Is There Enough Knowledge to Standardize a Cannabis sativa L. Medicinal Oil Preparation with a High Content of Cannabinoids?

Introduction: Cannabis sativa L. medicinal oils are good therapeutic options due to their wide spectrum of pharmacological applications and the easy adjustment of individual doses. The lack of standardization of methodology in the preparation of medicinal oil using the Cannabis crude extract results in elevated variability of cannabinoid concentration in the final product. The elevated variability impairs the understanding of beneficial and adverse effects related to dose-response pharmacological activities. Objective: This study aimed to conduct a review on the current methods of Cannabis oil preparation present in the literature, to demonstrate the most appropriate methodologies to ensure a product with high content of cannabinoids and terpenes. Results: The decarboxylation stage is essential for the conversion of acid cannabinoids into neutral cannabinoids, which are substances with the highest bioavailability. Lower temperatures for longer periods of time instead of high temperatures in less time are highly recommended to ensure that all the acidic cannabinoids have passed through decarboxylation. For the guarantee of a high terpene content, the separate addition of essential oil to the fixed oil prepared from the crude extract should be considered. Ultrasound-assisted extraction is one of the best performing methodologies because it is cheaper than other techniques, such as supercritical fluid extraction, besides that, ultrasound extraction is effective in short extraction times and uses small amounts of solvent when compared with other techniques. Conclusion: Although the literature about the methods of preparation of Cannabis medicinal oil is scarce, it is possible to standardize an optimized, low-cost, and effective Cannabis extractive methodology from the results found in the literature; however, this will depend on new research for methodological validation.

Apoptotic effect of ß-pinene on oral squamous cell carcinoma as one of the major compounds from essential oil of medicinal plant Piper rivinoides Kunth.

Oral squamous cell carcinoma (OSCC) is the most common type of head and neck malignancy. Research on essential oils (EOs) has shown important cytotoxic and anti-tumor properties, among others. Piperaceae species are rich in EOs and here we highlight Piper rivinoides Kunth. We investigated the crude EOs from P. rivinoides, their pure major constituents and an enriched fraction with the main EO compounds (EF) as cytotoxic and selective OSCC agents. EOs presented as main compounds (-)-α-pinene, (-)-ß-pinene and limonene. EOs showed an IC50 lower than all isolated compounds, except for (-)-ß-pinene in OSCC cells. The (-)-ß-pinene induced cell death with apoptotic characteristics. Commercial standards showed greater selectivity than EOs, and (-)-ß-pinene was the most selective among them. EF showed higher selectivity compared to crude EOs and carboplatin, turning it into a good candidate as an anticancer fraction. These results are important for the possible development of new treatments for OSCC.

Advanced Chemophenetic Analysis of Essential Oil from Leaves of Piper gaudichaudianum Kunth (Piperaceae) Using a New Reduction-Oxidation Index to Explore Seasonal and Circadian Rhythms.

The aromatic species Piper gaudichaudianum Kunth (Piperaceae) is widely used in Brazil for medicinal and ritualistic applications. In the current study, chemophenetic patterns were realized across season and circadian rhythm based on the chemical profile of essential oils (EOs) from leaves. Hydrodistilled essential oils were analyzed by GC-MS and GC-FID, and a new calculation of metabolite oxidation level, averaged for each individual molecule component of the EO, was used to explore the patterns of metabolism/biosynthesis. This new index used an intermediate calculation, the 'weighted average redox standard' (SRO), to enable a value for mixtures of metabolites to be generated, the 'general mixture redox index' (GMOR). The indices were subjected to a proof-of-concept approach by making comparison to outcomes from multivariate analyses, i.e., PCA and HCA. Chemical analysis demonstrated that the essential oils were dominated by sesquiterpenes, constructed of 15 classes of compound (C-skeletons), and 4 C-skeletons were recognized in the monoterpene group, giving a total of 19. The variation of chemical profiles was distinct at different phenological stages, but stronger chemical variation was evident between day and night as compared to season. Furthermore, due to comprehensive sampling across different regions, nine chemotypes were recognized, including those previously reported. The SRO and GMRO indices demonstrate that phenological variation of chemistry is mainly an outcome of redox fluctuations in terpene biosynthesis, changing from day to night. These indices also corroborate that chemical diversity is increased with oxidative metabolism. Lastly, the current study demonstrates pronounced phenotypic plasticity in P. gaudichaudianum, which makes it a suitable candidate to help further our understanding of chemophenetics and chemical ecology.

