Forensic Science International 296 (2019) 15–21

Contents lists available at ScienceDirect

Forensic Science International

journal homepage: www.elsevier.com/locate/forsciint

Application of differential scanning calorimetry in the analysis of

apprehended formulations of anabolic androgenic steroids
Lucas Moraes Berneiraa , Samantha Coelho de Freitasa , Caroline Carapina da Silvaa ,
Alexandre de Mattos Machadob , Claudio Martin Pereira de Pereiraa ,
Marco Aurélio Ziemann dos Santosa,*
a
Center of Chemical, Pharmaceutical and Food Sciences, Laboratory of Lipidomics and Bioorganic, Federal University of Pelotas, Campus Universitário Capão
do Leão s/n, 96900-010, Brazil
b
Scientific and Technical Division, Brazilian Federal Police, Pelotas, 96030-000, Brazil

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

Article history: Over the past ten years, there has been a significant increase in the amount of formulations containing
Received 5 April 2018 anabolic androgenic steroids apprehended worldwide. A considerable amount of these illicit
Received in revised form 22 December 2018 preparations is falsified imposing a series of challenges for the analytical identification of alleged
Accepted 31 December 2018
active ingredients due to the presence of interferers. In this sense, the aim of this work was to identify and
Available online 10 January 2019
quantify the active ingredient using cholesterol as internal standard in eight apprehended formulations
of anabolic androgenic steroids in either tablet, capsule or injectable forms employing visual inspection
Keywords:
and instrumental analysis of Fourier Transform Infrared Spectroscopy, Gas Chromatography — Mass
Anabolic androgenic steroids
Forensic identification
Spectrometry and Differential Scanning Calorimetry. The assessed samples were kindly provided by the
Falsified steroids Brazilian Federal Police as representative samples from an apprehension made in July of 2017.
Differential scanning calorimetry Qualitatively, 25% of the analyzed materials were determined to be falsified as they were composed of
Thermal analysis excipients only while the others had the alleged active ingredient confirmed. However, after quantitative
Infrared spectroscopy analysis, the majority of samples were placed as counterfeit materials as the active substance was found
Mass spectrometry in concentrations lower than stated in the label. Preliminary visual inspection provided important
information to distinguish genuine from falsified samples. It should be noted that this work was one of
the few available reports to employ Differential Scanning Calorimetry in the analysis of anabolic agents,
which proved to be an important complementary tool for the detection of the active ingredient, when
present, along with the calorimetric profile of the formulations studied. Fourier Transform Infrared
Spectroscopy and Gas–Chromatography — Mass Spectrometry were also efficient analytical tools in order
to identify and to characterize substances present in fraudulent preparations.
© 2019 Elsevier B.V. All rights reserved.

1. Introduction and so, their commercialization is strictly controlled or prohibited

in many countries [2,5].
Anabolic androgenic steroids (AASs) comprise a class of Over the past decade, the illicit consumption of anabolic agents
numerous synthetic compounds derived from testosterone that is increasing worldwide leading to an increase in the apprehension
were developed for the treatment of conditions associated to of these materials by law enforcement agencies. In Brazil, for
hypogonadism in males [1]. Nonetheless, these compounds are instance, apprehensions of AASs triplicated between the years of
known for its illicit use among professional and recreational 2008 and 2011. At the same time, falsifications involving
athletes in order to obtain enhanced sports performance and formulations have also been growing and reached roughly a third
muscle development [2]. The illegal usage of steroids is associated of the analyzed samples [6]. Fraudulent preparations can impose
with side effects such as cardiovascular system damage [3], liver health hazards associated with the abuse of anabolic agents since
intoxications [4] among other possible severe physical damage, the formulations are manufactured under poor sanitary conditions
[7], exposing illicit users to unpredictable side effects [5] or even
death [8].
Generally, analysis of AASs involves visual inspection for a
* Corresponding author. verification of signs of falsification followed by chemical determi-
E-mail address: marcsantoss@hotmail.com (M.A.Z. dos Santos). nation using a suitable analytical technique [9]. In this sense,

