Natural Products from Leaves of the Ancient Iranian Medicinal Plant Echium amoenum Fisch. & C. A. Mey.

For several millennia, leaves of Echium amoenum Fisch. & C. A. Mey., an important Iranian medicinal plant with nutritional value as nutraceutical, have been used as tea for the treatment of several conditions, including inflammation. The nutritional value of intake of E. amoenum tea has mainly been correlated to its rich content of mainly water-soluble antioxidants. Although the entire plant is utilized, only natural products of the flowers have previously been thoroughly investigated. The rare natural products bis(3-(3,4-dihydroxyphenyl)-1-methoxy-1-oxopropan-2-yl)-1-(3,4-dihydroxyphenyl)-6,7-dihydroxy-1,2-dihydronaphthalene-2,3-dicarboxylate, 4-Oxy-(E)-caffeoyl-2,3-dihydroxybutanoic acid methyl ester and 4-Oxy-(Z)-caffeoyl-2,3-dihydroxybutanoic acid methyl ester, in addition to the widely distributed compounds rosmarinic acid methyl ester and (E)-caffeic acid, were purified and characterized from leaves of Echium amoenum. The structures were determined by a combination of several 2D NMR spectroscopic techniques, circular dichroism spectroscopy and high-resolution mass spectrometry. The fact that bis(3-(3,4-dihydroxyphenyl)-1-methoxy-1-oxopropan-2-yl)-1-(3,4-dihydroxyphenyl)-6,7-dihydroxy-1,2-dihydronaphthalene-2,3-dicarboxylate belongs to a rare group of natural products which have previously been patented for their significant anti-inflammatory activity may rationalize the traditional treatment of inflammations with E. amoenum.

Design and in vitro effectiveness evaluation of Echium amoenum extract loaded in bioadhesive phospholipid vesicles tailored for mucosal delivery.

The Echium amoenum Fisch. and C.A. Mey. (E. amoenum) is an herb native from Iranian shrub, and its blue-violet flowers are traditionally used as medical plants. In the present study, an antioxidant phytocomplex was extracted from the flowers of E. amoenum by ultrasounds-assisted hydroalcoholic maceration. The main components, contained in the extract, have been detected using HPLC-DAD, and rosmarinic acid was found to be the most abundant. The antioxidant power of the extract along with the phenolic content were measured using colorimetric assays. The extract was loaded in liposomes, which were enriched adding different bioadhesive polymers (i.e., mucin, xanthan gum and carboxymethyl cellulose sodium salt) individually or in combination. The main physico-chemical properties (i.e. size, size distribution, surface charge) of the prepared vesicles were measured as well as their stability on storage. The viscosity of dispersion and the ability of vesicles to interact with mucus were evaluated measuring their stability in a mucin dispersion and mobility in a mucin film. The biocompatibility and the ability of the formulations to protect keratinocytes from damages caused by hydrogen peroxide and to promote the cell migration were measured in vitro.

Echiumin E, an Aryl Dihydronaphthalene Lignan from the Australian Invasive Plant Paterson's Curse (Echium plantagineum).

A new aryl dihydronaphthalene lignan, echiumin E (1), and four known compounds, echiumin A, globoidnan A, (-)-rabdosiin, and rosmarinic acid (2-5), were isolated from the Australian invasive plant Echium plantagineum (Paterson's curse) for the first time. Echiumin E (1) was characterized by 1D/2D NMR spectroscopy and MS spectrometry, with its absolute configuration assigned through comparison of experimental and TDDFT-calculated ECD data. Echiumin E (1) along with compounds 3-5 were screened in vitro against three cancer cell lines (SH-SY5Y, HeLa, and PC-3) and a prostate stromal (normal) cell line (WPMY-1) using a resazurin reduction assay. Echiumin E (1) was found to be active toward HeLa cells (IC50 0.21 µM).

Isolation of Echimidine and Its C-7 Isomers from Echium plantagineum L. and Their Hepatotoxic Effect on Rat Hepatocytes.

