Ameliorative effects of elderberry (Sambucus nigra L.) extract and extract-derived monosaccharide-amino acid on H2O2-induced decrease in testosterone-deficiency syndrome in a TM3 Leydig cell.

With aging, men develop testosterone-deficiency syndrome (TDS). The development is closely associated with age-related mitochondrial dysfunction of Leydig cell and oxidative stress-induced reactive oxygen species (ROS). Testosterone-replacement therapy (TRT) is used to improve the symptoms of TDS. However, due to its various side effects, research on functional ingredients derived from natural products that do not have side effects is urgently needed. In this study, using the mitochondrial dysfunction TM3 (mouse Leydig) cells, in which testosterone biosynthesis is reduced by H2O2, we evaluated the effects of elderberry extract and monosaccharide-amino acid (fructose-leucine; FL) on mRNA and protein levels related to steroidogenesis-related enzymes steroidogenic acute regulatory protein (StAR), cytochrome P450 11A1(CYP11A1, cytochrome P450 17A1(CYP17A1), cytochrome P450 19A1(CYP19A1, aromatase), 3ß-hydroxysteroid dehydrogenase (3ß-HSD), and 17ß-hydroxysteroid dehydrogenase(17ß-HSD). We analyzed elderberry extract and extract-derived FL for changes in ROS scavenging activity and testosterone secretion. Elderberry extract and FL significantly reduced H2O2-induced intracellular ROS levels, improved testosterone secretion, and increased the mRNA and protein expression levels of steroidogenesis-related enzymes (StAR, 3b-HSD, 17b-HSD, CYP11A1, CYp17A1). However, the conversion of testosterone to estradiol was inhibited by elderberry extract and extract-derived FL, which reduced the mRNA and protein expression of CYP19A1. In conclusion, elderberry extract and FL are predicted to have value as novel functional ingredients that may contribute to the prevention of TDS by ameliorating reduced steroidogenesis.

Sambucus williamsii Hance: A comprehensive review of traditional uses, processing specifications, botany, phytochemistry, pharmacology, toxicology, and pharmacokinetics.

OBJECTIVE: Sambucus williamsii Hance, belonging to the Sambucus L. family (Viburnaceae), possesses medicinal properties in its roots, stems, leaves, flowers, and fruits. It is recognized for its ability to facilitate bone reunion, enhance blood circulation, remove stasis, and dispel wind and dampness. This traditional Chinese medicine holds significant potential for development and practical use. Hence, this paper offers an in-depth review of S. williamsii, covering traditional uses, processing guidelines, botany, phytochemistry, pharmacology, toxicology, and pharmacokinetics, aiming to serve as a reference for its further development and utilization. MATERIALS AND METHODS: Information for this study was gathered from various books, bibliographic databases, and literature sources such as Google Scholar, Web of Science, PubMed, Chinese National Knowledge Infrastructure, Baidu Scholar, VIP Database for Chinese Technical Periodicals, and Wanfang Data. RESULTS: Phytochemical investigations have identified approximately 238 compounds within the root bark, stem branches, leaves, and fruits of S. williamsii. These compounds encompass flavonoids, sugars, glycosides, terpenoids, phenylpropanoids, alkaloids, phenols, phenolic glycosides, and other chemical constituents, with phenylpropanoids being the most prevalent. S. williamsii exhibits a wide range of pharmacological effects, particularly in promoting osteogenesis and fracture healing. CONCLUSION: This comprehensive review delves into the traditional uses, processing guidelines, botany, phytochemistry, pharmacology, toxicology, and pharmacokinetics of S. williamsii. It provides valuable insights into this plant, which will prove beneficial for future research involving S. williamsii.

Walnut By-Products and Elderberry Extracts-Sustainable Alternatives for Human and Plant Health.

