Int. J. Ion Mobil. Spec.
(2008) 11:61–69
DOI 10.1007/s12127-008-0006-5
ORIGINAL RESEARCH
A rapid analytical method for hair analysis using ambient
pressure ion mobility mass spectrometry with electrospray
ionization (ESI-IMMS)
Prabha Dwivedi & Herbert H. Hill Jr
Received: 13 September 2007 / Revised: 8 May 2008 / Accepted: 9 May 2008 / Published online: 10 June 2008
# Springer-Verlag 2008
Abstract Analysis of hair is often applied to assess drug
abuse history, exposure to environmental and industrial
pollutants, heavy metals, gestational drug exposure and
various other screening purposes. This manuscript reports
the application of ambient pressure ion mobility spectrom-
etry mass spectrometry (IMMS) with electrospray ionization
(ESI) source as a rapid analytical tool for hair analysis. The
study demonstrated that ion mobility spectrometry (IMS) as a
pre-separation technique prior to analysis by mass spectrom-
etry (MS) provides detection and determination of com-
pounds of interest present in hair at nano-molar concentration
level. After extraction of analytes from hair, the ESI-IMMS
method of analysis does not require the derivatization or
sample treatment that is often required for other separation
methods such as gas chromatography. One advantage of IMS
over chromatography separation is that resolving powers are
similar to those in GC and much greater than those possible
by liquid chromatography. In addition, separation speed is
faster than both gas and liquid chromatographic methods.
Four of the nine hair samples anonymously donated by
customers at a local hair salon tested positive for caffeine and
two of the four samples that tested positive for caffeine also
tested positive for nicotine. A positive response based on
mass analysis for methamphetamine was obtained for one of
the hair samples. Further investigation using the mobility
data demonstrated that the response was a false positive and
that it may have occurred from the use of a hair gel. This
article reports the potential of IMMS as an analytical
technique for rapid and routine screening of hair samples,
cosmetics, and pharmaceuticals.
P. Dwivedi : H. H. Hill Jr (*)
Department of Chemistry, Washington State University,
Pullman, WA 99164, USA
e-mail: hhhill@wsu.edu
Keywords Hair analysis . Ion mobility spectrometry-mass
spectrometry . Metabolomics . Drugs . Cosmetics .
Pharmaceuticals
Introduction
Analysis of hair is used in various toxicological, clinical, or
forensic investigations. The advantages of performing hair
analysis over other biological samples are that hair analysis
(1) is a less intrusive method, (2) provides a history of drug
use, (3) is more resistant to adulteration, (4) can be easily
collected and stored, (5) has long shelf life and (6) can be
easily decontaminated [1]. Determination of cause of death,
or assessment of drug abuse and exposure levels to
environmental and industrial pollutants [2–4] or monitoring
therapeutic compliance and treatment [5, 6] are achieved by
performing analysis of hair samples. Identification, detec-
tion and/or determination of metabolites present in hair can
also be used to predict, detect, and treat various diseases
[7–14]. In addition, segmental analysis of hair yields
information that can be related or associated with chronol-
ogy of diseases, poisonings, or drug use [15, 16].
The use of hair for forensic analysis is not a new concept.
In 1858, hair obtained from a dead body, exhumed after
11 years was analyzed for arsenic and cause of death by
arsenic poisoning was reported [17]. Hair analysis as a
means to determine histories of drug abuse gained popular-
ity only in 1970s and 1980s when methods to detect opiates
in digested hair samples were reported [18, 19].
Currently, analysis of hair to determine drugs of abuse is
performed by separation techniques such as gas chromatog-
raphy [20, 21], liquid chromatography [22–24] and capillary
electrophoresis [25–27] with or without mass spectrometry.
However, the steps required for these analyses such as
62
extraction, concentration, derivatization, separation and
detection, of targeted analytes reduce sample throughput.
Derivatization procedures are generally applicable to
certain chemical classes and often render chromatographic
methods suitable only for a specific class of analytes. For
example, preparation of hair sample for drug analysis
by gas chromatography-mass spectrometry (GC-MS)
involved acid hydrolysis of hair, followed by a three-step
liquid–liquid extraction, and derivatization with N,O-bis
(trimethylsilyl) trifluoroacetamide plus 1% trimethylchlor-
osilane [28]. Similarly, for sensitive determination of
histamine HA in hair by high pressure liquid chromatography–
electrospray ionization mass spectrometry (HPLC–ESI-MS),
HA was first extracted from hair by acid hydrolysis, followed
by fluorescence labeling for HPLC separation and then
detected by ESI-MS [29]. Thus, there is a need for develop-
ment of innovative approaches for hair analysis in order to
simplify the measurement, increase sample throughput and
minimize diversity in sample preparation.
Ion mobility spectrometry (IMS) with its millisecond
separation time provides 105 times greater number of plates
per second when compared to chromatographic separation
methods such as GC, LC, and CE [30]. Currently, IMS is
widely used as a rapid detection method for vapor phase
analytes such as chemical warfare agents, explosives, drugs
and air contaminants [31]. Pyrolysis-IMS and thermal
desorption-IMS have also been applied for analysis of hair
for drugs [32–35]. However, determination of non-volatiles