Echimidine is the main pyrrolizidine alkaloid of Echium plantagineum L., a plant domesticated in many countries. Because of echimidine's toxicity, this alkaloid has become a target of the European Food Safety Authority regulations, especially in regard to honey contamination. In this study, we determined by NMR spectroscopy that the main HPLC peak purified from zinc reduced plant extract with an MS [M + H]+ signal at m/z 398 corresponding to echimidine (1), and in fact also represents an isomeric echihumiline (2). A third isomer present in the smallest amount and barely resolved by HPLC from co-eluting (1) and (2) was identified as hydroxymyoscorpine (3). Before the zinc reduction, alkaloids (1) and (2) were present mostly (90%) in the form of an N-oxide, which formed a single peak in HPLC. This is the first report of finding echihumiline and hydroxymyoscorpine in E. plantagineum. Retroanalysis of our samples of E. plantagineum collected in New Zealand, Argentina and the USA confirmed similar co-occurrence of the three isomeric alkaloids. In rat hepatocyte primary culture cells, the alkaloids at 3 to 300 µg/mL caused concentration-dependent inhibition of hepatocyte viability with mean IC50 values ranging from 9.26 to 14.14 µg/mL. Our discovery revealed that under standard HPLC acidic conditions, echimidine co-elutes with its isomers, echihumiline and to a lesser degree with hydroxymyoscorpine, obscuring real alkaloidal composition, which may have implications for human toxicity.

Comparative analysis of the main medicinal substances and applications of Echium vulgare L. and Echium plantagineum L.: A review.

ETHNOPHARMACOLOGICAL RELEVANCE: Echium vulgare L. and Echium plantagineum L. originated in the Mediterranean, and were later domesticated in Africa, America, Asia, Europe and Oceania, where they were widely used to treat many diseases including cough, urinary tract infection, fever, inflammation and muscle strain. AIM OF THE STUDY: The purpose of this review is to provide scientific literature on the traditional uses, bioactive chemical components and pharmacological activities of two species of Echium, and to critically analyze the information provided, so as to understand the current work on these two species and explore the possible prospect of this plant in pharmaceutical research. METHODS: Systematic review and meta-analysis were conducted according to Prisma guidelines, and the related literatures searched on Google Academic, Science Direct, Baidu Scholars and China National Knowledge Infrastructure (CNKI) up to June 2021 were reviewed. The key words used are: Echium, E.vulgare, E.plantagineum, plant components, chemical components, pharmacological activities, pharmaceutical products and applications. Thereafter all eligible studies are analyzed and summarized in this review. The selection of manuscripts is based on the following inclusion criteria: the article has years of research or publication, is published in English, Portuguese or Spanish and Chinese, and there are keywords in the title, abstract, keywords or full text of the article. For the selection of manuscripts, first, select articles according to titles, then summarize them, and finally, analyze the full text of the publication. Elimination criteria: 1. Duplicate reports; 2. There are research design defects and poor quality; 3. Incomplete data and unclear ending effect; 4. The statistical method is wrong and cannot be corrected. RESULTS: The pharmacological characteristics of E.vulgare and E.plantagineum can basically support their traditional use, but the medicinal substances contained in them are quite different in composition and content, and the development and application of corresponding products are also different. CONCLUSIONS: At present, there is little clinical data about drugs related to the two species, and more research is needed in the future, especially human experiments and clinical trials, to evaluate the cellular and molecular mechanisms based on pharmacological, biological activity and safety studies, and to provide more powerful scientific basis for their traditional medicinal properties. In addition, the further application and development of the medicinal products of E.vulgare and E.plantagineum still need to be precise and identified, so as to give full play to their medicinal potential.

Evaluation of multilayer perceptron neural networks and adaptive neuro-fuzzy inference systems for the mass transfer modeling of Echium amoenum Fisch. & C. A. Mey.

