Drug Testing

Research article and Analysis

Received: 21 January 2013 Revised: 22 February 2013 Accepted: 22 February 2013 Published online in Wiley Online Library

(www.drugtestinganalysis.com) DOI 10.1002/dta.1474

Hair analysis for THCA-A, THC and CBN after

passive in vivo exposure to marijuana smoke
Bjoern Moosmann,a,b Nadine Rotha,b and Volker Auwrtera*
Condensation of marijuana smoke on the hair surface can be a source of an external contamination in hair analysis and may
have serious consequences for the person under investigation. 9-tetrahydrocannabinolic acid A (THCA-A) is found in
marijuana smoke and in hair analysis, but is not incorporated into the hair through the bloodstream. Therefore it might
be a promising marker for external contamination of hair and could facilitate a more accurate interpretation of analytical results.
In this study, three participants were exposed to the smoke of one joint every weekday over three weeks. Inhalation was
excluded by an alternative breathing source. Hair samples were obtained up to seven weeks after the last exposure and
analyzed for THCA-A, 9-tetrahydrocannabinol (THC) and cannabinol (CBN) by liquid chromatography-tandem mass
spectrometry (LC-MS/MS) analysis. Additionally 30 hair samples from various regions of the head were obtained seven weeks after
the exposure from one participant. The obtained results show that the degree of contamination depends on the hair length, with
longer hair resulting in higher THC and CBN concentrations (1300 pg/mg and 530 pg/mg at the end of the exposure period) similar
to the ones typically found after daily cannabis consumption. THCA-A could be detected in relatively low concentrations. Analysis
of the distribution of the contamination showed that the posterior vertex region was affected most. The relatively low THCA-A
concentrations in the samples suggest that most of the THCA-A found in forensic hair samples is not caused by sidestream
marijuana smoke, but by other sources. Copyright 2013 John Wiley & Sons, Ltd.

Keywords: hair analysis; contamination; tetrahydrocannabinol; tetrahydrocannabinolic acid A; LC-MS/MS

Introduction despite the presence of high THC concentrations even when

The exclusion of an external contamination of hair samples is one
of the most important aspects in forensic hair analysis. Besides
incorporation through the bloodstream during hair formation,
drugs can also get incorporated into the permanent hair through
external sources such as dust, smoke, contaminated hands,
sweat, or sebum. [15]
A common way applied for many drugs to avoid such
problems is to analyze at least one characteristic drug metabolite
proving body passage next to the parent compound. Even for
metabolites which are actually products of hydrolysis (e.g.
benzoylecgonine/cocaine), it is assumed with high probability
that no signicant external contamination occurred if the ratio
of the metabolite to the parent compound is above a certain
cut-off.[69]
However, for 9-tetrahydrocannabinol (THC) the detection of a
characteristic metabolite is difcult. Of the two main oxidative
metabolites of THC, 11-hydroxy-9-tetrahydrocannabinol and
9-carboxy-11-nor-9-tetrahydrocannabinol (THC-COOH) only
the latter can be found in hair samples of cannabis consumers
in very low but detectable amounts. However, even after
conrmed consumption, it is not detected in all cases even
when extremely sensitive methods are applied.[1015] This may
be due to the very low incorporation rate for THC-COOH into
hair, being the lowest of all the drugs tested in a rat model
by Nakahara [16] with an up to a 3600-fold difference in the
incorporation rate compared to hydrophilic basic drugs such
as cocaine or morphine (THC not tested). As a consequence,
concentrations of THC-COOH found in hair samples published
in the literature are in the low- or sub-picogram per mg
range.[10,12,13,17,18] In many cases it cannot be detected at all
extremely sensitive methods are applied [18,19] or after conrmed
consumption.[20] Furthermore, the detection and quantication
of THC-COOH requires hyphenated techniques and costly
equipment. Most forensic laboratories do not have the equipment
to reach such sensitivity and it is still common to detect THC alone
or in combination with cannabidiol and cannabinol (CBN).
However, all three compounds are present in the smoke of
cannabis products and do therefore not prove body passage.
Usually the concentration of THC is several orders of
magnitude higher than the concentration of THC-COOH and this
can be explained at least in part by the fact that acidic
compounds generally show lower incorporation rates due to
relatively poor afnity towards the acidic melanin.[4] However,
after smoking marijuana, the area under the curve of THC is
considerably smaller compared to the area under the curve of
THC-COOH.[21] Taking this fact into account, THC should be
incorporated to a smaller extent compared to THC-COOH. This
contradiction strongly suggests other factors playing a
signicant role. In 2004, Thorspecken et al. simulated a one-
time contamination under in vitro conditions by exposing hair
strands to marijuana smoke in a desiccator, showing that a single
* Correspondence to: Volker Auwrter, Institute of Forensic Medicine, Forensic
Toxicology Department, University Medical Center Freiburg, Albertstr. 9,
79104 Freiburg, Germany. E-mail: volker.auwaerter@uniklinik-freiburg.de
a Institute of Forensic Medicine, Forensic Toxicology Department, University
Medical Center Freiburg, Albertstr. 9, 79104 Freiburg, Germany
b Hermann Staudinger Graduate School, University of Freiburg, Hebelstrae 27,
79104 Freiburg, Germany

