Oregano and thyme by-products of olive oil aromatization process with microwave assisted extraction as a rich source of bio-active constituents.

Introduction: Processing of Medicinal Aromatic Plants (MAPs) results in the production of a significant amount of by-products, which are not commercially exploitable. Towards this direction, we studied extensively the by-products of oregano and thyme, remaining after the aromatization of olive oils with microwave assisted extraction (MAE). The purpose of the study was the exploitation of the "wastes" of these two economically significant herbs of Greece, for the potential development of innovative bioactive products. Methods: Hence, superior and inferior quality plant material from Origanum vulgare subsp. hirtum and Thymus vulgaris, were extracted with extra virgin olive oil using MAE. For the evaluation of raw plant material, beside the characterization of the essential oils (EOs), the hydroalcoholic extracts of superior and inferior plant material were afforded by ultrasound assistant extraction (UAE). In addition, the remaining plant material after the flavoring of olive oil by MAE, was extracted with c-Hex, MeOH, H2O:MeOH using UAE. All the extracts were evaluated for their DPPH free radical scavenging activity and total phenolic content (TPC) as well as their chemical profile was investigated by HPTLC. In parallel, the EOs, the olive oils and the c-Hex extracts were analyzed by GC-MS and Headspace Solid Phase Microextraction (HS-SPME)-GC-MS. Results and Discussion: The results showed that the composition of the EOs and the volatile fraction of the olive oil extracts were similar for the superior quality material whereas for the inferior the composition of the volatile fraction of olive oil extracts was not analogous to the respective EOs. GC-MS analyses of oregano and thyme by-products revealed the presence of carvacrol, thymol, γ-terpinene and p-cymene among the major constituents. Moreover, the hydroalcoholic extracts obtained from the plant material remaining after olive oil flavoring with MAE showed similar phenolic content and scavenging activity with the hydroalcoholic extracts of the corresponding raw plant materials underlying their potent use in the preparation of high-added value products such as nutraceuticals and cosmeceuticals as well as enriched animal nutrition products.

Taxonomic Identification and Molecular DNA Barcoding of Collected Wild-Growing Orchids Used Traditionally for Salep Production.

Molecular DNA barcoding combined with botanical taxonomy can be used for the identification and conservation of collected Greek orchids used for salep production as well as in the regulation of fair salep trade. A modified CTAB protocol was used for DNA extraction, amplification of barcoding regions (ITS, matK, rbcL, trnH-psbA), and sequencing. Sequencing data were assembled using Bioedit software, and the BLAST algorithm was used on the NCBI database for species identification at the genus level. Molecular barcoding data based on genetic similarity identification was in full coherence with taxonomic classification based on morphological data. The combination of ITS and matK exhibited a greater capacity to identify a species among the Greek salep samples. Out of the 53 samples examined, 52.9% were classified as Dactylorhiza spp. and 33.3% as Anacamptis spp., whereas only 6 samples were identified as Orchis spp. (11.8%). Given that a superior-quality salep beverage comes from tubers of the latter, the number of samples classified as such in northwestern Greece is unexpectedly low. A database of 53 original reference sequences from wild-growing samples of Greek origin was generated, providing a valuable resource for the identification of other salep samples from different regions. The DNA barcoding results unveiled that salep samples from northwestern Greece are related to nine members of four different genera of Orchidaceae. All species are nationally protected and covered by the CITES convention, while many of these orchids are included in the EU Directive 92/43/EEC appendix as "Other Important Species". Thus, expedited coordinated management actions are needed to ensure their survival in the future.

Sustainable Use of Greek Herbs By-Products, as an Alternative Source of Biologically Active Ingredients for Innovative Products.

The processing of medicinal and aromatic plants (MAPs) results in the production of a significant amount of plant by-products; herbal material of inferior quality and/or unusable plant parts that are not commercially exploitable. An extensive study of Greek native species was performed toward the production of innovative bioactive products using as raw materials the by-products obtained from the processing of cultivated MAPs. Origanum vulgare subsp. hirtum (oregano), Sideritis scardica (Greek mountain tea), Thymus vulgaris (thyme), and Matricaria recutita (chamomile) were selected due to their wide use for the preparation of beverages and culinary purposes. The determination of the percentage of the post-harvest processing by-products was performed for a 3 years period (2018-2020). Results showed that by-products derived from the above-mentioned species' processing constitute 64% (thyme), 54% (oregano), 37% (Greek mountain tea), and 24% (chamomile) of the total processed mass. To value the by-products as a potent source of bioactive ingredients, superior and inferior quality herbal material of the aforementioned plant species were extracted by an ultrasonic assisted extraction method. Hydroalcoholic extracts were chemically investigated using high-performance thin layer chromatography (HPTLC) and liquid chromatography-mass spectrometry (LC-MS) techniques. In addition, their free radical scavenging activity and total phenolic content (TPC) were estimated. Based on the results, herbs by-products revealed similar chemical content to the superior herbal material by the means of HPTLC and LC-MS analysis. In addition, strong free radical scavenging related to a high phenolic content was detected in the case of thyme, oregano, and Greek mountain tea. Moreover, the gas chromatography-mass spectrometry (GC-MS) analyses of the essential oils (EOs) of oregano and thyme by-products revealed the presence of carvacrol, thymol, γ-terpinene, and p-cymene among the major constituents. Finally, the LC-MS analyses of aqueous extracts of Greek mountain tea and chamomile by-products led to the identification of several bioactive compounds, such as flavonoids and phenylpropanoids. Overall, the presence of bioactive constituents in by-products, such as terpenes, phenolic compounds, and flavonoids underly their potent use as food antimicrobial and antioxidant additives, in the preparation of high added-value products, such as enriched aromatic edible oils, and innovative herbal teas, such as instant beverages.