BACKGROUND: Multilayer perceptron (MLP) feed-forward artificial neural networks (ANN) and first-order Takagi-Sugeno-type adaptive neuro-fuzzy inference systems (ANFIS) are utilized to model the fluidized bed-drying process of Echium amoenum Fisch. & C. A. Mey. The moisture ratio evolution is calculated based on the drying temperature, airflow velocity and process time. Different ANN topologies are examined by evaluating the number of neurons (3 to 20), the activation functions and the addition of a second hidden layer. Different numbers (2 to 5) and shapes of membership functions are examined for the ANFIS, using the grid partitioning method. The models with the best performance in terms of prediction accuracy, as evaluated by the statistical indices, are compared with the best fit thin-layer model and the available data from the experimental cases of 40 °C, 50 °C and 60 °C temperatures at 0.5, 0.75 and 1 ms-1 airflow velocity. RESULTS: The best performed ANFIS model, comprised by 5-2-2 of π-shaped andtriangular membership functions for time, temperature and airflow velocityinputs respectively, was able to describe the moisture ratio evolution of E. amoenum more precisely than the best ANN topology, achieving higher values of coefficientof determination (R2 ), root mean square error (RMSE) and sum of squared errors(SSE). The best thin-layer model involving six adjustable parameters, managedto describe experimental data most accurately with R2 = 0.9996, RMSE = 0.0057and SSE = 7.3·10-4 . CONCLUSION: The results of the comparative study indicate that empirical regression models with increased numbers of adjustable parameters, constitute a simpler and more accurate modeling approach for estimating the moisture ratio of E. amoenum Fisch. & C. A. Mey under fluidized bed drying. © 2021 Society of Chemical Industry.

Combination of natural strategies to improve the oxidative stability of echium seed oil.

Echium seed oil is an alternative source of omega 3 fatty acids but it is highly susceptible to oxidation. A combination of three natural strategies was proposed in this study aiming to improve the oxidative stability of echium oil obtained by pressing (PO) or solvent extraction (PSO), kept in the storage condition for 180 days or during the consumption for 30 days. Our results showed that the reduction of temperature was sufficient to keep the oil stable during storage for both samples. During the consumption time, the best stability was achieved by adding a mixture of antioxidants, composed of sinapic (500 ppm), ascorbic (250 ppm), and citric (150 ppm) acids, and/or 20% of high oleic sunflower oil. The combined strategies promoted a 34 to 80% reduction of peroxide value and 0 to 85% reduction of malondialdehyde concentrations in the samples, showing to be a feasible and natural alternative to improve the oxidative stability of echium oil. PRACTICAL APPLICATION: Our study successfully applied an optimized combination of simple and low-cost strategies to enhance the chemical stability of echium seed oil. As the use of echium oil expands around the world, the oil industry and final consumers may benefit from our results to increase the oil shelf-life.

Pyrrolizidine alkaloids and beehive products: A review.

Pyrrolizidine alkaloids (PA) are secondary metabolites of plants, which are mostly found in the genus Senecio, Echium, Crotalaria, and Eupatorium. The presence of 1,2-unsaturated PA in foods is a concern to food regulators around the world because these compounds have been associated to acute and chronic toxicity, mainly in the liver. The intake foods with PA/PANO usually occur through accidental ingestion of plants and their derivatives, besides to products of vegetal-animal origin, such as honey. PA/PANO are transferred to honey by their presence in nectar, honeydew, and pollen, which are collected from the flora by bees. In addition to honey, other beekeeping products, such as pollen, royal jelly, propolis, and beeswax, are also vulnerable to PA contamination. In this context, this review provides information about chemical characteristics, regulation, and toxicity, as well as summarizes and critically discusses scientific publications that evaluated PA in honeys, pollens, royal jelly, and propolis.

The Toxicity Effect of Echium amoenum on the Liver and Kidney of Mice.

AIMS: The aim of this study was to investigate the toxic effect of Echium amoenum plants on the liver and kidney of the animal model. BACKGROUND: Echium amoenum is one of the medicinal plants containing pyrrolizidine alkaloids with several properties which has widely consumed among different communities. OBJECTIVE: The toxic effects of Echium amoenum on the liver and kidney were investigated in this study. METHODS: Sixty mice were kept for 28 days under the appropriate laboratory conditions. Echium amoenum extract (25, 12.5, 50 mg / kg, ip.) was administered for 28 days. At the end of the experiment, blood samples were drawn and liver and kidneys were removed for evaluating hepatotoxicity and nephrotoxicity of extract. Additionally, experiments were conducted to assay the enzymatic and oxidative activities. RESULTS: There was no significant difference in the levels of copper ion in the liver and kidneys among all groups. There was a significant difference in the levels of lipid peroxidation in the liver of treated groups versus the control group. The significant difference was not observed in the levels of glutathione of the liver of all groups. However, the levels of glutathione of the kidney significantly decreased in the treated groups versus the control group. There was no significant difference in the liver enzymes, including ALP, SGOT, and SGPT, between all groups. This indicates that damage increases with enhancing the time and concentrations of the extract. Biochemical analysis showed the creatinine and urea levels did not change in the treated groups versus the control group. CONCLUSION: According to the present findings, it is suggested that Echium amoenum causes hepatotoxicity and nephrotoxicity effects in dose and time-dependent manner.