present in hair is not feasible using pyrolysis-IMS and
thermal desorption-IMS. With the introduction and devel-
opment of electrospray ionization (ESI) as the ion source
for IMS [36, 37], application of IMS have been extended to
the analysis of biological samples and structural studies of
non-volatile ionic species [38–41]. Thus, ESI-IMMS seems
to be viable analytical option that can be used for rapid
determination of volatile and non-volatile species present in
hair. Two dimensional information (mobility and m/z)
acquired through an IMMS experiment allows the analysis
of complex biological samples. In a typical IMMS
experiment, ions produced at atmospheric pressure by an
ion source are injected in the drift region of an IMS where
ions are separated based on their size-to-charge ratio. These
ions are then introduced into the MS through a pin-hole
leak, guided by einzel lenses to the mass analyzer at
pressure of ∼10−5 torr, and are separated based on their
mass-to-charge ratio. Due to high velocity of ions in
vacuum compared to that at ambient pressures, time spent
by an ion in the mass analyzer is considered negligible with
respect to the time that the ion spends in the drift region of
the ambient pressure IMS.
Hair, as any other biological sample, is expected to be
complex in nature with multiple constituents. The objective
of this study was to investigate if ion mobility spectrometry
Int. J. Ion Mobil. Spec. (2008) 11:61–69
coupled to ESI as ion source and mass spectrometry (ESI-
IMMS) can be applied for rapid analysis of hair samples.
As a “proof of concept study” the study focused on
determination of caffeine and nicotine present in hair
samples that were obtained as donations from customers
at a local hair salon. These two drugs were selected as the
target compounds for this study because caffeine and
nicotine are the most common drugs that can be found in
hair samples from the general population.
Materials and methods
Samples and solvents
High performance liquid chromatography grade solvents
(methanol, water and acetic acid) were purchased from J. T.
Baker (Phillipsburg, NJ, USA). Caffeine and nicotine were
purchased from Sigma-Aldrich (Sigma Aldrich Chemical
Co., St. Louis, MO, USA). Hair samples were anonymous-
ly donated by customers at a local hair salon.
Sample preparation
Hair samples collected at a local hair salon from nine
customers were cut into about 5 mm lengths, washed three
times with 50:50 methanol water solution, and left to dry
overnight at room temperature. After the sample was
evaporated to dryness, 50 mg of each hair sample was
extracted with 4 ml of methanol in a screw cap culture tube
by heating in a water bath for 8 h at ∼80 °C and then cooled
to room temperature. The extract were then filtered using a
0.2 μm size syringe filter, and diluted with HPLC grade
water and acetic acid to make solutions of composition
47.5:47.5:5 (methanol–hair extract/water/acetic acid). The
extract was diluted to approximately half its original
concentration (110% dilution). Samples were stored at
−4 °C until analysis.
ESI-IMMS analyzer
The ESI-IMMS analyzer comprised of an electrospray
ionization source (ESI), an ion mobility analyzer (IMS)
and a quadrupole mass analyzer (MS), the schematic of
which is shown in Fig. 1.
ESI Source For the generation of electrospray ions, sample
was infused into a 30 cm long, 50 μm inner diameter silica
capillary by a KD Scientific 210 syringe pump (New Hope, PA,
USA) at a flow rate of 5 μl/min. The capillary was then inserted
into a water-cooled 22-gauge stainless steel needle, the tip of
which was centered ∼1.0 cm from the target screen of the IMS
Int. J. Ion Mobil. Spec. (2008) 11:61–69
Fig. 1 Schematics of the ion
mobility mass spectrometer with
electrospray ionization source.
The mass spectrometer was a
quadrupole mass analyzer with
electron multiplier as detector.
The ESI and the IMS were
operated at ambient pressure and
the MS was at a pressure of
10−5 torr. The IMS and MS were
coupled through a 40 μm pin-
hole interface
analyzer and the capillary protruded ∼2 mm out of the stainless
steel needle tip. The ESI needle was maintained at a potential of
+3.0 kV with respect to that on the target screen of the IMS.
IMS analyzer The IMS analyzer was constructed at
Washington State University, and used a stacked-ring
design that has been described in previous publications
[30]. In summary, the IMS tube was divided into a
desolvation region (7.2 cm in length) and a drift region
(21.8 cm in length) separated by a Bradbury–Nielsen-style
ion gate. Both regions consisted of alternating alumina
spacers and stainless steel rings with high temperature
resistors connecting the stainless steel rings (Caddock
Electronics, Riverside, CA, USA; 500 kΩ resisters for the
desolvation region, 1 MΩ resisters for the drift region,
250 °C, 0.1% tolerance). The temperature of the drift
region, desolvation region, and the drift gas was maintained
at 250 °C, the ion gate held at a potential of 7.51 kV, and
the instrument was operated at ambient pressure (696 to
703 torr in Pullman, WA, USA). With a potential of 9.5 kV
at the IMS target screen, electric field in the desolvation
region was ∼276 V/cm and ∼344 V/cm in the drift region of
the IMS (E/N between 2–3 Td). Preheated counter flowing
drift gas (nitrogen) at a flow rate of ∼1,300 ml/min was
introduced at the end of the drift region (preheated to the
temperature of the IMS). The ions were injected into the
IMS drift region in pulses of 0.2 ms using an ion gate
operating at a frequency of 25 Hz.
MS analyzer The IMS analyzer was interfaced to a model