A current alternative for sustainable development through green chemistry is the replacement of synthetic compounds with natural ones through the superior capitalization of natural resources, with numerous applications in different fields. The benefits of walnuts (Juglans regia L.) and elderberries (Sambucus nigra L.) have been known since ancient times, due to the presence of phytochemicals such as flavonoids, polyphenols, carotenoids, alkaloids, nitrogen-containing compounds, tannins, steroids, anthocyanins, etc. These active compounds have multiple biological activities for human health, including benefits that are antibacterial, antioxidant, anti-inflammatory, antidiabetic, hepatoprotective, antihypertensive, neuroprotective, etc. Like other medicinal plants, the walnut and the elderberry possess important phytosanitary properties (antibacterial, antifungal, and insecticidal) and their extracts can also be used as environmentally safe biopesticides, with the result that they constitute a viable and cheap alternative to environmentally harmful synthetic products. During recent years, walnut by-products and elderberries have attracted the attention of researchers, and investigations have focused on the species' valuable constituents and active properties. Comparing the information from the literature regarding the phytochemical profile and biological activities, it is highlighted that, apart from the predominant specific compounds, the walnut and the elderberry have common bioactive compounds, which come from six classes (phenols and derivatives, flavonoids, hydroxycinnamic acids, tannins, triterpenoids, and phytosteroids), and act on the same microorganisms. From this perspective, the aim of this review is to provide an overview of the bioactive compounds present in the different constitutive parts of walnut by-products and elderberries, which present a specific or common activity related to human health and the protection of agricultural crops in the context of sustainable development.

Elderberry Extracts: Characterization of the Polyphenolic Chemical Composition, Quality Consistency, Safety, Adulteration, and Attenuation of Oxidative Stress- and Inflammation-Induced Health Disorders.

Elderberry is highly reputed for its health-improving effects. Multiple pieces of evidence indicate that the consumption of berries is linked to enhancing human health and preventing or delaying the onset of chronic medical conditions. Compared with other fruit, elderberry is a very rich source of anthocyanins (approximately 80% of the polyphenol content). These polyphenols are the principals that essentially contribute to the high antioxidant and anti-inflammatory capacities and the health benefits of elderberry fruit extract. These health effects include attenuation of cardiovascular, neurodegenerative, and inflammatory disorders, as well as anti-diabetic, anticancer, antiviral, and immuno-stimulatory effects. Sales of elderberry supplements skyrocketed to $320 million over the year 2020, according to an American Botanical Council (ABC) report, which is attributable to the purported immune-enhancing effects of elderberry. In the current review, the chemical composition of the polyphenolic content of the European elderberry (Sambucus nigra) and the American elderberry (Sambucus canadensis), as well as the analytical techniques employed to analyze, characterize, and ascertain the chemical consistency will be addressed. Further, the factors that influence the consistency of the polyphenolic chemical composition, and hence, the consistency of the health benefits of elderberry extracts will be presented. Additionally, adulteration and safety as factors contributing to consistency will be covered. The role of elderberry in enhancing human health alone with the pharmacological basis, the cellular pathways, and the molecular mechanisms underlying the observed health benefits of elderberry fruit extracts will be also reviewed.

Could elderberry fruits processed by modern and conventional drying and extraction technology be considered a valuable source of health-promoting compounds?

The influence of drying and extraction processes on the phytochemical composition and biopotential of elderberry is challenging for the food industry. For this reason in this research two drying techniques (lyophilization and natural convection) and three extraction techniques (ultrasound (UAE), microwave (MAE), and conventional (maceration (MAC)) was applied using two"green" solvents (water and 50 % ethanol). Results of the research showed that lyophilization was a better way of drying than natural convection, while MAE extraction was the most efficient technique for the isolation of secondary metabolites. The most abundant phenolic compounds established by LC-MS/MS analysis, were chlorogenic acid and rutin, identified in extracts of lyophilized elderberries. Elderberry extracts achieved great antioxidant (CUPRAC: 2.30-5.13 mg TE/mL) and enzyme inhibitor potential (α-amylase: 0.51-8.34 mg ACAE/mL). The results suggest that elderberry is a rich natural source of bioactive compounds and could be used for the future development of dietary supplements and functional foods.

