Journal of Chromatography B 1195 (2022) 123202

Contents lists available at ScienceDirect

Journal of Chromatography B
journal homepage: www.elsevier.com/locate/jchromb

Detection of mescaline in human hair samples by UPLC-MS/MS:

Application to 19 authentic forensic cases
Shuo Yang a, b, 1, Yan Shi b, 1, Zhuonan Chen a, b, Mobing Chen a, b, Xinze Liu a, b, Wei Liu b, *,
Mengxiang Su a, *, Bin Di a
a
School of Pharmacy, China Pharmaceutical University, Nanjing 210009, PR China
b
Academy of Forensic science, Shanghai Key Laboratory of Forensic Medicine, Shanghai 200063, PR China

A R T I C L E I N F O A B S T R A C T

Keywords: Mescaline, a natural alkaloid found in the peyote cactus (Lophophora williamsii) in the Americas, has gradually
Mescaline become a drug of abuse in China because of its psychedelic properties. Its intake may lead to hallucinations and
Hallucinogen confusion or even be life-threatening. Mescaline is classified as a class I psychotropic drug in China, which means
Illicit drug
its use in medicine or scientific research is under strict control of the government. However, studies on sur­
Ultra-high performance liquid
veillance of mescaline abuse in the Chinese population are lacking.
chromatography–tandem mass spectrometry
Hair analysis A rapid and sensitive liquid chromatography–tandem mass spectrometry (LC-MS/MS) method was developed
and validated for the determination and quantification of mescaline in hair. The method had good linearity in the
range from 10 to 1000 pg/mg, with the limit of detection (LOD) of 3 pg/mg and the limit of quantitation (LOQ)
of 10 pg/mg. The total runtime was 5 min. Acceptable intraday and interday precision (RSD < 15%) and ac­
curacy (bias, − 11.2% ~ 6.8%) were achieved. The recovery was 85.0–101.0%, and the matrix effect was
92.0–105.0%. The validated method was successfully applied to 19 real forensic cases. The concentrations of
mescaline in hair ranged from 10 to 784 pg/mg. The method has the benefits of simple sample preparation, high
sensitivity, and short running time, making it suitable for large-scale quantitative surveillance analysis of
mescaline in forensic toxicology.

1. Introduction are similar to those caused by other psychoactive substances, such as

Mescaline (3,4,5-trimethoxyphenethylamine) is a natural alkaloid
found in the peyote cactus (Lophophora williamsii) endemic to the
Americas. Its abuse history can be traced back to prehistoric times, and it
has been used in religious rituals for thousands of years due to its psy­
chedelic properties [1]. In addition, since the 1960s, hallucinogens has
been used widely for recreational purposes [2]. Even now, the usage of
mescaline has become more widespread by the Native American Church
during religious ceremonies [3,4]. Possession of peyote is illegal, but
buying cacti seeds and growing them in a greenhouse together with
other plants is possible considering cacti are common ornamental plants,
increasing the potential for abuse.
The drug effects of mescaline in humans are well studied. Its phar­
macodynamic mechanisms are primarily generated from the interaction
with the serotonergic 5-HT2A-C receptors, which means its clinical effects
lysergic acid diethylamide (LSD) and psilocybin. The psychoactive ef­
fects include euphoria, hallucinations, depersonalization, and psychoses
[5]. Four cases of death related to mescaline have been described [6–9]
though the toxicity of mescaline is lower than that of other alkaloids,
such as morphine (and its derivative heroin), cocaine, and the non-
illegal alkaloids caffeine and nicotine [10]. Cases of fights among drug
abusers or self-harm had been reported due to the severe effects of the
hallucinations [6]. In China, mescaline has been classified as a class I
psychotropic drug, which means that its application in medicine or
scientific research is strictly controlled by the government. Over the past
few years, mescaline-related smuggling cases have increased in China.
Therefore, the need to monitor the potential diffusion of mescaline
among people with a history of drug addiction and abuse represents an
important task, as excess mescaline intake may lead to hazardous social
events.

* Corresponding authors at: Academy of Forensic science, Shanghai Key Laboratory of Forensic Medicine, No. 1347 Guangfuxi Road, Shanghai 200063, PR China
(W. Liu) and School of Pharmacy, China Pharmaceutical University, No. 24 Tongjiaxiang Road, Nanjing 210009, PR China (M. Su).
E-mail addresses: liuw@ssfjd.cn (W. Liu), sumengxiang@cpu.edu.cn (M. Su).
1
These authors contributed equally to this work.