Drug Test. Analysis (2013) Copyright 2013 John Wiley & Sons, Ltd.
Drug Testing
and Analysis
exposure may produce a positive cannabinoid nding in
hair.[3] After extensive washing with methanol, negative test
results could be obtained. The downside of this approach is
that an extraction of the analytes from the hair may take
place, especially when organic solvents such as methanol,
which are commonly used as an extraction solvent in hair
analysis, are used for extensive washing. In addition to that,
the in vitro model does not reect real-life conditions. Once
condensed on the hair, the contamination can be distributed
over the entire hair shaft via the sebum. Particularly during
night-time, when the head rests on a pillow and the local
temperature may rise to >30  C with high humidity due to
sweating and movement, a distribution and diffusion into
the cell membrane complex of the hair may take place.
Furthermore, unlike in the experiment (one-time exposure),
an accumulation of the compounds has also to be taken into
consideration, as most regular consumers smoke marijuana a
couple of times a week.[22]
To differentiate between an external contamination and
active consumption, a marker compound being present in
the marijuana sidestream smoke but not getting incorporated
through the bloodstream would facilitate the interpretation of
positive hair analysis results. One compound with such potential
is the biogenetic precursor of THC, 9-tetrahydrocannabinolic
acid A (THCA-A).
In 2005, Dussy et al. published an article showing that
THCA-A is not completely decarboxylated to THC when
heated and small amounts of THCA-A are still present in the
smoke.[23] Therefore, THCA-A can be detected in blood and
urine samples of cannabis smokers.[24] Additionally it has
been shown that THCA-A is not incorporated signicantly into
the hair through the bloodstream after oral intake of THCA-A
daily doses of 10 mg for 30 consecutive days [25] (applied limit
of detection: 50 pg/mg). In many forensic hair samples,
however, the concentrations of THCA-A are considerably
higher than the concentrations of THC. This nding suggests
that the THCA-A found in forensic hair samples must arise
from some other origin like condensation of smoke on the
head and diffusion of the compound into the hair.
Besides being a promising marker for contamination, THCA-A
in hair samples may be decarboxylated during extraction or
analysis (e.g. high GC temperature) and articially increase the
measured THC concentration. If alkaline hydrolysis of the hair is
applied before extraction as in many routine methods,[15,2528]
an almost quantitative decarboxylation can be assumed. A
consequence of the articially elevated THC concentrations could
be an overestimation of consumption frequency/intensity.
Further useful information which would facilitate the
interpretation of positive test results would be whether or not
there are differences in the extent of the contamination through
sidestream smoke on various regions of the head. In this respect,
the posterior vertex region, which is recommended for
sampling by the Society of Hair Testing (SoHT) and the German
Society of Toxicological and Forensic Chemistry (GTFCh), [29,30]
is of special interest.
The objective of the present study was to investigate if THCA-A
could act as a marker for an external contamination of hair and
to determine the amount of THC and CBN in hair caused by
external contamination of hair through repeated exposition to
sidestream marijuana smoke under realistic conditions. In
addition, the regional difference in the degree of contamination
was to be evaluated on various areas of the head.
wileyonlinelibrary.com/journal/dta Copyright 2013 John Wiley & Sons, Ltd.
B. Moosmann, N. Roth and V. Auwrter
Material and methods
Materials
THC and CBN (1 mg/ml each) were purchased from Cerilliant (Round
Rock, TX, USA). THCA-A was obtained from Lipomed (Arlesheim,
Switzerland) and dissolved in methanol resulting in a stock
solution of 0.1 mg/ml. THC-D3 (0.1 mg/ml) was obtained from
Cerilliant, CBN-D3 (0.1 mg/ml) from Lipomed. THCA-A-D3 was
synthesized by an in-house procedure [31] yielding a mixture of
THCA-A-D3 (> 90%), THC-D3 (approximately 9%) and THCA-A (< 1%).
Acetonitrile and methanol (both HPLC grade) were purchased
from J.T. Baker (Deventer, the Netherlands) formic acid
(ROTIPURANW 98%, p.a.) and petroleum ether (ROTIPURANW
4060  C) from Carl Roth (Karlsruhe, Germany). Acetone (p.a.,
ACS) was obtained from Sigma Aldrich (Steinheim, Germany)
and deionized water was prepared using a cartridge deionizer
from Memtech (Moorenweis, Germany). Lecithin (egg phosphati-
dylcholine, batch 108064-03/968) was obtained from Lipoid
(Ludwigshafen, Germany). Blank hair for the calibration was
supplied from volunteers and analyzed for the target cannabinoids
prior to use.
Study design
All three participants gave informed consent prior to the study.
The participants had untreated dark blonde or brown hair and
hair lengths of three (participant 1), six (participant 2) and eleven
(participant 3) centimetres. The hair was exposed to the smoke of
one marijuana joint in the morning of every weekday for a three-
week period leading to a total exposure to the smoke of 15 joints.
The joints were prepared from 500 mg marijuana owers with
a total THC content of 9.2% and 500 mg tobacco each. To avoid
any contamination of the hands, latex gloves were worn during
the entire preparation and exposure period.
The exposure took place in a room measuring 2.5 by 2 by
2.5 metres and the joint was smoked using a water-jet
vacuum pump. Inhalation of the smoke was excluded by
breathing compressed air through SCUBA regulators. The
three participants sat in a small circle facing each other and
the joint was held in front of the mouth of one participant
for 1015 s, afterwards connected to the vacuum for one puff
and then passed on to the next participant. The whole
procedure was repeated until the joint was completely
burned down (1520 min). The participants followed their
daily routine until next morning. The hair was washed daily
using commercially available hair shampoo. The washing
procedure and brand of shampoo were not standardized.
Sampling
Prior to the exposure period, hair samples were obtained from
each participant. During the exposure period two strands
were obtained every week after the weekend shortly before
exposure to the next joint. Participant 3 continued to provide
four strands per week for the next seven weeks, while the other
two participants provided two strands four weeks after the
exposure period. Finally, 30 strands of hair from various regions
of the head were obtained seven weeks after the exposure
period from participant 3. All the samples were taken as close
to the scalp as possible from the posterior vertex region of
the head (remaining hair length at the scalp ~ 1 mm). The
proximal end of the hair was labelled and the samples stored
Drug Test. Analysis (2013)
 Drug Testing
Hair analysis for THCA A, THC and CBN after passive in vivo exposure to marijuana smoke and Analysis