Novel insights on extraction and encapsulation techniques of elderberry bioactive compounds.

BACKGROUND: Elderberry (Sambucus nigra L.) has been used in traditional medicine and as a supplement in many beverages and meals. Elderberry is a good source of bioactive flavonoids like quercetin, kaempferol, and rutin, as well as other phenolic compounds. Extraction techniques significantly influence the efficiency of extraction of bioactive compounds. Green chemistry elements such as safety, environmental friendliness, run-down or at least minimal contaminants, efficiency, and economic criteria should all be addressed by an effective bioactive extraction process. Furthermore, micro/nanoencapsulation technologies are particularly effective for increasing bioavailability and bioactive component stability. SCOPE AND APPROACH: This review article comprehensively describes new developments in elderberry extraction and encapsulation. Elderberry is largely employed in the food and pharmaceutical industries due to its health-promoting and sensory characteristics. Elderberry has traditionally been used as a diaphoretic, antipyretic, diuretic, antidepressant, and antitumor agent in folk medicine. KEY FINDINGS AND CONCLUSIONS: Conventional extraction methods (e.g. maceration and Soxhelt extraction) as well as advanced green techniques (e.g. supercritical fluids, pulsed electric field, emulsion liquid extraction, microwave, and ultrasonic extraction) have been used to extract bioactives from elderberry. Over the other protective measures, encapsulation techniques are particularly recommended to protect the bioactive components found in elderberry. Microencapsulation (spray drying, freeze drying, extrusion, emulsion systems) and nanoencapsulation (nanoemulsions, solid lipid nanoparticles and nanodispersions, nanohydrogels, electrospinning, nano spray drying) approaches for elderberry bioactives have been examined in this regard.

Comparison of the Chemical Composition of Selected Varieties of Elderberry with Wild Growing Elderberry.

Elderberries of wild-growing shrubs are most often used; however, various cultivated varieties of this shrub appear more and more often. The aim of this research was to compare the fruit composition of specific varieties with those grown wild in urban and ecologically clean conditions. Six varieties of elderberry grown on one experimental farm and two wild-growing samples from the city center and the landscape park were assessed. The content of vitamin C, antioxidant activity, sugar and organic acid content, triterpenes and carotenoids was marked in the tested fruits. The analyses show that there were significant differences in the content of the tested ingredients between the varieties tested, while the place of cultivation was of less importance. Apart from organic acids and triterpenes, fruits from wild-growing shrubs were more abundant in other compounds determined. The white variety of 'Albida' turned out to be the poorest in bioactive compounds.

Effects of Sambucus ebulus Extract on Cell Proliferation and Viability of Triple- Negative Breast Cancer: An In Vitro and In Vivo Study.

BACKGROUND: Triple-negative breast cancer (TNBC) accounts for 15% of all breast cancer (BC) cases and is a severe type of BC. Since medicinal herbs containing biocompatible substances that are accepted by patient more than chemical therapeutics, they can be considered a safe option for treating BC. OBJECTIVE: This study evaluated the effect of Sambucus Ebulus (S. ebulus) extract on a model of TNBC. METHODS: S. ebulus extract was prepared using petroleum ether, ethyl acetate, and methanol. The petroleum ether extract was fractionated and analyzed using vacuum liquid chromatography and GC-MS, respectively. MDAMB- 231 and MCF-10A were used as TNBC and normal breast cells, respectively. Flowcytometry and MTT assays were performed to evaluate cell cycle, apoptosis, and viability of the cells. Gene expression analysis was performed using RT-qPCR. Nude mouse allograft tumor models were used, and pathological sections were evaluated. RESULTS: The findings indicated that S. ebulus extract remarkably decreased cell proliferation and viability. The extract had no toxicity to the normal breast cells but efficiently killed the cancer cells. Cell cycle- and apoptosisrelated gene expression showed that fraction 4 of S. ebulus extract significantly increased the expression of Bax, Bak, P53, and c-MYC. CONCLUSION: This study showed satisfactory results of the effect of S. ebulus extract on clearing BC cells both in vitro and in vivo. Thus, S. ebulus extract may be a safe herbal compound for eliminating BC cells without toxicity to host cells.

