampling vol-
ume followed by simultaneous SIM mode GCMS analysis. Using cryofocusing conditions for the head-
space extraction of Hodgdon smokeless powder, a 2938% improvement in the analytical performance
of NG was achieved with 18 %RSD precision, by sampling at 2.5 C to 10 C, in comparison to room tem-
perature at 20 C.
2017 Elsevier B.V. All rights reserved.

1. Introduction example. The list of VOCs thought to be toxic organic compounds

The detection of volatile organic compounds (VOCs) present in
ambient air is of great concern to air quality monitoring programs
because of the potential hazards to human health and the environ-
ment [1,2], but also of interest to forensic scientists interested in
detecting VOCs associated with smokeless powders, gunshot resi-
dues (GSR), explosives, drugs, and ignitable liquid residues for
Corresponding author.
E-mail addresses: atarifa@fiu.edu (A. Tarifa), nkreital@fiu.edu (N.M. Kreitals),
jmull008@fiu.edu (J. Mulloor), sgura@fiu.edu (S. Gura), almirall@fiu.edu (J.R.
Almirall).
is extensive and has been compiled by the Environmental Protec-
tion Agency (EPA) to indicate chemicals of concern that may be
detectable in areas where air pollution is present, such as indus-
trial sites [3]. While the severity of the hazard posed by different
contaminants varies, many VOCs present in ambient air have the
potential to act as mutagens and carcinogens [1,2]. Therefore, the
detection and quantitation of VOCs is important to managing and
mitigating health impacts from toxic compounds in ambient air.
In an effort to address this issue, the EPA has published the Com-
pendium of Methods for Toxic Organic Air Pollutants since 1984
(TO-1TO-17). These are a series of reports describing the most

http://dx.doi.org/10.1016/j.forc.2017.02.007
2468-1709/ 2017 Elsevier B.V. All rights reserved.
82 A. Tarifa et al. / Forensic Chemistry 3 (2017) 8189
current methods and guidelines to be followed for the monitoring
of VOCs in ambient air or polluted environments.
The analysis of VOCs in ambient air is currently performed with
sorbent tubes following the guidelines from the EPA method TO-
17. The commercially available sorbent tubes consist of a thin
cylinder that can be made out of glass or stainless steel and packed
with sorbent material. The commonly used sorbent materials
include: several variations of Tenax, Carbotrap, and Car-
bopackTM, with the possibility of using multiple sorbents in a single
tube [4]. The sorbent material selected depends largely on the tar-
get compounds properties, specifically the volatility or vapor pres-
sure of the molecules of interest. Each sorbent material is classified
according to its strength, which is described as the affinity of the
compounds to the sorbent. A strong sorbent will allow greater
sampling volumes for all or most of the targeted VOCs. The
strength of a sorbent tube is related to the surface area of the sor-
bent material. In general, a weak sorbent has a surface area less
than 50 m2/g (e.g. Tenax TA), a medium sorbent has a surface area
in the range of 100500 m2/g, and a strong sorbent has a surface
area around 1000 m2/g [3]. In general, stronger sorbents are used
for highly volatile compounds. Some of the limitations observed
for the analysis of VOCs with sorbent tubes include: long head-
space extraction times (1 h) with low flow rates, and the use of
expensive thermal desorption units coupled with a transfer line
to the gas chromatography-mass spectrometry (GCMS) instru-
ment, which can result in poor recoveries [57].
This study describes for the first time the use of capillary
microextraction of volatiles (CMV), an innovative headspace
extraction technique [8], while being coupled to a cryofocusing
device (a Peltier-cooled module, Cryo), to enhance the sampling
and pre concentration capabilities of the CMV. The CMV has
already demonstrated to result in improved sensitivity and selec-
tivity for the extraction of volatiles in the headspace of explosives
compared to SPME [9], for the detection of gunshot residues on
swab samples from the hands of shooters [10], and for marijuana
markers from air and small plastic bags [11]. Herein, the enhanced
performance of the Cryo-CMV is demonstrated for air monitoring
of VOCs with its potential in forensic applications.
The CMV consists of an open-ended 2 cm glass capillary tube,
packed with approximately seven 2 cm  2 mm glass filter strips,
each coated with vinyl-terminated polydimethylsiloxane
(vt-PDMS) [8,12], at overall average weight of 0.23 g (USPTO patent
Fig. 1. The dimensions of a CMV device (left) with an inner diameter of 2 mm and length of 2 cm, and (right) a CMV device inserted into the thermal separation probe for
injection into the GCMS.
No. 61/779,690) [9] (see Fig. 1). PDMS is a non-polar polymer that
provides a hydrophobic coating over the glass filter, thus improv-
ing extraction of VOCs in humid conditions [7,8].
In addition, the use of PDMS has been demonstrated to be more
effective than Tenax TA for the analysis of large injection volumes
and retention efficiency with good precision [7]. With a total sur-
face area of 0.05 m2 and a phase volume of 100 mm3, the CMV
device offers 5000 times greater absorption capacity compared to
SPME [9]. It also facilitates dynamic sampling, thus increasing sam-
pling efficiency and decreasing sampling time.
In order to further improve the trapping efficiency of the target
compounds vapor, a cryofocusing CMV technique was developed
by cooling the device down to 10 C during the dynamic extrac-
tion, using a newly designed single-stage Peltier-cooled module
with a temperature controller (see Fig. 2, patent pending). Similar
thermoelectric devices have been reported in analytical applications
for the preconcentration of analytes using cold fiber SPME [13], cool-
ing assisted liquid extraction, and thin film microextraction [14].
The aim of this work was to demonstrate the utility of CMV
samplers, including while being coupled to the novel cryofocusing
device, for the enhancement in extraction capabilities of VOCs in
indoor air, with its potential in forensic chemistry applications
demonstrated herein with smokeless powder. The performance
of Cryo-CMV for VOCs was evaluated following the reported guide-
lines of the EPA Compendium Method TO-17 based on four perfor-
mance criteria: 1) a method detection limit of 0.5 ppbv or less, 2)
an analytical precision of 20%, 3) a precision for distributed volume
pair of 25% or less and 4) an audit accuracy within 30% for concen-
trations expected in contaminated ambient air (0.525 ppbv). In
addition to these 4 criteria, the EPA TO-17 mentions that the break-
through of the sorbent tubes should be less than 5% during the
sampling time.
2. Materials and methods
2.1. Materials
Standard compounds for 17 hazardous air contaminants
(Table 1) were sourced from Fisher Scientific (Pittsburgh, PA),
Sigma-Aldrich (St. Louis, MO), TCI America (Tokyo, Japan), Acros
(New Jersey, USA), and Aldrich (Milwaukee, WI). Purities of
stock standards exceeded 97.0% with the exception of
 A. Tarifa et al. / Forensic Chemistry 3 (2017) 8189 83