https://doi.org/10.1016/j.jchromb.2022.123202
Received 21 November 2021; Received in revised form 26 February 2022; Accepted 28 February 2022
Available online 2 March 2022
1570-0232/© 2022 Elsevier B.V. All rights reserved.
S. Yang et al.
Fig. 1. Chemical structures of mescaline (a) and 25B-NBOMe-D3 (b).
As a monitoring method, hair analysis has become a hot topic in
forensic science. Hair is considered as a valuable specimen for its
accessibility, preservation and longer detection window [11]. Before
metabolism, the drug molecules in hair are combined with the keratin
matrix from sweat, bloodstream, or sebum. Hence, the target analytes in
hair are usually represented by the parent drugs [12]. Therefore,
establishing a reliable hair analysis method is important for application
to authentic forensics cases.
There are many previous analytical methods we can use to detect
mescaline due to its increasing popularity for recreational use. The
methods applied in analysis of the mescaline content in peyote cactus
include liquid–chromatography photodiode array detection (LC-PAD)
[13], direct analysis in real time-high resolution mass spectrometry
(DART-HRMS) [14], and capillary electrophoresis–electrospray ioniza­
tion mass spectrum (CE-ESI-MS) [15]. A gas chromatography–mass
spectrometry (GC–MS/MS) [16] method and a liquid chromatogra­
phy–mass spectrometry (LC-MS/MS) [17] method have also been
developed for mescaline determinations in urine and oral fluid samples,
respectively. The challenge that arises when attempting to quantify
mescaline in hair is associated with the complex hair matrices and very
low concentrations. As a result, due to its high sensitivity, mass spec­
trometry (MS) is recommended for accurate quantification of low drug
concentrations in biological samples. GC–MS [16,18] and LC-MS/MS
[12,17,19] methods have been validated for quantification of mesca­
line in hair samples. Nevertheless, previous LOQ’s might not be suffi­
ciently low to detect low mescaline levels in real cases.
The aim of the present work was to develop and validate a sensitive
UPLC-MS/MS method for the quantification and identification of
mescaline in hair. This method was applied for the determination of the
mescaline content in 19 real positive hair samples and to provide
reference values for hair concentrations for forensic cases.
2. Methods
2.1. Chemicals and reagents
Mescaline was purchased from Cerilliant (Texas, USA), and 25B-
NBOMe-D3 (used as an internal standard [IS]) with an isotopic purity of
99.72% also purchased from Cerilliant (Texas, USA). The chemical
structures of mescaline and 25B-NBOMe-D3 are shown in Fig. 1. Acetone
was obtained from Shanghai Lingfeng Chemical Reagent Co. (Shanghai,
China), and formic acid (98%, w/v) were purchased from Fluka (Buchs,
Switzerland). Methanol and acetonitrile for mass spectrometry were
purchased from Sigma-Aldrich (St. Louis, MO, USA). Ultra-purified
water was generated with a Milli-Q water system (Millipore, MA,
USA). Unless stated otherwise, all of the aforementioned materials were
of HPLC grade.
All standards were provided at a concentration of 1 mg/mL. Working
standard mixtures were prepared by appropriate dilution of the stock
standards in methanol. All solutions were stored at − 20℃ in the dark.
2
Journal of Chromatography B 1195 (2022) 123202
2.2. Hair samples
Drug-free hair obtained from eight healthy volunteers was used to
prepare the matrix for the control and calibration samples. Suspected
users in arrest cases provided their hair as samples. The entire length of
hair shafts were collected from the scalp, and the hair 0–3 cm away from
the root was used for detection. This study was approved by the Acad­
emy of Forensic Science Ethics Committee for research on human sub­
jects. All individuals provided written informed consent.
2.3. Sample preparation
The hair samples were washed three times with acetone and deion­
ized water and then dried at room temperature before extraction. The
hair was cut into 1–2 mm pieces, 20 mg of hair was weighed into a 2 mL
tube with ceramic beads, followed by addition of 40 µL IS (10 ng/mL)
and 660 µL methanol were added. Then, all samples were placed in the
JXFSPRP-CLN freeze-grinder (Shanghai Jing Xin Industrial Develop­
ment Co., Ltd., China) and pulverized below − 35℃ using the following
procedure: grinding speed: 18 m/s; run time: 40 s; speed: 2500 rpm;
interval time: 60 s; number of repetitions: 15 cycles. The mixture was
ultrasonicated for 10 min and then centrifuged at 13,500 rpm for 5 min.
After centrifugation, 300 μL of supernatant was filtered through a 0.22
μm filter membrane (Sinopharm Chemical Reagent Co., Ltd., China),
and a volume of 5 µL filtrate was injected into the chromatographic
system.
For fresh flowers, the stamens, calyxes, and petals were separated
from each other with scalpel and divided into three parallel samples
equally. After that, these materials were weighed into a 2 mL tube with
ceramic beads, followed by addition of 40 µL IS (10 ng/mL) and 660 µL
methanol were added. The remaining steps were the same as above.
2.4. lC-MS/MS
The LC-MS/MS analyses were performed with a Sciex 6500 Q-
trapTM quadrupole mass spectrometer (AB Sciex, Foster City, USA)
equipped with an AcquityTM Ultra Performance LC (Waters Corpora­
tion, USA). Analyst 1.6.3 software was used to control the system and
collect the data, and MultiQuant 3.0.2 was used to analyze the data. The
mass spectrometer was operated in positive electrospray ionization (ESI
+ ) and multiple reaction monitoring (MRM) modes.
2.4.1. LC conditions
Chromatographic separation was achieved on an ACQUITY UPLC
BEH C18 column (2.1 mm × 100 mm, 1.7 μm i.d., Waters, USA) at room
temperature. The mobile phase consisted of a mobile phase A (0.1%
formic acid in water) and a mobile phase B (acetonitrile). The following
gradient was used: A:B; 95:5(0 min)-95:5(0.5 min)-85:15(1 min)-85:15
(2 min)-50:50(2.5 min)-50:50(4 min)-95:5(5 min). The flow rate was
0.25 mL/min. The total run time was 5 min and the sample injection
volume was 5 μL per injection. The autosampler temperature was held at
S. Yang et al. Journal of Chromatography B 1195 (2022) 123202