in aluminium foil in the dark at room temperature until analysis. Results

Additionally, twice a week urine samples were obtained from
the participants.
Sample preparation
For sample preparation the hair strands of participants 2 and
3 were segmented into segments of 3 cm length. Due to
the short hair length of participant 1, segmentation was not
necessary. For washing and extraction, a previously
published method was applied.[32] In brief, the hair was
washed by shaking for four minutes with 4 ml of water
followed by 4 ml acetone and 4 ml petroleum ether,
respectively. The hair was then dried for 24 h and 50 mg were
cut into 12 mm pieces. Extraction was performed by the
addition of 2 ml of methanol and 20 ml of internal standard
solution (~3 ng THCA-A-D3, 20 ng THC-D3, 20 ng CBN-D3).
The analytes were extracted for 4 h under shaking. After
centrifugation, 1.5 ml of methanol was transferred into an
LC-vial and evaporated to dryness at 40  C under a gentle
stream of nitrogen. The sample was then reconstituted in
100 ml acetonitrile with 0.1% formic acid and 0.25% lecithin
prior to liquid chromatography-tandem mass spectrometry
(LC-MS/MS) analysis.
LC-MS/MS method
A fully validated LC-MS/MS method for the quantication of
cannabinoids in hair described by Roth et al. was applied [32].
The lower limits of quantication (LOQ) of the method were
2.5 pg/mg for THCA-A as well as 20 pg/mg for THC and CBN
and the MS parameters of the analytes and internal standards
are listed in Table 1.
Immunoassay
All the urine samples obtained, tested negative for THC-COOH in
the immunoassay.
THC and CBN concentrations in the hair samples
In the hair samples of all three participants obtained prior to the
exposure period, no THC, CBN or THCA-A was detected. The
results of the THC and CBN concentrations measured in the hair
samples are shown in Table 2 for participant 1 and 2 and in
Table 3 for participant 3.
Time prole
Figure 1 shows the time prole of the measured concentrations in the
hair samples of participant 3 over the 10-week period of the study.
The growth rate of the hair of participant 3 was 1 cm per month.
Distribution
Figure 2 shows the THC concentration found in the 30 samples
obtained seven weeks after the exposure period. The CBN
concentrations showed a similar behaviour. THCA-A was detectable
but could not be quantied. The individual segments showed no
difference in the distribution compared to the total hair.
THCA-A concentrations
The highest concentration of THCA-A measured in the hair sample
of participant 1 was 19 pg/mg at the end of the exposure period.
Participant 2 also had the highest concentration at the end of the
exposure period measuring 30.1 pg/mg. In the hair of participant
3 the highest concentration of THCA-A was 2.9 pg/mg.

