Reliable Methods for Analyses of Volatile Compounds of Copaifera Oleoresins Combining Headspace and Gas Chromatography.

Two analytical methods were developed in this study for direct and fast chemical investigation of authentic Copaifera oleoresins (COR) and commercial products. Polydimethylsiloxane microfiber coupled to gas chromatography-mass spectrometry (HS-SPME-GC/MS) showed the best results for oleoresin qualitative analysis, setting the following extraction conditions: equilibrium time of 15 min, extraction time of 30 min, extraction temperature at 60 °C and constant stirring of 400 rpm. Sesquiterpenes α-copaene, ß-elemene, ß-caryophyllene and trans-α-bergamotene were found in all investigated samples. Quantitative analysis by gas chromatography coupled with flame ionization detector (GC-FID) measured the content of the four sesquiterpenes in all samples. Qualitative and quantitative results showed important differences between COR of distinct species and commercial products. Data regarding the volatile composition of C. oblongifolia and C. trapezifolia oleoresins were first presented in this study and two new analytical methods were reported for direct and fast qualitative and quantitative analysis of COR.

Leishmanial activity of Brazilian brown propolis and its diterpenes.

Propolis is a natural product widely used in folk medicine. Among its various applications, its antiparasitic properties stand out. Due to its great biodiversity, Brazil is a major producer of several types of propolis. This study proposes to evaluate the leishmanicidal properties of the hydroalcoholic extract of propolis collected in the southern region of Brazil (Brown propolis - HEBP) and its main isolated compounds: abietic acid (1), 13-epi-cupressic acid (2), 13-epi-torulosol (3), dehydroabietic acid (4), cis-communic acid (5) and ent-agatic acid (6). In general, the diterpenes did not show activity against the promastigotes of Leishmania (Leishmania) amazonensis at the evaluated concentrations. However, the HEBP was very active with an inhibition concentration of 50% at 8.32 µg/mL. Moreover, transmission electron microscopy (TEM) and scanning electron microscopy (SEM) assays showed morphological and structural alterations in promastigote forms of L. (L.) amazonensis when incubated with HEBP.

An insight into the botanical origins of propolis from permanent preservation and reforestation areas of southern Brazil.

Brown propolis from permanent preservation and reforestation areas of southern Brazil have attracted international commercial interest and have a unique composition, although little is known about their botanical origins, which are the plant resins used by bee foragers to produce propolis. Hence, the volatile profiles of organic and non-organic brown propolis and resins of suspected botanical origins-Araucaria angustifolia, Pinus elliott and Pinus taeda-were determined using static headspace gas chromatography coupled to mass spectrometry (SHS-GCMS) and compared. Nighty nine volatiles were tentatively identified, and monoterpenes and sesquiterpenes were the most abundant classes. Principal component analysis (PCA) showed similarity between organic propolis and A. angustifolia volatile profiles (p < 0.05). Hierarchical clustering analysis showed singularities among propolis, even between propolis produced 1 km away from each other. Heatmaps were used to identify peaks present in similar relative intensities in both propolis and conifer resins. Hence, the approach using volatile profiles shed light to propolis botanical origins, which is important for authentication and traceability purposes.

Effect of light, oxygen and temperature on the stability of artepillin C and p-coumaric acid from Brazilian green propolis.

Brazilian Green Propolis (BGP) is an important bee product, which displays important biological activities, making it valuable in the international market. The major prenylated phenolic compound in BPG is (E)-artepillin C, along with its precursor (E)-p-coumaric acid, both contributing to the biological effects of BGP. Taking that into account, it was evaluated the effect of light, temperature and air oxygen in their content to establish the best storage and transport conditions for crude BGP and the pure compounds. For that, (E)-artepillin C and (E)-p-coumaric acid were initially submitted to degradation for five days under sunlight and high temperature (50 °C), furnishing three major (E)-Artepillin C isomers and one from (E)-p-coumaric acid. Then, it was developed and validated a Reverse Phase High Performance Liquid Chromatography (RP-HPLC) method for quantifying these compounds in crude BGP and in its extracts. In the stability studies, it was used a Full Factorial and Central Composite Design to establish the desirable storage conditions. (E)-Artepillin C, both pure and in BGP should be kept protected from light and storage below -2.5 °C. (E)-p-Coumaric acid can be stored at room temperature. Therefore, the best storage and transport conditions to keep the content of both compounds in BGP are protection from light at low temperatures.

Development of a Validated High-Performance Liquid Chromatography Method and Optimization of the Extraction of Lignans from Piper cubeba.

Piper cubeba L. f. is a food seasoning, which contains secondary metabolites displaying several biological properties, such as cytotoxic, anti-inflammatory, and antiparasitic activities. The lignans (+)-dihydroclusin, (-)-clusin, (-)-cubebin, (-)-yatein, and (-)-haplomyrfolin were isolated, with (-)-haplomyrfolin reported for the first time in P. cubeba seeds. Chromatographic standards were used to develop a reliable reverse-phase high-performance liquid chromatography analytical method according to the Agência Nacional de Vigilância Sanitária and International Conference on Harmonization guidelines to quantitate these lignans in both P. cubeba seeds and their extracts. The extraction of the lignans was also optimized, with the best conditions being ultrasound-assisted extraction, with 84% aqueous ethanol for 38 min in a single extraction. This procedure allows for the extraction of more than 80% of the total lignans, which is better in comparison to other techniques, such as maceration and Soxhlet extraction.