Tratamento da fístula anal com extrato de Baccharis dracunculifolia. Estudo experimental em ratos / Treatment of anal fistula with Baccharis dracunculifolia extract. Experimental study in rats

RESUMO Objetivo: avaliar a eficácia do extrato de Baccharis dracunculifolia no tratamento de fístulas anais em ratos. Métodos: vinte ratos Wistar machos foram submetidos à confecção de fístula anal e, após 30 dias, foram distribuídos em três grupos: Grupo Controle, com cinco animais; Grupo Carbopol, com cinco animais; e Grupo Baccharis dracunculifolia, com dez animais. No Grupo Controle, não se realizou nenhum tratamento. No Grupo Carbopol, realizou-se infusão diária de carbopol, e no Grupo Baccharis dracunculifolia, infusão de extrato de Baccharis dracunculifolia com carbopol, ambos por 30 dias. Foram retirados espécimes para análise histológica após a eutanásia. Resultados: não houve fechamento completo do trajeto fistuloso em nenhum dos animais. A média da área do trajeto resultante foi de 847,2µm2, 565,6µm2 e 372,7µm2, respectivamente, nos Grupos Controle, Carbopol e Baccharis dracunculifolia (p=0,001). A média do escore de processo inflamatório foi de 2,4, 2,4 e 2,1, respectivamente, nos Grupos Controle, Carbopol e Baccharis dracunculifolia (p=0,285), enquanto a média de congestão vascular foi de 1,6, 1,4 e 1,1, respectivamente, nos Grupos Controle, Carbopol e Baccharis dracunculifolia (p=0,031). Conclusão: o extrato de Baccharis dracunculifolia foi capaz de reduzir o lúmen dos trajetos fistulosos e a congestão vascular, sem reduzir, no entanto, o processo inflamatório local ou fechar totalmente os trajetos fistulosos.

ABSTRACT Objective: to evaluate the efficacy of Baccharis dracunculifolia extract in the treatment of anal fistulas in rats. Methods: twenty male Wistar rats were submitted to anal fistula and, after 30 days, were divided into three groups: Control Group, with five animals; Carbopol Group, with five animals; and Baccharis dracunculifolia Group, with ten animals. In the Control Group, no treatment was performed. In the Carbopol Group, a daily infusion of Carbopol was performed for 30 days. In the Baccharis dracunculifolia Group, a daily infusion of Carbopol plus Baccharis dracunculifolia extract was performed for 30 days. Specimens were taken for histological analysis after euthanasia. Results: there was no complete closure of the fistulous tract in any of the animals. The mean area of the remaining tract was of 847.2µm2, 565.6µm2 and 372.7µm2, in the Control Group, Carbopol Group, and Baccharis dracunculifolia Group, respectively, (p=0.001). The mean of the inflammatory process score was of 2.4, 2.4, and 2.1, in the Control Group, Carbopol Group, and Baccharis dracunculifolia Group, respectively, (p=0.285), while the mean values of vascular congestion were of 1.6, 1.4, and 1.1, in the Control Group, Carbopol Group, and Baccharis dracunculifolia Group, respectively, (p=0.031). Conclusion: Baccharis dracunculifolia extract was able to reduce the lumen of the fistulous tracts and the degree of vascular congestion, without, however, reducing the local inflammatory process or totally closing the fistulous tracts.

A newly validated high-performance liquid chromatography method with diode array ultraviolet detection for analysis of the antimalarial drug primaquine in the blood plasma

Abstract INTRODUCTION: Primaquine (PQ) diphosphate is an 8-aminoquinoline antimalarial drug with unique therapeutic properties. It is the only drug that prevents relapses of Plasmodium vivax or Plasmodium ovale infections. In this study, a fast, sensitive, cost-effective, and robust method for the extraction and high-performance liquid chromatography with diode array ultraviolet detection (HPLC-DAD-UV ) analysis of PQ in the blood plasma was developed and validated. METHODS: After plasma protein precipitation, PQ was obtained by liquid-liquid extraction and analyzed by HPLC-DAD-UV with a modified-silica cyanopropyl column (250mm × 4.6mm i.d. × 5μm) as the stationary phase and a mixture of acetonitrile and 10mM ammonium acetate buffer (pH = 3.80) (45:55) as the mobile phase. The flow rate was 1.0mL·min-1, the oven temperature was 50OC, and absorbance was measured at 264nm. The method was validated for linearity, intra-day and inter-day precision, accuracy, recovery, and robustness. The detection (LOD) and quantification (LOQ) limits were 1.0 and 3.5ng·mL-1, respectively. The method was used to analyze the plasma of female DBA-2 mice treated with 20mg.kg-1 (oral) PQ diphosphate. RESULTS: By combining a simple, low-cost extraction procedure with a sensitive, precise, accurate, and robust method, it was possible to analyze PQ in small volumes of plasma. The new method presents lower LOD and LOQ limits and requires a shorter analysis time and smaller plasma volumes than those of previously reported HPLC methods with DAD-UV detection. CONCLUSIONS: The new validated method is suitable for kinetic studies of PQ in small rodents, including mouse models for the study of malaria.