A Pilot, Phase II, Observational, Case-Control, 1-Month Study on Asthenopia in Video Terminal Operators without Dry Eye: Contrast Sensitivity and Quality of Life before and after the Oral Consumption of a Fixed Combination of Zinc, L-Carnitine, Extract of Elderberry, Currant and Extract of Eleutherococcus.

The aims of the study were to investigate the ability and effectiveness of an oral intake of a fixed combination of zinc, L-carnitine, elderberry extract, black currant and Eleutherococcus extract in controlling the symptoms of eyestrain in videoterminal (VDT) users and to record its effects on contrast sensitivity. A single-center, phase II, observational, case-control, 1-month study in VDT workers without dry eye disease was carried out. Demographics and number of actual hours at VDT/day were taken into account. All subjects underwent a complete ophthalmic examination, including assessment of contrast sensitivity, and completed the computer vision symptom scale questionnaire at baseline and one month later. A total of 30 Caucasian subjects adhered to the required inclusion criteria and completed the study; 15 subjects were treated (T) and 15 were controls (C). All clinical data at baseline were similar in both groups (p > 0.05): after one month, all subjects had stable visual acuity, refractive defect and intraocular pressure (IOP); screen exposure time was unchanged. Regarding symptoms, at randomization, the groups had a similar score: 33.1 ± 3.3 in T and 32.8 ± 5.6 in C. One month later, the computer vision symptom scale (CVSS) questionnaire score decreased by -14.1 ± 3, 1 (p = 0.000) and -2.3 ± 1.8 (p = 0.568), respectively. Regarding contrast sensitivity, in group C the values of spatial frequencies remained unchanged, while they improved in almost all the cycles per degree stimuli in the treated group. Oral intake of a fixed combination of zinc, L-carnitine, elderberry extract, black currant and eleutherococcus extract can significantly improve contrast sensitivity and symptoms in VDT workers with no signs of dry eye disease.

Physicochemical, Antioxidant, Microstructural Properties and Bioaccessibility of Dark Chocolate with Plant Extracts.

In this study, dark chocolates (DCh) containing zinc lactate (ZnL) were enriched with extracts from elderberries (EFrE), elderflowers (EFlE), and chokeberries (ChFrE) to improve their functional properties. Both dried plant extracts and chocolates were analyzed for antioxidant capacity (AC) using four different analytical methods: 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), cupric ion-reducing antioxidant capacity (CUPRAC), and ferric-reducing antioxidant power (FRAP), while total phenolic content (TPC) was determined by Folin-Ciocalteu (F-C) assay. An increase in antioxidant properties of fortified chocolates was found, and the bioaccessibility of their antioxidants was evaluated. The highest AC and TPC were found in ChFrE and chocolate with chokeberries (DCh + ChFrE) before and after simulated in vitro digestion. Bioaccessibility studies indicated that during the simulated digestion the AC of all chocolates reduced significantly, whereas insignificant differences in TPC results were observed between chemical and physiological extracts. Moreover, the influence of plant extracts on physicochemical parameters such as moisture content (MC), fat content (FC), and viscosity of chocolates was estimated. Furthermore, scanning electron microscopy with dispersive energy spectroscopy (SEM-EDS) was used to analyze surface properties and differences in the chemical composition of chocolates without and with additives.

Leishmanicidal activities of biosynthesized BaCO3 (witherite) nanoparticles and their biocompatibility with macrophages.