Fig. 2. Prototype single-stage Peltier-cooled cryofocusing device with a programmable temperature controller (top) showing the CMV module connected to the sampling
tube and a pump, and inserted into a Peltier-cooled aluminum block cooled down to 10 C (bottom).

Table 1
Compound list in order of elution time (tR) for the headspace extraction at 20 C, the quantifier and qualifier ions, method limits of detection (MDL), method quantitation limits
(MQL), precision (as % RSD, n = 3) for both direct spike onto a CMV and the headspace extraction using CMV, and recoveries (as %) for the headspace extraction using the CMV.

Compounds tR min Quantifier ion Qualifier Direct spike on CMV %RSDa Headspace %RSDa %Recoverya
ions extraction with CMV
Q1 Q2 MDL ng MQL ng MDL ng MQL ng
Methylene chloride 2.61 84 49 86 4.1 14 25 2.7b
Benzene 3.44 78 77 51 4.6 15 19 37 123 13 11
Pyridine 4.35 79 52 78 3.3 11 13 10b
Toluene 4.54 91 92 65 2.9 10 23 17 55 25 18
Furfural 5.31 96 95 39 3.7 12 15 74 247 30 8.0
Ethylbenzene 5.67 91 106 77 2.0 6.6 11 33 110 7 26
m-Xylene/p-Xylene 5.77 91 106 77 3.0 10 15 23 75 15 18
o-Xylene 6.05 91 106 105 2.9 10 12 24 81 14 17
Benzaldehyde 6.87 105 106 77 2.5 8.4 12 52 173 20 16
Phenol 6.92 94 66 65 3.1 10 15 0.6b
Benzonitrile 7.09 103 76 50 2.6 8.8 14 53 177 16 11
1,2,4-Trimethylbenzene 7.19 105 120 91 2.4 8.1 12 47 155 7 14
1,2,3-Trimethylbenzene 7.47 105 120 91 2.6 8.7 14 88 292 4 12
Acetophenone 7.88 105 77 120 2.8 9.3 11 83 276 29 4.5
Nonanal 8.15 57 67 81 5.6 19 9 95 315 33 1.2
Naphthalene 8.88 128 127 102 2.5 8.4 12 56 186 15 11
a
Precision (%RSD) from the calibration curve at 15 ng (direct spike) and 200 ng (headspace), %Recovery from headspace extraction at 1 min equilibrium of 200 ng inside a
quart can.
b
%Recoveries were calculated for these compounds at 500 ng.

