Water extract of Cryphaea heteromalla (Hedw.) D. Mohr bryophyte as a natural powerful source of biologically active compounds

Bryophytes comprise of the mosses, liverworts, and hornworts. Cryphaea heteromalla, (Hedw.) D. Mohr, is a non-vascular lower plant belonging to mosses group. To the date, the most chemically characterized species belong to the liverworts, while only 3.2% and 8.8% of the species belonging to the mosses and hornworts, respectively, have been investigated. In this work, we present Folin–Ciocalteu and oxygen radical absorbance capacity (ORAC) data related to crude extracts of C. heteromalla obtained by three different extraction solvents: pure water (WT), methanol:water (80:20 v/v) (MET), and ethanol:water (80:20 v/v) (ETH). The water extract proved to be the best solvent showing the highest content of biophenols and the highest ORAC value. The C. heteromalla-WT extract was investigated by HPLC-TOF/MS (High Performance Liquid Chromatography-Time of Flight/Mass Spectrometry) allowing for the detection of 14 compounds, five of which were phenolic compounds, derivatives of benzoic, caffeic, and coumaric acids. Moreover, the C. heteromalla WT extract showed a protective effect against reactive oxygen species (ROS) generation induced by tert-butyl hydroperoxide (TBH) on the murine NIH-3T3 fibroblast cell line

HPLC-ESI-TOF-MS chemical characterization of comfrey root extract obtained by subcritical water extraction

In this work, a study on phytochemical profiles of comfrey (Symphytum officinale L.) root extract obtained by subcritical water extraction (SWE) has been carried out. Chemical composition was assessed by high-performance liquid chromatography coupled with electrospray time-of-flight mass spectrometry (HPLC-ESI-TOF-MS) identifying 22 compounds including organic acids, phenolic acids, flavonoids and fatty acids. Great number of phenolic acids and flavonoids were found in the extract obtained by SWE, with citric acid, caffeic acid and derivative, salvianolic acid B, hydroxybenzoic acid, syringetin-3-O-glucoside and quercetin 3-O-malonylglucoside as the most abundant compounds. Moreover, quercitin-3- malonylglucoside isomers, hydroxybenzoic acid glucoside, cirismaritin isomers, p-coumaric acid, hydroxycoumarin and methylcoumarin, among others, were identified for the first time in S. officinale root. Overall, the results indicate the potential of SWE for the production of highquality plant extracts from S. officinale root

HPLC-DAD-ESI-QTOF-MS and HPLC-FLD-MS as valuable tools for the determination of phenolic and other polar compounds in the edible part and by-products of avocado

Avocado is a tropical fruit increasingly cultivated around the world due to global interest and rising consumption. Thus, there is also a surge in avocado by-products that needs assessment. The aim of this work is to compare the phenolic profile of avocado pulp, peel and seed when the fruit is at optimal ripeness for consumption and when overripe. Two analytical techniques were used: (1) HPLC-DAD-ESI-QTOF-MS was used for the first time to determine phenolic and other polar compounds in avocado peel and seed. Phenolic compounds quantified with these methods were in higher concentration in overripe than in pulp and seed of optimally ripe fruit. (2) HPLC-FLD-MS was used to specifically determine flavan-3-ols. Procyanidins to degree of polymerization 13 have been quantified singularly here for the first time. In addition, A- and B-type procyanidins from the degree of polymerization 2 to 6 were differentiated and quantified. The procyanidin concentration increased after ripening probably due to the release of tannins linked to cell-wall structures. Because of this situation and the presence of A-type procyanidins, avocado peel and seed from overripe fruit, the main by-products of avocado processing, hold interest for developing functional foods, nutraceuticals and cosmetics

Structure–Biological Activity Relationships of Extra-Virgin Olive Oil Phenolic Compounds: Health Properties and Bioavailability