The aim of this study was cost-effective and greener synthesis of barium carbonate (BaCO3 or witherite) nanoparticles with economic importance, and to evaluate their therapeutic potentials and biocompatibility with immune cells. Barium carbonate nanoparticles were biosynthesized using black elderberry extract in one step with non-toxic precursors and simple laboratory conditions; their morphologies and specific structures were analyzed using field emission scanning electron microscopy with energy dispersive X-ray spectroscopy (FESEM-EDX). The therapeutic capabilities of these nanoparticles on the immune cells of murine macrophages J774 and promastigotes Leishmania tropica were evaluated. BaCO3 nanoparticles with IC50 = 46.6 µg/mL were more effective than negative control and glucantium (positive control) in reducing promastigotes (P < 0.01). Additionally, these nanoparticles with a high value of cytotoxicity concentration 50% (CC50) were less toxic to macrophage cells than glucantime; however, they were significantly different at high concentrations compared to the negative control.

Probiotic Yoghurts with Sea Buckthorn, Elderberry, and Sloe Fruit Purees.

Elderberries, sea buckthorn, and sloe berries are fruits of wild-grown bushes, valued in folk medicine for their health-promoting properties but still rarely applied in food. The aim of the present study was to produce probiotic yoghurts with a 10% addition of sweetened purees prepared from elderberries (EPY), sea buckthorn (SBPY), and sloe berries (SPY) and to assess their chemical composition, acidity, content of polyphenols and anthocyanins, ferric reducing antioxidant power (FRAP) and antiradical power (ARP), level of starter microbiota, concentration of acetaldehyde and diacetyl, syneresis, instrumentally measured color and texture parameters, and sensory acceptance. The results were compared to those obtained for plain probiotic yoghurt (PPY) and the changes tracked during 1 month of cold storage at 2 week intervals. The addition of elderberry and sloe berries significantly increased the antioxidant capacity of probiotic yoghurts, probably due to a high content of polyphenols, especially anthocyanins. However, anthocyanins were more stable in the EPY when compared to the SPY. All yoghurt treatments were characterized by good sensory quality and viability of starter microorganisms, including probiotic strains during cold storage. Elderberries promoted the evolution of diacetyl in yoghurts during storage and, together with sloe berries, produced increased syneresis and the greatest changes in color profile compared to PPY.

The lignan-rich fraction from Sambucus Williamsii Hance ameliorates dyslipidemia and insulin resistance and modulates gut microbiota composition in ovariectomized rats.

Menopausal women are susceptible to have high risk of cardiovascular diseases, type II diabetes and osteoporosis due to the metabolic disorder caused by estrogen deficiency. Accumulating evidence supports that gut microbiota is a key regulator of metabolic diseases. Our previous metabolomics study interestingly demonstrated that the anti-osteoporotic effects of lignan-rich fraction (SWCA) from Sambucus wialliamsii Hance were related to the restoration of a series of lipid and glucose metabolites. This study aims to investigate how SWCA modulates lipid and glucose metabolism and the underlying mechanism. Our results show that oral administration of SWCA (140 mg/kg and 280 mg/kg) for 10 weeks alleviated dyslipidemia, improved liver functions, prevented glucose tolerance and insulin actions, attenuated system inflammation and improved intestinal barrier in OVX rats. It also induced a high abundance of Actinobacteria, and restored microbial composition. We are the first to report the protective effects of the lignan-rich fraction from S. williamsii on dyslipidemia and insulin resistance. Our findings provide strong evidence for the application of this lignan-rich fraction to treat menopausal lipid disorder and insulin resistance-related diseases.

Fluorescent Products of Anthocyanidin and Anthocyanin Oxidation.