1,2,3-trimethylbenzene (90.0%), benzonitrile (95.0%), and nonanal (Fisher Scientific, Pittsburgh, PA). The standard mixture solution
(95.0%). A 300 ng mL 1 mixture of the standard compounds was was further diluted to concentrations ranging from 1.0 to
prepared in methanol and stored in a 10 mL amber glass vial 250 ng mL 1.
84 A. Tarifa et al. / Forensic Chemistry 3 (2017) 8189
Standard compounds for smokeless powders, nitroglycerin (NG)
at 1 mg mL 1 acetonitrile and diphenylamine (DPA) at 5 mg mL 1
methanol, were sourced from Sigma-Aldrich (St. Louis, MO). Both
standards were diluted in methanol to 2100 ng mL 1 for NG and
0.525 ng mL 1 for DPA. For each solution, 1 ml was spiked directly
on the CMV device for the generation of GCMS calibration curves.
The double-base classification smokeless powder, BLC-2, was
sourced from Hodgdon (KS, USA).
2.2. Fabrication of the cryofocusing device
A newly designed 5 V single-stage Peltier-cooled module was
attached to a heat sink assembly on its hot surface (Adafruit,
New York, NY), and to an aluminum block configured with a tem-
perature controller and thermocouple probe on its cold surface.
The latter was attached to an orifice large enough to fit the CMV
(see Fig. 2). Two small vacuum pumps capable of drawing 0.2L/
min for VOCs (Bailey Nurture III) or 1 L/min for smokeless powders
(Escort Elf Air Sampling Pump, Zefon International Inc, Ocala, FL)
were used to sample the air through the CMV devices. A commer-
cially available thermal separation probe (Agilent, Santa Clara, CA)
was used to insert the CMV into the inlet port (Fig. 3) of the GCMS
(Agilent, Santa Clara, CA).
2.3. Instrumentation
All analyses were conducted using a 7890A gas chromatograph
coupled to a 5975C inert XL MSD with a triple axis detector (Agi-
lent Technologies, Santa Clara, CA). The CMV was introduced
directly into the GC injection port for thermal desorption using
an Agilent Thermal Separation Probe (TSP) with a Sky 4 mm ID
single taper inlet liner (Restek, Bellefonte, PA). A schematic of the
Fig. 3. Schematic of the CMV device inserted in the injection port of a GC using the
thermal separation probe.
TSP with the CMV in the injection port of the GC system is shown
in Fig. 3.
VOCs analysis: A 29.17 m  0.25 mm  0.25 mm DB-5 ms Ultra
Inert column (Agilent Technologies, Santa Clara, CA) was used for
chromatographic separation. The GC oven ramp temperature
started at 35 C with a 1 min hold. The oven was then ramped to
120 C at 15 C/min, to 220 C at 30 C /min with a 1.5 min hold,
and subsequently to a final temperature of 280 C at 30 C /min
with a 1 min hold. The total time for the chromatographic separa-
tion was 14.50 min. The injector temperature was set at 180 C in
split mode (split ratio 5:1) with a column flow of 1.2 mL/min. The
EI source was kept at 230 C, the transfer line to the mass spec-
trometer was set to 280 C and the quadrupoles were maintained
at 150 C. The scan mass range was set at 45300 amu. The resolu-
tion of the mass analyzer is 0.1 amu. A list of the target compounds
with their respective retention times and ion peaks is presented in
Table 1 and used to confirm the presence of the target compounds
in the field samples.
Smokeless powder analysis: A 5.8 m  0.25 mm  0.25 mm DB-
5 ms Ultra Inert column (Agilent Technologies, Santa Clara, CA)
was used for chromatographic separation. The GC oven ramp tem-
perature started at 40 C with a 0.5 min hold, then to 240 C at
15 C/min and to 280 C at 30 C/min with a 1 min hold. The total
time for the chromatographic separation was 21.16 min. The injec-
tor temperature was set at 180 C in split mode (split ratio 5:1)
with a column flow of 1.2 mL/min. The EI source was kept at
230 C, the transfer line to the mass spectrometer was set to
280 C and the quadrupoles were maintained at 150 C. The total
ion chromatogram (TIC) scan mass range was set at 45500 amu
and at selected ion monitoring (SIM) simultaneously. Applying this
method respective retention times of 6.1 min. and 8.1 min. were
obtained for NG (46, 76 m/z) and DPA (169 m/z) and the SIM
response was used for quantification.
The instrument was tuned before each experiment using the
autotune feature as recommended by the manufacturer.
2.4. Method development
Prior to sampling, conditioning of the CMV was performed in an
oven at 250 C for 30 min. The CMV was then desorbed as a blank
sample in the GCMS. Liquid standards containing a known mass
of the analytes were spiked (1 lL) directly onto the CMV and ther-
mally desorbed in the GC inlet (see Fig. 3). Thermal desorption of
the CMV devices was performed inside the GC inlet preheated at
180 C and through the entire runtime. For all cryofocusing mea-
surements, the CMV device was suspended 2 min inside the mod-
ule, to cool down prior to sampling, as was confirmed by the
thermocouple probe.
The headspace extractions were performed using a portable
vacuum pump connected to a flow meter with a sampling rate of
0.2 L/min for VOCs or 1 L/min for smokeless powder applications.
For VOCs 1 lL standard containing a known amount of the target
analyte(s) was spiked onto a Kimwipe (Kimberly-Clark Global Sales
LLC, Roswell, GA) that was placed inside a quart can (0.95 L) (All-
American Containers, Miami, FL, USA) and sealed. For smokeless
powder, 100 mg of powder was placed inside a quart can and
sealed. All cans were previously baked to 250 C in an oven over
three days prior to use, to remove any volatile contaminants from
the manufacturing process. Holes were fashioned on the can lids
and sealed using a rubber septum (Capitol Scientific Inc. Austin,
TX) to facilitate sampling through the holes. The extraction results
are reported in total mass extracted (ng), with values for the head-
space calibration curve reported as the calculated amount of mass
extracted.
The evaluation of CMV devices for the detection of VOCs using
the EPA compendium TO-17 method was performed using the stan-
 A. Tarifa et al. / Forensic Chemistry 3 (2017) 8189
dard solutions with the following optimized parameters: 1 min
equilibrium time, 10 min extraction time, 0.2 L/min sampling flow
and total sampling volume of 2 L at a room temperature of 20 C.
The headspace calibration curves were also constructed using those
parameters. Optimization of the method using the cryofocusing
device at 10 C and at room temperature (20 C) was tested for
30 min equilibrium time at different extraction times (15 s, 30 s,
1, 1.5, 2.5, 5, 10, 15, 20, 25, 30 min) to determine recovery and vol-
ume extraction capability at a sampling flow of 0.2 L/min. The
experiments showed that 10 min extraction was optimum for most
VOCs. Recovery experiments were performed at room temperature
(20 C) and using the cryofocusing device ( 10 C) applying the
same sampling and pumping parameters for comparison.