Transfer of pyrrolizidine alkaloids from ragwort, common groundsel and viper's bugloss to milk from dairy cows.

To investigate the transfer of pyrrolizidine alkaloids (PAs) from feed to milk, rumen-cannulated dairy cows were intra-ruminally fed with 200 g/day of dried plant material of either ragwort (mixture of Jacobaea vulgaris and Senecio inaequidens), common groundsel (Senecio vulgaris) or viper's bugloss (Echium vulgare) for a period of 4 days. PA levels in the plant materials were 3767, 2792 and 1674 µg g-1 respectively. Feed intake, milk yield and several blood parameters indicative for liver function were not influenced by the treatment. When fed ragwort, increased levels of PAs were detected in the milk, in particular jacoline and an unidentified cyclic diester, possibly a hydroxylated metabolite from retrorsine. The latter was the most important PA in milk from cows fed common groundsel. For viper's bugloss, echimidine was the most abundant identified PA but in addition several hydroxylated PA metabolites were detected. For ragwort, the overall PA transfer was estimated at 0.05% and 1.4% for jacoline (N-oxide). Transfer rates were similar for viper's bugloss (0.05%) but lower for common groundsel (0.01%). Only a small portion of the administered PAs was quantified in milk, urine and faeces, with an overall balance of 4.5%, 2.9% and 5.8%, for ragwort, common groundsel and viper's bugloss, respectively. Samples taken from the rumen indicated that the N-oxides were converted into the free bases, which was confirmed by in vitro studies with the same plant species incubated with ruminal fluid. These results confirm that the transfer of PAs to milk is relatively low but may be of concern for human health regarding the genotoxic and carcinogenic properties of these compounds. The transfer rate depends on the type of PAs present in the weeds. The incomplete balance of input vs output stresses the need to further investigate the metabolism and the potential transfer of metabolites into edible products.

The role of hippocampal GABAA receptors on anxiolytic effects of Echium amoenum extract in a mice model of restraint stress.

Echium amoenum (EA), a popular medicinal plant in Persian medicine, has anxiolytic, antioxidant, sedative, and anti-inflammatory effects. This study examined whether GABA-ergic signaling is involved in the anxiolytic effects of EA in mice. Sixty BALB/c mice (25-30 g) were divided into six groups (n = 10) as follows: the (I) control group received 10 ml/kg normal saline (NS). In the stress groups, the animals underwent 14 consecutive days of restraint stress (RS), and received following treatments simultaneously; (II) RS + NS; (III) RS + Diaz (Diazepam); (IV) RS + EA; (V) RS + Flu (Flumazenil) + EA; (VI) RS + Flu + Diaz. Behavioral tests including the open field test (OFT) and elevated plus maze (EPM) were performed to evaluate anxiety-like behaviors and the effects of the regimens. The plasma level of corticosterone and the hippocampal protein expressions of IL-1ß, TNF-α, CREB, and BDNF, as well as p-GABAA/GABAA ratio, were also assessed. The findings revealed that chronic administration of EA alone produced anxiolytic effects in both behavioral tests, while diazepam alone or in combination with Flu failed to decrease the anxiety-like behaviors. Furthermore, the p-GABAA/GABAA and p-CREB/CREB ratios, and protein levels of BDNF were significantly increased in the EA-received group. On the other hand, plasma corticosterone levels and the hippocampal IL-1ß and TNF-α levels were significantly decreased by EA. However, pre-treatment with GABAA receptors (GABAA Rs) antagonist, Flu, reversed the anxiolytic and molecular effects of EA in the RS-subjected animals. Our findings confirmed that alternation of GABAAR is involved in the effects of EA against RS-induced anxiety-like behaviors, HPA axis activation, and neuroinflammation.