We found that the oxidation of cyanidin with hydrogen peroxide, tert-butyl hydroperoxide, 2,2'-azobis(2-methylpropionamidine), 3-morpholinosydnonimine hydrochloride, sodium hypochlorite, Fe3+, and Fe2+ induced the appearance of a new fluorescence band, centered at 525-540 nm when excited at 470-490 nm. The intensity of this fluorescence was related to the oxidant concentration. The same fluorescence was induced by the oxidation of other anthocyanidins, an anthocyanin (cyanidin-3-O-glucoside) as well as anthocyanin-rich red cabbage extract, and blackcurrant and elderberry juices. Peroxidized blood plasma also induced the appearance of cyanidin fluorescence. We also showed that the formation of the fluorescent product of pelargonidin can be observed in a culture of the MRC-5 human fetal lung fibroblast cell line and the DU-145 human prostate carcinoma cell line. Our results suggest that this new anthocyanidin/anthocyanin fluorescence may be an indicator of oxidation, especially of food products, where these compounds are present or added as colorants, and can also be useful to detect oxidation in biomedical experiments.

Seven recommendations to rescue the patients and reduce the mortality from COVID-19 infection: An immunological point of view.

Now COVID-19 is causing a severe public health emergency and the mortality is rapidly increasing all over the world. In the current pandemic era, although there have been many efforts to diagnose a number of patients with symptoms or close contacts, there is no definite guideline for the initial therapeutic approach for them and therefore, many patients have been dying due to a hyperinflammatory immunological reaction labeled as "cytokine storm". Severe patients are hospitalized and the treatment is done, though they have not been established yet. Currently, however, no treatment is provided for those who are isolated at home or shelter until they get severe symptoms, which will increase the harms to the patients. In this review, we discuss some important points dedicated to the management of patients with COVID-19, which should help reducing morbidity and mortality. In this era, we suggest 7 recommendations to rescue the patients and to reduce the morbidity and mortality due to COVID-19 based on the immunological point of view.

Effect of harvesting year and elderberry cultivar on the chemical composition and potential bioactivity: A three-year study.

Sambucus nigra is one of the richest sources of anthocyanins and other polyphenols being used industrially as a source of antioxidants, colorants, and bioactives. Although cultivars can influence elderberry composition, no study has addressed the effect of harvesting year on elderberries composition and bioactivity. The composition of the main Portuguese cultivars, "Sabugueiro", "Sabugueira" and "Bastardeira", were evaluated during three consecutive years. Harvesting year had a stronger influence on the chemical composition than cultivars, including total sugars, anthocyanins, and phenolic compounds, being related to the different climatic conditions, especially water status. "Bastardeira" was the best cultivar concerning total soluble solids, anthocyanins, polyphenols, and antioxidant activity, but containing a lower total free sugar content compared to "Sabugueiro". The results obtained in this study provide novel information from a nutritional perspective and for breeding programs aiming to select cultivars with enhanced levels of health-promoting compounds or for other industrial applications of elderberries.

HPLC-DAD-ESI-MS Analysis for Simultaneous Quantitation of Phenolics in Taiwan Elderberry and Its Anti-Glycation Activity.

Sambucus formosana is most commonly used as a traditional herb medicine in Taiwan. In this study, high performance liquid chromatography equipped with photodiode array detection-mass (HPLC-DAD-ESI-MS) method was developed for the identification and quantification of bioactive phenolics. The developed method was also validated for accuracy, precision, limit of detection, and quantification. In this method, chlorogenic acid, rutin, isoquercetrin, nictoflorin, astragalin, and quercetin were quantified in linearity range of 10-100 (µg/mL) with a correlation coefficient of greater than 0.996. High recovery (86.5-93.1%) and good reproducibility were obtained for six phenolics with the relative standard deviation ranging from 1.7-3.1%. Therefore, the proposed method for simultaneous quantification of six bioactive phenolics in the extract and fractions of S. formosana using HPLC-DAD-ESI-MS detection under the optimized conditions is accurate and validated. Among the results, methanol extract showed the greatest values of total phenolic content (93.1 mg gallic acid equivalent/g). Additionally, the methanol extract revealed best antioxidant capacity based on the DPPH scavenging activity and anti-glycation activity (IC50 was observed at 97.1 and 77.9 µg/mL, respectively).