https://doi.org/10.1016/j.forsciint.2018.12.022
0379-0738/© 2019 Elsevier B.V. All rights reserved.
16 L.M. Berneira et al. / Forensic Science International 296 (2019) 15–21

several analytical tools require an extraction procedure which may unknown, a fraction of 50 mg of the AASs was used for analysis. For
be performed employing methanol in ultrasonic bath [2,7,9]. For water suspensions, the formulation was dried under nitrogen flow
Gas Chromatography — Mass Spectrometry (GC–MS), a derivati- and the extraction was carried similarly to the other formulations.
zation step may be required to enable the detection and increase After the extraction procedure, one mL of the samples was
the resolution of the chromatogram, which requires the use of injected in split mode (1:25) in a GC–MS QP2010SE (Shimadzu,
derivatizing agents [8]. However, this process is optional and, in Kyoto, Japan) equipped with an Rtx-5MS capillary column (Restek,
several reports [7,10,11], derivatization is not employed since it is Bellefonte, USA) employing helium as the carrier gas. The
expensive and time consuming [11]. temperature of the injection port and the transfer line were
According to previous research works, formulations of AASs are 280  C and 300  C, respectively. In the oven, the programmed
generally analyzed using GC–MS [10], High Performance Liquid temperature was initially 100  C for 2 min increasing 30  C min-1 to
Chromatography (HPLC) [2] or Fourier Transform - Infrared 280  C and gradually increasing 2  C min-1 to a final temperature of
Spectroscopy (FT-IR) [7]. There are still other techniques that 310  C, holding for 10 min. The mass spectrometer operated with
can be employed such as Direct Analysis in Real Time Ionization — ion electron ionization at 70 eV in the ion source scanning from m/
Mass Spectrometry (DART-MS) [11] and Thin-Layer Chromatogra- z 30 to m/z 550. Identification of compounds was performed by
phy [12]. It should be noted that the application of novel analytical comparison of the resulting mass spectra with the NIST-08 library
tools with reduced sample preparation, cost and analysis time is while the quantitation of the analysis was performed using a
crucial for the characterization of illicit compounds. In this sense, calibration curve of cholesterol in concentrations of 1.25 mg/mL,
Differential Scanning Calorimetry (DSC) is a promising technique 1 mg/mL, 0.75 mg/mL, 0.50 mg/mL and 0.25 mg/mL in methanol.
rarely used for the forensic determination of apprehended drugs,
which may be worth exploring as it can be used to measure several 2.3. FT-IR analysis
physico-chemical properties of a sample with minimum work-up
[13–15]. For the FT-IR analysis, tablets and capsules had their content
As previously discussed, the higher incidence of apprehensions extracted following the procedure of Neves and Caldas (2017).
of AASs formulations demands solid forensic analysis in order to Briefly, 50 mg of the grounded sample was dissolved in 5 mL of
determine the chemical fingerprint of the material as well as to methanol, vortexed for 10 s, then sonicated in an ultrasonic bath
detect falsifications. Nonetheless, the analysis of anabolic agents is (USC-1800A, Unique) for 10 min, and centrifuged for 5 min at 3000
still limited to chromatographic and spectroscopic techniques, RPM. Finally, the organic phase was recovered and evaporated
which can hamper forensic determination in countries with under reduced pressure [9]. FT-IR analysis were performed on an
different infrastructure, making it necessary to develop alternative Attenuated Total Reflection spectrometer with Fourier Transform
techniques. Therefore, the aim of this work was to identify and (Shimadzu Prestige 21, Shimadzu, Kyoto, Japan) with a resolution
quantify the active ingredients of eight formulations of AASs in oily of 4 cm-1 ranging from 4000 cm-1 to 600 cm-1. Approximately 3 mg
solutions, water suspensions, tablets and capsules forms appre- of the extracted material from tablets or capsules and the crude
hended by the Brazilian Federal Police by means of visual oily formulations or dried fraction of water suspensions were
inspection, GC–MS, FT-IR and DSC. analyzed. For water suspensions, a preliminary drying under
nitrogen flow was required.
2. Method
2.4. DSC analysis
2.1. Chemicals and materials
DSC analysis was carried out on a 200 F3 Maia equipment (Netzsh,
HPLC-grade methanol was obtained from J.T. Baker (Phillips- Selb, Germany) previously calibrated with a standard containing
burg, USA), cholesterol (
99%) was purchased from Sigma-Aldrich indium. Initially, 3 mg of the extracted samples from tablets or
(St. Louis, USA) and aluminum crucibles were acquired from capsules as well as the crude material of oily based or water
Netzsh (Selb, Germany). The Brazilian Federal Police provided the suspensions were placed in aluminum crucibles and the analysis was
samples (Table 1) from an apprehension made in July of 2017 in the conducted from 30 to 300  C under constant heating of 10  C min-1 and
city of Rio Grande, localized in the south region of Brazil. nitrogen flow at 50 mL min-1. The extraction of tablets and capsules
followed the same procedure described in the FT-IR Analysis section.
2.2. GC–MS analysis
3. Results and discussion
GC–MS analysis was performed following the procedure
described by Coopman and Cordonnier [10]. Briefly, an aliquot 3.1. Visual inspection
of tablets finely grounded or injectables was diluted in methanol in
order to achieve a nominal concentration of 1 mg/mL [10]. In cases Visual inspection of the apprehended samples (Table 2) was an
in which the concentration of the tablets or the capsules was important preliminary procedure in order to distinguish genuine