Analysis of urine samples Discussion

The obtained urine samples were tested for THC-COOH by
Fluorescence Polarization Immunoassays using an Axsym
4602 instrument (software version 9.0) and cannabinoid-
calibrators (both Abbott Laboratories, Abbott Park, IL, USA).
The assay was validated to report concentrations of THC-COOH
greater than 10 ng/ml as positive (assessed after alkaline hydrolysis
and gas chromatographymass spectrometry (GC-MS) analysis).
The hair samples taken before the exposure period showing
negative results proved that neither of the participants had
contact with cannabis products prior to the study. In addition
to that, any active uptake of the smoke as well as any cannabis
consumption on other occasions during the study could be ex-
cluded since the urine samples collected twice a week were
all tested negative for THC-COOH. The exclusion of any active

Table 1. Summary of MRM transitions and corresponding voltages for the mass spectrometric analysis. (Q1) mass-to-charge ratio of the
precursor ion (Q3) mass-to-charge ratio of the fragment ion (DP) declustering potential, (EP) entrance potential, (CE) collision energy, (CXP) collision
cell exit potential
Analyte MRM-Mode Q1 [amu] Q3 [amu] dwell time [ms] DP [V] EP [V] CE [V] CXP [V]

THCA-A Quantier neg. 357.2 313.2 20 45 10 34 7

Qualier neg. 357.2 245.2 10 45 10 43 5
THC Quantier pos. 315.2 193.2 20 60 10 34 3
Qualier pos. 315.2 259.3 10 60 10 28 5
CBN Quantier pos. 311.2 223.2 20 60 10 30 4
Qualier pos. 311.2 241.2 10 60 10 28 6
Internal standard
D3-THCA-A neg. 360.3 316.4 20 95 10 34 7
D3-THC pos. 318.2 196.2 20 60 10 34 3
D3-CBN pos. 314.3 223.2 20 60 10 30 4

Drug Test. Analysis (2013) Copyright 2013 John Wiley & Sons, Ltd. wileyonlinelibrary.com/journal/dta
Drug Testing
and Analysis B. Moosmann, N. Roth and V. Auwrter

Table 2. THCA-A, THC and CBN concentrations measured in the hair of participant 1 and in the segmented hair of participant 2 over the study
period (LOQ: THC and CBN 20 pg/mg, THCA-A 2.5 pg/mg)

End of Week Time point Segment THCA [pg/mg] THC [pg/mg] CBN [pg/mg]

Participant 1
0 before the study 02 cm 0 0 0
1 beginning of week 2 02 cm 3.2 < LOQ < LOQ
2 beginning of week 3 02 cm 3.1 < LOQ < LOQ
3 end of exposure period 02.3 cm 19 140 51
7 4 weeks after the exposure 03.5 cm 0 0 0

Participant 2
0 before the study 03 cm 0 0 0
36 cm 0 0 0
Total hair 0 0 0
1 beginning of week 2 03 cm 9.6 16 < LOQ
36 cm 21 33 < LOQ
Total hair 15 24 < LOQ
2 beginning of week 3 03 cm 3 240 100
36 cm 12 390 190
Total hair 7.7 320 140
3 end of exposure period 03 cm 35 950 330
36.5 cm 25 990 350
Total hair 30 970 340
7 4 weeks after the exposure 03 cm 0 41 31
37 cm 4.4 140 92
Total hair 2.5 100 66

