A Comparative Evaluation of Antioxidant Activity of Extract and Essential Oil of Origanum onites L. In Vivo.

In the present study, the effects of Origanum onites L. extract and essential oil of O. onites L. on the antioxidant status of the liver and brain of mice were investigated. Due to certain disadvantages of essential oils, such as poor solubility, high volatility and sensitivity to UV light and heat, formulation of liposomes with Oregano essentials (OE) was optimized and used in this study. The results demonstrated that the best composition of the lipid carriers and OE were conducted in terms of the polydispersity index (PDI), mean particle size and encapsulation efficiency (EE). For further study the LE4 formulation was used, which contained Lipoid S100 at 45 mg, Lipoid S75 at 45 mg and 90 mg of EO. The administration of O. onites L. extract to mice for 21 days significantly decreased the glutathione (GSH) level in the livers and brains of the mice as well as the malondialdehyde (MDA) concentration in the livers. In the brains of the mice, MDA level significantly increased after this extract consumption. Whereas liposomes with OE significantly decreased GSH concentration in the mouse brain and MDA concentration in the mouse liver, there was an increased (p > 0.05) GSH level in the liver and MDA concentration in the brain of mice compared with the control group. It was found that both O. onites. ethanolic extract as well as liposomes with OE of this plant material affect the antioxidant status in the livers and brains of mice.

Investigation of Organic Acids in Saffron Stigmas (Crocus sativus L.) Extract by Derivatization Method and Determination by GC/MS.

The beneficial health properties of organic acids make them target compounds in multiple studies. This is the reason why developing a simple and sensitive determination and investigation method of organic acids is a priority. In this study, an effective method has been established for the determination of organic (lactic, glycolic, and malic) acids in saffron stigmas. N-(tert-butyldimethylsilyl)-N-methyltrifluoroacetamide (MTBSTFA) was used as a derivatization reagent in gas chromatography combined with mass spectrometric detection (GC/MS). The saffron stigmas extract was evaporated to dryness with a stream of nitrogen gas. The derivatization procedure: 0.1 g of dried extract was diluted into 0.1 mL of tetrahydrofuran, then 0.1 mL MTBSTFA was orderly and successively added into a vial. Two different techniques were used to obtain the highest amount of organic acid derivatives from saffron stigmas. To the best of our knowledge, this is the first report of the quantitative and qualitative GC/MS detection of organic acids in saffron stigmas using MTBSTFA reagent, also comparing different derivatization conditions, such as time, temperature and the effect of reagent amount on derivatization process. The identification of these derivatives was performed via GC-electron impact ionization mass spectrometry in positive-ion detection mode. Under optimal conditions, excellent linearity for all organic acids was obtained with determination coefficients of R2 > 0.9955. The detection limits (LODs) and quantitation limits (LOQs) ranged from 0.317 to 0.410 µg/mL and 0.085 to 1.53 µg/mL, respectively. The results showed that the highest yield of organic acids was conducted by using 0.1 mL of MTBSTFA and derivatization method with a conventional heating process at 130 °C for 90 min. This method has been successfully applied to the quantitative analysis of organic acids in saffron stigmas.

Development of extraction technique and GC/FID method for the analysis of cannabinoids in Cannabis sativa L. spp. santicha (hemp).

INTRODUCTION: Nowadays, the interest in industrial Cannabis sativa L. herb has been increasing in the world. As a result, it is becoming one of the most studied plants due to its multifunctional benefits. OBJECTIVES: To the best of our knowledge, no study has been conducted so far to determine the impact of extraction methods and conditions on the extraction yields of CBD and CBG from the Cannabis sativa L. ssp. Santhica. Therefore, we aimed to investigate a simple and sensitive GC-FID method to determine CBD and CBG in hemp extract. METHODS: As regards sample preparation, three extraction techniques were compared, including maceration (ME), ultrasound-assisted extraction (UAE) and reflux-heat extraction (RHE), in order to obtain a high recovery of the CBD of interest from the plant material. The GC-FID method developed in this study represents a powerful tool for the extraction and analysis of non-psychoactive cannabinoids from hemp varieties to be used for the preparations of extracts with a high content of bioactive compounds for both pharmaceutical and nutraceutical applications. RESULTS: A simple extraction procedure for CBD and CBG from hemp was also optimized in this work, by using ultrasound assisted extraction method with 96% ethanol, material/solvent ratio 1:10 and extraction time 10 min at room temperature. CONCLUSION: The overall analytical method was fully validated in agreement with international guidelines. Therefore, proving a powerful and reliable tool for both the selection of hemp varieties with a high content of bioactive compounds and the quality control of its derivatives.

Formation and Investigation of Electrospun Eudragit E100/Oregano Mats.

An electrospun mat of Eudragit E100 (EE100) (a cationic copolymer based on dimethylaminoethyl methacrylate, butyl methacrylate, and methyl methacrylate) was used as a delivery system for oregano ethanolic extract (OEE). Oregano is a biologically active material which is widely used because of the antibacterial and antifungal activity. The oregano herb consists of phenolic compounds, the main of which are rosmarinic acid and from essential oil-carvacrol. Such a material could be an ideal candidate for oral drug systems. The influence of the EE100 concentration in the OEE on the structure of electrospun mats, encapsulation efficiency, dissolution profile, release kinetics and the stability of biologically active compounds was investigated. The concentration of the solution is a critical parameter for the structure and properties of electrospun mats. The diameter of electrospun fibers increased with the increase of EE100 concentration in the OEE. Electrospun mats obtained from 24% to 32% EE100 solutions showed high encapsulation efficiency, quick release and high stability of rosmarinic acid and carvacrol. Dissolution tests showed that 99% of carvacrol and 80% of rosmarinic acid were released after 10 min from electrospun nano-microfiber mats and capsules obtained from such formulations. The stability tests showed that physicochemical properties, dissolution profiles, and rosmarinic acid and carvacrol contents of the formulations were not significantly affected by storage.

Impact of Gelatin Supplemented with Gum Arabic, Tween 20, and ß-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 ß-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 ß-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 ß-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.

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.

Optimization of carvacrol, rosmarinic, oleanolic and ursolic acid extraction from oregano herbs (Origanum onites L., Origanum vulgare spp. hirtum and Origanum vulgare L.).

The aim of our study was to increase the extraction efficiency of carvacrol, rosmarinic, oleanolic and ursolic acid from the different species of oregano herbs (Origanum onites L., Origanum vulgare spp. hirtum and Origanum vulgare L.). Various extraction methods (ultrasound-assisted, heat-reflux, continuous stirring, maceration, percolation) and extraction conditions (different solvent, material:solvent ratio, extraction temperature, extraction time) were used, and the active substances were determined by HPLC. The lowest content of carvacrol, rosmarinic, oleanolic and ursolic acid was obtained by percolation. During heat-reflux extraction, the content of active substances depended on the solvent used: ethanol/non-aqueous solvent (glycerol or propylene glycol) mixture was more effective compared with ethanol alone. The results showed that for each species of oregano the most optimal extraction method should be selected to maximize the content of biologically active substances in the extracts.