The evaluation of the cryofocusing for the detection of smoke-
less powders was performed using previously reported optimized
parameters for headspace sampling [9,10]: 24 h equilibrium time
at ambient conditions, 2 min extraction time and 1 L/min sampling
flow for a total sampling volume of 2 L air. Optimization of the
method using the cryofocusing device was performed at different
extraction temperatures, 10 C, 2.5 C and 20 C for enhanced
pre-concentration, all at three replicates.
Equivalent blank samples were analyzed, through all method
development stages and headspace samplings, both for VOC and
smokeless powder applications. Blank samples were prepared fol-
lowing the exact parameters and sample treatments while exclud-
ing target compounds.
2.5. Field sampling
To demonstrate the utility of air monitoring in the field, tripli-
cate samples were collected in a chemical laboratory, a lecture hall
and a nail and hair salon. CMVs were conditioned and analyzed
prior to field sampling to ensure no previous contamination or car-
ryover. All samples were extracted for 10 min at a flow rate of
0.2 L/min, at 20 C. The GCMS chromatographic peaks of the
detected compounds in the samples were first identified using
the NIST mass spectral reference library and confirmed by compar-
ing the mass spectra of the individual standard compounds, under
the same instrumental conditions.
3. Results and discussion
3.1. CMV and Cryo-CMV for VOCs sampling
3.1.1. Method validation
A rapid GCMS temperature program (14.5 min) was developed
for the detection of 17 volatile organic compounds commonly
found in the air of polluted environments (Table 1) that represent
a wide range of boiling points (40.0 C217.9 C) [15]. The reten-
tion time for each compound was determined by injecting 1 mL of
a 20 ppm of the standard into the GCMS. Chromatographic sepa-
rations were achieved for all the compounds except for m-xylene
and p-xylene therefore, a combined quantitation was used for
these compounds.
The parameters and performance of the developed method as
an extraction technique for air sampling was established by follow-
ing the criteria specified in the EPA TO-17 [3]. The criteria include
method detection limit (MDL) of 0.5 ppbv (35 ng), analytical
precision of replicate measurements within 20%, precision for the
distributed volume pair of 25% or less, and an audit accuracy of
30% or better for the expected concentration range 0.525 ppbv
(5300 ng).
Calibration curves for direct liquid injections into the GC, direct
injection on the CMV, and headspace extraction were developed
with the standard mixture across a range of 1 ppm-300 ppm to
85
correspond with the highest concentration of compounds that
are commonly detected in polluted air [3]. The linearity of the cal-
ibration curves was determined by plotting concentrations with a
response of a signal-to-noise ratio (SNR) of 10 or larger. The linear-
ity obtained for the calibration curves were greater than 0.958,
0.969, and 0.951 for direct liquid injection, direct spike on CMV
and headspace extraction, respectively.
The sensitivity of the method was tested by determining the
method detection limits (MDL) and the method quantitation limits
(MQL) for all analytes by direct spike on CMV and by headspace
extraction with the CMV device (Table 1). The reported MDL and
MQL in Table 1 from the calibration curves, represent the minimal
sample concentration that can be spiked on a can for the signal-to-
noise ratio (SNR) of the response to be at least 3. Calibration curves
for headspace extraction of methylene chloride, pyridine, and phe-
nol were not feasible due to their low detection. These results can
be attributed to different physicochemical properties of each com-
pound [2931]. Methylene chloride is the most volatile compound
(vapor pressure = 414 Torr at 25 C) in the mixture, which lead to a
tendency to incur breakthrough. Pyridine is the most polar com-
pound (log P = 0.65 and m = 2.2 Debye) in the mixture, which may
have led to an incompatibility in its physical interaction with the
hydrophobic PDMS adsorption phase of the CMV sampler. Overall,
an extremely low extraction recovery of only 0.6% was obtained for
phenol, which may be attributed to a combination of low volatility
(vapor pressure = 0.325 Torr at 25 C) and the polarity of the com-
pound (m = 1.2 Debye) when sampling the headspace. In a separate
experiment a spiked amount of 500 ng yield extracted amounts of
13 ng, 50 ng, 3 ng for methylene chloride, pyridine and phenol,
respectively.
Detection limits for each compound were confirmed using two
methods. In the first method analysis of 10 replicates of a methanol
blank sample was performed. The standard deviation of the inte-
grated signal (or noise) at the known analyte retention times was
multiplied by 3. The second method (described in EPA TO-17) con-
sisted of conducting an analysis on 7 replicates at a concentration
close to the expected MDL. The concentrations selected were 1 ng
and 5 ng. The standard deviation of the replicates was multiplied
by 3.14 (Students t value for confidence interval of 99%). The mass
detected for headspace MDL and MQL were calculated using the
regression line results from the direct spike calibration curves.
The percent recovery for headspace extraction using a 200 ng stan-
dard ranged from 2.726%, with the exception of phenol (0.6%) and
nonanal (1.2%).
Table 2 summarizes the results obtained for the target com-
pounds following the additional EPA TO-17 performance criteria:
analytical precision, distributed volume pair, accuracy and break-
through. The analytical precision calculated demonstrates lower
performance for acetophenone and nonanal. However, all other
compounds have an analytical precision of 20% or less. The dis-
tributed volume pair precision is calculated by performing several
measurements at different volumes. The amount (ng) of VOCs
extracted at different sampling volumes and sampling flow
(extraction time constant) should result in a linear response. For
this purpose, two different sampling volumes (2 L and 3 L) were
evaluated. The distributed volume pair precision obtained ranged
from 136% for all compounds. Factors that can affect the precision
are artifact formation, and breakthrough of the compounds [3]. The
compounds with distributed volume pairs higher than the
expected (25%) were benzonitrile and acetophenone. The % audit
accuracy was less than the expected percentage value (30%) for
the compounds. Calculations for % audit accuracy take into account
the % recovery for each compound to properly represent the
extraction efficiency of the CMV. All compounds were found to
meet the four method criteria specified in the EPA method TO-17
except benzonitrile, nonanal, and acetophenone.
86 A. Tarifa et al. / Forensic Chemistry 3 (2017) 8189