Improvement of thermal stability and antioxidant activity of anthocyanins of Echium amoenum petal using maltodextrin/modified starch combination as wall material.

To protect anthocyanins of Iranian borage extract (IBE) by encapsulation, maltodextrin (MD)/modified maize starch (MMS) combinations were used at four ratios (MD/MMS: 1/0, 1/0.25, 1/0.5, 1/1, w/w%). The microencapsulated powders with different ratios of wall material presented encapsulation efficacy between 93.1 and 97.4%. FTIR confirmed the cross-linking interaction between extract and MD/MMS wall. Increased thermal stability of extract was also indicated by DSC analysis. The stability of microstructures and crude extract at two conditions (5 and 40 °C) was evaluated by UV and HPLC methods during 60 days. The results of stability showed that encapsulation by treatment 4 (MD/MMS 1:1) improved the stability of anthocyanins 43.8% compared to crude extract. Additionally, microcapsule created in this treatment provided great antioxidant stability. The release in the simulated gastrointestinal tract revealed that the anthocyanins were chiefly retained inside the microparticles in the stomach, and were released in the intestine. The results demonstrated that the microencapsulation with MD and MMS by the spray-drying method can improve the stability, antioxidant and functional characteristics of borage extract, which could be useful in the food and pharmaceutical industry.

Effect of sinapic acid ester derivatives on the oxidative stability of omega-3 fatty acids rich oil-in-water emulsions.

Oil-in-water (O/W) emulsions are important delivery systems of omega-3 fatty acids (n-3 FA). We investigated the effect of sinapic acid esters concentration and chain length, the electrical charge of the emulsifier and emulsion pH on the oxidative stability of n-3 FA rich O/W emulsions. Echium oil was applied as n-3 FA source. A 24 factorial design was used to simultaneously evaluate these factors. Peroxide value, malondialdehyde, 2,4-heptadienal and 2,4-decadienal were measured in the emulsions. pH and the electrical charge of the emulsifier modulated the antioxidant effectiveness of sinapic acid esters, while concentration was not relevant. The combination of positively charged emulsifier with neutral pH provided the best oxidative stability for echium oil emulsions. Our results also suggested that the increase of length chain of sinapic acid, from C4 to C12, reduced the secondary products of oxidation, when echium oil emulsions were prepared using negatively charged emulsifier under acidic conditions.

Partial replacement of pork fat by Echium oil in reduced sodium bologna sausages: technological, nutritional and stability implications.

BACKGROUND: The present research aimed to evaluate the nutritional, technological, microstructural and sensory characteristics of bologna sausages made with pork fat replaced with Echium oil. Three different treatments were processed, all of them with approximately 35% less sodium than a regular bologna-type sausage: Control (only pork backfat addition) and T25 and T50 (replacement of 25 and 50% of the added animal fat with Echium oil respectively). RESULTS: Proximate composition, texture profile analysis, fatty acid profile and microstructure were evaluated to characterize the products. In refrigerated storage, the following characteristics were assessed: thiobarbituric acid-reactive substances (TBARS) index, objective color, pH value, microbiological counts and sensory acceptance. Both samples with Echium oil (T25 and T50) showed a healthier lipid profile, with high omega-3 content, mainly from α-linolenic and stearidonic fatty acids, consequently presenting better (P ≤ 0.05) values of nutritional indices than the Control. However, the replacement of 50% animal fat with Echium oil negatively affected (P ≤ 0.05) most of the technological characteristics and the sensory acceptance of bolognas. In contrast, the treatment with 25% replacement did not show significant differences in most of the evaluated parameters compared with the Control sample. CONCLUSION: Thus the replacement of 25% of pork backfat with Echium oil in bologna sausages can be recommended, representing a good balance between nutritional gains and sensory impairment. © 2019 Society of Chemical Industry.

Development of a Selective Adsorbing Material for Binding of Pyrrolizidine Alkaloids in Herbal Extracts, Based on Molecular Group Imprinting.

