Development of the nutraceutical and pharmaceutical applications of plants selected from Portugal and Iran with presumptive health potentials

Abstract

All over the years scientists have studied an extensive range of medicinal and aromatic plants species which exist in nature and contain an unlimited category of bioactive compounds that could potentially be used within an extensive range of products including nutraceutical; pharmaceutical and even cosmetic. This seems to be more substantial since scientists have detected an extensive range of negative adverse effects among many of the synthetic compounds commonly used today. In this research, we have selected plants which, are particularly well known among people as being healthy and could have therapeutic for chronic and acute diseases. Therefore, after the precise researches and discussions with native residents of the selected regions-including the northwest of Iran and south Portugal, with focus on the Algarve, about 14 different plant species were selected and identified for this research. Among them, different individual parts of plants including seeds (Portugues and Iranian Pimpinella anisum L., with Coriandrum sativum L., and Levisticum officinale W. D. J. Koch., from Iran); flowers (Echium amoenum Fisch & C.A. Mey., Echinacea angustifolia DC., Matricaria chamomilla L., and Althaea officinalis L., from Iran); leaves (Thymus vulgaris L., Rosmarinus officinalis L., Salvia officinalis L., and Alisma plantago-aquatica L. subsp. orientale (Sam.)Sam., from Iran and Olea europaea L. var. sylvestris., from Portugal) have been studied. Moreover the aerial parts of Lavandula pedunculata L. subsp. pedunculata., from Portugal were selected and studied in this research. An extensive range of in vitro chemical and microbial experiments as well as optimization of different variables in the extraction process and encapsulation efficiency of the bioactive compounds have been performed on the different species of plants. Initially, the research intended to study the encapsulation of two of the selected plants’ infusions including Olea europaea., and Alisma plantago-aquatica., with the preparation of calcium-alginate beads in combine of potato starch as a filling substance to study and optimise the release kinetics and encapsulation efficiency of the bioactive compounds in deionised water and simulated gastric fluid (SGF) (as solvents) respectively for each plant. The results confirmed the positive role of the potato starch, in a particular concentration, in the monitoring of release kinetics of phenols from the CAS (Calcium Alginate Starch) beads. On the other hand, the performance of the applied models of Response Surface Methodology (RSM) and Artificial Neural Networks (ANN) on the optimisation of the release kinetics and encapsulation efficiency of the bioactive compounds from the encapsulated infusion in simulated gastric fluid (SGF) as the solvent was compared, and the ANN design was reported as the more accurate tool compared to RSM in the prediction of the optimum range of the selected independent parameters including potato starch as one of the variables. Portugues P. anisum., seeds was selected to optimise the extraction process of the bioactive compounds regarding the two selected independent variables, including the time and temperature of extraction using water as a solvent. Maximum desirability was obtained for 𝐷𝑃𝑃𝐻.− and TMA parameters. Gas Chromatography-Mass Spectrometry (GC-MS) analysis was performed on P. anisum., seeds’ infusion to determine the major compounds profile. The obtained results demonstrated that, fatty acids with indicating 47.68% of the whole detected compounds are the most detected predominant compounds found in the tested extracts followed by triterpenoids; among fatty acids, linoleic acid in combine of oleic and palmitic acids were detected as major compounds. In the current research, the authors also detected bioactive compound content alongside different anti-oxidant parameters of 10 different plants’ infusions picked from Iran. In this regard, total phenolic content (TPC); total flavonoid content (TFC); total chlorophyll content (TCC) values along with total anti-oxidant activity (TAA); reducing power (RP); ferric reducing anti-oxidant power (FRAP); ABTS free radical inhibition and the DPPH free radical scavenging activity of different plants species were determined. The extraction of the bioactive compounds was performed using different pathways; firstly: hot aqueous infusions were obtained at 90 ºC within 5 minutes’ extraction time and cold aqueous extracts were obtained at room temperature within 2 hours blending. Most of the achieved values for the hot infusions were significantly higher compared to cold aqueous extracts. In some cases, a few exceptions were found in P. anisum., and L. officinale., species (seeds) justified by their