Table 1
MRM parameters and retention times for mescaline and 25B-NBOMe-D3.
Substance Precursor ion (m/z) Product ion (m/z) Declustering potential (V) Collision energy (eV) Retention time (min)

Mescaline 212.0 180.0* 50 30 2.75

212.0 165.2 53 23 2.75
25B-NBOMe-D3 383.1 124.0* 47 22 3.9
383.1 92.0 47 46 3.9
*
Represents quantifier ions.

Fig. 2. Standard Mescaline in ESI mode.

matrix [20].
Table 2
Recovery and matrix effects of different volumes of methanol.
Volume (mL) Recovery Matrix effects 2.6. Method validation

0.5 83.1% 141.5%

0.7 97.3% 100.9%
1.0 93.8% 110.4%
4 ◦ C, and the autosampler needle was washed thoroughly between
injections.
2.4.2. Mass spectrometer conditions
The mass spectrometer system was operated using electrospray
ionization in positive ionization with multiple reaction monitoring
(MRM) mode. The optimum mass spectrometric conditions were as
follows: ion spray voltage (ISV), 5500 V; entrance potential (EP), 10 V;
collision cell exit potential (CXP), 13 V; source temperature (TEM),
500 ◦ C; curtain gas (CUR), 30 psi (nitrogen); ion source gas 1 (GS1), 35
psi; ion source gas 2 (GS2), 40 psi; and collision activation dissociation
(CAD) gas, middle. Characteristic ions (qualifier and quantifier ions) and
other experimental parameters for mescaline and IS are listed in Table 1,
and the MS/MS spectra related to mescaline are shown in Fig. 2.
2.5. Method optimization
Two different extraction solvents, methanol and EM (a mixture of
methanol/acetonitrile/2 mmol/L ammonium formate, 25:29:46, v/v/v,
pH 5.3), were compared for recovery and matrix effects. After this
comparison, the volume of extraction solvent required further optimi­
zation. The matrix effect and recovery were tested with five replicates of
drug-free hair samples spiked with 100 pg/mg of mescaline. Matrix ef­
fect values close to 100% (80–120%) indicated that the quantitative
analysis of the samples was not significantly affected by the sample
Validation experiments were performed following the recommen­
dations of international guidelines[7,20,21]. The validations applied in
the present study included selectivity, limit of detection (LOD), limit of
quantification (LOQ), linearity, precision, accuracy, matrix effect, re­
covery, and stability. Eight blank human hair samples and eight samples
spiked with IS were prepared and analyzed to check for possibly inter­
fering peaks.
The LOD in MRM mode was evaluated by decreasing the analyte
concentration until it fulfilled the requirement of the signal-to-noise (S/
N) ratio of greater than 3:1. The LOQ was the lowest concentration of
which the accuracy (bias) and permitted precision (relative standard
deviation [RSD]) were within ± 20%. It also fulfilled the requirement of
a S/N ratio of at least 10. Calibration standards containing LOQ con­
centration, 20, 100, 500, 800, and 1000 pg mescaline per mg hair
samples were prepared daily. Linear calibration curves (Y = aX + b)
were constructed by plotting the peak area ratios between mescaline and
the IS. A weighted linear regression model (1/x) was selected and the
squared correlation coefficients (r) were always better than 0.995 in all
cases.
The accuracy and precision were determined at the four QC sample
concentrations [LOQ (10 pg/mg), low (20 pg/mg), medium (100 pg/
mg) and high (800 pg/mg)] by analyzing five replicates per level on four
different days. When the accuracy (bias) and precision (RSD) were
within ± 15% of the accepted reference value, within ± 20% near LOQ,
the standard was considered acceptable.
The matrix effect and recovery were assessed using the method
proposed by Matuszewski et al. [22] Set A and B were the mean peak
areas of five replicates of blank hair pre-extraction and post-extraction
spiked samples, respectively. Set C were the mean peak areas of each