Table 2
Compounds listed in order of elution time following the EPA TO-17 criteria for the headspace extraction with CMV: analytical precision (n = 3), distributed volume pair (as %),
audit accuracy (as %) and calculated breakthrough (as %). In addition recoveries (as %) for headspace extraction above 30 ng target compounds, at sampling flow of 0.2 L/min and
10 min extraction time at room temperature (RT) and 10 C using the cryofocusing device.

Compounds Analytical precision % Distributed volume pair % Accy % Breakthrough % Recovery %RT Recovery% 10 C
Methylene chloride 16 21
Benzene 15 8.9 46 20
Pyridine
Toluene 20 12 8.2 35 5 37
Furfural 4 9.2
Ethylbenzene 3 16 7.4 27 20 25
m-Xylene/p-Xylene 18 17 8.2 22 24 29
o-Xylene 14 11 8.3 21 18 36
Benzaldehyde 1 11 8.4 10 2 8
Phenol
Benzonitrile 2 36 8.9 7 1 3
1,2,4-Trimethylbenzene 3 6 8.6 12 7 19
1,2,3-Trimethylbenzene 3 10 8.8 15 3 8
Acetophenone 42 34 9.5 28 2
Nonanal 60 1.3 9.9 19 12 5
Naphthalene 18 10 8.9 5 1 1

Analytical precision in percent, precision for the distributed volume pair and audit accuracy for a 200 ng standard solution. Breakthrough at 0.2 L/min and 10 min extraction
time at ambient temperature, 1 min equilibrium time for a 30 ng standard solution. Percent recoveries at ambient temperature and 10 C, 30 min equilibrium time for a
30 ng standard solution and 2 L extraction volume. Values not reported are from concentrations below the method detection limit.

