Cannabidiol (CBD) and tetrahydrocannabinol (THC) are the main active principles in the Cannabis extracts. Rich-CBD extracts are indicated to control seizures in epileptic patients, THC is indicated as antiemetic to patients in chemotherapy treatment and extracts with THC and CBD in proportions of approximately 1:1 are indicated to control spasticity in patients with multiple sclerosis. In Brazil epileptic pediatric patients are treated with imported Cannabis extracts registered as food supplement and with national not regulated Cannabis extracts. With the medical Cannabis sanitary regulation in Brazil, quality control methods will be required. A High Performance Liquid Chromatography (HPLC) method was developed and validated for five cannabinoids quantification in oily extracts and resins. The method was applied to 101 Cannabis extracts consumed by patients attended by a risk assessment program. Linearity, selectivity and precision met the validation criteria of national and international sanitary agencies. Cannabis imported oily extracts showed high CBD levels (72.53±60.63 mg mL-1) and low THC levels (1.21±1.32 mg mL-1), while national Cannabis extracts showed mainly two profiles, first composed by negative or cannabinoids traces and the second composed by THC rich extracts (9.81±13.49 mg mL-1). The HPLC method is useful to pharmaceutical production and risk assessment purposes.

Resumo A partir da iminente regulação da Cannabis para uso médico, métodos para o controle de qualidade desta matéria-prima serão necessários. Além de testes para contaminantes e potência, há também interesse farmacêutico e forense na determinação do perfil de terpenos nas diferentes variedades de Cannabis como método complementar de identificação. Um método HS-SPME GC-MS simples e não destrutivo foi usado para identificar o conteúdo de terpenoides em doze amostras de Cannabis, quatro delas do tipo cânhamo (Harle-tsu), sete diferentes do tipo maconha e uma do tipo intermédio. Todas foram previamente analisadas por HPLC para medir a potência através dos teores de THC e CBD. Comparação com a biblioteca de espectros foi usada para identificar os terpenos. Trinta compostos tipo terpeno foram detectados, nove deles estavam presentes em todas as amostras em níveis significativos, os quais foram usados para definir o perfil de terpenos: humuleno, cariofileno, D-limoneno, α-pineno, β-pineno, β-mirceno, linalool, terpinoleno e α-bergamoteno. Três deles, cariofileno, α-pineno e β-mirceno resultaram em maior quantidade na maioria das amostras. De acordo com a análise de componentes principais (PCA), as quatro amostras da variedade Harle-Tsu (perfil de cânhamo com THC/CBD < 1) apresentaram dois perfis diferentes, em duas amostras o componente majoritário foi o cariofileno, enquanto nas outras duas amostras o β-mirceno foi o componente majoritário. As amostras de maconha (THC/CBD > 1) mostraram perfis mais amplos, sem padrões claros que pode ser justificado pelo reduzido tamanho amostral. O método mostrou-se simples e viável para determinação do perfil de terpenos em matéria-prima vegetal de espécies do gênero Cannabis. Por outro lado, a atribuição de perfis químicos às variedades específicas necessita de mais estudos com maior tamanho amostral.

Abstract With the upcoming medical Cannabis regulation, quality control methods on raw material will be required. Besides testing for contaminants and potency, there are also pharmaceutical and forensic interests in the determination of the terpene profile in different strains of Cannabis as complementary identification methods. A simple non-destructive HS-SPME GC-MS method was used to identify the terpene content in twelve Cannabis samples, four of them were of the hemp type (Harle-tsu), seven from various marihuana types and one of the intermediate type. They all were previously analyzed by HPLC to determine the potency (THC and CBD content). Spectral library matching was used to identify the terpenes compounds. Thirty terpenes compounds were detected, nine of them were present in all Cannabis samples and used to find their terpene profile: α-pinene, β-pinene, β-myrcene, D-limonene, terpinolene, linalool, caryophyllene, α-bergamotene and humulene. Three of them, caryophyllene, α-pinene and β-myrcene were found as larger components in most of samples. A principal components analyses (PCA) was performed. The four hemp type samples showed two different profiles, two samples showed caryophyllene as main component and the others two with β-myrcene as such. The marihuana type samples showed wider profiles with no clear patterns at all, which is not surprising because of the low number of samples. The simple methodology shows viable to set the terpenes profile for analyses of raw Cannabis material. Suitability for differentiation between different sorts of types needs more studies, with increasing numbers of samples.