3
S. Yang et al. Journal of Chromatography B 1195 (2022) 123202

Fig. 3. Chromatograms of blank hair and multiple reaction monitoring (MRM) chromatograms for mescaline at the LOQ concentration.

standard solution. The matrix effects were determined by dividing set B at 4℃ for 24 h in the autosampler.
by set C, and recovery was obtained as the percentage ratios of Set A/B.
The stability of samples in autosampler was assessed by testing three
replicates of the low, medium, and high mescaline contents after storage

4
S. Yang et al. Journal of Chromatography B 1195 (2022) 123202

Table 3 study than in the previously published LC-MS/MS studies (10 min [12],
Concentrations of Mescaline in hair samples of 19 Authentic Cases. 12.40 min [17] and 16.5 min [19]). And the shorter total runtime can
Case Sex Age Overall Hair color Concentration save batch testing time which will optimize the turnover rate of samples.
number length (cm) (pg/mg)

1 male 37 5 brown 119.1 3.2. Method validation

2 male 32 6 black 59.5
3 male 43 7 black and 63.7
3.2.1. Selectivity
white
4 female 30 16 brown < 10 (LOQ) No significant peaks or responses were detected at the retention
5 male 51 4 brown 145.2 durations of the mescaline and IS in blank hair, as shown in Fig. 3.
6 female 51 36 brown < 10 (LOQ) Therefore, neither the analytes nor the IS showed disrupted elution due
7* male 31 3 brown 24.4 to endogenous substances in the hair samples. The analytes and the IS
8* male 34 1 black 76.8
9 female 40 50 brown 10.2
also did not interfere with each other’s elution patterns.
10* female 50 52 black 242.4
11* female 46 28 black 116.8 3.2.2. LOD, LOQ and linearity
12* male 38 3 black 160.5 The result showed that the LOD and LOQ of mescaline were 3 pg/mg
13 male 42 1 brown 42.6
and 10 pg/mg respectively. These LOD and LOQ values are lower than
14 male 39 1 brown < 10 (LOQ)
15* female 38 12 brown 341.3 those previously published for other LC-MS/MS studies [12,17,19]. As
16 female 46 28 black 784.1 shown in the present study (Table 3), the concentrations of positive
17* female 39 42 black 243.8 samples can range from 10 to 784 pg/mg. Therefore, the limit of
18* female 36 3 black 107.5 quantification (LOQ, 100 pg/mg [16]) of the GC–MS method and the
19 female 32 4 black 18.6
LOQ (50 pg/mg [19]) of the UPLC-MS method would not seem to qualify
*
Positive for both mescaline and dimethyltryptamine (DMT). for the detection of hair samples with low mescaline concentrations.
Only the LOQ (13 pg/mg [17]) of LC-MS/MS methods appears to meet
2.7. Analysis of authentic hair samples the requirement. However, the previously published LC-MS/MS method
suffered from a lack of validation of authentic forensic cases [17]. The
The authentic hair samples (n = 19) were collected from known linear range of the calibration curves (Y = 0.01015x + 0.00348) was
mescaline drug abusers from September 2020 to March 2021. Hair root 10–1000 pg/mg with a linear regression coefficient of r greater than
samples were collected from the vertex posterior region of the head, and 0.995 for the hair samples. Fig. 3 displays the multiple reaction moni­
the hair 0–3 cm away from the root was washed, cut, and stored at room toring (MRM) chromatograms which showed the quantifier transitions
temperature. Hair samples were pretreated as described in section 2.3, for the analytes in a blank hair sample spiked at LOQ concentrations.
and 5 µL of the filtrate was then injected into the LC-MS/MS system in
ESI mode. 3.2.3. Accuracy and precision
Intra-day and inter-day variations met the criteria of the interna­
3. Results and discussion tional guidelines [23]. So did the accuracy ranging from –11.2% to
6.8%. The intra-day precision RSD values were < 8.8%, while the inter-
3.1. Method optimization day precision RSD values were between 1.6% and 8.8%. In summary, all
the QC samples, except the LOQ samples, met the validation criteria of
In this study, two different extraction solvents (methanol and EM) RSD with a bias of < 15%. The accuracy and intra-day and inter-day
were compared. However, the matrix effect of the EM solution was precision data are presented in Table 4.
128%, which did not meet the range of 80–120% recommended by
Matuszewski et al. [22]. Therefore, the EM solution was abandoned as a 3.2.4. Matrix effect and relative recovery
solvent and methanol was used exclusively as the extraction solvent. The Table 4 showed the results of the extraction recovery and matrix
extraction effects of different volumes of methanol (0.5, 0.7, and 1.0 mL) effect. The range of recovery values are 85.0–101.0%. The matrix effect
were also investigated (Table 2). As shown in Table 2, volumes of 0.7 mL values ranged from 92.0% to 105.0%, with the RSD of<12.8% for all
and 1.0 mL were sufficient for the extraction of mescaline from the hair
samples and were deemed suitable. These two volumes of methanol (0.7 Table 5
mL and 1.0 mL) were investigated for further optimization using an Stability data for mescaline in hair after storage at 4 ◦ C for 24 h in the auto­
authentic mescaline-positive hair sample (Case 11). The use of 0.7 mL sampler (n = 5).
methanol as the extraction solvent gave a larger area response for the Substance Concentrations (pg/mg) Determination results (pg/ RSD (%)
mescaline peak; therefore, 0.7 mL methanol was selected as the mg)
extraction solvent volume for all subsequent analyses. The washing steps 0h 24 h
were also studied in this work, and the last cleaning solution was
Mescaline 10 (LOQ) 10.7 11.0 2.2
negative for mescaline, indicating that the cleaning steps used as part of 20 20.4 19.6 3.0
this method removed external contaminants well. 100 88.7 94.9 4.7
Furthermore, we also report a far shorter total runtime (5 min) in our 800 825.9 864.6 3.2

