Cannabis Farmacocinetica

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Farmacannabis-UFRJ: The first laboratory in Brazil to analyze therapeutic


products derived from Cannabis

Article in Brazilian Journal of Analytical Chemistry · January 2017

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Virginia M Carvalho
Federal University of Rio de Janeiro
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Br. J. Anal. Chem., 2017, 4 (16), pp 44-49

Feature

Farmacannabis-UFRJ: The first laboratory in Brazil to


analyze therapeutic products derived from Cannabis

Virgínia Martins Carvalho


Associate Professor
Faculty of Pharmacy, Federal University of Rio de Janeiro, RJ, Brazil
Farmacannabis-UFRJ Project Coordinator
virginiamc@pharma.ufrj.br

Therapeutic treatment using Cannabis extracts is sometimes the only alternative in the control of
intractable epilepsy and other serious illnesses. Cannabis sativa is a banned plant in Brazil, but the import
of Cannabis extracts has been authorized for medical use since 2015, and the first safeguards to cultivate
Cannabis for medicinal purposes were obtained in 2016.
Although sanitary regulations created the demand for the chemical analysis of Cannabis products, all
laboratories in Brazil that analyze Cannabis and Cannabis products have forensic purposes, and
laboratories focused on the monitoring of therapies are a medical necessity.
To assess the safety of Cannabis derivative therapies,
the Laboratory of Analytical Toxicology at the Faculty of
Pharmacy, Federal University of Rio de Janeiro (LATOX-
FAR-UFRJ) was structured, and is the first laboratory in
Brazil to monitor medical treatment using
phytocannabinoids. Through the project Farmacannabis,
the LATOX-FAR-UFRJ analyzes Cannabis and its
products to monitor the safety of therapies, and to
minimize the risk of improper preparation of its extracts
by people without pharmaceutical training. Most of the
patients are children with intractable epilepsy and autism,
but it also serves patients with cancer, chronic pain, Researchers at the LATOX-FAR-UFRJ
Alzheimer's, Parkinson's and other diseases.
The medicinal properties of the plant from the Cannabis genus have been described in the Chinese
Pharmacopoeia Pen-Ts'ao Ching for 2000 years, considered the first pharmacopoeia known in the world,
and described by the Assyrians for 3000 years [1]. The book “Dicionário de Plantas Úteis do Brasil e das
Exóticas Cultivadas”, a compendium of references in the pharmaceutical area in Brazil, published by M.
Pio Côrrea in 1926 [2], describes the botanical characteristic and medicinal properties of Cannabis sativa
L. being named generically by "true hemp". The “Farmacopeia Brasileira” published in 1929 [3] describes
the Cannabis sativa Linnaeus var. indica, with the generic names "maconha", "meconha", "diamba" and
"cannabis", indicating the flowered summits as a raw material in preparing the Cannabis officinalis extract
or fluid extract of India hemp, hemp powder and the Indian hemp dye.
In forensic analysis guidelines, Cannabis is considered to be monospecific (Cannabis sativa L.) however,
Cannabis is divided into several subspecies (C. sativa subsp. sativa, C. sativa subsp. indica, C. sativa
subsp. ruderalis, C. sativa subsp. spontanea, C. sativa subsp. kafiristanca). The chemical and morphological