150-QC ABB Extrel (Pittsburgh, PA, USA) quadrupole
analyzer (m/z range of 0–4,000 amu) via a 40-μm pinhole
interface. Earlier publications from our research group
document the detailed description and schematics of the
IMMS analyzer [30, 42]. The output signal from the
multiplier was amplified by a Keithley model 427 amplifier
63
(Keithley Instruments, Cleveland, OH, USA). The ampli-
fied signal was then sent to either the MS data acquisition
system or IMS data acquisition system.
Instrument control, data acquisition and processing Merlin
software (ABB Extrel, Pittsburgh, PA, USA) was utilized for
all MS data acquisition, analysis and control. For the IMS
gating and data acquisition, the electronic controls were built
at Washington State University. Labview (National Instru-
ments, Austin, TX, USA) based software developed at
Washington State University, Pullman WA, USA; was used
for IMS data acquisition and IMS gate-control. Two types of
ion mobility spectrum were generated through ESI-IMMS
experiments: non selected ion mobility (NSIM) spectrum
and selected ion mobility (SIM) spectrum. In the first case,
the MS was operated in the “scan mode” with the DC voltage
turned off and served as an ion transfer line between the IMS
and the detector and provided a non-selected ion mobility
(NSIM) spectrum for the analyte(s). In the second case, the
MS was operated in the “single ion monitoring mode” and
served as a filter that allowed transfer of ions with a
preselected m/z value reach the detector. Under these
conditions, a selected ion mobility spectrum (SIM) was
obtained. Operation of IMMS in SIM mode allows one to
monitor the drift time(s) of ions associated with preselected
m/z values.
Calculations
Reduced mobility value for ions (Ko in the units of square
centimeter per volt second) was calculated using the
following equation:
 2   
L 273:15 P
K0 ¼
Vtd T 760
64
where, L is the length of the drift region in centimeter, V is
the voltage applied to the ion-gate in volts, td is the ion drift
time in seconds, T is the drift region temperature in Kelvin,
and P is the ambient pressure in torr.
Results and discussion
Analysis of hair samples
Non selective ion mobility (NSIM) spectra of hair samples
identified as A, B, C, D, E, F, and G were acquired by
operating the MS in the “scan mode”. Figure 2 shows the
superimposed NSIM spectra of (1) negative control sample
(m/z 10–400 Da), (2) hair sample A (m/z 100–400 Da, and
(3) hair sample E (m/z 100–400 Da). Since most of the ions
detected in the negative control sample were of m/z values
less than 100 Da, the cutoff m/z value for NSIM spectra of
hair samples was set at 100 Da to exclude background ion
peaks from the NSIM spectra. Depending on the identity of
the hair sample, number of ions detected in the m/z range of
100–400 Da varied between 30 and 55. Peak patterns
(NSIM spectra) and number of peaks detected were
reproducible for each hair sample in two replicate measure-
ments. Qualitative differences of the hair samples were
observed by comparing their respective NSIM spectra. For
example, Fig. 2 shows the difference in peak pattern/number
1.40
Negative control
Hair Sample A
Hair Sample E
1.15
IMS Response (nA)
0.90
0.65
Negative
0.40 A E control
0.15
-0.10
0 10 20 30 40
Drift time (ms)
Fig. 2 Non selective ion mobility (NSIM) spectra of negative control,
hair sample A and hair sample E. Each of the hair samples produced
characteristic and reproducible IMS spectrum as illustrated for hair
samples A and E. The IMS spectra were obtained in the scanning
mode of the MS. The negative control NSIM spectrum was obtained
in the mass range of 10–400 Da and the NSIM spectra of hair samples
were obtained in the mass range of 100–400 Da
Int. J. Ion Mobil. Spec. (2008) 11:61–69
of ions detected between hair samples A and E. Absence of
IMS peaks in the drift time range of 15–30 ms in the NSIM
spectrum of the control and their presence in the NSIM
spectra of hair extracts shows that various analytes were
extracted from the hair samples and were detected by ESI-
IMS. Incomplete resolution of IMS response peaks
corresponding to 30–55 m/z ions that were detected by MS
resulted in a broad “hump” like peak shape in the NSIM
spectra of each hair sample. Selected ion mobility (SIM)
spectrum provided specific details (m/z, mobility, and
intensity) for each of the detected analytes. SIM spectra of
few target analytes detected in the extracts of hair samples
are discussed in the following sections. Ability to rapidly
analyze hair samples without any further sample treatment
after extraction, allows one to use ESI-IMS as a screening
tool for rapid analysis of hair samples. ESI-IMMS can be
used to rapidly characterize complex biological samples such
as hair and determine the identity of each constituent using
two dimensional IMMS data.
Targeted component analysis of hair samples
Since the hair samples collected were anonymous and thus
provided no prior information about the subjects, the
experiments were directed towards detection of most
common drugs that can be potentially present in hair samples
(e)
Hair sample A
(d)
IMS Response
Hair sample E
(c)
Hair Sample B
(b)
Hair sample G
(a)
Negative control
50
0 10 20 30 40 50
Drift Time (ms)
Fig. 3 Selected ion mobility (SIM) spectra obtained at m/z 195 of (a)
negative control, (b) hair sample G, (c) hair sample B, (d) hair sample
E, and (e) hair sample A. The drift time of ion at m/z 195, the (MH)+
ion of caffeine, was monitored in each sample. Variation in intensity
of the IMS peak at m/z 195 in each hair sample was observed
Int. J. Ion Mobil. Spec. (2008) 11:61–69 65