Table 4
Intra-day and inter-day accuracy and precision, matrix effect and recovery for mescaline in hair.
Substance Concentrations (pg/mg) Intra-day (n = 5) Inter-day (5 × 4 days) Matrix effect (n = 5, %) Recovery (%)
(n = 5)
Accuracy (%) Precision (RSD/%) Accuracy (%) Precision (RSD/%) Mean (%) RSD (%)

Mescaline 10 (LOQ) 6.8 5.6 0.3 8.8 92.0 3.7 101.0

20 2.1 5.3 1.3 1.6 105.0 12.8 85.0
100 − 11.2 4.0 − 8.9 4.3 84.0 2.0 91.0
800 3.2 8.8 3.5 2.5 95.0 2.1 95.0

5
S. Yang et al.
Fig. 4. Chromatogram of mescaline in positive hair samples (case 10).
samples at both levels.
3.2.5. Stability
The good stability of substances in hair underwent demonstration
and was recognized as an asset for drug testing[24]. Stability of working
standard solutions and processed samples was tested in consideration of
the reasons above. Autosampler stability was evaluated using three
replicates of the LOQ, low, medium, and high content samples after
storage at 4 ◦ C for 24 h in the autosampler. The mescaline present in hair
extracts was stable and showed a precision RSD of 2.2–4.7%
（Table 5）.
3.3. Application to authentic hair samples
To our knowledge, only two reports have appeared for the quanti­
fication of mescaline in real positive hair samples. Those studies re­
ported concentrations of mescaline in hair samples of 1000 pg/mg [16]
6
Journal of Chromatography B 1195 (2022) 123202
and 80–130 pg/mg [19]. The developed UPLC-MS/MS method was
validated and applied to quantify mescaline in 19 authentic user hair
samples. The concentrations of mescaline in the hair segments of the 19
users ranged from 10 to 784 pg/mg, with a median of 112.2 pg/mg
(Table 3). The chromatogram for Mescaline Case 10 is shown in Fig. 4.
3.4. Case discussion
The 19 mescaline user individuals ranged in age from 30 to 51 years,
with a mean age of 40 years and a median of 39 years. People of this age
may possibly use the internet more than people of other age groups and
can therefore access the drug more readily, while also being able to
afford it. These persons were almost equally divided between males and
females (9 males and 10 females), suggesting that the use of mescaline
shows little or no gender preference. However, from a total of 31 cases
reported to the California Poison Control System (CPCS) between 1997
and 2008 [25], for peyote and mescaline exposure that met the study
S. Yang et al.
inclusion criteria, the patient ages ranged from 14 to 59 years, with a
mean of 23 years and median of 21 years, and twenty-six patients (84%)
were male. In that case series, the vast majority of exposures occurred in
adolescents and young adults. It’s a bold conjecture that the proportion
of female mescaline users may have increased, while the proportion of
young adult mescaline users may have decreased. However, there are
limitations in the conjecture since the pattern of drug abuse is different
in different localities, especially countries. Here suggestion is limited
and further study is needed.
In the 19 cases examined here, there were 3 cases with mescaline
levels lower than 10 pg/mg (the LOQ), although all 3 cases admitted
taking mescaline. In addition, the color of hair of those people is brown.
The mescaline concentrations in persons with black hair ranged from
18.6 pg/mg to 784.1 pg/mg (mean 201.1 pg/mg, median 116.8 pg/mg)
and in persons with brown hair from ＜10 pg/mg (the LOQ) to 341.3 pg/
mg (mean 79.2 pg/mg, median 24.4 pg/mg). Obviously, the concen­
tration of mescaline was higher in persons with black hair than in those
with brown hair. This suggests that the accumulation of drugs in hair
roots may relate to the color of the hair significantly. Previous research
studies have shown that melanin also has a substantial effect on drug
incorporation[26–28]. Steven et al. [28] considered that the melanin
pigments, which are polyanionic polymers with many negatively
charged carboxyl groups, are incorporated into the keratin fibrils of the