Table 1
Details of the confiscated anabolic androgenic steroids.

Sample Sample form Sample appearance Declared compound Batch number

Product #1 Tablet Round, white and marked with an “L” Oxymetholone 15135
Product #2 Tablet White and blue capsules Unknown Unknown
Product #3 Tablet Round, light-pink and marked with an “L” Unknown Unknown
Product #4 Water suspension White liquid Stanozolol 416092
Product #5 Water suspension White liquid Stanozolol Unknown
Product #6 Oily solution Light yellow viscous liquid Nandrolone decanoate 416087
Product #7 Oily solution Light yellow viscous liquid Testosterone propionate 0141725
Product #8 Oily solution Dark yellow viscous liquid Trenbolone acetate TRE081094
 L.M. Berneira et al. / Forensic Science International 296 (2019) 15–21 17

Table 2
Visual inspection of the apprehended samples.

Parameter Product #1 Product #2 Product #3 Product #4 Product #5 Product #6 Product #7 Product #8

Container Sealed Sealed Sealed Sealed Sealed Sealed Sealed Sealed
Legible information Yesb n.a.a n.a.a Yesb Yesb Yesb Yesb Yesb
Registration of trade name Noc n.a.a n.a.a Noc Noc Noc No No
Registration of manufacturer Noc n.a.a n.a.6 Noc Noc Noc No No
Spelling of the active ingredient Correctb n.a.a n.a.a Correctb Correctb Correctb Correctb Correctb
Status of the active ingredient Legald n.a.a n.a.a Legald Legald Legald Legald Legald
Logotype and name of Yes n.a.a n.a.a Yes Yes Yes Yes Yes
manufacturer
Hologram of manufacturer No n.a.a n.a.a Yes Yes Yes No No
Address of manufacturer Yes n.a.a n.a.a Yes Yes Yes No No
Statement of drug strength Yes n.a.a n.a.a Yes Yes Yes Yes Yes
Statement of dosage form Yes n.a.a n.a.a Yes Yes Yes No No
Statement of number of units Yes n.a.a n.a.a n.a.a n.a.a n.a.a n.a.a n.a.a
Batch number and expiry date Yes n.a.a n.a.a Yes Yese Yes Yes No
Size of solid materials Round Ellipsoid n.a.a n.a.a n.a.a n.a.a n.a.a n.a.a
Shape of solid materials Uniform Uniform Uniform n.a.a n.a.a n.a.a n.a.a n.a.a
Damaged or empty solid No No No n.a.a n.a.a n.a.a n.a.a n.a.a
materials
Content Uniform Uniform Uniform Uniform Uniform Uniform Uniform Uniform
solid solid solid suspension suspension solution solution solution
a
Non-available (n.a.).
b
Information written in other language (English, Spanish or German) rather than in Portuguese.
c
Products and trade names not registered in Brazil, but registered in Paraguay.
d
Active ingredients legal, but placed under controlled substances list in Brazil.
e
Product had expiry date, but did not have batch number.