RESUMO A malária é uma doença de distribuição tropical, sendo endêmica em mais de 90 países, responsável por cerca de 212 milhões de casos reportados ao redor do mundo em 2016. As estratégias de erradicação dessa doença são ineficazes até o presente, sem medidas de prevenção específica, como vacinas. Atualmente, o arsenal terapêutico existente é limitado e vem se tornando ineficaz frente à expansão de plasmódios resistentes a artemisinina, evidenciando a necessidade de estudos que viabilizem o desenvolvimento de novos compostos contra a doença. Nesse contexto, estudamos o óleo essencial obtido de rizomas de Cyperus articulatus (VOCA), uma espécie vegetal comumente encontrada na região amazônica, utilizada popularmente como alternativa terapêutica para o tratamento de malária. Visamos confirmar o potencial antimalárico da planta através de testes in vitro e in vivo. Utilizamos cepas de Plasmodium falciparum W2 (cloroquina-resistente) e 3D7 (cloroquina-sensível) cultivadas em hemácias e expostas ao VOCA em microplacas de 96 poços. A atividade antimalárica in vivo foi testada em camundongos da linhagem BALB/c infectados com aproximadamente 106 eritrócitos parasitados por Plasmodium berghei. O VOCA apresentou alto potencial antimalárico (IC50 < 10 µg ml-1) frente às duas cepas de P. falciparum testadas (IC50=1,21 µg ml-1 para W2 e 2,3 µg ml-1 para 3D7). Além disso, houve redução significativa da parasitemia induzida por P. Berghei em camundongos tratados com EOAC, e também observamos diminuição da anemia, uma sintomatologia provocada pela infecção. Nossos resultados confirmam o potencial antimalárico do óleo essencial de Cyperus articulatus.

ABSTRACT Malaria is a disease of global tropical distribution, being endemic in more than 90 countries and responsible for about 212 million cases worldwide in 2016. To date, the strategies used to eradicate this disease have been ineffective, without specific preventive measures such as vaccines. Currently, the existing therapeutic arsenal is limited and has become ineffective against the expansion of artemisinin-resistant Plasmodium, demonstrating the need for studies that would allow the development of new compounds against this disease. In this context, we studied the volatile oil obtained from rhizomes of Cyperus articulatus (VOCA), a plant species commonly found in the Amazon region and popularly used as a therapeutic alternative for the treatment of malaria, in order to confirm its potential as an antimalarial agent by in vitro and in vivo assays. We cultured Plasmodium falciparum W2 (chloroquine-resistant) and 3D7 (chloroquine-sensitive) strains in erythrocytes and exposed them to VOCA at different concentrations in 96-well microplates. In vivo antimalarial activity was tested in BALB/c mice inoculated with approximately 106 erythrocytes infected with Plasmodium berghei. VOCA showed a high antimalarial potential against the two P. falciparum strains, with IC50 = 1.21 μg mL-1 for W2 and 2.30 μg mL-1 for 3D7. VOCA also significantly reduced the parasitemia and anemia induced by P. berghei in mice. Our results confirmed the antimalarial potential of the volatile oil of Cyperus articulatus.

Derivatives of steviol, isosteviol and betulinic acid were prepared and tested in vitro against Plasmodium falciparum W2 (chloroquine-resistant) and Trypanosoma cruzi. The best results against P. falciparum were found for the betulinic acid derivatives 3-[(2,4-dinitrophenyl)hydrazono]lup-20(29)-en-28-oic acid and 3-hydroxy-20-[(2,4-dinitrophenyl)hydrazono]-29-norlupan-28-oic acid (values concentration in that the growth of the parasites is 50% inhibited, IC50 = 9.96, 6.45 and 21.06 µg mL-1, respectively). Isosteviol was inactive but its 4-nitro- and 2,4-dinitrophenylhydrazone showed moderate activity (IC50 = 17.50-22.58 µg mL-1). The derivatives of betulinic acid were more selective to the parasite when compared with hepatoblastoma HepG2 A16 cells. The results showed that nitrophenylhydrazone derivatives of lupane and beyerane terpenes are promising prototypes of antimalarial agents. Among the compounds tested against T. cruzi the oxime of isosteviol was active against both epimastigote and trypomastigote forms of the parasite.

This study describes a simple, fast and reproducible method using RP-HPLC-UV, in a gradient system, for quantification of reserpine in Rauvolfia sellowii stem bark. The analysis were carried out on a C18 column; mobile phase was water and acetonitrile, and separations were carried out in 10 min, flow rate of 1.0 mL min-1, 25 ºC and 268 nm. The validation data showed that the method was specific, accurate, precise and robust. Results were linear over a range of 0.625-40.0 μg mL-1, and the mean recovery was 95.1%. The amount of reserpine found in the dried stem bark was 0.01% (m/m).

Himatanthus lancifolius (Müll. Arg.) Woodson é um arbusto nativo do Brasil, latescente, popularmente conhecido como agoniada e utilizado principalmente para distúrbios menstruais. O presente trabalho teve como objetivo caracterizar morfoanatomicamente a folha, o caule e a casca caulinar dessa planta medicinal, a fim de contribuir para o controle de qualidade e a autenticidade dessa espécie. O material vegetal foi fixado e submetido às microtécnicas usuais. A folha é simples, glabra e obovado-lanceolada. A epiderme é uniestratificada, revestida por cutícula estriada e possui estômatos anisocíticos na face abaxial. O mesofilo é dorsiventral. A nervura central é biconvexa e o pecíolo é circular, ambos apresentando feixes vasculares bicolaterais. Laticíferos, amiloplastos e idioblastos fenólicos estão presentes no parênquima fundamental da nervura central e do pecíolo. O sistema vascular do caule é tipicamente bicolateral. Laticíferos e idioblastos fenólicos ocorrem no córtex, no floema e na medula. Esses caracteres morfoanatômicos, em conjunto, podem ser utilizados como parâmetros para o controle de qualidade dessa espécie.