Antiviral activity of Sambucus FormosanaNakai ethanol extract and related phenolic acid constituents against human coronavirus NL63.

Human coronavirus NL63 (HCoV-NL63), one of the main circulating HCoVs worldwide, causes respiratory tract illnesses like runny nose, cough, bronchiolitis and pneumonia. Recently, a severe respiratory illness outbreak of HCoV-NL63 has been reported in a long-term care facility. Sambucus FormosanaNakai, a species of elderberry, is a traditional medicinal herb with anti-inflammatory and antiviral potential. The study investigated the antiviral activity of Sambucus FormosanaNakai stem ethanol extract and some phenolic acid constituents against HCoV-NL63. The extract was less cytotoxic and concentration-dependently increased anti-HCoV-NL63 activities, including cytopathicity, sub-G1 fraction, virus yield (IC50 = 1.17 µg/ml), plaque formation (IC50 = 4.67 µg/ml) and virus attachment (IC50 = 15.75 µg/ml). Among the phenolic acid constituents in Sambucus FormosanaNakai extract, caffeic acid, chlorogenic acid and gallic acid sustained the anti-HCoV-NL63 activity that was ranked in the following order of virus yield reduction: caffeic acid (IC50 = 3.54 µM) > chlorogenic acid (IC50 = 43.45 µM) > coumaric acid (IC50 = 71.48 µM). Caffeic acid significantly inhibited the replication of HCoV-NL63 in a cell-type independent manner, and specifically blocked virus attachment (IC50 = 8.1 µM). Therefore, the results revealed that Sambucus Formosana Nakai stem ethanol extract displayed the strong anti-HCoV-NL63 potential; caffeic acid could be the vital component with anti-HCoV-NL63 activity. The finding could be helpful for developing antivirals against HCoV-NL63.

NMR Applications for Botanical Mixtures: The Use of HSQC Data to Determine Lignan Content in Sambucus williamsii.

Lignans found in the botanical extract of the Traditional Chinese Medicine Sambucus williamsii Hance exhibit protective effects on trabecular bone mass and mechanical strength of cortical bone of ovariectomized rats. A novel approach was adapted using HSQC NMR methods to estimate the total amount of these bioactives in a complex mixture. It was determined that lignans possessing the hydroxy- or oxybenzyl carbon signal were bioactive. These compounds were readily identified and assigned in a defined region of the 13C NMR spectrum at 80-90 ppm and calculated as 10-15% of the lignan-rich fraction of S. williamsii. Comparison of the peak heights of the oxybenzyl-substituted carbon resonance signals of the lignans in the botanical extract was made against those of a standard lignan pinoresinol. The application of this simple and reliable NMR method can be used to estimate amounts of related compounds and chemical families in complex mixtures or botanical extracts and offers measurable scientific evidence in quality processes. This is of particular importance for registration requirements of botanical drugs and in complex mixtures of botanical extracts.

Changes in the phenolic compositions of Elaeagnus umbellata and Sambucus lanceolata after in vitro gastrointestinal digestion and evaluation of their potential anti-diabetic properties.

In this work, the phenolic composition of E. umbellata leaves and berries is reported. Berries were rich in flavonols, whereas leaves presented abundant flavonols and ellagitannins. Then, the enzyme-inhibitory properties, anti-glycation and antioxidant activities of E. umbellata and Sambucus lanceolata (its phenolic profile has been already established in a previous work) were tested by several in vitro assays and compared. The simulated gastrointestinal digestion resulted in a decrease on their phenolic composition. Nevertheless, both species still had the ability to inhibit aldose reductase activity and protein glycation and scavenge free radicals at the end of the process. Thus, the potential beneficial effects of E. umbellata and S. lanceolata seems to be kept to some extent after passage throughout the digestive system. Altogether, this study provides further insight into investigation of these species as dietary sources of bioactive compounds to lower the risk of type-2 diabetes and obesity.