Pyrrolizidine alkaloids are secondary plant constituents that became a subject of public concern because of their hepatotoxic, pneumotoxic, genotoxic, and cytotoxic effects. Due to disregardful harvesting and/or contamination with pyrrolizidine alkaloid-containing plants, there is a high risk of ingesting these substances with plant extracts or natural products. The limit for the daily intake was set to 0.007 µg/kg body weight. If contained in an extract, cleanup methods may help to minimize the pyrrolizidine alkaloid concentration. For this purpose, a material for depleting pyrrolizidine alkaloids in herbal preparations was developed based on the approach of molecular imprinting using monocrotaline. Molecular imprinted polymers are substances with specific binding characteristics, depending on the template used for imprinting. By means of group imprinting, only one molecule is used for creating selective cavities for many molecular pyrrolizidine alkaloid variations. Design of Experiment was used for the development using a 25 screening plan resulting in 64 polymers (32 MIPs/32 NIPs). Rebinding trials revealed that the developed material can compete with common cation exchangers and is more suitable for depleting pyrrolizidine alkaloids than C18- material. Matrix trials using an extract from Chelidonium majus show that there is sufficient binding capacity for pyrrolizidine alkaloids (80%), but the material is lacking in selectivity towards pyrrolizidine alkaloids in the presence of other alkaloids with similar functional groups such as berberine, chelidonine, and coptisine. Beyond this interaction, the selectivity could be proven for other structurally different compounds on the example of chelidonic acid.

Pyrrolizidine Alkaloids: The Botanical Origin of Pollen Collected during the Flowering Period of Echium vulgare and the Stability of Pyrrolizidine Alkaloids in Bee Bread.

Previous studies have shown that pollen products sold as nutritional supplements and used in apitherapy may contain toxic pyrrolizidine alkaloids (PAs) if bees collect pollen from PA-containing plants, such as Echium vulgare. In this study, the botanical origin of pollen from two observation sites was studied. Despite a high PA content in pollen samples that bees collected during E. vulgare's flowering period, bees were found to collect relatively few Echium pollen loads. Thus, the monitoring of pollen loads collected at the apiaries is unviable to estimate the risk of PA contamination in pollen or bee bread. In a second step, the stability of PAs in bee bread samples containing PAs at concentrations of 2538 ng/g and 98 ng/g was assessed over a period of five or six months, respectively. No significant PA reduction was observed in bee bread stored at 15 °C, but there were overall PA reductions of 39% and 33% in bee bread stored at 30 °C, reflecting hive conditions. While PA N-oxides decreased over time, other types of PAs remained relatively stable. Monitoring PAs in pollen products remains important to ensure consumer safety and should include echivulgarine (and its N-oxide), the major PA type found in pollen from E. vulgare.

Protective effect of hydroxytyrosol and rosemary extract in a comparative study of the oxidative stability of Echium oil.

The oxidative stability of Echium oil has been studied by two accelerated oxidation methodologies namely Rancimat and differential scanning calorimetry at selected temperatures in the range 50-110 °C. Kinetic analyses have provided Arrhenius parameters, oxidative stability indexes (OSI), temperature coefficients (tcoef), Q10 and OSI at 20 °C (OSI20) for both methodologies focussing on secondary and tertiary oxidation products. Peroxide, p-anisidine and polar compounds values have been measured to characterize from primary to tertiary oxidation products. In general, susceptibility of this oil to oxidation has been found to be larger than for other vegetable oils, according to the OSI20 values estimated by Rancimat and differential scanning calorimetry. However, the values of tcoeff and Q10 are found to be similar to those described for vegetable oils. In addition, a study of the influence of two antioxidants, namely hydroxytyrosol and rosemary extract on the oxidative stability of Echium oil has been carried out using both methodologies. Protective effects over oxidation by up to a threefold increase of OSI20 values have been found for both antioxidants at the highest dose utilized.

Elucidation of Infusion-Induced Changes in the Key Odorants and Aroma Profile of Iranian Endemic Borage ( Echium amoenum) Herbal Tea.

