The responsible beneficiaries provided the consortium with the necessary by products and raw materials of plants with great interestsuch as R. damascene, L. vera, M. Officinalis, G. macrorrhizum, M. Camomilla, S. scardica etc. Mastic gum by-product named colophony and by-products of the mastic gum hydrodistillation were also collected. Collection of stems and leaves from Pistacia species was also performed. The by-products were transferred to the responsible participants in order to be investigated. A great number of extracts and enriched fractions were prepared using environmentally friendly technologies and extraction protocols were developed for each collected raw material. Depending on the raw material, “green” techniques such as Supercritical Fluid extraction, Microwave Assisted Solvent Extraction, Subcritical water extraction and highly cross-linked adsorptive resins were used. Additionally, the optimal extraction conditions for most of the raw material were also determined. All the preparations were forwarded for the initial assessment of their chemical content. Total Phenolic Content (TPC), Total Flavonoids Content (TFC) and HPTLC analysis using both general and specific reagents for the visualization of the chromatograms were performed. The results of that initial chemical profiling revealed the most promising extracts. These extracts were submitted to Centrifugal Partition Chromatography technology in order for the main components to be isolated. For the identification of the volatile components an analytical procedure on GC-MS was developed. The qualitative characterization of the extracts was also performed with analytical processes based on HPTLC. Antioxidant activities of extracts and enriched fractions were investigated using DPPH antioxidant assay and ABTS to measure the radical scavenging capacities of the samples. The extracts were also evaluated for their skin-whitening activity using an established tyrosinase assay. The evaluation of the anti-inflammatory activity, the cytotoxicity and their ability to inhibit enzymes involved in the degradation of elastin, collagen and hyaluronic acid was performed. In total, more than 150 preparations (extracts, enriched fractions, essential oils) have been prepared, 12 relative extraction protocols have been developed and more than 70 ingredients purified. Pure compounds were characterized with the use of chromatographic and spectroscopic features using LC-HRMS/MS combined with structural information deriving from 1D and 2D NMR experiments. Moreover, biotransformation of selected extracts using enzymes in several conditions and chemical transformations such as hydrolysis was carried out in order to obtain extracts enriched in bioactive compounds. In total, a library of extracts, enriched fractions and purified compounds was created. The pilot scale production of selected extracts was based on the laboratory scale extraction protocol. For the optimization of the methods, several parameters were taken under consideration. Furthermore, aiming to the smooth transition into industrial implementation all the respective steps of the process as well as the corresponding results were evaluated for their compliance with the demands from industry and regulatory authorities.
