Elucidation and quantification health-promoting phenolic compounds, antioxidant properties and sugar levels of ultrasound assisted extraction, aroma compositions and amino acids profiles of macroalgae, Laurencia papillosa.

Currently, sustainability is one of the most critical issues confronting society today. The growing of macroalgae in ocean farms appears more sustainable than agriculture on land due to it does not require any fresh water, chemical fertiliser, or soil. Macroalgae have been shown to be a sustainable marine source of amino acids, novel bioactive phenolic and aroma compounds that can be exploitation in food, cosmetic, nutraceuticals, pharmacological applications. Despite starting the huge cultivation of macroalgae in world, bioactive compounds in the edible macroalgae have not been well characterized. Ultrasound assisted extraction (UAE) and conventional extraction (CE) techniques were compared and red macroalgae, L. papillosa extracts were characterized. The highest amount of amino acid was glutamic acid (GLU) and composed of 35% was essential amino acids. UAE at 10% amplitude for 15 min showed significantly highest (p < 0.05) phenolic (212.03±3.03 mg gallic acid equivalent/100 g) as well as antioxidant activity determined by DPPH (105.69±3.02 µmol Trolox/100 g), ABTS (238.69±2.23 µmol Trolox/100 g) radical assay and FRAP value (72.47±3.13 µmol Trolox/100 g) when in comparison with CE. Furthermore, bioactive compounds in extracts were indicated as phlorotannins, flavonoids, phenolic acids and other polyphenols using liquid chromatography coupled to diode array detection and electrospray ionisation tandem mass spectrometry (LC-DAD-ESI-MS/MS). This result confirmed higher antioxidant capacity detected with the UAE. A total of 46 volatile organic compounds were identified and quantified by GC-FID/MS with HS-SPME system. This study emerges as first report to novel extraction method used and deeply characterization of L papillosa. The results seem that significant potential application in the functional food, active packaging and nutraceuticals industry.

HPLC-DAD-ESI/MS profiles of bioactive compounds, antioxidant and anticholinesterase activities of Ephedra alata subsp. alenda growing in Algeria.

Ephedra (Ephedraceae) is used in medicine for various purposes as having, antioxidant, anticarcinogen, antibacterial, anti-inflammatory hepatoprotective, anti-obesity, antiviral and diuretic activities. In this study the aim was to investigate chemical constituents of Ephedra alata and understand the possible effects of those constituents in antioxidant activity and alzheimer's disease essay. For this purpose, natural compounds from E.alata were characterized by LC-DAD-ESI-MS/MS using negative and positive ionization modes, while the bioactivity was assessed by acetylcholinesterase (AChE) inhibition study and determining of antioxidant activity; DPPH radical scavenging and ß-carotene bleaching assays were used to assess the antioxidant potential. The proposed method of spectrometry provided tentative identification of 27 compounds including alkaloids and phenolic compounds as flavonoids. The methanolic extract showed high contents of total phenolic and exhibited an important antioxidant potential and demonstrated a potent inhibitory effect against acetylcholinesterase (IC50: 11,25 ± 0,25 µg/mL). The results showed that the plant possesses a therapeutic effect.

LC-DAD/ESI-MS/MS characterization of phenolic constituents in Rosa canina L. and its protective effect in cells.

In an aim to prove the efficiency of polyphenols of Rosa canina fruits in promoting human health. A methanolic extract of R. canina fruits was prepared by successive maceration with solvents of increasing polarity. The polyphenol composition was analyzed by HPLC-DAD-ESI-MS. The biological activity of this extract on SH-SY5Y cells and HepG2 cells was then studied. The antioxidant activity was tested by various in vitro tests such as DPPH-radical-scavenging activity, FRAP assay, hydroxyl radical scavenging assay and total antioxidant capacity. The subacute toxicity of R. canina was tested on female rats by repeated intraperitoneal administration of various doses. The phenolic profiles showed 25 antioxidants distributed into three classes of phenolic compounds: glycosylated and agglomerated flavonoids/isoflavonoids, tannins and phenanthrenes. Qualitative phytochemical analyses showed that this extract lacks alkaloids. The methanolic extract of R. canina fruits has a total antioxidant capacity of 82.69 ± 1.18 µg EAA/mg of methanol extract and the IC50 of the methods used is in the following increasing order: FRAP assay (61.88 µg/ml), then hydroxyl radical scavenging assay (67.45 µg/ml) and then DPPH radical-scavenging activity (129.81 µg/ml). The extract of R. canina did not cause any phenotypic signs of toxicity or mortality during and after treatment. The LD50 was >5,000 mg/kg, hence, R. canina was considered nontoxic. An in vivo study proved the protective effect of R. canina against cardiac and hepato-renal toxicities. These results drew the importance of a healthy diet, where diets rich in R. canina fruits can be used as a rich natural source of antioxidants and anticarcinogenic phenolic compounds.

LC-DAD-ESI-MS/MS-based phenolic profiling and antioxidant activity in Turkish cv. Nizip Yaglik olive oils from different maturity olives.

