Development of Water-in-Oil Emulsions as Delivery Vehicles and Testing with a Natural Antimicrobial Extract.

Water-in-oil (W/O) emulsions have high potential for several industrial areas as delivery systems of hydrophilic compounds. In general, they are less studied than oil-in-water (O/W) systems, namely in what concerns the so-called fluid systems, partly due to problems of instability. In this context, this work aimed to produce stable W/O emulsions from a natural oil, sweet almond oil, to be further tested as vehicles of natural hydrophilic extracts, here exemplified with an aqueous cinnamon extract. Firstly, a base W/O emulsion using a high-water content (40/60, v/v) was developed by testing different mixtures of emulsifiers, namely Tween 80 combined with Span 80 or Span 85 at different contents. Among the tested systems, the one using a 54/46 (v/v) Span 80/Tween 80 mixture, and subjected to 12 high-pressure homogenizer (HPH) cycles, revealed to be stable up to 6 months, being chosen for the subsequent functionalization tests with cinnamon extract (1.25-5%; w/v; water-basis). The presence of cinnamon extract leaded to changes in the microstructure as well as in the stability. The antimicrobial and antioxidant analysis were evidenced, and a sustained behavior compatible with an extract distribution within the two phases, oil and water, in particular for the higher extract concentration, was observed.

Mushroom ethanolic extracts as cosmeceuticals ingredients: Safety and ex vivo skin permeation studies.

Mushrooms are important sources of natural bioactive compounds that are increasingly used as cosmeceutical ingredients. In this context, ethanolic extracts were prepared from Ganoderma lucidum and Pleurotus ostreatus and tested by incorporation into base cosmetic creams. In vitro safety evaluation of the extracts and cosmetic formulations prepared therefore was carried out using the MTT and LDH assays in keratinocyte (HaCaT) and fibroblast (HFF-1) cell lines. Moreover, they were submitted to ex vivo skin permeation studies using a Franz diffusion apparatus with pig ear skin as permeation membrane. The results showed the absence of toxicity for keratinocytes and fibroblasts in a concentration dependent manner, which is indicative of the safety of these extracts for cosmeceutical ingredients purposes. Protocatechuic and syringic acids were the only compounds permeating from G. lucidum extract in the first 8 h of study, no penetration was observed for phenolic acids found in P. ostreatus extract and formulation. These results pointed out for the suitability of using mushroom extracts as skin care ingredients and may contribute for the valorisation of wastes generated by the mushroom processing industry, which can serve as raw-materials for the obtainment of the extracts.

A comparative study between conventional and non-conventional extraction techniques for the recovery of ergosterol from Agaricus blazei Murrill.

Response surface methodology was applied to optimize the recovery of ergosterol from an agroindustrial residue of Agaricus blazei Murrill, by comparing conventional (heat-assisted extraction, HAE) and more sustainable non-conventional approaches (ultrasound and microwave-assisted extractions, UAE and MAE, respectively). A circumscribed central composite design was employed to optimize and describe the interactive effects among variables of the HAE, MAE (time and temperature) and UAE systems (time and ultrasound power), and their performances compared with the standard soxhlet extraction system. The optimum conditions were predicted as 150 min, 81.6 °C for HAE, 30 min, 400 W for UAE, and 25 min, 134.6 °C, for the MAE system. The UAE system was identified as the most appropriate technique to obtain extracts with the highest amount of ergosterol, followed by MAE and HAE. Overall, these results demonstrate the potential of using the fruiting bodies of A. blazei as a suitable source to obtain ergosterol-rich extracts, which can be used as an ingredient for various bio-based industrial applications.

Agaricus blazei Murrill from Brazil: an ingredient for nutraceutical and cosmeceutical applications.

Operations for sorting mushrooms at the industrial level usually generate large amounts of bio-residues not conforming to strict morphological criteria for commercial purposes, even though their biological content is not compromised. In this context, the present work aimed at evaluating the potential for reutilizing industrially discarded Agaricus blazei Murill (ABM). Thus, the content of essential nutrients and the chemical composition were determined, and MTT and LDH assays were used to evaluate the viability and cell death of Caco-2 and HT29 cell lines of an ethanolic extract prepared from ABM (preliminary safety tests for nutraceutical applications). The extract was incorporated into a semi-solid base cosmetic cream and cell viability effects of the extract, and of the final cream formulation, on a keratinocyte cell line (HaCaT) were studied (preliminary safety tests for cosmeceutical applications). Essential nutrients, such as proteins and carbohydrates, and a low fat content were determined for ABM. Twenty-two fatty acids were detected, with polyunsaturated fatty acids (PUFA) (∼53%) being the most abundant fraction. The cell viabilities of Caco-2 and HT29 cells were maintained up to 100 µg mL-1. After incorporation into the base cream, a formulation with a pale yellow colour and favourable pH was obtained. The cell viability of HaCaT cells in the presence of the extract and the final cream formulation was maintained in a concentration dependent manner, which indicates the safety of this extract for cosmeceutical applications. The results suggest that ABM residues can be used as an inexpensive and sustainable source of nutraceutical and cosmeceutical ingredients.

Extraction of triterpenoids and phenolic compounds from Ganoderma lucidum: optimization study using the response surface methodology.

The extraction of triterpenoids and phenolic compounds from Ganoderma lucidum was optimized by using the response surface methodology (RSM), using heat and ultrasound assisted extraction techniques (HAE and UAE). The obtained results were compared with that of the standard Soxhlet procedure. RSM was applied using a circumscribed central composite design with three variables (time, ethanol content, and temperature or ultrasonic power) and five levels. The conditions that maximize the responses (extraction yield, triterpenoids and total phenolics) were: 78.9 min, 90.0 °C and 62.5% ethanol and 40 min, 100.0 W and 89.5% ethanol for HAE and UAE, respectively. The latter was the most effective, resulting in an extraction yield of 4.9 ± 0.6% comprising a content of 435.6 ± 21.1 mg g-1 of triterpenes and 106.6 ± 16.2 mg g-1 of total phenolics. The optimized extracts were fully characterized in terms of individual phenolic compounds and triterpenoids by HPLC-DAD-ESI/MS. The recovery of the above-mentioned bioactive compounds was markedly enhanced using the UAE technique.

The potential of Ganoderma lucidum extracts as bioactive ingredients in topical formulations, beyond its nutritional benefits.

Ganoderma lucidum was characterized in terms of nutritional value and chemical composition. Thereafter, ethanolic Soxhlet extracts were evaluated for antioxidant, anti-inflammatory, antityrosinase, antimicrobial and cytotoxic effects, and further characterized in terms of phenolic acids, polysaccharides and triterpenoids. Finally, the obtained extracts were tested as cosmeceutical ingredients. G. lucidum proved to be a source of macronutrients and important bioactive compounds such as terpenoids, specially triterpenoids, and polysaccharides. In the extracts, ganoderic acids C2, A and H were the most abundant triterpenic acids and protocatechuic, p-hydroxibenzoic and syringic acids the identified phenolics. The developed cosmeceutical formulation preserved the extract bioactivities, presented a light-yellow colour and a pH of 4.6, which is considered appropriate for cosmeceutical's design. Behind the important nutritional/bioactive composition of G. lucidum, a potential towards its valorisation in the field of cosmeceuticals is foreseeable, as deduced from the bioactivities of its ethanolic extract and preservation in the tested formulation.