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distinctions of the subspecies are often not readily discernible and are not required for forensic purposes
(UNODC, 2009). The main active compounds present in the plant are the cannabinoid substances, of which
the more abundant being Δ9 and Δ8-tetrahydrocannabinol (THC), cannabinol (CBN) and the cannabidiol
(CBD) that allow the identification of the plant chemistry. The THC and CBD can still be identified in its acidic
forms (CBDA and THCA) which are converted to the neutral forms by heating. After the elucidation of the
CBD structure [4] and identification of Δ9-THC, one of the substances responsible for their psychoactive and
euphoric effects [5], several scientific papers have been developed showing the pharmacological and clinical
properties of its compounds [6-9]. The pharmacological effects are attributed to the interaction of
cannabinoids with cannabinoid receptors mainly distributed in the central nervous system (CNS) known as
CB1, and peripheral nervous system and immune system known as CB2 [10]. CBD offers paradoxical
pharmacological effects in relation to the effects of Δ9-THC, while the former acts as a depressant and
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anxiolytic, the second acts as a stimulant and hallucinogen [8]. The Δ -THC binds to the CB1 and CB2
receptors acting as a partial agonist, and seems to have a mixed neural activity, excitatory and inhibitory, in
9
different areas of the brain, showing they do not only act in specific cannabinoid receptors [11]. Unlike Δ -
THC, CBD has low affinity for CB1 and CB2 receptors, its action seems to result from an increase in
anandamide, the endogenous cannabinoid neurotransmitter [12], shows anxyolytic [13] and anticonvulsant
effects [14,15]. The pharmacological potential of cannabinoids in the treatment of Parkinson's disease [16],
schizophrenia [17,18], Alzheimer's [19-21], neuropathic pain present in multiple sclerosis [9], rheumatoid
arthritis [22], and epilepsy [15,23,24] has been demonstrated.
Probably, the first scientific work which has shown the effectiveness of the CBD in the control of seizures,
was published in 1980 by the group of Elizaldo Carlini, professor at the Federal University of São Paulo,
together with Raphael Mechoulam, professor at the Hebrew University [14]. Currently extracts rich in CBD
are being used at the clinic in the management of refractory epilepsy.
After a long period in which the production of Cannabis based medicines was banned in Brazil,
international experience stimulated the search for treatment of epilepsy with extracts from hemp, a flagship
experience was the case of British girl, Charlotte Figi, who suffered from multiple episodes of convulsions
since the first months of life, and diagnosed with Dravet syndrome when she was 2.5 years old. Her family,
discouraged with the ineffectiveness of traditional pharmaceutical drugs, started administrating a Cannabis
extract in Colorado, United States of America (USA). The Cannabis treatment controlled the epilepsy in the
first week. In the USA, hemp extract has been approved by the Food and Drug Administration (FDA) as a
food supplement, and the medical use of Cannabis in California was approved in 1996.
Charlotte's Cannabis treatment encouraged mothers, parents, patients and activists to put pressure on
the regulation of medical Cannabis use in Brazil. The “Conselho Federal de Medicina” authorized the
compassionate prescription of CBD in December 2014, and the “Agência Nacional de Vigilância Sanitária”,
Brazilian regulatory agency (ANVISA) authorized its importation for personal medical use in 2015. In March
2016, by determination of Federal prosecutors to answer the plea of a patient, ANVISA has authorized the
importation for personal medical use of the plant Cannabis sativa L, parts of the plant and THC, and
partially corrected an inconsistency contained in regulating controlled substances (Portaria 344/98) from
the year 2000, update dronabinol (Marinol™) on the list of stimulant drugs. Dronabinol is the synthetic THC
structure identical to phytocannabinoid THC indicated primarily to the control of nausea and vomiting in
patients undergoing through chemotherapy treatment. In addition, the first Cannabis extract named
Mevatyl™ (Sativex™ in other countries) was registered as a pharmaceutical drug by ANVISA in 2017,
while kept the plant and its active ingredients on the proscribed list (of prohibited drugs).
Imported extracts are classified as “hemp”, that is plants of the genus Cannabis rich in CBD and poor in
THC usually used as fiber. International law determined that the concentration of THC is less than 0.3%