collected from general population. The focus of the study
was to investigate if drugs such as nicotine (m/z=162) and or
caffeine (m/z=194) in the collected hair samples could be
extracted and detected by ESI-IMMS. Out of nine hair
samples analyzed,
 based on m/z values
of protonated
 ions of
nicotine ðMHÞþ
ðm=z ¼ 163Þ and caffeine ðMHÞþ

ðm=z ¼ 195Þg, peaks corresponding to caffeine were
detected in hair samples identified as A, B, E, and G and
peaks corresponding to nicotine were detected in hair
samples identified as A and E. The peaks detected at m/z
values of 163 and 195 were identified as that of nicotine and
caffeine extracted from hair because (1) the m/z values and
reduced mobility values (Ko) of these peaks matched those
measured for standard solutions of nicotine and caffeine
analyzed by ESI-IMMS and (2) these peaks were not
detected in the negative control. The Ko values of the ions
at m/z 163 and m/z 195 in hair sample A also matched those
measured in hair sample E.
Figure 3 shows selected ion mobility (SIM) spectra of
hair samples A, B, E, and G at m/z value of 195 acquired by
operating the MS in the single ion monitoring mode. The
drift times of the IMS peak at m/z 195 was measured to be
19.84, 19.81, 19.88, and 19.86 ms in samples A, B, E, and
G, respectively, with Ko values of 1.53±0.02 cm2 V−1 s−1.
With three consecutive measurements, the standard devia-
tion in drift time for all samples was measured to be less
than ±0.06 ms. The Ko for standard solution of caffeine at
m/z 195 was measured to be 1.53±0.01 cm2 V−1 s−1 which
compared well with the reduced mobility measured for the
detected ion of m/z 195 in each of the hair samples. SIM
spectra for the negative control at m/z 195 is also shown.
Hair samples containing both caffeine and nicotine also
produced a peak at m/z value of 231 Da and drift time of
21.26±0.04 ms. The ion was tentatively identified as water
adduct peak of caffeine fM 2ðH2 OÞHgþ
ðm=z ¼ 231Þ. By
comparing mobility values measured for the standard