uptake of sidestream smoke was particularly important, since
massive exposure can lead to blood and urine cannabinoid levels
similar to the levels measured after active consumption.[33]
By comparing the THC and CBN concentrations in the hair of
the three participants at the end of the exposure period and
one month thereafter, it could be shown that the degree of
contamination differs depending on the length of the hair with
shorter hair being less affected. Some reasons for the low
concentrations in the short hair could be that the daily hair
wash with shampoo is more efcient in short hair. Furthermore
with shorter hair sebum may be removed more effectively
with every hair wash from the skin surface, reducing the total
amount of sebum available to store and distribute the
cannabinoids along the hair shaft.
The individual segments obtained from participant 2 show that
with 6-cm-long hair there is a small difference between the
distal and proximal segment, indicating that the cannabinoids
are distributed along the entire hair shaft and diffuse into the
hair. Because of the fact that the contamination extends over
the entire hair length, the interpretation of segmental hair
analysis for retrospective determination of changing habits of
use has to be doubted. In case of the 12-cm-long hair of
participant 3 there is a larger difference between the individual
segments which could be explained by the proximal hair
being covered by the distal parts of the longer hair, leaving it less
exposed to the smoke and thus less affected. While the distal
segments (9 cmend) of participant 3 showed the highest THC
and CBN concentrations during the exposure period, they show
a larger decline in the concentration than the other segments
once the exposure stopped. This is probably caused by the fact
that in hair tips the cuticula tends to be more damaged than in
other regions along the hair shaft and therefore analytes may
be washed out more easily. However, due to the small number
of participants as well as the non-standardization of shampoo
products and washing procedures, further investigations need
to be carried out for an in-depth interpretation of the inuencing
factors of the analyte concentrations in the wash-out period.
Even though the THC concentration begins to drop once the
exposure stops, it stays above the cut-off applied in Germany
for driving license issues of 20 pg/mg THC [34] for up to seven
weeks (week 10), when the last hair sample was obtained.
At this time the hair has been washed almost 50 times since
the last exposure. Towards the last weeks there is a uctuation
in the concentrations of the individual samples, showing for
example an increase from week 7 to week 8. One explanation
for this result could be inhomogeneities in the distribution of
the analytes. Although all the samples were obtained from
the posterior vertex region of the head, there is a slight change
in the area, as previous sampling made it necessary to obtain
samples further away from the centre of this region. To
substantiate the suspicion that the sampling site makes a
difference in the degree of contamination, the 30 samples
obtained from various regions of the head seven weeks after
the exposure period (week 10) from participant 3 were
analyzed. The results demonstrate that the posterior vertex
region of the head suffers from the highest degree of contam-
ination and marked differences occur depending on the
sampling region. This is especially critical since most of the hair
samples in routine analysis are taken from this particular
region, as it shows a relatively constant hair growth rate and
leads to fewer cosmetic problems. The observed distribution
can be the result of various factors. One factor could be that
the head was slightly facing the ground during smoke
exposure, resulting in the smoke passing by the side of the
head and unifying in the posterior vertex region. It has to be
noted that in this study set-up, each participant held the joint

wileyonlinelibrary.com/journal/dta Copyright 2013 John Wiley & Sons, Ltd. Drug Test. Analysis (2013)
 Drug Testing
Hair analysis for THCA A, THC and CBN after passive in vivo exposure to marijuana smoke and Analysis

Table 3. THCA-A, THC and CBN concentrations measured in the segmented hair of participant 3 over the entire study period (LOQ: THC and CBN
20 pg/mg, THCA-A 2.5 pg/mg)

End of week Time point Segment THCA-A [pg/mg] THC [pg/mg] CBN [pg/mg]