Infusion-induced changes in the aroma and key odorants and their odor activity values of Iranian endemic herbal (Gol-Gavzaban) tea obtained from shade-dried violet-blue petals of borage ( Echium amoenum) were studied for the first time. Two hot teas and one cold tea were investigated and coded as 4MN (4 min/98 °C), 16MN (16 min/98 °C), and 24HR (24 h/ambient temperature), respectively. Aromatic extracts of the tea samples were isolated by the liquid-liquid extraction method and analyzed by gas chromatography-mass spectrometry-olfactometry (GC-MS-O) for the first time. According to the results of the aroma profiling, a total of 35 common aroma compounds comprising alcohols, acids, volatile phenols, lactones, aldehydes, ketone, pyrroles, and furans were identified and quantified in the tea samples. Indeed, it is worth noting that the aroma profiles of the borage teas were similar. However, the effects of the infusion techniques were clearly different as observed on the content of each individual and total compounds in the samples. The highest mean total concentration was detected in 24HR (266.0 mg/kg), followed by 16MN (247.1 mg/kg) and 4MN (216.1 mg/kg). 1-Penten-3-ol was the principal volatile component in all borage teas. On the basis of the result of the flavor dilution (FD) factors, a combined total of 22 different key odorants was detected. The potential key odorants with regard to FD factors in all samples were prevailingly alcohols, acids, and terpenes. The highest FD factors were observed in 2-hexanol (2048 in 4MN and 24HR; 1024 in 16MN) and 1-penten-3-ol (2048 in 24HR; 1024 in 4MN and 16MN) in samples providing herbal and green notes. Principal component analysis (PCA) showed that the tea samples could clearly be discriminated in terms of their aroma profiles and key odorants. The findings of the current study demonstrate that the tea preparation conditions have a significant impact on the organoleptic quality of borage tea.

Production of stearidonic acid-rich triacylglycerol via a two-step enzymatic esterification.

The aim of this study was to synthesize stearidonic acid (SDA)-rich triacylglycerol (TAG) via a two-step lipase-catalyzed esterification under vacuum. SDA-rich fatty acid, which was prepared from echium oil via Candida rugosa lipase-catalyzed selective esterification, was used as the substrate. Two different immobilized lipases, Novozym 435 from Candida antarctica and Lipozyme TL IM from Thermomyces lanuginosus, were employed for the synthesis of SDA-rich TAG. In the first step, Novozym 435-catalyzed esterification of the SDA-rich fatty acid with glycerol was carried out for 2 h. In the second step, Lipozyme TL IM-catalyzed esterification of the reaction mixture from the first step was performed for an additional 10 h. The optimal reaction conditions for the second step were a temperature of 65 °C, an enzyme loading of 20%, and a vacuum of 0.7 kPa. Consequently, the maximum TAG conversion of ca. 86.4 wt% was obtained after 12 h via a two-step lipase-catalyzed esterification.

Stearidonic Acid Concentration by Urea Complexation from Echium Oil.

Concentration of polyunsaturated fatty acids ethyl esters (FAEE) by urea complexation from Echium oil was studied. Different variables involved in the process were investigated: amount and particle size of urea, solvent volume and ratio (hexane/ethanol), load of FAEE and reaction time. Hence, the main goal was to optimize SDA concentration (%) and yield (%) of stearidonic acid (SDA, 18:4 ω-3) and other bioactive FAEE. Similar behaviors were observed in fractionation between α-linolenic (ALA)-linoleic (LA), and γ-linolenic (GLA)-stearidonic (SDA) acids, attributed to similarities on their chemical structures, due to the position of the double bonds. At laboratory scale, the optimal conditions were 3 g urea (powder), 3.6 mL of hexane, 0.54 mL of ethanol and 800 mg of FAEE, during 20 h at 25°C. A scaling-up at pilot plant was carried out twice, obtaining more than 100 g of a final product, with ~29% SDA concentration and ~78% yield. Besides, after two washings with water, ethyl carbamates (urethanes) were not detected in the final product. Thus, a mixture of FAEE with about 85% of bioactive fatty acids with anti-inflammatory properties was obtained, which can be a high added-value product with great potential for the synthesis of functional lipids and nutraceuticals.