Impact of Gelatin Supplemented with Gum Arabic, Tween 20, and β-Cyclodextrin on the Microencapsulation of Turkish Oregano Extract

Microencapsulation protects core materials from deteriorating due to environmental conditions, such as moisture or oxidation, and improves the bioavailability of active compounds, allowing one to make solid formulations from oils and increase their solubility. Wall and core material properties determine the microencapsulation efficiency and the best results are achieved when a wall material mixture is used to prepare the microcapsules. In this work, we optimized the wall material composition (gelatin supplemented with gum Arabic, Tween 20, and β-cyclodextrin) of Turkish oregano microcapsules prepared by spray-drying technology to increase the product yield, the encapsulation efficiency, and to achieve narrower particle size distribution. When the wall material solution contained 10 g of gelatin, 7.5 g of gum Arabic, 1.99 g of Tween 20, 1.98 g of β-cyclodextrin, and 20 g of ethanolic oregano extract, the encapsulation efficiency of oregano’s active compounds, rosmarinic acid and carvacrol, were 96.7% and 99.8%, respectively, and the product yield was 85.63%. The physicochemical properties, microscopic morphology, and in vitro release of the prepared microcapsules were characterized in the study. The use of gelatin as the main coating material, in supplementation with gum Arabic, Tween 20, and β-cyclodextrin, not only improved the encapsulation efficiency, but also increased the in vitro release of both main active compounds of Turkish oregano extract—rosmarinic acid and carvacrol

In Vitro and Clinical Safety Assessment of the Multiple W/O/W Emulsion Based on the Active Ingredients from Rosmarinus officinalis L., Avena sativa L. and Linum usitatissimum L.

The multiple W/O/W emulsion supplemented with the extracts of Rosmarinus officinalis L., Avena sativa L. and Linum usitatissimum L. was prepared in the study, its active compounds were determined by HPLC and its safety was evaluated in vitro by the means of reconstituted human skin model EpiDerm™ for the assessment of its irritation, phototoxicity and early skin inflammation effects and by the 48 h human skin patch test for its skin irritation and allergenic potential. The microbiological challenge test of W/O/W emulsion was performed to ensure its preservation efficiency. The results showed that the W/O/W emulsion loaded with self-preserving plant-based bio-actives had no irritant potential, was not phototoxic and did not provoke skin inflammation or sensitization and thus could be used as a safe base for cosmetic products. Furthermore, our results demonstrate that the safety evaluation of cosmetic ingredients of natural or organic origin could be easily performed using reconstructed human skin model EpiDerm™ similar to the well-defined chemicals used in the cosmetics industry

Development and Evaluation of Ginkgo biloba L. Extract Loaded into Carboxymethyl Cellulose Sublingual Films

Oral bioavailability of flavonoids, including G. biloba extract, is limited due to their chemical complexity, which determines slow dissolution in vitro behavior of the extract. The overall research objective was to compare the effect of increasing freeze-dried G. biloba extract (GFD) concentrations in carboxymethyl cellulose (CMC) films on their mechanical properties, release profile of flavonoid glycosides, stability and disintegration time. Physicochemical evaluation of films was performed by SEM and FTIR. The mechanical properties and in vitro release profile of flavonoid glycosides from the prepared films were characterized in the study. The higher elongation at break and tensile strength values, quick release of flavonoids and good stability were observed in formulation, coded FRG—15 (the film contained 0.4 g of GFD, 0.3 g of glycerol and 2 g of 2% CMC), (p < 0.05). Dissolution rate tests showed that approximately 85% of loaded flavonoid glycosides had been released; the release profile of flavonoid glycosides from FRG-15 had levelled off after only 15 min. The results could lay the groundwork for further studies, concerning the development of sublingual films as G. biloba extract-based dosage forms, which might increase the multifunctional properties and pharmacological activity closer to the desired level

Effect of the Amount of Polysorbate 80 and Oregano Essential Oil on the Emulsion Stability and Characterization Properties of Sodium Alginate Microcapsules

Essential oils have a high volatility that leads to evaporation and loss of their pharmacological effect when exposed to the environment. The objectives of the present work were to prepare microcapsules with oregano essential oil by extrusion using sodium alginate as a shell material and non-ionic surfactant polysorbate 80 as an emulsifier to stabilize the emulsion. The present study was aimed to evaluate the physical parameters of microcapsules and to compare the influence of the amount of emulsifier and the essential oil-to-emulsifier ratio on the capsules’ physical parameters and encapsulation efficiency; to our knowledge, the existing research had not yet revealed whether unstable emulsion affects the encapsulation efficiency of oregano essential oil. This study showed that increasing the emulsifier amount in the formulation significantly influenced encapsulation efficiency and particle size. Moreover, increasing the emulsion stability positively influenced the encapsulation efficiency. The emulsion creaming index depended on the emulsifier amount in the formulation: the highest creaming index (%) was obtained with the highest amount of polysorbate 80. However, the essential oil-to-polysorbate 80 ratio and essential oil amount did not affect the hardness of the microcapsules (p > 0.05). In conclusion, the obtained results could be promising information for production of microcapsules. Despite the fact that microencapsulation of essential oils is a promising and extremely attractive application area for the pharmaceutical industry, further basic research needs to be carried out