Fig. 4 Ion mobility spectra il- 150 163 195 231

lustrating the detection of nico-
0.8
tine in hair samples. On the left
panel are shown the NSIM
spectra of hair samples A, and E 0.05 Hair sample A
with highlighted nicotine peak at m/z 163 amu
m/z value of 163. Also shown 0.6
drift time 19.24 ms
IMS Response (nA)

are the peaks at 150, 195, and

231 Da for reference. The right
hand panel shows the SIM 0.03
spectra obtained at m/z 163 with 0.4
drift times of 19.24 and
19.26 ms in hair samples A and
E, respectively
0.01
0.2
Hair sample A

0.0 -0.01
17 18 19 20 21 22 23 0 10 20 30 40 50

0.8

0.05 Hair sample E

m/z 163 amu
0.6
drift time 19.26 ms
IMS Response (nA)

0.03
0.4

0.01
0.2

Hair sample E

0.0 -0.01
17 18 19 20 21 22 23 0 10 20 30 40 50
Drift Time (ms) Drift time (ms)
66 Int. J. Ion Mobil. Spec. (2008) 11:61–69

caffeine solution and the peak observed in the hair samples

A A, B, E, and G at m/z 195 it was concluded that the samples
Hair Sample A; m/z 150
Drift time 18.14 ms contained caffeine and can be determined by ESI-IMMS
method.
NSIM spectra of hair sample extracts A and E in the drift
time range of 17–22 ms are shown in Figure 4, left hand
side panel. The IMS peaks at m/z values of 150, 163, 195
and 231 were identified in the two spectra for reference. As
shown, a peak at m/z value of 163 was detected in both
samples. From the acquired SIM spectra the drift times of
IMS Response