hair. Positively charged xenobiotics are therefore easily bound to
melanin by electrostatic forces, which cause the higher content of drug
in black hair. This could explain why the concentrations of mescaline
were higher in black hair than in brown hair in the present study.
However, there are limitations in the interpretation of hair analyses
considering the interpretation can be complicated by many factors, such
as poor dose-hair concentration relationships, variable growth rates of
the hair, inter-individual variance, and cosmetic treatment of the hair
[24,29–31]. Moreover, the sample size is too small to justify further
interpretation of this result. Further study is needed to verify the
apparent positive correlation between the hair root mescaline concen­
tration and hair color.
As mentioned above, there were 3 cases with mescaline levels lower
than 10 pg/mg (the LOQ). According to the self-portrait of those in­
dividuals, they had admitted taking mescaline only once in the 2 months
prior to the analysis. Moreover, the suspect user (case 19) consumed
peyote tea once a month before detection but information of amount
taken is unknown. The dose-hair concentration relationships remain
unclear.
During that period, abuse of DMT became more common in cases
processed. Therefore, the hair samples were also tested for DMT using
standard laboratory methods [32]. In total, 8 of the 19 authentic hair
samples shown in Table 3 were positive for both mescaline and
dimethyltryptamine (DMT). DMT has agonist activity at serotonin 5-
HT2A receptor sites, and is found in Ayahuasca, a plant-based ceremo­
nial potion with hallucinogenic effects. The binding affinities to the 5-
HT2A receptor are lower for mescaline than for DMT [33]. This in­
dicates that these suspects may smoke both drugs together to achieve
better hallucinogenic effects or that they have previously used mesca­
line/DMT and then chose another hallucinogen (DMT/mescaline)
instead. However, this conjecture has limitations because feedback
about mescaline/DMT consumption patterns is rare, and evidence to
prove that users combined the two drugs is inconclusive. Thus, further
study is needed to elucidate the frequency of the combined use of
mescaline and DMT among mescaline users.
3.5. Application to cactus samples
A sample of peyote cactus was also seized by the police from one
suspect. The validated UPLC-MS/MS method was also applied to quan­
tify mescaline in the fresh flower of the peyote cactus, Lophophora wil­
liamsii. As a result, the content of mescaline in the stamen, calyx, and
petal were 0.00526%, 0.00524%, and 0.00342% respectively. Peyote
7
Journal of Chromatography B 1195 (2022) 123202
contains a large spectrum of phenylethylamine (or phenethylamine or
β-phenylethylamine) alkaloids, with the principal form being mescaline.
The content is about 0.4% in fresh (undried) and 0.053–4.7% in dried
material, according to the literature by Olabode Ogunbodede et al. [34],
who examined samples of chlorenchyma from the green outer cortex of
the stem of several Echinopsis species. In another literature by Hulsey et
al [35], the mescaline content is 2.77–3.52% in dried stem of Lophophora
williamsii. Compared with the previously published data, the mescaline
content is much lower in the peyote flower of Lophophora williamsii than
in the stem of Echinopsis species. However, the lack of methodological
validation in a cactus matrix and parallel determination of cactus
chlorophyll tissue means that the specific distribution of mescaline
needs further study.
4. Conclusion
A fast and reliable ultra-high-pressure liquid chromatography tan­
dem mass spectrometry method was established and deemed suitable for
the qualitative and quantitative analysis of mescaline in hair in forensic
practice. The validated method was applied to 19 practical cases. The
mescaline concentrations in the hair samples of the suspected users