Himatanthus lancifolius (Müll. Arg.) Woodson is a Brazilian native shrub, laticiferous, popularly known as "agoniada" and it is mainly used for uterine disorders. The present work aimed to study the leaf, stem and stem bark morpho-anatomy of this medicinal plant, in order to contribute to its quality control and identification. The plant material was fixed and submitted to standard microtechniques. The leaf is simple, glabrous and obovate-lanceolate. The epidermis is uniseriate, coated with striated cuticle and it has anysocitic stomata on the abaxial surface. The mesophyll is dorsiventral. The midrib is biconvex and the petiole is circular, both presenting bicollateral vascular bundles. Laticiferous ducts, amyloplasts and phenolic idioblasts are found in ground parenchyma of the midrib and petiole. The vascular system of the stem is tipically bicollateral. Laticiferous ducts and phenolic idioblasts are present in the cortex, phloem and pith. These morpho-anatomical characters, all together, can be used as quality control parameters of this species.

Rauvolfia sellowii Müll. Arg. (Apocynaceae), uma árvore nativa brasileira rica em alcaloides indólicos, é conhecida como "pau-pra-tudo" e utilizada popularmente como hipocolesterolêmica, hipoglicêmica e anti-hipertensiva. O presente trabalho teve como objetivo caracterizar anatomicamente o caule jovem e a casca caulinar dessa planta medicinal, a fim de contribuir para a identificação e autenticidade da droga. O material vegetal foi fixado e submetido às microtécnicas usuais. O caule jovem possui epiderme remanescente, porém uma periderme suberificada é observada. O felogênio instala-se na região cortical, formando súber externamente. Subjacentes ao felogênio são observadas a feloderme e uma região colenquimática. No córtex, há numerosos laticíferos e algumas fibras. Há uma bainha esclerenquimática incompleta, constituída de vários grupos de fibras e células pétreas. O caule mostra floema interno organizado em grupos isolados lado a lado. Numerosos laticíferos, cristais de oxalato de cálcio, idioblastos e amiloplastos são encontrados no córtex, no floema, no xilema e na medula. A casca do caule possui várias camadas de súber e parênquima cortical, uma bainha composta de fibras e células pétreas totalmente lignificadas, e floema externo. Estes caracteres anatômicos, quando analisados em conjunto, podem ser utilizados como parâmetros para controle de qualidade desta espécie.

Rauvolfia sellowii Müll. Arg. (Apocynaceae), a Brazilian native tree rich in indole alkaloids, is known as "pau-pra-tudo" and popularly used as hypocholesterolemic, hypoglycemic and antihypertensive. The aim of this work was to study the anatomy of the young stems and stem barks of this medicinal plant, in order to contribute to the identification of the species as a drug. The plant material was fixed and prepared according to standard microtechniques. The young stems have remaining epidermis, but a suberified peridermis is evident. The phellogen is located in the cortical region, forming suber externally. Underneath the phellogen, lies the phelloderm and collenchymatic region. In the cortex, there are numerous laticifers and some fibers. There is an incomplete sclerenchymatic sheath, consisting of several groups of fibers and stone cells. The stem has internal phloem ordered as isolated groups side by side. Numerous laticifers, calcium oxalate crystals, idioblasts and amyloplasts are found in the cortex, phloem, xylem and pith. The stem bark has many layers of suber and cortical parenchyma, a sheath composed of fibers and stone cells totally lignified, and external phloem. These anatomical characteristic, taken together, can be used as quality control parameters for this species.

Os extratos etanólicos de folhas de guaco (Mikania laevigata) cultivado tradicionalmente no solo ou por hidroponia foram avaliados quanto às atividades alelopática e antimicrobiana. Para a atividade alelopática foi utilizado o ensaio de inibição da germinação de sementes de alface (Lactuca sativa), enquanto que para a atividade antimicrobiana utilizouse a técnica de difusão em disco. Observou-se um notável efeito alelopático dos extratos de ambos os cultivos, em especial do extrato etanólico do guaco tradicional, que mesmo na menor concentração testada inibiu completamente a germinação das sementes. Nenhum dos extratos etanólicos do guaco apresentou atividade antibacteriana significativa para as linhagens de Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Enterococcus faecalis e Enterococcus faecium.

The ethanolic extract of "guaco" (Mikania laevigata) traditionally cultivated in the soil or in a hydroponic system were tested for allelopathic and antimicrobial activities. Allelopathic activity was evaluated by the inhibition of germination assay using lettuce seeds, and antimicrobial activity by the disc diffusion assay. A notable allelopathic effect was observed for both extracts, although a more expressive activity of traditional "guaco" was verified, since the inhibition of seeds germination was 100% even in the lower concentration. None ethanolic extract of "guaco" presented significant antimicrobial activity against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Enterococcus faecalis and Enterococcus faecium.