0 before the study 03 cm 0 0 0

36 cm 0 0 0
69 cm 0 0 0
911 cm 0 0 0
Total hair 0 0 0
1 beginning of week 2 03 cm < LOQ 37 < LOQ
36 cm < LOQ 34 < LOQ
69 cm < LOQ 140 50
911 cm < LOQ 260 140
Total hair < LOQ 100 43
2 beginning of week 3 03 cm < LOQ 140 46
36 cm < LOQ 200 72
69 cm < LOQ 500 190
911 cm < LOQ 890 320
Total hair < LOQ 390 140
3 end of exposure period 03 cm < LOQ 480 180
36 cm < LOQ 1300 510
69 cm < LOQ 1600 700
912 cm < LOQ 1700 720
Total hair < LOQ 1300 530
4 1 week after the exposure 03 cm < LOQ 270 100
36 cm 3.2 930 370
69 cm 4.1 620 310
911.5 cm 4.6 550 280
Total hair 2.9 590 260
5 2 weeks after the exposure 03 cm 0 140 64
36 cm < LOQ 380 180
69 cm < LOQ 360 180
911.5 cm < LOQ 220 160
Total hair < LOQ 280 140
6 3 weeks after the exposure 03 cm 0 170 75
36 cm < LOQ 580 240
69 cm < LOQ 270 140
912 cm < LOQ 220 140
Total hair < LOQ 310 150
7 4 weeks after the exposure 03 cm < LOQ < LOQ < LOQ
36 cm < LOQ 77 57
69 cm 2.9 64 61
912 cm < LOQ 42 50
Total hair < LOQ 50 44
8 5 weeks after the exposure 03 cm 0 26 23
36 cm < LOQ 150 120
69 cm < LOQ 260 210
912 cm 3.5 200 250
Total hair < LOQ 160 150
9 6 weeks after the exposure 03 cm 0 < LOQ < LOQ
36 cm 0 42 29
69 cm 0 64 41
913 cm 0 57 68
Total hair 0 44 38
10 7 weeks after the exposure 03 cm 0 < LOQ < LOQ
36 cm 0 64 51
69 cm 0 110 87
913 cm 0 51 68
Total hair 0 58 54

Drug Test. Analysis (2013) Copyright 2013 John Wiley & Sons, Ltd. wileyonlinelibrary.com/journal/dta
Drug Testing
and Analysis B. Moosmann, N. Roth and V. Auwrter

Figure 1. Time prole of the measured THC concentration in the hair of participant 3. End of week 3 marks the end of the exposure period. The dotted
horizontal line indicates the cut-off currently applied for driving liability cases in Germany (20 pg/mg).

Figure 2. Distribution of the THC concentration measured seven weeks after the last exposure (week 10). Concentrations below 20 pg/mg are
approximate values (LOQ = 20 pg/mg).

for about one-third of the time in front of his mouth. People
sitting in a room without such direct exposure might show a
different distribution of concentrations.
From comparing the THCA-A concentrations detected in
the forensic samples measured in our laboratory (own
unpublished data, [25]) with the concentrations measured in
this study it can be concluded that the high THCA-A
concentrations of up to 10 000 pg/mg in the forensic samples
cannot result from external contamination through sidestream
marijuana smoke, although this was suggested as a hypothe-
sis in our previous work. [25] While the analyzed forensic
samples often show THCA-A to THC ratios of 2:1 or more,
the highest ratio found in the hair samples from this study
was 1:1.5 in one sample. In all other samples the ratio was
not higher than approximately 1:8. Therefore, other sources
must account for the relatively high THCA-A concentrations
found in hair samples of cannabis users. An explanation for
the high concentrations in forensic samples could be that
cannabis users touch their hair with contaminated ngers
after direct contact with drug material (e.g. when rolling a
joint). This seems plausible, as contamination by this route
has been shown previously for heroin.[35]
for THC and not for THC-COOH is applied, positive results in
the range of concentrations typically measured in excessive
cannabis users may originate solely from sidestream smoke.
Furthermore it could be shown that the posterior vertex region
of the head, which is the preferred sampling site in routine
analysis, suffers from the highest degree of contamination after
repeated cannabis smoke exposition. However, THCA-A, which
is found in high concentrations in many forensic hair samples,
is detected only in low concentrations in the chosen experimen-
tal setup and can therefore not act as a marker for an external
contamination of hair through marijuana smoke. Nevertheless
it may be a marker for the contamination through direct transfer
of cannabinoids by contaminated surfaces or ngers.
These ndings underline that hair-ndings in general have to
be interpreted with utmost care, particularly in case of drugs
which are smoked or when cross-contamination via hand
contact is likely to occur. Considering the severe consequences
people concerned may experience, contamination issues in hair
analysis still do not seem to be investigated to an extent that
would be desirable.
Acknowledgements

Conclusion
In conclusion, the study demonstrated that sidestream
marijuana smoke can lead to a considerable and persisting THC
and CBN contamination in hair samples. If a method only testing
The authors would like to thank Dr. Andrea Jacobsen-Bauer from
the State Ofce of Criminal Investigation Baden-Wrttemberg for
providing the marijuana and Lee Perry-Smith for providing the
3D-headmodel data le. The project was funded by the Deutsche
Forschungsgemeinschaft (DFG-AU: 324/3-1)

wileyonlinelibrary.com/journal/dta Copyright 2013 John Wiley & Sons, Ltd. Drug Test. Analysis (2013)