ranged from 10 to 784 pg/mg.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Acknowledgements
This work was supported by the National Natural Science Foundation
of China (No.81971789, 81872833); the Research Institute Projects (No.
GY2021G-6), the Shanghai Key Laboratory of Forensic Medicine (No.
21DZ2270800) and Shanghai Forensic Service Platform (No.
19DZ2292700).
References
[1] H.R. El-Seedi, P.A.G.M.D. Smet, O. Beck, G. Possnert, J.G. Bruhn, Prehistoric
peyote use: alkaloid analysis and radiocarbon dating of archaeological specimens
of Lophophora from Texas, J. Ethnopharmacol. 101 (1-3) (2005) 238–242, https://
doi.org/10.1016/j.jep.2005.04.022.
[2] J.H. Halpern, Hallucinogens and dissociative agents naturally growing in the
United States, Pharmacol. Ther. 102 (2) (2004) 131–138, https://doi.org/10.1016/
j.pharmthera.2004.03.003.
[3] R.K. Bullis, Swallowing the scroll: legal implications of the recent Supreme Court
peyote cases, J. Psychoactive Drugs 22 (3) (1990) 325–332, https://doi.org/
10.1080/02791072.1990.10472556.
[4] J.H. Halpern, A.R. Sherwood, J.I. Hudson, D. Yurgelun-Todd, H.G. Pope, Jr.
Psychological and cognitive effects of long-term peyote use among Native
Americans, Biol. Psychiatry 58 (8) (2005) 624–631, https://doi.org/10.1016/j.
biopsych.2005.06.038.
[5] R.J. Dinis-Oliveira, C.L. Pereira, D.D. da Silva, Pharmacokinetic and
Pharmacodynamic Aspects of Peyote and Mescaline: Clinical and Forensic
Repercussions, Curr. Mol. Pharmacol. 12 (3) (2019) 184–194, https://doi.org/
10.2174/1874467211666181010154139.
[6] J.L. Henry, J. Epley, T.P. Rohrig, The analysis and distribution of mescaline in
postmortem tissues, J. Anal. Toxicol. 27 (6) (2003) 381–382, https://doi.org/
10.1093/jat/27.6.381.
[7] H.H. Maurer, Liquid chromatography-mass spectrometry in forensic and clinical
toxicology, J. Chromatography B Biomedical Sci. Appl. 713 (1) (1998) 3–25.
[8] K.B. Nolte, R.E. Zumwalt, Fatal peyote ingestion associated with Mallory-Weiss
lacerations, West. J. Med. 170 (1999) 328.
[9] P.C. Reynolds, E. Jindrich, J.A mescaline associated fatality, J. Anal. Toxicol. 9
(1985) 183–184, https://doi.org/10.1093/jat/9.4.183.
[10] J. Beyer, O.H. Drummer, H.H. Maurer, Analysis of toxic alkaloids in body samples,
Forensic Sci. Int. 185 (1-3) (2009) 1–9, https://doi.org/10.1016/j.
forsciint.2008.12.006.
[11] M. Shen, P. X.Fundamentals and Applications of Hair Analysis. China Science,
Beijing China (2010).
[12] V.A. Boumba, M. Di Rago, M. Peka, O.H. Drummer, D. Gerostamoulos, The analysis
of 132 novel psychoactive substances in human hair using a single step extraction
S. Yang et al.
by tandem LC/MS, Forensic Sci. Int. 279 (2017) 192–202, https://doi.org/
10.1016/j.forsciint.2017.08.031.
[13] R. Casado, I. Uriarte, R.Y. Cavero, M.I. Calvo, I.LC-PAD Determination of Mescaline
in Cactus “Peyote” (Lophophora williamsii), Chromatographia 67 (7-8) (2008)
665–667, https://doi.org/10.1365/s10337-008-0553-2.
[14] C.M. Longo, R.A. Musah, An Efficient Ambient Ionization Mass Spectrometric
Approach to Detection and Quantification of the Mescaline Content of Commonly
Abused Cacti from the Echinopsis Genus, J. Forensic Sci. 65 (1) (2020) 61–66,
https://doi.org/10.1111/1556-4029.14134.
[15] Skácelová, L. Development of an analytical method for determination and identification
of mescaline in plant and biological material, (2017).
[16] C. Gambelunghe, R. Marsili, K. Aroni, M. Bacci, Rossi, R.GC-MS and GC-MS/MS in
PCI mode determination of mescaline in peyote tea and in biological matrices,
J. Forensic Sci. 58 (2013) 270–278, https://doi.org/10.1111/j.1556-
4029.2012.02249.x.
[17] M. Sergi, S. Napoletano, C. Montesano, R. Iofrida, R. Curini, D. Compagnone,
Pressurized-liquid extraction for determination of illicit drugs in hair by LC-MS-
MS, Anal. Bioanal. Chem. 405 (2-3) (2013) 725–735, https://doi.org/10.1007/
s00216-012-6072-x.
[18] J.Y. Kim, K.S. Jung, M.K. Kim, J.I. Lee, M.K. In, Simultaneous determination of