The Effects of Cannabis sativa L. Extract on Oxidative Stress Markers In Vivo

In recent decades, a lot of attention has been paid to Cannabis sativa L. due to its useful applications, including in fibers, oil, food for humans and animals, and therapeutics. The present study aimed to determine antioxidant activity of cannabinoids in Cannabis sativa L. in vivo, evaluating the possible antioxidative effect of Cannabis sativa L. extract (CE) on malondialdehyde (MDA) and glutathione (GSH) concentrations as well as on catalase (CAT) activity in BALB/c mice. In total, 40 mice were divided into five equal groups: the aluminum group (7.5 mg AlCl3/kg/d (0.15 LD50), the saline group, the 10% ethanol group (an appropriate amount of the solution for mouse weight), the CE group (1.6 mg CE/g/day), and the aluminum-CE group (7.5 mg AlCl3 plus 1.6 mg CE/g/day). The results of the study showed that CE significantly decreased (by 26.81%, p < 0.05) the concentration of GSH in blood of the mice and the concentration of MDA in the brain (by 82.12%) and liver (by 53.5%) of the mice compared to the respective concentrations in the AlCl3 group. CE significantly (p < 0.05) increased CAT activity in the brain (by 64.79%) and liver (by 72.37%) of the mice after the AlCl3-induced prooxidant effect. The results showed the antioxidant activity of cannabidiolic acid (CBDA) in vitro. The findings in vivo indicate that Cannabis sativa L. is a good source of natural antioxidants and can be used in the management of oxidative stress

Optimization and Validation of the GC/FID Method for the Quantification of Fatty Acids in Bee Products

To the best of our knowledge, so far, no study has been conducted about the comparison of the total fatty acid concentration in the four bee products (honey, bee pollen, bee bread, and propolis) collected from Lithuania. Therefore, we aimed to optimize the derivatization parameters and to investigate a simple and sensitive gas chromatography-flame ionization detection (GC-FID) method to determine fatty acids. The optimal derivatization parameters were used to analyze fatty acids in the bee products. Regarding sample preparation, three derivatization parameters were compared (temperature and extraction time with BF3/MeOH reagent) in order to obtain a high amount of the total fatty acids of interest from the fatty acid methyl ester (FAME) standard. The results showed that the highest total yield of fatty acids was conducted by using the conventional heating process at 70 °C for 90 min. Under optimal conditions, there was obtained excellent linearity for fatty acids with determination coefficients of r2 > 0.9998. The LODs and LOQs ranged from 0.21 to 0.54 µg/mL and 0.63 to 1.63 µg/mL, respectively. This method has been successfully applied to the qualitative analysis of fatty acids in bee products. The above findings might provide a scientific basis for evaluating the nutritional values of bee products

The Influence of Different Oregano Species on the Antioxidant Activity Determined Using HPLC Postcolumn DPPH Method and Anticancer Activity of Carvacrol and Rosmarinic Acid

The aim of this study was to evaluate concentration-dependent antioxidant and anticancer activities of CA and RA in ethanol extracts of three different Oregano species (Origanum onites L., Origanum vulgare L., and Origanum vulgare ssp. hirtum). The study revealed the highest RA antioxidant activity in O. vulgare ssp. hirtum (9550±95 mmol/g) and the lowest in O. vulgare L. (2605±52 mmol/g) (p0.05) but they had different activities against MDA-MB231 cell line

Liposomal Incorporation to Improve Dissolution and Stability of Rosmarinic Acid and Carvacrol Extracted from Oregano (O. onites L.)

The potential antimicrobial benefit of high levels of rosmarinic acid (RA) and carvacrol (CA) in oregano (O. onites L.) extract has been limited until now by poor bioavailability arising from the low aqueous-phase solubility and slow dissolution behaviour of the lyophilized extract (E). To address this issue, various ratios of phospholipon 90H (P90H) and 1,2-dimyristoyl-sn-glycero-3-phospho-(1'-rac-glycerol), sodium salt (DMPG) were sonicated, yielding four empty liposomes (L1, L2, L3, and L90). After an initial selection process, Turkish oregano extract was internalized into the more promising candidates. Each empty liposome, extract-loaded liposome (LE1, LE2, and LE3), and freeze-dried control (E) was assessed in terms of structure, composition, RA and CA dissolution profile, storage stability, and, when relevant, zeta potential. Empty liposome L1, which was prepared using P90H and DMPG in a 1:1 ratio, displayed the most convenient encapsulation traits among the four unloaded types. Loaded liposome LE1, obtained by combining oregano extract and L1 in a 1:1 ratio, proved superior as a vehicle to deliver RA & CA when compared against control freeze-dried E and test liposomes LE2 and LE3. Dissolution profiles of the active compounds RA and CA in loaded liposomes were determined using a semi-automated dissolution tester. The basket method was applied using artificial gastric juice without pepsin (AGJ, 50rpm, 500mL). The pH value was maintained at 1.5 (37 ± 0.5°C). Aliquots (5ml) were manually extracted from parallel dissolution vessels at 1, 3, 5, 7, 10, 15, 20, 25, 30, 45, and 60-minute time points. Dissolution tests, run to completion on LE1, showed that approximately 99% of loaded CA and 88% of RA had been released. Shorter dissolution times were also noted in using LE1. In particular, the release profile of CA and RA had levelled off after only 25 minutes, respectively, depicting an impressive 3.0–3.3 and 2.3-2.6 rate increase compared to the freeze-dried control extract. The improved dispersibility of RA and CA in the form of LE1 was supported by particle size and zeta potential measurements of the liposome, yielding 234.3nm and −30.9mV, respectively. The polydispersity index value was 0.35, indicating a reasonable particle size distribution. To study storage stability, liposomes were stored (4°C, 6 months) in amber coloured glass containers (4 oz.). Each container held 30 capsules, which were stored according to the ICH guidelines prescribed for long-term storage (25°C±2°C; 60%±5% RH). Triplicate samples were withdrawn after 0, 3, 6, 9, and 12 months for analysis. Lastly, LE1 displayed good storage stability. The results imply that RA and CA can be conveniently and routinely delivered via oral and mucosal routes by first internalizing oregano extracts into appropriately engineered liposomes