Anatomical aspects, chemical analysis and cytotoxic effect of the essential oil from leaves of Casearia arborea (Salicaceae) / Aspectos anatómicos, análisis químico y efecto citotóxico del aceite esencial de Casearia arborea (Salicaceae)

The genus Casearia (Salicaceae) is found in sub-tropical and tropical regions of the world and comprises about 200 species. In Brazil, there are about 48 species and 12 are registered in the State of Rio de Janeiro; including Casearia arborea (Rich.) Urb. Essential oil was obtained from the fresh leaves by hydrodistillation and analyzed by GC-MS and GC-FID. The cytotoxic effect was determined by WST-1 assay. Chemical analysis of the essential oil revealed a very diversified (n = 37 compounds) volatile fraction composed mainly of non-oxygenated sesquiterpenes (90.2%). These sesquiterpenes included byciclogermacrene (18.7%), germacrene D (12.1%) and α-humulene (11.5%). In addition, the essential oil demonstrated cytotoxic effects against A549 tumor cells in the concentration of 4 µg/mL (EC50) (p < 0.05).

El género Casearia (Salicáceas) se encuentra en las regiones tropicales y sub-tropicales del planeta y comprende alrededor de 200 especies. En Brasil existen 48 especies, 12 de las cuales fueron registradas en el Estado de Río de Janeiro incluyendo Casearia arborea (Rich.) Urb. El aceite esencial fue extraído de hojas frescas por hidrodestilación y analizado por GC-MS y GC-FID. El efecto citotóxico fue determinado por ensayo WST-1. Las cavidades secretorias fueron ocasionalmente encontradas tanto en la lámina foliar como en el pecíolo. El análisis químico del aceite esencial reveló una muy diversa fracción volátil (n = 37 compuestos) formada principalmente por sesquiterpenos no oxigenados (90,2%). Estos sesquiterpenos incluyen biciclogermacreno (18,7%), germacreno D (12,1%) y α-humuleno (11,5%). Además, el aceite esencial demostró efectos citotóxicos contra las células tumorales A549 en una concentración de 4µg/mL (EC50) (p < 0.05).

An Overview of Neolignans of the Genus Piper L.: Isolation Methods and Biological Activities.

BACKGROUND: The genus Piper L. has the shikimic acid pathway predominantly expressed, biosynthesizing many cinnamic acid derivatives (CAD). OBJECTIVE: Neolignans comprise an important class of CAD that exhibit a wide range of pharmacological properties such as antibacterial, antitumor, insecticidal, anti-inflammatory, antioxidant, smooth muscle relaxant, neuroprotective, antiprotozoal and against platelet aggregation factor. These substances have been extracted and isolated from Piper species using different technics. RESULTS: The present review aims to summarize extraction and isolation methods and biological activities of the different types of neolignans covering the period from 1968 to January 2016.

Chemical composition and efficacy in the egg-hatching inhibition of essential oil of Piper aduncum against Haemonchus contortus from sheep

Piper aduncum L., Piperaceae, has been used to treat mainly inflammatory diseases and has shown several biological activities such as insecticidal and larvicidal. The aim of this study was to analyze the chemical composition of essential oil of P. aduncum and its efficacy to egg-hatching inhibition of Haemonchus contortus from sheep. The essential oil was obtained from leaves and analysed by gas chromatography coupled to flame ionization detector and gas chromatography coupled to mass spectrometry. It was possible to characterize 22 different substances, among them monoterpenes (80.6%) and sesquiterpenes (13.9%). The major compound was identified as 1,8-cineole (55.8%). Eggs of the nematode were exposed to four concentrations of the essential oil. Levamisole phosphate was used as positive control. The essential oil showed to be effective in inhibiting H. contortus hatchability and the LC90 was calculated as 8.9 mg.ml-1. These results can point out the P. aduncum essential oil and its chemical components as potential alternative to control of H. contortus .

Substâncias fenólicas com atividade antioxidante de Pseudopiptadenia contorta (Leguminosae-Mimosoideae)

O extrato bruto em acetona/ água das folhas de Pseudopiptadenia contorta foi purificado rendendo frações e substâncias puras. A atividade antioxidante foi avaliada para o extrato e frações, empregando-se o teste químico do DPPH. As frações contendo taninos condensados e substâncias fenólicas de baixo peso molecular mostraram ser as responsáveis pela atividade antioxidante do extrato bruto das folhas de P. contorta.