from falsified samples. Generally, the format of the label which could give the user a false impression that the formulation
information vary worldwide but, according to the World Health was imported. However, similarly to Product #8, the manufacturer
Organization (WHO), it should contain, for instance, trade name, of Product #7 does not exist leading to the conclusion that the
active ingredient, manufacture’s name, logotype and full address, sample was counterfeit despite having an adequate label and
batch and expiration date. In counterfeit samples, such data may be recipient.
absent, adulterated or incomplete indicating that the product Product #2 and #3 were not labeled as the samples were found
under review may be falsified. Moreover, other components of the in a blank recipient. Notwithstanding the absence of labeling,
sample including the recipient and its content can also be visual inspection of the recipient and its contents did not reveal
evaluated assisting in this process [16]. particularities found in counterfeit formulations such as damages,
Throughout visual inspection, it was noted that Product #1, #4, cracks or unevenness of the constituents. Product #3 had an “L”
#5 and #6 were manufactured by a Paraguayan laboratory that did inscription on its surface, which is characteristic of a Paraguayan
not have a health license to distribute anabolic agents in Brazil, manufacturer that also produced Product #1, #4, #5 and #6, while
which means that these preparations were illegal, even if the the Product #2 did not have any distinguishable features that
formulation was genuine [17]. According to the Brazilian Penal could indicate its possible origin. Nonetheless, a chemical
Code, the illicit importation and distribution of these materials can characterization was conducted in order to confirm the presence
be typified as a crime punishable from 10 to 15 years of or absence of anabolic agents in Product #2 and #3 as well as in the
incarceration [18]. Observation of the physical presentation of other samples visually inspected [16].
the aforementioned AASs did not show signs of falsification as Table 2. Visual inspection of the apprehended samples.
several required information typical of genuine formulations were As it could be observed in the results of visual inspection, the
found in the packaging. It should be noted that the informed batch procedure allowed an adequate initial screening to discriminate
numbers, expiration date and physical form of the samples agreed possible falsified products from genuine formulations. However,
with those found in the website of the Paraguayan laboratory visual inspection is not commonly used in the analysis of AASs
indicating that the formulations were genuine. among the literature as labeling of counterfeit and veridical
On the other hand, Product #8 lacked several information in its samples can be very similar as well as some apprehended materials
label such as expiration and manufacturing date, composition as are not labeled hampering the procedure. These obstacles were
well as origin, indicating that the sample was falsified. These observed in the current work as well as in previous research works
preliminary observations were reinforced since the manufacturer that dealt with the visual inspection of apprehended anabolic
does not officially exist and, therefore, the formulation was from a agents. Nonetheless, a clear distinction between falsified and
clandestine laboratory. It is worth noting that there are no genuine materials was possible for most samples of this work
manufacturers with a health license to distribute the alleged active based in the information of the label which was not observed in the
ingredient trenbolone acetate in Brazil and, thus, the potentially previous publications found in the literature [19,20].
falsified product is illegal within the country [17].
Nevertheless, identifying falsified pharmaceuticals throughout 3.2. Chemical characterization
visual inspection may not be an easy task because fraudulent
preparations can be meticulously designed to look like as genuine FT-IR analysis of Product #7 and #8 oil droplets (Fig. 1S) showed
medicine [16]. This was the case of Product #7 whose label had the same patterns of bands with similar intensities that correspond
several information common to non-falsified products. Its label to = CH (3008 cm1), C¼O (1743 cm-1), CO stretching (1159 cm-
1
was written in German while the logotype and name of the ) and CH2 rocking vibration (721 cm-1). The identified chemical
supposed manufacturer were very similar to the Bayer laboratory, groups matched several of the possible molecular vibrations of the
18 L.M. Berneira et al. / Forensic Science International 296 (2019) 15–21