tough and wooden textures (abnormality in extraction). The Pearson correlation coefficient established that the highest anti-oxidant parameters are related to phenols; chlorophylls and eventual further bioactive compound existence such as triterpenoids; fatty acids; sterols or others rather than flavonoids and further bioactive compounds. On the other hand, synergistic activity hypothesis among at least two or more compounds were proposed. Mostly, the examined leaves, particularly T. vulgaris., presented higher bioactivity than the other studied seeds and flowers species in this research. Gas Chromatography-Mass Spectrometry (GC-MS) analysis was used in the identification of the bioactive compounds in the leaves and two seeds. The achieved results demonstrated an extensive range of compounds with eventual bioactivity including: fatty acids; fatty alcohols; sterols; triterpenoids; monoacylglycerols; aromatics; alkanes; waxes; di-acids; α-hydroxy fatty acids and sugars. It has been observed that in the leaves triterpenoids combined with sterols are the most abundant compounds demonstrating 63.42; 47.64 and 33.51% of the total characterised compounds respectively in R. officinalis., S. officinalis., and T. vulgaris., while in the seeds, fatty acids were discovered as the most predominant compounds, expressing 80.22, 49.04 and 47.68% of the specified compounds in Iranian C. sativum., and P. anisum from Iran and from Portugal., respectively. Betulinic, maslinic, ursolic and oleanolic acids were the most predominant triterpenoids in the leaves and oleic, palmitic and linoleic acids were identified as the most abundant fatty acid content in the seeds. The obtained results proposed that in the plants, the synergistic effects among different bioactive compounds might be expected. According to the characterised and identified bioactive compounds of the plants’ infusions in this study and their established potential health benefits, it could be expected that these plants might demonstrate further potential health benefits for human body such as anti-carcinogenic, anti-diabetic, anti-Alzheimer’s, anti-Parkinson’s, or others. Therefore, this research expanded the research and studied the potential health benefits including anti-bacterial and in vitro anti-diabetic, as well as anti-Alzheimer’s disease, along with anti-radical and further anti-oxidant properties of the plants. Finally the relations between the achieved in vitro results with the characterised compounds of the Gas Chromatography-Mass Spectrometry (GC-MS) were interpreted. Selected leaves’ water infusions also were detected as stronger inhibitors in supressing of the enzymes attributed in revealing of diabetes and Alzheimer’s diseases. The anti-bacterial potential of some of the plants against five different bacterial species showed that the selected Iranian plants, except E. angustifolia., (flower) inhibited E. faecalis in MIC value ≤0.156 mg. mL-1, while other bacterial species showed activities in MIC values ≤10 mg. mL-1. Among flowers M. chamomilla., and among leaves S. officinalis., alongside R. officinalis., revealed slightly higher anti-bacterial activities than others. Alongside the abovementioned studies this research also studied the modelling of the extraction of monomeric anthocyanins using microwave assisted extraction (MAE) from L. pedunculata., picked in Algarve-Portugal. Three independent variables were selected for two responses including TAA and TMA values, among variables microwave power showed 465 W as the optimized value for the extraction of monomeric anthocyanins. In the final of this study, the inhibitory activities of Portugues P. anisum., seeds’ infusion against α-Amylase and Acetylcholinesterase (AChE) were determined, confirming the eventual anti-oxidant; anti-diabetic and anti-Alzheimer’s activities of this plant. Free fatty acids as well as terpenoids and sterols respectively, have demonstrated higher quantities in Portugues P. anisum L., seeds. With respect to the obtained results of inhibitory activities of this infusion against tested enzymes and free radicals, it has been approved that compounds with free radical suppressing activities might not able to participate in competitive enzymes suppressive reactions with substrates. Therefore the needs to in vivo tests for obtaining most reliable results are revealed. This research aims to introduce novel and natural sources of bioactive compounds that might be used in promoting good health, in general, and in particular in the treatment of chronic disorders such as Alzheimer’s, diabetes, cancers as well as Parkinson’s disease along with microbial infections. This research revealed that the tested plants from Portugal and Iran could be potential sources of compounds with an extensive range of application for health benefits as well as in nutraceutical, pharmaceutical and cosmetic products.