for the registration of products as dietary supplements and cosmetics. On the other hand, in Brazil emerges
the use of handmade products and trade in clandestine products that are prepared with different varieties
of Cannabis have not yet been characterized scientifically.
Many people carry out experiments to produce Cannabis products for recreational and medical use,
and have empirical knowledge in their own language of that which has been identified as "Cannabis culture",
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they claim to differ in the macroscopic feature, organoleptic characteristics, develop agronomic methods
and taxonomic classifications based on feeling and personal experience during use, and describe
compositions and levels of active ingredients. Although this empirical knowledge is being an instrument for
the remedies of those who need these medicinal extracts, research and development is fully needed,
along with scientific methodology, from the description of the varieties of Cannabis available in the national
market, to the standard raw vegetal material.
Cannabis features at least 50 phytocannabinoids and that, in addition to this variety of substances, the
effect can be determined by the acidic or neutral form of the same cannabinoids such as THCA which is
converted to THC by heating. The preparation technique of the extract (type of solvent extraction and
temperature) is decisive in the final concentration of THC and THCA [25] since its concentrations can be
modulated by heating. If forensic toxicology gas chromatography is sufficient to analyzing cannabinoid
compounds, for medical purposes the decarboxylation of acid forms is a technical limitation easily solved
by the use of liquid chromatography.
To monitor Cannabis therapies and to give pharmaceutical support to patients, the Farmacannabis
project was created at LATOX-FAR-UFRJ, where Cannabis products are analyzed and the production of
homemade extracts is assisted, and medicinal extracts and plants used in its preparation are analyzed to
determine the cannabinoids, toxic metal levels and microbiological quality. For structuring the project,
many barriers were overcome with the persuasion of professional peers, that with professional ethics and
a commitment to public health, agreed that it is necessary to brave prejudices in order to study and
evaluate the safety of Cannabis treatments.
Currently, Farmacannabis has ten PhD. Professors
from pharmacy, neurosciences, botanic, natural
products and juridical areas, several students and
technical professionals, partnerships with public
institutions like “Instituto de Tecnologia em Fármacos,
Farmanguinhos, Fiocruz” (Far-Fiocruz), and “Instituto
Nacional de Controle de Qualidade em Saúde,
Fiocruz” (INCQS-Fiocruz), and partnerships with
non-governmental Organizations (NGOs), such as
“Associação Brasileira para Cannabis” Some researchers of the Farmacannabis project
(ABRACANNABIS) and “Associação de Apoio à Pesquisa e Pacientes de Cannabis Medicinal” (APEPI).
Financial support was obtained by collective funding (crowdfunding) and more than 800 people backed the
project, with the collection of about US $ 30,000. Furthermore, Farmacannabis received financial support
from the Manserv Company, a resident enterprise from Technological Park in UFRJ, and the technical
support of the Brazilian company Nova Analítica. With strategically worked idea, the first laboratory in
Brazil that officially analyzes medical Cannabis extracts was structured.
Farmacannabis' preliminary results, obtained by the high-performance liquid chromatographic method
development, showed that the commercial imported extracts have high CBD content with a chemical profile
very different from homemade and illicit trade products (Tables I and II). In general, the homemade
products showed low cannabinoid concentrations and are rich in THC, except for the Harle-Tsu variety with
hemp profile introduced between patients by ABRACANNABIS. Hemp varieties are probably rare in Brazil
because of the Cannabis cultivations which have been turned over to recreational use, and the agronomic
techniques have been employed for the production of varieties rich in THC. The illegal Cannabis samples
seized in the United States of America from 1995 to 2014 increased THC content of 5% to 15% on average
and the THC/CBD ratio increased from 14 times in 1995 to about 80 times in 2014 [26].
The development of phytocannabinoid medicine in Brazil depends on the studies of available varieties
of Cannabis that can become active pharmaceutical ingredients, and in this way, analytical chemistry
laboratories and toxicological analyzes need to be prepared for the Cannabis pharmaceutical market that
requires technical specificities different from the forensic application.