Hair analysis for THCA A, THC and CBN after passive in vivo exposure to marijuana smoke
References
[1] G.L. Henderson. Mechanisms of drug incorporation into hair. Forensic
Sci. Int. 1993, 63, 19.
[2] F. Pragst, V. Auwaerter, F. Sporkert, K. Spiegel. Analysis of fatty acid
ethyl esters in hair as possible markers of chronically elevated
alcohol consumption by headspace solid-phase microextraction
(HS-SPME) and gas chromatographymass spectrometry (GC-MS).
Forensic Sci. Int. 2001, 121, 76.
[3] J. Thorspecken, G. Skopp, L. Ptsch. In vitro contamination of hair by
marijuana smoke. Clin. Chem. 2004, 50, 596.
[4] F. Pragst, M.A. Balikova. State of the art in hair analysis for detection
of drug and alcohol abuse. Clin. Chim. Acta 2006, 370, 17.
[5] P.R. Stout, J.D. Ropero-Miller, M.R. Baylor, J.M. Mitchell. External
contamination of hair with cocaine: Evaluation of external cocaine
contamination and development of performance-testing materials.
J. Anal. Toxicol. 2006, 30, 490.
[6] R. Wennig. Potential problems with the interpretation of hair
analysis results. Forensic Sci. Int. 2000, 107, 5.
[7] Society of hair testing. Forensic Sci. Int. 1997, 84, 3.
[8] Y. Gaillard, G. Ppin. Simultaneous solid-phase extraction on C18
cartridges of opiates and cocainics for an improved quantitation in
human hair by GC-MS: One year of forensic applications. Forensic
Sci. Int. 1997, 86, 49.
[9] E. Han, H. Chung, J.M. Song. Segmental hair analysis for 11-Nor-9-
tetrahydrocannabinol-9-carboxylic acid and the patterns of cannabis
use. J. Anal. Toxicol. 2012, 36, 195.
[10] M.J. Baptista, P.V. Monsanto, E.G. Pinho Marques, A. Bermejo, S. vila,
A.M. Castanheira, C. Margalho, M. Barroso, D.N. Vieira. Hair analysis
for 9-THC, 9-THC-COOH, CBN and CBD, by GC/MS-EI: Comparison
with GC/MS-NCI for 9-THC-COOH. Forensic Sci. Int. 2002, 128, 66.
[11] V. Cirimele, H. Sachs, P. Kintz, P. Mangin. Testing human hair for
cannabis. III. rapid screening procedure for the simultaneous
identication of 9-tetrahydrocannabinol, cannabinol, and
cannabidiol. J. Anal. Toxicol. 1996, 20, 13.
[12] J.Y. Kim, J.C. Cheong, J.I. Lee, M.K. In. Improved gas chromatography-
negative ion chemical ionization tandem mass spectrometric
method for determination of 11-nor-9-tetrahydrocannabinol-
9-carboxylic acid in hair using mechanical pulverization and bead-
assisted liquid-liquid extraction. Forensic Sci. Int. 2011, 206, e99.
[13] T. Mieczkowski. Assessing the potential of a color effect for hair
analysis of 11-nor-9-carboxy-9-tetrahydrocannabinol: Analysis of a
large sample of hair specimens. Life Sci. 2003, 74, 463.
[14] M. Uhl, H. Sachs. Cannabinoids in hair: Strategy to prove marijuana/
hashish consumption. Forensic Sci. Int. 2004, 145, 143.
[15] D. Wilkins, H. Haughey, E. Cone, M. Huestis, R. Foltz, D. Rollins.
Quantitative analysis of THC, 11-OH-THC, and THCCOOH in human
hair by negative ion chemical ionization mass spectrometry. J. Anal.
Toxicol. 1995, 19, 483.
[16] Y. Nakahara, K. Takahashi, R. Kikura. Hair analysis for drugs of abuse.
X. Effect of physicochemical properties of drugs on the incorporation
rates into hair. Biol. Pharm. Bull. 1995, 18, 1223.
[17] M. Uhl. Determination of drugs in hair using GC/MS/MS. Forensic Sci.
Int. 1997, 84, 281.
[18] H. Sachs, U. Dressler. Detection of THCCOOH in hair by MSD-NCI
after HPLC clean-up. Forensic Sci. Int. 2000, 107, 239.
[19] T. Mieczkowski. A research note: The outcome of GC/MS/MS
conrmation of hair assays on 93 cannabinoid (+) cases. Forensic
Sci. Int. 1995, 70, 83.