Extra-virgin olive oil is regarded as functional food since epidemiological studies and multidisciplinary research have reported convincing evidence that its intake affects beneficially one or more target functions in the body, improves health, and reduces the risk of disease. Its health properties have been related to the major and minor fractions of extra-virgin olive oil. Among olive oil chemical composition, the phenolic fraction has received considerable attention due to its bioactivity in different chronic diseases. The bioactivity of the phenolic compounds could be related to different properties such as antioxidant and anti-inflammatory, although the molecular mechanism of these compounds in relation to many diseases could have different cellular targets. The aim of this review is focused on the extra-virgin olive oil phenolic fraction with particular emphasis on (a) biosynthesis, chemical structure, and influence factors on the final extra-virgin olive oil phenolic composition; (b) structure–antioxidant activity relationships and other molecular mechanisms in relation to many diseases; (c) bioavailability and controlled delivery strategies; (d) alternative sources of olive biophenols. To achieve this goal, a comprehensive review was developed, with particular emphasis on in vitro and in vivo assays as well as clinical trials. This report provides an overview of extra-virgin olive oil phenolic compounds as a tool for functional food, nutraceutical, and pharmaceutical applications.Junta de Andalucía B-AGR-466-UGR1

Uptake and metabolism of olive oil polyphenols in human breast cancer cells using nano-liquid chromatography coupled to electrospray ionization–time of flight-mass spectrometry

Polyphenols from extra virgin olive oil (EVOO), a main component of the Mediterranean diet, have demonstrated repeatedly anti-tumor activity in several in vitro and in vivo studies. However, little is known about the efficiency of the absorption process and metabolic conversion of these compounds at cellular level. In this study, a nano liquid chromatography–electrospray ionization–time of flight mass spectrometry (nanoLC–ESI–TOF MS) method was developed to study the cellular uptake and metabolism of olive oil phenols in JIMT-1 human breast cancer cells. After incubation for different time periods with EVOO-derived phenolic extracts, culture media, cytosolic fraction and solid particles fraction were separated and analyzed. Most of the free phenols, mainly hydroxytyrosol, its secoiridoid derivatives, and the flavonoid luteolin, disappeared in the culture media in different ways and at different times. Besides, several metabolites were detected in the culture media, fact that may indicate absorption and intracellular metabolism followed by rapid cellular export. Low intracellular accumulation was observed with only traces of some compounds detected in the cytosolic and solid particles fractions. Methylated conjugates were the major metabolites detected, suggesting a catalytic action of catechol-O-methyl transferase (COMT) in cancer cells.The authors are very grateful to Ministry of Education and Science (FPU, AP2005-4356) and Junta de Andalucia (project P09-FQM-5469, project P07-AGR-02619 and AGL 2008-05108- CO3-03/ALI). Besides, this work was supported in part by Instituto de Salud Carlos III (Ministerio de Sanidad y Consumo, Fondo de Investigación Sanitaria – FIS, Spain, Grants CP05-00090, PI06-0778 and RD06-0020-0028)

Exploratory analysis of human urine by LC–ESI-TOF MS after high intake of olive oil: understanding the metabolism of polyphenols

Olive oil polyphenols have important biological properties which closely depend on their bioavailability, therefore it is essential to understand how polyphenols are absorbed, metabolized and eliminated from the body. An analytical methodology based on rapid resolution liquid chromatography (RRLC) coupled to mass spectrometry detection with a time of flight analyzer (RRLC-ESI-TOF MS) was developed for the determination of the main olive oil phenolic compounds and their metabolites in human urine. Urine samples from ten healthy volunteers were collected before and 2, 4 and 6 h after the intake of 50 mL of extra-virgin olive oil. The proposed method includes liquid-liquid extraction with ethyl acetate that provides extraction recoveries of the phenolic compounds studied between 35 and 75% from spiked urine samples. Good repeatability was obtained, since the relative standard deviations (RSDs) of peak areas in the intra- and inter-day studies were 4.3 and 6.5%, respectively. Statistical studies allowed us to discriminate between the urine samples before and after the intake, and facilitated to find out the m/z values responsible of this discrimination. Based on the very accurate mass information and the isotopic pattern provided by TOF-MS analyzer, together with other available information, ten of these biomarkers and more than 50 metabolites, obtained through phase I and phase II biotransformation reactions, were tentatively identified. Additionally, kinetic studies of the metabolites identified as possible biomarkers were developed, obtaining maximal values in the first two hours for most compounds.The authors are very grateful to Ministry of Education and Science (FPU, AP2005-4356 and Project AGL 2008- 05108-613 CO3-03/ALI), and Junta de Andalucía (P07-AGR-02629 and P09-FQM-5469)

Incorporation of lippia citriodora microwave extract into total-green biogelatin-phospholipid vesicles to improve its antioxidant activity