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Table I. Preliminary results from commercial imported products analyzed by HPLC-DAD

Pure CBD Concentration [mg/mL or mg/g]


Labeled
Origin Form/vehicle CBDA CBD THCA CBN THC
content
USA Oil 200 mg/mL ND 210.45 ND ND ND
USA Oil 50 mg/mL ND 13.17 ND 0.45 ND

USA Oil 50 mg/mL ND 77.45 ND ND ND


USA Oil 50 mg/mL ND 41.17 1.10 0.40 ND
USA Oil 100 mg/mL ND 104.13 ND ND ND

USA Oil 200 mg/mL ND 284.90 ND ND ND


USA Oil 100 mg/mL ND 88.40 ND ND ND

USA Oil 100 mg/mL ND 90.70 ND ND ND

CBD Enriched Extracts CBDA CBD THCA CBN THC

USA Oil 50 mg/mL ND 47.88 ND 0.75 3.30


USA Oil 60 mg/mL ND 39.34 ND ND 1.44
USA Oil 61 mg/mL ND 40.67 ND ND 1.74

Canada Oil 24.97 mg/mL 1.15 21.14 0.56 0.30 2.16


USA Resin 170 mg/g ND 121.93 1.00 0.42 2.31

160 mg/g
USA Resin 4.49 142.48 ND 1.32 8.15
THC < 9 mg/g
USA Resin 240 mg/g 1.97 118.81 ND ND 1.60

USA Capsule ND ND 80.87 ND ND 4.12


USA Capsule 50 mg/cap ND 95.88 ND ND 4.33

ND: Not detected (Limit of Detection = 0.01 mg/mL or 0.01 mg/g for resin); USA: United States of America.

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Table II. Preliminary results from homemade or clandestine products analyzed by HPLC-DAD

Concentration [mg/mL or mg/g]


THC enriched extracts or THC-CBD (1:1)

CBDA CBD THCA CBN THC


Origin Form/vehicle Cannabis strain
RJ Oil Cronic ND 0.10 2.36 ND 1.64
RJ Oil Pur Kush ND 0.10 3.65 ND 1.39
RJ Oil Cinderela ND ND 0.80 ND 2.19
RJ Oil Cinderela ND ND 1.16 ND 2.64
RJ Oil Wildfire ND 0.97 ND ND 6.39
RJ Oil Harletsu oil + Cinderela resin 3.35 2.59 0.74 0.40 2.83
RJ Oil Hybrid:CBD skunk haze + LSD 3.81 ND 2.39 ND 0.10
SC Oil UN 0.10 0.61 2.65 0.11 12.90
SC Oil UN ND 2.20 0.90 1.03 44.85
SC Oil UN 0.10 1.36 0.71 0.77 29.60
RJ Resin Cinderela 0.10 0.33 0.14 8.42 28.30
SC Resin UN ND 16.67 6.32 5.43 338.22
SC Resin UN 0.75 14.59 0.70 4.20 314.12
SC Resin UN ND 14.05 1.02 5.84 303.25
CBD enriched extracts
RJ Oil Harletsu 2.32 1.06 0.10 ND ND
RJ Oil Harletsu 2.67 1.21 0.10 ND 0.10
RJ Oil Harletsu 4.33 3.30 0.11 ND 0.25
RJ Oil Harletsu 0.24 4.77 ND 0.10 0.40
RJ Oil Harletsu 0.10 7.95* ND ND 0.33
RJ Oil Harletsu 4.25 17.43* ND 0.15 0.73
Extracts with cannabinoids at trace level
PB Glicerin UN ND ND 0.20 ND 0.14
PB Glicerin UN ND ND 0.20 ND 0.14
PB Glicerin UN 0.30 0.10 0.12 ND 0.10
PB Glicerin UN 0.40 0.14 0.10 ND 0.12
PB Glicerin UN 0.21 0.10 0.10 ND 0.10
PB Glicerin UN 0.21 0.10 0.10 ND 0.10
PB Glicerin UN ND ND 0.19 ND 0.10
PB Glicerin UN ND 0.10 0.21 0.05 0.16
PB Glicerin UN 0.04 0.03 0.04 ND 0.02
PB Water/spray UN 0.05 ND 0.20 ND 0.06
ND: Not detected (Limit of Detection = 0.01 mg/mL or 0.01 mg/g for resin); PB: Paraíba, BR; RJ: Rio de Janeiro,
BR; SC: Santa Catarina, BR; UN: unknown; *Prepared by alcoholic extraction with patients in LATOX with
pharmaceutical supervision.
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ACKNOWLEDGMENTS
The Farmacannabis' coordinator is grateful to Dr. Ernesto Diaz Rocha, MS. Fábio Luiz Costa de Souza
Release
and Andrey Fabiano Lourenço de Aguiar for the laboratorial and analytical help.

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