All in-text references underlined in blue are linked to publications on ResearchGate, letting you access and read them immediately.
Drug Test. Analysis (2013) Copyright 2013 John Wiley & Sons, Ltd.
Drug Testing
and Analysis
[20] M.A. Huestis, R.A. Gustafson, E.T. Moolchan, A. Barnes, J.A. Bourland,
S.A. Sweeney, E.F. Hayes, P.M. Carpenter, M.L. Smith. Cannabinoid
concentrations in hair from documented cannabis users. Forensic
Sci. Int. 2007, 169, 129.
[21] M.A. Huestis, J.E. Henningeld, E.J. Cone. Blood cannabinoids. I.
Absorption of THC and formation of 11-OH-THC and THCCOOH
during and after smoking marijuana. J. Anal. Toxicol. 1992, 16,
276.
[22] EMCDDA. 2011 Annual Report of the State of the Drugs Problem in
Europe, Publications Ofce of the European Union, Luxembourg,
2011.
[23] F.E. Dussy, C. Hamberg, M. Luginbuhl, T. Schwerzmann, T.A.
Briellmann. Isolation of 9-THCA-A from hemp and analytical
aspects concerning the determination of 9-THC in cannabis
products. Forensic Sci. Int. 2005, 149, 3.
[24] J. Jung, J. Kempf, H. Mahler, W. Weinmann. Detection of 9-
tetrahydrocannabinolic acid A in human urine and blood serum by
LC-MS/MS. J. Mass Spectrom. 2007, 42, 354.
[25] V. Auwrter, A. Wohlfarth, J. Traber, D. Thieme, W. Weinmann. Hair
analysis for 9-tetrahydrocannabinolic acid A--new insights into
the mechanism of drug incorporation of cannabinoids into hair.
Forensic Sci. Int. 2010, 196, 10.
[26] F. Musshoff, H.P. Junker, D.W. Lachenmeier, L. Kroener, B. Madea.
Fully automated determination of cannabinoids in hair samples
using headspace solid-phase microextraction and gas chromatogra-
phymass spectrometry. J. Anal. Toxicol. 2002, 26, 554.
[27] T. Nadulski, F. Pragst. Simple and sensitive determination of 9-
tetrahydrocannabinol, cannabidiol and cannabinol in hair
by combined silylation, headspace solid phase microextraction
and gas chromatographymass spectrometry. J. Chromatogr.
B 2007, 846, 78.
[28] A. Salomone, E. Gerace, F. DUrso, D. Di Corcia, M. Vincenti.
Simultaneous analysis of several synthetic cannabinoids, THC, CBD
and CBN, in hair by ultra-high performance liquid chromatography
tandem mass spectrometry. Method validation and application to
real samples. J. Mass Spectrom. 2012, 47, 604.
[29] Society of Hair Testing. Recommendations for hair testing in forensic
cases. Forensic Sci. Int. 2004, 145, 83.
[30] F. Muhoff, G. Skopp, F. Pragst, H. Sachs, D. Thieme. Anhang C
zur Richtlinie der GTFCh zur Qualittssicherung bei forensisch-
toxikologischen Untersuchungen. Toxichem. Krimtech. 2009, 76,
209.
[31] N. Roth, A. Wohlfarth, M. Mller, V. Auwrter. Regioselective
synthesis of isotopically labeled 9-tetrahydrocannabinolic acid A
(THCA-A-D3) by reaction of 9-tetrahydrocannabinol-D3 with
magnesium methyl carbonate. Forensic Sci. Int. 2012, 222, 368.
[32] N. Roth, B. Moosmann, V. Auwrter. Development and validation of an
LC-MS/MS method for quantication of 9-tetrahydrocannabinolic
acid A (THCA-A), THC, CBN and CBD in hair. J. Mass Spectrom. 2013,
48, 227.
[33] E.J. Cone, R.E. Johnson. Contact highs and urinary cannabinoid
excretion after passive exposure to marijuana smoke. Clin. Pharm.
Ther. 1986, 40, 247.
[34] W. Schubert, R. Mattern. Beurteilungskriterien: Urteilsbildung in der
Medizinisch-Psychologischen Fahreignungsdiagnostik, Vol. 2, Kirschbaum
Verlag, Bonn, 2008.
[35] G. Romano, N. Barbera, G. Spadaro, V. Valenti. Determination of
drugs of abuse in hair: Evaluation of external heroin contamination
and risk of false positives, Forensic Sci. Int. 2003, 131, 98.
wileyonlinelibrary.com/journal/dta