Breakthrough experiments were conducted applying 0.2 L/min
pumping flow through two CMV devices connected in series, to
extract the headspace above 30 ng of the target compounds main-
tained at 20 C, and the two CMVs were analyzed separately. As
shown in Table 2, the breakthrough calculated as a percentage
from the second CMV, under the present sampling conditions,
was higher than that specified in the EPA TO-17 method (5%).
The only compounds with similar breakthroughs were benzonitrile
(7%) and naphthalene (5%). Nonetheless, the compounds were
detected at the expected detection limits.
In order to improve breakthrough and recoveries of the target
compounds, a thermoelectric cooler was constructed to accommo-
date the CMV devices and perform cryofocusing of the extracted
headspace. Breakthrough experiments were conducted using the
cryofocusing device operated at 10 C and at room temperature
(20 C) at 0.2 L/min. Compared to room temperature results, the
calculated breakthrough at 10 C were generally lower for most
compounds. In a different experiment, a higher flow rate of (1 L/
min) was used and resulted in an increase in breakthrough and
reduced recoveries, for some compounds. However, a sampling
flow of 1 L/min at 10 C favors the extraction of the compounds
eluting after 6.8 min (Table 1).
Recovery experiments (Table 2) were conducted using the cry-
ofocusing device operated at 10 C and at room temperature
(20 C) at 0.2 L/min and 30 min equilibrium time. Compared to
room temperature results, well defined higher recoveries were
measured for the Cryo-CMV extraction for all volatile compounds,
excluding naphthalene and nonanal. A 25%640% enhanced recov-
eries were achieved with 10 C extraction temperature over
20 C, for sampling the headspace generated inside a quart can
containing 30 ng of target compounds, indicating on the significant
higher detection sensitivities attributed to the novel cryofocusing
add-on CMV device (Table 2). Notably, the detection of the volatile
compounds benzene and acetophenone, became feasible in this
experimental concentration with cryo extraction, although for pyr-
idine, phenol and furfural higher concentrations are required. For
the less volatile compounds, naphthalene and nonanal, the Cryo-
CMV advantage was less apparent.
Fig. 4 shows the response curves of % recovery with different
extracted volumes (0.05, 0.1, 0.2, 0.3, 0.5, 1, 2, 3, 4, 5, 6 L) sampled
at 10 C (fig. 4 top) in comparison to the response curves at 20 C
(fig. 4 bottom). The figure shows that, at the extraction tempera-
ture of 10 C, up to 4 L of sampling volume can be drawn through
the CMV while recovering 14% and up to 67% for the BTEX com-
pounds, as well as for most detected compounds except naph-
thalene (1%). At 5 and 6 L sampled air the percent recovery starts
to plateau for some compounds and show a gradual decrease to
7% up to 44% for the latest. However, at an extraction temperature
of 20 C, sampling volume can be drawn through the CMV up to 2 L
volume only, followed by a gradual decrease for BTEX down to 6 L,
and complete loss of benzene is observed at 3 L due to break-
through. An improvement between 35100% in recoveries resulted
in the use of the novel Cryo-CMV device with lower temperature
extraction for all compounds, notably for benzene, allowing its
detection in the tested experimental concentration even at high
sampling volumes. Generally, the decrease in recoveries with the
increased extracted volume of air, while a continuous vapor source
is sampled, is attributed to saturation of the sampler adsorption
phase leading to breakthrough. The experimental setup developed
herein, is based on a finite amount of compound generated inside a
semi-closed system, and up to 37% of BTEX compounds were sam-
pled in a short time scale, followed by a moderate loss with
increased extracted air. These results demonstrate the efficient
irreversible trapping of the CMV adsorption phase for the volatile
compounds, at 20 C and all the more at -10 C extraction
conditions.
3.1.2. Dynamic headspace sampling of indoor air with CMV
As a proof of concept, CMV sampling was applied for collection
of indoor air VOCs using the optimized parameters: sampling flow
rate (0.2 L/min) and 10 min extraction time. Three replicates of
indoor air samples were extracted from different rooms including
a laboratory (918 ft2), a classroom (1694 ft2), and a nail and hair
salon (1053 ft2). For air extraction in the laboratory the portable
pump and the CMV were placed on top of a table in the center of
the room, with the CMV positioned in an upward direction. The
air extraction in the classroom was performed in one of the corners
of the room. The air extraction in the hair and nail salon was per-
formed towards the middle of the room between the hair and nail
sections. All replicates were performed in the same location for
each room. The extraction was performed at room temperature
(21.0 C).
An example of the chromatogram obtained from the hair and
nail salon is shown in Fig. 5. At all three indoor air spaces, the com-
pounds identified were detected at least in two of the three repli-
cates obtained from each room. From the compounds calibrated in
 A. Tarifa et al. / Forensic Chemistry 3 (2017) 8189 87

70% -10 C CMV extraction

temperature
60%
50%

% Recovery
40% Benzene
Toluene
30%
Ethylbenzene
20%
m-Xylene/p-Xylene
10%
o-Xylene
0%
0 1 2 3 4 5 6 7
Extracted volume, L

70% 20C CMV extraction

temperature
60%
50%
% Recovery

Benzene
40%
Toluene
30%
Ethylbenzene
20% m-Xylene/p-Xylene
10% o-Xylene
0%
0 1 2 3 4 5 6 7
Extracted volume, L

Fig. 4. Response curves for the recovery (as %) at 30 min equilibrium time and different extracted volumes at 10 C (top) and 20 C (bottom) for the BTEX compounds
showing improvements of up to 100% in the recovery of the compounds of interest.