indicated active ingredients of both samples. However, the in several health hazards to a user that include, for instance,
indicated vibrations may also be related to the oily excipient, generalized infections and muscular atrophy [2].
which is in the form of long-chain alkyl esters normally found in On the other hand, spectroscopic analysis of Product #3, #4 and
vegetable oils [21]. Therefore, it was not possible to confirm or #5 (Fig. 2S) indicated the presence of stanozolol due to the
exclude the presence of the active ingredient employing FT-IR and occurrence of characteristic vibrations associated with the
further analysis were required in order to identify the formulation compound including N H (3473 cm-1 and 1523 cm-1), C ¼ N
properly. Infrared spectrum of all samples can be seen in the -1
(1660 cm ), O H (3112 cm-1) and ¼ C H (3014 cm-1). The
Supplementary Information section. Based upon the spectroscopic presence of stanozolol in the samples was also observed in DSC
analysis that did not clearly established the presence of the analysis (Fig. 2), in which it was possible to observe the MP of one
supposed active ingredients, DSC analysis of both samples (Fig. 1) of its crystals forms (151.2  C) in agreement with the literature
was conducted. The use of DSC allowed the identification of a certain (155  C). Other calorimetric characteristics such as dehydration
compound due to its distinctive melting point (MP) and other (85.9  C) and degradation of the molecule (219.4  C) were also
physico-chemical properties in the thermogram [13]. observed in the thermogram [22].
Similarly to the FT-IR results, thermograms from both Product GC–MS analysis of Product #3, #4 and #5 showed only one
#7 and #8 were identical, strongly indicating that these materials peak that was identified as stanozolol according to the library of
were constituted of the same compounds. As shown in Fig. 1, there the equipment. In the mass spectra of these AASs (Fig. 3S) the
were no events related to the MPs of testosterone propionate (118– molecular ion of m/z 328.25 proximate to the exact mass of
122  C) or trenbolone acetate (94–97  C) indicating that these stanozolol (m/z: 328.25) was identified and, thus, confirming its
compounds were not present in any of the samples [22]. The first presence in the samples. GC–MS results of all analyzed materials
exothermic event occurred at 175.0  C, which was probably related can be seen in the Supplementary Information section. Stanozolol
to the partial oxidation of the oily formulation while the entered the US market in the 1960s as aqueous suspensions or
exothermic event at 232.1  C can be associated with the vaporiza- tablets and was mainly used for the treatment of osteoporosis and
tion of the sample. These results are very similar to a previous work protein deficiency disorders [12]. The anabolic agent was later
reported in the literature [23] related to the analysis of long-chain banned in the US due to its hepatotoxicity, however its illicit use
alkyl esters of soybean oil performed under identical DSC continued since it cannot be converted to estrogen in the organism
conditions. Comparison of the results strongly indicated that and, thus, avoid some common anabolic disorders such as water
none of the samples had active ingredients but were composed retention and ginecomastia [25].
only of vegetable oil. For Product #6, FT-IR analysis of the oily formulation (Fig. 4S)
GC–MS analysis confirmed the results indicated by visual indicated the presence C¼O (1719 cm1), C O (1174 cm-1), C¼C
inspection and DSC as both testosterone propionate and trenbo- (1673 cm ), C¼C related to the presence of monosubstituted
-1