The current study was designed to find out how olive maturity indices (2.5, 3.5, and 4.5) affect the individual phenolic compounds and antioxidant potencies of olive oils produced from cv. Nizip Yaglik olives. Liquid chromatography coupled to diode array detection and electrospray ionization tandem mass spectrometry in multiple reaction monitoring mode was utilized for the determination of phenolic composition qualitatively and quantitatively. Findings asserted a quite similar phenolic profile (14 phenols) depending on the various phenolic groups in all oils, while the concentration of total and individual phenolic compounds revealed significant differences between the samples statistically (p < 0.05). Among the individual phenolic classes in all samples, secoiridoids were the most prevailing group and their total content showed a clear significant decline as the olive fruits get ripened. Antioxidant potency values showed a clear diminution attitude during the maturation of the olives. The principal component analysis revealed that oils were discriminated from each other according to phenolic compounds and antioxidant potencies. Moreover, oils obtained from the unripe and medium-ripe fruits possessed a very good quality marked by their elevated phenolic levels.

Effect of hulling methods and roasting treatment on phenolic compounds and physicochemical properties of cultivars 'Ohadi' and 'Uzun' pistachios (Pistacia vera L.).

The aim of this study was to identify phenolic compounds and measure the physicochemical properties of two pistachio (Pistacia vera L.) cultivars ('Uzun' and 'Ohadi'). For this purpose, the pistachios were subjected to four different hulling methods (wet hulling-WT, brine hulling-BR, wet-dry hulling-WD, and dry-dry hulling-DD) and a standard roasting treatment. Phenolic compounds were analyzed by HPLC-DAD-ESI-MSn. Twelve phenolics were identified and quantified in the samples, 10 of which were flavonoid and two non-flavonoid phenolic acids. In both cultivars, the major compounds were found to be gallic acid and catechin followed by eriodictyol-7-O-glucoside and eriodictyol. The dry-dry hulling method treated sample had higher antioxidant capacity, total phenol content, and phenolic compounds than the other hulling methods in both pistachio cultivars. After the roasting treatment, the antioxidant capacity, phenolic content, and a∗ value increased while the L∗ and b∗ values decreased.

Optimization and validation of reverse phase HPLC method for qualitative and quantitative assessment of polyphenols in seaweed.

A simple reverse phase-high performance liquid chromatography (RP-HPLC) coupled to a diode array detector (DAD) and negative ion electrospray mass spectrometer (ESI-MS) method was developed for simultaneous identification and quantification of phenolic antioxidants in seaweed. The proposed method was validated in terms of linearity, limits of detection (LOD), limits of quantification (LOQ), recovery and intermediate precision. The calibration curves were linear with correlation coefficient ranging from 0.9909 to 0.9997 while the values of LOD (0.26-0.82mg/L), LOQ (0.77-2.50mg/L), recovery (≥97.2%) and precision in terms of retention time (%RSD ≤2.27) and peak area (% RSD ≤5.11) were satisfactory. Brown seaweed Himanthalia elongata used in this study was extracted with 60% methanol and the crude extract was cleaned with SPE (Solid Phase Extraction) cartridge. HPLC-DAD-MS/MS analysis of the SPE fraction allowed the identification of 7 phenolic compounds comprising phlorotannins, hydroxybenzoic acid, hydroxycinnamic acid and flavonols subclasses of polyphenols. Quantitative analysis of these compounds revealed the presence of phloroglucinol (394.1±4.33µg/g), gallic acid (96.3±3.12µg/g), chlorogenic acid (38.8±1.94µg/g), caffeic acid (44.4±2.72µg/g), ferulic acid (17.6±0.85µg/g), myricetin (8.6±0.85µg/g) and quercetin (4.2±0.15µg/g), in the extract. The SPE fraction were tested for antioxidant capacity which were significantly (P <0.05) higher (EC50; 14.5±0.57mg/g) than the ascorbic acid (EC50; 35.8±0.59mg/g) and the crude extract (EC50; 46.3±0.48mg/g). The occurrence of all these phenolic antioxidant compounds in H. elongata extract suggested that the developed method is sensitive enough and reproducible and could be used for qualitative and quantitative assessment of polyphenols in seaweed.

HPLC-DAD-ESI-MS/MS analysis of fruits from Firmiana simplex (L.) and evaluation of their antioxidant and antigenotoxic properties.