this study, toluene, m, p-xylenes and o-xylenes were detected
below the MQL in the laboratory and the classroom. In the hair
salon, the amount of toluene detected was only 3.25  10 3 mg/
m3 (overall 6.5 ng). According to the Occupational Safety & Health
Administration (OSHA), the permissible exposure limit (TWA) for
toluene is 37 mg/m3, which is orders of magnitude higher than
the amount detected in the nail salon [16].
The presence of other compounds detected in the samples
including hexane, camphor, tridecane, tetradecane, pentadecane,
and diethyl phthalate were confirmed by obtaining the retention
time and mass spectrum of a 10 ppm standard solution. Quantita-
tion of these compounds was not conducted. Other compounds
such as acetone, ethyl acetate, ethyl methacrylate, butyl ester
acetic acid, 1-chloro-4-(trifluoromethyl)benzene, and limonene
were identified using the mass spectra by comparison with the
NIST library. Presence of acetone in nail salon is expected since this
is the main ingredient of nail polish remover. Ethyl acetate, ethyl
methacrylate, and butyl ester acetic acid are also ingredients of nail
polish, acrylic nails, and acrylic removers [1720]. In addition,
1-chloro-4-(trifluoromethyl)benzene is used as a solvent in com-
mercial surface finishes [21]. Limonene is a common monoterpene,
detected in indoor air and is found in products such as, citrus fruits,
air refresheners, household cleaning agents, hair dye, and waxes
[2227].
3.2. CMV and Cryo-CMV for smokeless powders sampling
Calibration curves for direct injection on the CMV, across the
range of 2 ppm100 ppm for nitroglycerine (NG) and 0.5 ppm
25 ppm for diphenylamine (DPA), were generated to correspond
with the concentration range detected above the smokeless pow-
der samples. The linearity obtained for the calibration curves were
greater than 0.976 and 0.998 for NG and DPA, respectively. The
percent recovery for headspace extraction, generated by 1000 ng
(spiked in 10 mL methanol) standard solutions in a quart can after
10 min. incubation time at ambient conditions, was determined
to be 1% or less for 2 L extraction air at a flow rate of 1 L/min,
which corresponds with previously reported and expected results

4.5E+05 1 4 5 13
10 11 CMV blank
3.0E+05 7 9
8 12 Hair and nail
salon
1.5E+05
2 3 6
0.0E+00
2 3 4 5 6 7 8 9 10 11 12
Retention time, min