Phytochemicals from Lippia citriodora leaves were extracted by applying an innovative technology based on the use of microwaves, which represents an alternative method to extract bioactive substances. The obtained extract was incorporated into phospholipid vesicles in order to promote the antioxidant effect of the bioactive molecules present in L. citriodora extract. The extract was analyzed by High Performance Liquid Chromatography coupled to Time-Of-Flight mass spectrometer by electrospray (HPLC-ESI-TOF-MS) and different phytochemicals were detected and quantified. The whole extract was incorporated in liposomes, glycerosomes (liposomes modified with glycerol) and propylene glycol-containing vesicles (PG-PEVs). Moreover, a biopolymer obtained from fish by-product, that is Thunnus albacares skin, was added to improve the bioactivity of the formulations. The in vitro biocompatibility and the antioxidant efficacy of the extract in solution or loaded in the vesicles were tested in primary mouse embryonic fibroblasts (3T3). The results showed the superior bioactivity of the vesicle formulations over the aqueous solution of the extract, which points to an interesting strategy for the treatment of skin disorders

Extraction of the antioxidant phytocomplex from wine-making by-products and sustainable loading in phospholipid vesicles specifically tailored for skin protection

The present study is aimed at valorizing grape pomace, one of the most abundant winery-making by-products of the Mediterranean area, through the extraction of the main bioactive compounds from the skin of grape pomace and using them to manufacture innovative nanoformulations capable of both avoiding skin damages and promoting skincare. The phytochemicals were recovered through maceration in hydroethanolic solution. Catechin, quercetin, fisetin and gallic acid, which are known for their antioxidant power, were detected as the main compounds of the extract. Liposomes and phospholipid vesicles modified with glycerol or Montanov 82® or a combination of both, were used as carriers for the extract. The vesicles were small (~183 nm), slightly polydispersed (PI ≥ 0.28), and highly negatively charged (~−50 mV). The extract was loaded in high amounts in all vesicles (~100%) irrespective of their composition. The antioxidant activity of the extract, measured by using the DPPH (2,2-Diphenyl-1-picrylhydrazyl) test, was 84 ± 1%, and slightly increased when loaded into the vesicles (~89%, P < 0.05). The grape pomace extract loaded vesicles were highly biocompatible and able to protect fibroblasts (3T3) from the oxidative stress induced by hydrogen peroxide

Recovering Bioactive Compounds from Olive Oil Filter Cake by Advanced Extraction Techniques

The potential of by-products generated during extra-virgin olive oil (EVOO) filtration as a natural source of phenolic compounds (with demonstrated bioactivity) has been evaluated using pressurized liquid extraction (PLE) and considering mixtures of two GRAS (generally recognized as safe) solvents (ethanol and water) at temperatures ranging from 40 to 175 °C. The extracts were characterized by high-performance liquid chromatography (HPLC) coupled to diode array detection (DAD) and electrospray time-of-flight mass spectrometry (HPLC-DAD-ESI-TOF/MS) to determine the phenolic-composition of the filter cake. The best isolation procedure to extract the phenolic fraction from the filter cake was accomplished using ethanol and water (50:50, v/v) at 120 °C. The main phenolic compounds identified in the samples were characterized as phenolic alcohols or derivatives (hydroxytyrosol and its oxidation product), secoiridoids (decarboxymethylated and hydroxylated forms of oleuropein and ligstroside aglycones), flavones (luteolin and apigenin) and elenolic acid derivatives. The PLE extraction process can be applied to produce enriched extracts with applications as bioactive food ingredients, as well as nutraceuticals

The role of high-resolution analytical techniques in the development of functional foods

The approaches based on high-resolution analytical techniques, such as nuclear magnetic resonance or mass spectrometry coupled to chromatographic techniques, have a determining role in several of the stages necessary for the development of functional foods. The analyses of botanical extracts rich in bioactive compounds is one of the fundamental steps in order to identify and quantify their phytochemical composition. However, the compounds characterized in the extracts are not always responsible for the bioactive properties because they generally undergo metabolic reactions before reaching the therapeutic targets. For this reason, analytical techniques are also applied to analyze biological samples to know the bioavailability, pharmacokinetics and/or metabolism of the compounds ingested by animal or human models in nutritional intervention studies. In addition, these studies have also been applied to determine changes of endogenous metabolites caused by prolonged intake of compounds with bioactive potential. This review aims to describe the main types and modes of application of high-resolution analytical techniques in all these steps for functional food development