lone acetate were not detected in the formulations of Product #7 aromatic rings (709 cm-1), and stretches of long carbon chains
and #8, respectively. In this order, both apprehended samples were (606 cm-1) which would be in agreement either with the presence
considered falsifications categorized as formulations only com- of active ingredient or its excipients. DSC analysis of the oily
posed of excipients according to the WHO guidelines. It should be solution (Fig. 3) revealed one endothermic event at the onset
noted that this is one of the most common form of counterfeit temperature of 201.8  C that was related to the melting and
accounting for approximately 40% of falsifications involving vaporization of the oily formulation. It was not possible to observe
pharmaceutical products [24]. Although only composed of the MP of nandrolone decanoate (118  C) as well as other particular
vegetable oil, the samples were manufactured in inadequate physico–chemical properties due to the high affinity of the
sanitary conditions and poor quality control which could implicate components in the formulation [22].

Fig. 1. DSC thermogram of the oily formulation of Product #7 and #8. Fig. 2. DSC thermogram of the content from Product #3, #4 and #5.
 L.M. Berneira et al. / Forensic Science International 296 (2019) 15–21 19

Fig. 3. DSC thermogram of the oily formulation of Product #6. Fig. 4. DSC thermogram of the extracted content of Product #1.

GC–MS analysis (Fig. 5S) detected eight peaks for Product #6 [22]. It can also be observed that in the endothermic event found at
with a predominance of benzyl benzoate, ethyl oleate and 222.8  C the solid begins to suffer a degradation process.
nandrolone decanoate, and a peak confirming (Fig 6S) the presence GC–MS analysis of Product #1 showed the presence of one peak
of the active ingredient (nandrolone decanoate; m/z 428) and the whose mass spectra (Fig 8S) revealed a molecular ion of 332.25
excipients. Qualitatively, the sample was not considered a related to the active ingredient. Oxymetholone belongs to the 17α
counterfeit, which was further confirmed by a quantitative alkylated group of AASs and, therefore, cannot be inactivated by
analysis. Nandrolone decanoate acts as a prodrug releasing the first-pass liver metabolism when consumed orally. However,
nandrolone in the organism up to 3 weeks after injection and is the alkylation in the 17α position also enhanced the risk of
therapeutically used to treat osteoporosis and sarcopaenia. hepatotoxicity of oxymetholone which made the compound
However, due to a higher ratio of anabolic: androgenic activity controlled worldwide [26,28].
compared to other AASs, nandrolone decanoate is widely used Lastly, for Product #2 bands such as O H (3512 cm-1) and
illicitly [26,27]. C ¼ O (1716 cm-1) were observed (Fig 9S). Since the apprehended
Benzyl benzoate is extensively used as an excipient in injectable sample was unlabeled, the observed bands could not be associated
anabolic agents in order to decrease the viscosity of the oily to the stated anabolic agent as these vibrations can be seen in
solution as well as to prevent crystallization of steroids. The innumerous AASs as well as other compounds. DSC analysis (Fig. 5)
substance is also used in food preservatives, perfume solvents, showed an initial exothermic event at the onset temperature of
flavorings, repellents and insecticides, but it can cause serious 82.2  C related to the crystallization of the solid. Furthermore, an
adverse effects such as skin irritation, dermatitis, anaphylactic endothermic event at 238.0  C was related to a MP that was close to
shock and seizures when used in high concentrations. Ethyl oleate the reported in the literature for oxandrolone (235  C) indicating
was found among other substances in the formulation, which is a the presence of the active ingredient in the sample [22]. GC–MS
common vehicle for injectables although not approved by the Food analysis (Fig 10S) confirmed oxandrolone (m/z 306).
and Drug Administration since there are no studies on its biosafety The overall results of the spectroscopic analysis showed that the
[27]. In addition to this compound, other fatty acid ethyl esters identification of the active ingredients was difficult due to the
such as ethyl palmitate, stereate and laurate are commonly used in interference of excipients. Moreover, in cases in which the active
oily-based AASs in order to prolong the effect of the steroid in the ingredient was not stated in the label, it was not possible to
organism [26]. associate the bands to a particular anabolic agent. In this sense, it
FT-IR analysis of the extracted content from Product #1 (Fig 7S) was necessary to complement the results with an evaluation by
indicated the presence of oxymetholone due to distinguishing DSC in which it was possible to identify most of the AASs stated or
bands of O H (3331 cm-1), C¼O (1613 cm-1), C O (1199 cm-1) and not in the label with little sample preparation, low cost and quick
C ¼ C (931 cm-1) bands. It should be noted that the C¼O stretch of analysis [29]. Given the results, the association of DSC with another
this compound is very characteristic since it undergoes tautome- screening technique can be used to identify the active ingredient in
rization, which shifts the appearance of the carbonyl group to questioned formulations which could serve as an is important tool
lower wavenumbers. DSC analysis (Fig. 4), indicated endothermic for a wider range of laboratories with different instrumentation
events at 65.5  C and 117.2  C associated with solid-solid tran- capacity [16].
sitions followed by an exothermic event related to crystallization For quantitation purposes, an analytical curve containing
(82.5  C) as well as endothermic events related to the MP at cholesterol (R2 = 0.9999) was employed for the procedure. The
164.7  C and 187.0  C possibly due to the polymorphism in the use of cholesterol can be a feasible approach to quantify synthetic
material. These results were close to the reported in the literature androgens due to their chemical structure similarity overcoming
(172–180  C) and also indicated the presence of oxymetholone certain limitations or the inability to acquire pure standards. It is
20 L.M. Berneira et al. / Forensic Science International 296 (2019) 15–21