OBJECTIVES: The secondary metabolites of the fruits of Firmiana simplex (L.) were analysed by LC-DAD-ESI-MS/MS; furthermore, we evaluated their antioxidant and antigenotoxic properties. METHODS: The antioxidant activity was investigated using the 2,2'-diphenyl-1-picrylhydrazyl radical (DPPH), the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS) and the ferric reducing antioxidant power (FRAP) assays. The antigenotoxic potential was determined via the comet assay. KEY FINDINGS: The ethyl acetate fraction (EtOAc) was analysed by LC-DAD-ESI-MS/MS: phenolic acids and flavonoids were the main polyphenols of the fruits. The EtOAc fraction yielded the highest content of polyphenols with 314.61 mg GAE/g extract, followed by 297.51, 153.75, 101.47, 97.19 for dichloromethane, butanol, methanol and water extracts, respectively. As expected, a strong correlation exists between the antioxidant activity of the investigated extracts and their total phenolic content. In the DPPH assay, the IC50 value of the most active EtOAc fraction was 6.79 µg/ml, relative to 2.92 µg/ml of the standard ascorbic acid. ABTS and FRAP assays supported the results of DPPH assay. Moreover, using the comet assay, we could show that the phenol-rich EtOAc extract exhibits an antigenotoxic potential in human liver cancer cells (Hep-G2) treated with hydrogen peroxide (H2 O2 ) as a genotoxic agent. CONCLUSIONS: The fruits of Firmiana simplex may be a good natural source of antioxidant and antigenotoxic agents.

Characterization of phenolic compounds in chia (Salvia hispanica L.) seeds, fiber flour and oil.

The consumption of chia seeds products has increased recently and it has been suggested that the inclusion of this functional food in a daily human diet could contribute to improve consumers' health. However, a better knowledge about the composition of these products is mandatory. In this work, the phenolic compounds from commercial samples of chia seed, fiber flour and oil were extracted using an ultrasound-assisted methodology and were separated and identified by high-performance liquid chromatography coupled to a mass spectrometer. Methanol:water extracts were prepared and submitted to an acidic hydrolysis. Crude and hydrolyzed extracts were analyzed and phenolic compounds found were mainly caffeic acid and danshensu and its derivatives, such as rosmarinic and salvianolic acids. TPC was higher in the hydrolyzed extracts. These results supply new information about the main phenolic compounds presents in chia, which are important dietary sources of natural antioxidants for prevention of diseases caused by oxidative stress.

LC-MS phenolic profiling combined with multivariate analysis as an approach for the characterization of extra virgin olive oils of four rare Tunisian cultivars during ripening.

In this work, the phenolic composition of four rare cultivars grown under the same agronomical and environmental conditions was studied. This is to test the effects of cultivars and ripening index essentially on phenolic composition in olive oils as well as tocopherols composition, organoleptic profiling and oxidative properties. Furthermore, some agronomical traits were determined in which a general increase in the size of the fruit and oil contents were recorded for all cultivars. The phenolic fractions were identified and quantified using liquid chromatography coupled to diode array detection and electrospray ionization tandem mass spectrometry (LC-DAD-ESI-MS/MS) in multiple reaction monitoring mode (MRM). A total of 13 phenolic compounds belonging to different chemical families were determined. Qualitative and quantitative differences in phenolic composition were observed among cultivars and also among sampling times. On the contrary to the agronomical traits, a general decrease (p<0.05) of total phenolic compounds was observed during maturation. Likewise, a decrease in tocopherols concentrations and oxidative properties was observed.

Contribution to the characterization of Opuntia spp. juices by LC-DAD-ESI-MS/MS.

Opuntia spp. fruits are considered as health promoting foods due to the diversity of bioactive molecules found in these fruits. The composition in organic acids, flavonols and betalains in the Opuntia ficus-indica juice from a region of Portugal was accomplished for the first time by liquid chromatography and tandem mass spectrometry using an electrospray ionization source operating in negative and positive mode. The methodology used allowed the detection of 44 compounds, from which 32 were identified. Isorhamnetin derivatives were the dominant flavonol glycosides. A total of 9 betalains including 6 betaxanthins and 3 betacyanin were also detected in the fruit juice samples and indicaxanthin, betanin and isobetanin were the major pigments. Phenolic acid and phenylpyruvic acid derivatives were also identified. To our knowledge, it is the first time derivative compounds from piscidic acid, phenolic compounds and betalains are characterized in cactus pear juice using a single LC-DAD-ESI-MS/MS method.

Fructus Psoraleae contains natural compounds with potent inhibitory effects towards human carboxylesterase 2.

Fructus Psoraleae (FP) is an edible Chinese herbal which is widely used in Asia for the treatment of various diseases including asthma, diarrhea, and osteoporosis. This study aimed to investigate the inhibitory effects of the crude ethanol extract from FP on human carboxylesterase 2 (hCE2), as well as to identity and characterize the naturally occurring inhibitors of hCE2 in FP. Our results demonstrated that the ethanol extract of FP displayed potent inhibitory effects towards hCE2, while five major bioactive constitutes in FP were efficiently identified by LC-DAD-ESI-MS/MS, with the aid of LC-based activity profiling. The identified bioactive compounds including neobavaisoflavone, isobavachalcone, bavachinin, corylifol A and bakuchiol were found to be naturally occurring potent inhibitors of hCE2, with low Ki values ranging from 0.62µM to 3.89µM. This is the first report of the chemical constitutes in FP as potent inhibitors of hCE2.