B the peak at m/z 163 was measured to be 19.24 and 19.26 ms

Methamphetamine standard
m/z 150; Drift time 18.66 ms
C
Methamphetamine
in Hair Sample A; m/z 150
Drift time 18.11 and 18.65 ms
10 15 20 25
Drift Time (ms)
Fig. 5 SIM spectra at m/z 150 showing the separation of isobaric
ions. SIM spectra of hair sample A (a), standard solution of
methamphetamine (b) and hair sample A spiked with methamphet-
amine (c) are shown
in the samples A and E respectively as shown in the right
hand panel of Fig. 4. The reduced mobility for nicotine in
the standard solution was determined as 1.60±0.02 cm2
V−1 s−1. This peak was tentatively identified as that of
nicotine by comparing the mobility of the m/z 163 peak
detected in hair samples to the m/z 163 peak detected for a
standard solution of nicotine. A Ko value of 1.61±0.01 cm2
V−1 s−1 was measured for the standard nicotine solution.
Figure 4, left hand side panel, also shows an intense
peak at m/z value of 150 that was detected in hair sample A.
Since methamphetamine (m/z=149), a commonly used drug
of abuse, produces a peak at m/z 150 Da ðMHÞþ

30 35 40
ðm=z ¼ 150Þg, IMMS measurements were performed to
tentatively identify the peak. Figure 5 shows the SIM
spectra at m/z 150 obtained for (a) hair sample A, (b)
standard methamphetamine solution, and (c) hair sample A
spiked with methamphetamine. The reduced mobility value

Fig. 6 NSIM spectra of vitamin

C (a), vitamin B (b), aspirin (c), 3.4
hair gel (d), and hair sample A (a)
(e). IMS peak with a drift time Vitamin B
of 18 ms at m/z 150 observed in
hair sample A is aligned with
that observed in the NSIM (d)
1.6
spectrum of the hair gel Hair gel
IMS Response (nA)

-0.2
(b)
Vitamin C 3.4

(e)
1.6 Hair sample A
(c)
Aspirin

-0.2

10 15 20 25 30 10 15 20 25 30
Drift time (ms) Drift time (ms)
Int. J. Ion Mobil. Spec. (2008) 11:61–69 67

Calibration curve for caffeine

of the m/z 150 peak in the hair sample was measured to be 4.0
1.63±0.02 cm2 V−1 s−1. However, the standard solution of 3.5
methamphetamine produced a peak at m/z 150 with a y = 0.721x -0.000

IMS Response (nA)

3.0
reduced mobility of 1.69±0.01 cm2 V−1 s−1 was produced. 2.5
R2 = 0.999

Mass-mobility comparison between standard and hair 2.0

sample A suggested that the m/z 150 peak detected in the 1.5
hair sample A was not methamphetamine. The separation of 1.0
isobaric ions based on mobility measurements also demon- 0.5
strates one of the advantages of using IMS as a rapid 0.0
separation method prior to MS analysis. 0 1 2 3 4 5 6
In an effort to identify the m/z 150 peak detected in hair Mixing Ratio of Caffiene in ppm
extract, various commonly used hair cosmetics such as gels,
Calibration curve for nicotine
shampoos and creams, and over the counter drugs and 1.6
health supplements that were suspected to be the source of 1.4
y = 0.138x + 0.0231
the peak at m/z 150, were analyzed by ESI-IMMS.