Fig. 5. Chromatogram obtained by CMV-GCMS showing the compounds identified in the hair and nail salon. Peak identities: (1) acetone; (2) ethyl acetate; (3) toluene; (4)
ethyl methacrylate; (5) butyl ester acetic acid; (6) 1-chloro-4-(trifluoromethyl)benzene; (7) benzaldehyde; (8) limonene; (9) camphor; (10) tridecane; (11) tetradecane; (12)
pentadecane; (13) diethyl phthalate.
88 A. Tarifa et al. / Forensic Chemistry 3 (2017) 8189
[9]. These two compounds, known to be detected in the headspace
of smokeless powder samples [9], were also the only organic com-
ponents detected in the headspace of swabs collected from the
hands of 50 gun shooters [10].
To reach enhanced sensitivity in the detection of smokeless
powders and gunshot residues, the novel Cryo-CMV device devel-
oped in this research, was applied for the detection of NG and
DPA vapors. Fig. 6 demonstrate the amounts of NG detected at
three different extraction temperatures using the operated tem-
perature Cryo-CMV at 10 C, 2.5 C and 20 C for comparison.
All measurements were performed for the headspace extraction
of a quart can incubated at ambient conditions for 18 h with
100 mg double-base smokeless powders, sampled for 2 min at
1 L/min flow rate. Enhanced signals were measured for the cryofo-
cusing extractions. An average (3 replicates) of 38% and 29% rela-
tive increase in recoveries was measured at the 2.5 C and
10 C, respectively, compared to ambient temperature extraction.
For NG, 90 15.5 ng was extracted at 2.5 C and 84 12.5 ng at
10 C, compared to 65 12 ng at ambient extraction for 2 min
sampling time. Although a minor overlap in amount extracted
was observed between the ambient and both cooled extraction
temperatures, improved recoveries were still measured for the
Cryo-CMV. The amounts detected are at least 30-fold higher than
the MDL. The higher recovery achieved for 2.5 C extraction over
10 C, can be attributed to the reduced condensation, while still
keeping the CMV cooled during sampling. The condensation
observed inside the CMV at 10 C, modify the performance of
the adsorption surface while sampling. In order to address conden-
sation effects, using the same experimental setup, silica gel desic-
cant were placed inside the quart can during incubation. Compared
to ambient temperature, at 10 C a 62% higher recoveries were
observed for NG, confirming the contribution of the cryo extraction
for enhanced recoveries. This apparent increase suggests that fur-
ther improvements in the setup will lead to better results for cry-
ofocused extraction of smokeless powders.
Generally, higher sampling flows are preferred to enhance the
detection sensitivity of semi-volatile compounds, while avoiding
the condensation effects generated from cooling the device at
sub-ambient temperatures. Unlike the sampling flow rate (0.2 L/
min) used for the extraction of VOCs to yield higher recoveries
and avoid breakthrough, the sensitivity of NG was lower at 0.2 L/
min than when extraction was performed at a higher sampling
flow of 1 L/min, while sampling the same volume of air for VOCs
Fig. 6. NG headspace extraction above 100 mg BLC-2 double base smokeless
powder incubated for 18 h inside a quart can using Cryo-CMV device operated at
2.5 C and 10 C in comparison to 20 C (room temperature).
and NG (2 L). Therefore, optimization of the extraction parameters
is crucial to obtain higher recoveries.
The well-known marker of smokeless powder, DPA, was
detected in all the smokeless powder samples at the different
extraction temperatures tested. The amount of DPA detected at
room temperature was 1.6 ng and the amount extracted at
2.5 C and 10 C was not significantly higher for this semi-
volatile compound (vapor pressure 6.4 4 torr, 25 C). These results
indicate on the low availability of DPA vapors inside the cans for
extraction with the CMV and the short time available during
extraction for the vapors to rebuild at the experimental setup
applied.
4. Conclusions
The method proposed facilitates a more rapid sampling and
detection method for VOCs using a novel technique, CMV-GC
MS. The CMV extraction method has been shown to be a fast and
sensitive technique for the headspace extraction of organic vola-
tiles present in the air. One of the major advantages of CMV devices
over sorbent tubes for air monitoring purposes is the ability to con-
duct dynamic headspace sampling in less than 10 min, compared
to 1 h for sorbent tubes. The CMV is also more cost effective, cost-
ing less than $1 to manufacture compared to ($100/each) for
sorbent tubes. The low cost facilitates the disposable nature of
the CMV when multiple uses are not feasible. However, the CMV
is reusable after desorption in the GC injection port with experi-
mental results demonstrating a life span of the CMV used for more
than 100 injections and headspace extractions, comparable to sor-
bent tubes such as Tenax TA [27]. Furthermore, the CMV can be
directly introduced into the injection port of the GC using a low-
cost thermal separation probe. The advantages of direct injection
in the GC port include reduced sample loss and faster elution com-
pared to other thermal desorption units that contain a transfer line.
The significance of this study was to demonstrate the utility of
CMV devices for the extraction of volatile organic compounds in
ambient air through method validation following a comparison
to the EPA method TO-17. The results indicate that the CMV
method meets the criteria described in the EPA method, depending
on the analyzed compound. The four criteria consisted of: method
detection limit (MDL) of 0.5 ppbv (5 ng), analytical precision
within 20%, precision for the distributed volume pair of 25% or less,
and an audit accuracy of 30% or better for the expected concentra-
tion range 0.525 ppbv (5300 ng) of the TO-17 method outlined
by EPA [28]. All compounds were found to meet the four method
criteria specified in the EPA method TO-17 except benzonitrile,
nonanal, and acetophenone. The data also suggests that improving
the chemistry of the CMV sorbent coatings will enhance the molec-
ular weight range of compounds that can be extracted as well as
improve on the extraction performance, further enhancing the ana-
lytical figures of merit described in the EPA TO-17 method.
A cryofocusing device consisting of a single-stage Peltier cooled
module was designed and constructed for this study, to perform
headspace extraction of target compounds by cooling the CMV
device to 10 C during the extraction. The application of the
Cryo-CMV for VOCs sampling improved recoveries for all com-
pounds and remained similar for methylene chloride, toluene,
and naphthalene. Similarly, the application of the Cryo-CMV for
double-base smokeless powder sampling improved the extraction
recovery of NG, suggesting this additional application within
forensic chemistry as well as many other potential applications
such as fire debris analysis, volatiles in clandestine laboratories
and others. Further improvement of the cryofocusing device sam-
pling setup to include insulating the sampling tube and to the
CMV sampling geometry, both of which are currently pursued as
 A. Tarifa et al. / Forensic Chemistry 3 (2017) 8189
lines of investigation in our group, may provide significant
improvements in the extraction capability to retain high volatility
compounds and small molecules, while minimizing condensation
effects of water when sampling at sub-zero temperatures.
The use of CMV at 20 C has already been demonstrated by Fan
and Almirall for the analysis of explosives in air [9], by Tarifa and
Almirall for the analysis of organic GSR from the hands of shooters
[10] and by Wiebelhaus et al. for the analysis of marijuana markers
[11]. It is anticipated that cryofocusing CMV extraction operated at
sub-ambient temperatures will further improve these applications
as well as other forensic chemistry applications. Additional foren-
sic applications for cryofocusing CMV headspace extraction under
study in our research group include the detection of VOCs from fire
debris, either from collected samples in a can or directly at the
scene of a fire, in order to detect VOCs associated with the presence
of ignitable liquid residues.
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