psychotropic phenylalkylamine derivatives in human hair by gas chromatography/
mass spectrometry, Rapid Commun. Mass Spectrom. 21 (11) (2007) 1705–1720,
https://doi.org/10.1002/rcm.3010.
[19] S. Pichini, E. Marchei, O. García-Algar, A. Gomez, R. Di Giovannandrea, R. Pacifici,
Ultra-high-pressure liquid chromatography tandem mass spectrometry
determination of hallucinogenic drugs in hair of psychedelic plants and mushrooms
consumers, J. Pharm. Biomed. Anal. 100 (2014) 284–289, https://doi.org/
10.1016/j.jpba.2014.08.006.
[20] E.M. Agency, Guideline on bioanalytical method validation, Drug Evaluation
Research 35 (2012) 396–398.
[21] F.T. Peters, O.H. Drummer, F. Musshoff, Validation of new methods, Forensic Sci.
Int. 165 (2-3) (2007) 216–224, https://doi.org/10.1016/j.forsciint.2006.05.021.
[22] B.K. Matuszewski, M.L. Constanzer, C.M. Chavez-Eng, Strategies for the
Assessment of Matrix Effect in Quantitative Bioanalytical Methods Based on HPLC-
MS/MS, Anal. Chem. 75 (13) (2003) 3019–3030, https://doi.org/10.1021/
ac020361s.
[23] Scientific Working Group for Forensic Toxicology (SWGTOX) Standard Practices
for Method Validation in Forensic Toxicology. J. Anal. Toxicol. 37 (2013)452-474.
doi:10.1093/jat/bkt054.
Journal of Chromatography B 1195 (2022) 123202
[24] F. Pragst, M.A. Balikova, State of the art in hair analysis for detection of drug and
alcohol abuse, Clin. Chim. Acta 370 (1-2) (2006) 17–49, https://doi.org/10.1016/
j.cca.2006.02.019.
[25] S.D. Carstairs, F.L. Cantrell, Peyote and mescaline exposures: a 12-year review of a
statewide poison center database, Clinical toxicology (Philadelphia, Pa.) 48 (4)
(2010) 350–353, https://doi.org/10.3109/15563650903586745.
[26] S.J. Green, J.F. Wilson, The effect of hair color on the incorporation of methadone
into hair in the rat, J. Anal. Toxicol. 20 (1996) 121–123, https://doi.org/10.1093/
jat/20.2.121.
[27] S.P. Gygi, R.E. Joseph Jr., E.J. Cone, D.G. Wilkins, D.E. Rollins, Incorporation of
codeine and metabolites into hair. Role of pigmentation, Drug Metab. Dispos. 24
(1996) 495–501.
[28] D.E. Rollins, D.G. Wilkins, G.G. Krueger, M.P. Augsburger, A. Mizuno, C. O’Neal, C.
R. Borges, M.H. Slawson, The effect of hair color on the incorporation of codeine
into human hair, J. Anal. Toxicol. 27 (2003) 545–551, https://doi.org/10.1093/
jat/27.8.545.
[29] J. Barbosa, J. Faria, F. Carvalho, M. Pedro, O. Queirós, R. Moreira, R.J. Dinis-
Oliveira, Hair as an alternative matrix in bioanalysis, Bioanalysis 5 (8) (2013)
895–914, https://doi.org/10.4155/bio.13.50.
[30] M. Barroso, E. Gallardo, D.N. Vieira, M. López-Rivadulla, J.A. Queiroz, Hair: a
complementary source of bioanalytical information in forensic toxicology,
Bioanalysis 3 (1) (2011) 67–79, https://doi.org/10.4155/bio.10.171.
[31] P. Kintz, Issues about axial diffusion during segmental hair analysis, Ther. Drug
Monit. 35 (2013) 408–410, https://doi.org/10.1097/FTD.0b013e318285d5fa.
[32] M. Liu, H. Yang, J. Hu, B. Shen, P. Xiang, H. Qiang, H. Deng, Z. Yu, Y. Shi, Analysis
of 28 hair samples from users of the hallucinogenic beverage ayahuasca, Forensic
Sci. Int. 323 (2021) 110790, https://doi.org/10.1016/j.forsciint.2021.110790.
[33] A. Rickli, O.D. Moning, M.C. Hoener, M.E. Liechti, Receptor interaction profiles of
novel psychoactive tryptamines compared with classic hallucinogens, Eur.
Neuropsychopharmacol. 26 (8) (2016) 1327–1337, https://doi.org/10.1016/j.
euroneuro.2016.05.001.
[34] O. Ogunbodede, D. McCombs, K. Trout, P. Daley, M. Terry, New mescaline
concentrations from 14 taxa/cultivars of Echinopsis spp. (Cactaceae) (“San Pedro”)
and their relevance to shamanic practice, J. Ethnopharmacol. 131 (2) (2010)
356–362, https://doi.org/10.1016/j.jep.2010.07.021.
[35] Hulsey, D., Kalam, M. A., Daley, P., Fowler, N. & Terry, M.CLINAL GEOGRAPHIC
VARIATION IN MESCALINE CONCENTRATION AMONG TEXAS POPULATIONS
OF LOPHOPHORA WILLIAMSII (CACTACEAE). Journal of the Botanical Research
Institute of Texas 5 (2011)677-683.