IMS Response (nA)

1.2
R2 = 0.996
Although all samples produced distinguishable ion mobility 1.0
patterns, the drift time of the m/z 150 peak observed in the 0.8
hair sample A matched only with a peak observed in one of 0.6
the hair gels analyzed. Non selective ion mobility spectra of 0.4
vitamin B, vitamin C, aspirin and hair gel are shown in 0.2
Fig. 6. On the right hand side panel of the figure is shown 0.0
the NSIM spectrum of the hair sample A aligned with the 0 2 4 6 8 10 12
NSIM spectrum of hair gel. The presence of a m/z 150 peak Mixing Ratio of Nicotine in ppm

in the hair gel at a drift time that matched with that
measured in the hair sample suggested that the peak
observed in the hair sample at m/z 150 could have
originated from the hair gel either as adsorbed or absorbed
hair gel component. In the absence of MS-MS capability
the identity of the m/z 150 peak was not confirmed.
However, the basic inference drawn from the experiment
was that IMS can rapidly provide a confirmatory piece of
data to distinguish ions with same m/z values present in
complex samples.
Determination of concentration of extractible target
compounds in hair samples
To determine the extractible concentration of caffeine and
nicotine incorporated in hair, calibration curves for both
were constructed by spiking hair samples in which peaks
for caffeine and/or nicotine were not detected with known
amount of caffeine and nicotine. Standard solutions of
caffeine and nicotine in mixing ratio range of 0.01–5.0 and
0.01–10.0 ppm respectively were analyzed by ESI-IMMS.
Intensity of IMS spectra obtained from an average of 2,500
IMS spectra was measured. Average signal intensity of
three consecutive measurements was plotted against the
concentration of the standard caffeine and nicotine solu-
tions (Fig. 7). In the concentration range studied, linear
relationship between the concentration and IMS signal
intensity was observed. Concentration of caffeine in hair
sample A was measured to be ∼8.2 ng of caffeine in 1 mg
of hair. Caffeine concentration in hair sample G was
Fig. 7 Calibration curve for caffeine and nicotine. Standard solutions
of caffeine and nicotine in the mixing ratio range of 0.01–5.0 ppm for
caffeine and 0.05–10.0 ppm for nicotine were used. Measured
concentration of caffeine in the hair samples A, B, E, and G varied
between ∼8.2 to ∼0.5 ng/mg. Nicotine concentration was measured to
be ∼12 and ∼ 6 ng/mg in hair samples E and A respectively
measured to be 0.5 ng/mg of hair sample. Concentration
of nicotine in hair sample E was measured to be ∼12 ng of
nicotine in 1 mg of hair whereas concentration of nicotine
in hair sample A was measured to be ∼6 ng/mg of hair. For
more accurate quantification of analytes, calibration curves
should be generated using control hair samples with known
amount of target compounds, and recovery data. The study
illustrates the basic concept that ESI-IMMS method of
analysis has the ability to rapidly analyze hair samples for
either target compound measurement or comprehensive
determination.
Conclusions
Ion mobility spectrometry in tandem with mass spectrom-
etry and electrospray ionization (ESI-IMMS) can be used as
a rapid analytical tool for hair analysis. The ability of ion
mobility spectrometry to rapidly provide confirmatory
information in a second dimension allows one to exclude
false positives and increase confidence in the identification
of ions of interest. Also, the method does not require
derivatization or extensive sample preparation. This method
can be used either for measurement of target compound or
68
for the determination of all constituents extracted from hair
samples. ESI-IMMS method can thus be extended to target
metabolite analysis or studies investigating the whole hair
metabolome. With ability to analyze hair as a biological
matrix and detect caffeine and nicotine at low nano-molar
concentrations demonstrated, further modifications in sam-
ple preparation methods and/or improvement in instrument
sensitivity can facilitate analysis of hair for determination
of constituents (metabolites, drugs) present in sub nano-
molar concentrations. Additionally, ESI-IMMS or ESI-IMS
as a stand-alone method can be exploited for rapid screen-
ing of hair samples, cosmetics and pharmaceuticals.
Acknowledgements The authors would like to thank Fantastic Sams
of Pullman, Washington for providing hair samples. Also, the authors
would like to thank the Alcohol and Drug Abuse Research Program
(ADARP) of Washington State University (WSU) and National
Institutes of Health (NIH) for providing financial support. This work
was supported by an ADARP grant and a Road Map Grant from NIH
the (R21 DK 070274).
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