the application of sequential analytical tools was required for the

adequate identification of the samples. It was demonstrated that
visual inspection allied with an instrumental characterization of
the AASs were crucial procedures in order to characterize and
detect falsifications. Finally, DSC proved to be a feasible
complementary analysis by providing events related to character-
istic physico-chemical properties that assisted in the identification
of the alleged active ingredients in the samples. Preliminary visual
inspection associated to FT-IR, DSC or GC–MS successfully allowed
for the detection of anabolic agents and, therefore, could be used in
laboratories of different infrastructure for the forensic analysis of
apprehended formulations.

Conflict of interest

The authors declare that they have no conflict of interest.

CRediT authorship contribution statement

Lucas Moraes Berneira: Conceptualization. Samantha Coelho

de Freitas: Conceptualization. Alexandre de Mattos Machado:
Conceptualization, Methodology. Claudio Martin Pereira de
Pereira: Conceptualization, Funding acquisition. Marco Aurélio
Ziemann dos Santos: Conceptualization, Supervision.
Fig. 5. DSC thermogram of the content of Product #2.
Acknowledgments
worth noting that there could be minor differences in the obtained
concentration of the studied active ingredients as cholesterol and The authors are thankful to the Brazilian Federal Police (Pelotas,
anabolic agents do not share the same chemical structure and Rio Grande do Sul State) for providing the samples, to the Forensic
chromatographic characteristics. National Institute of Science and Technology (Grant number
Quantification of active ingredients in the samples (Table 3) 465450/2014-8), Research Support Foundation of the State of Rio
showed that most of the apprehended formulations of AASs were Grande do Sul (FAPERGS) and Coordination of Improvement of
counterfeit as Product #6 was the only formulation to achieve the Higher Education Personnel (CAPES) for research funding.
concentration stated in the label. For the other anabolic agents, the
active substance, when present, was generally found in concen- Appendix A. Supplementary data
trations equal to half or less than declared on the label. Neves and
Caldas (2017) also observed that, after a quantitation step, the rate Supplementary material related to this article can be found, in the
of counterfeit formulations reached approximately 50% and was online version, at doi:https://doi.org/10.1016/j.forsciint.2018.12.022.
higher than the falsifications detected by qualitative analysis that
were found in 25% of the preparations [9]. It should be noted that References
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