Identification Of Phenolic Acids Research Articles

Investigation the biological activities and the metabolite profiles of endophytic fungi isolated from Gundelia tournefortii L.

Endophytic fungi are microorganisms that are considered as a potential source of natural compounds, and can be applied in various industries. The aims of this research were molecular identification of endophytic fungi isolated from the Gundelia tournefortii stems, and investigation their biological activities as well as phenolic and fatty acid profile. Surface sterilized stems of G. tournefortii were placed on potato dextrose agar (PDA) to isolate the fungal endophytes. Genomic DNA was extracted by CTAB method, and PCR amplification was performed by ITS 1 and ITS 4 as primers. The enzyme production of endophytic fungi was determined based on the formation of a clear zone that appeared around the colonies of fungus. The anti-oxidant activity was evaluated by measuring the amount of free radicals DPPH. Also, the total phenol and flavonoid contents were measured obtained by Folin-Ciocalteu and aluminum chloride colorimetric methods, respectively. Moreover, the separation and identification of phenolic acids and fatty acids were done by HPLC and GC, respectively. Phylogenetic analysis was done based on the Internal Transcribed Spacer (ITS) region, and five isolates were identified as following: Aspergillus niger, Penicillium glabrum, Alternaria alternata, A. tenuissima, and Mucor circinelloides. Evaluation of the enzymatic properties showed that P. gabrum (31 ± 1.9 mm), and A. niger (23 ± 1.7) had more ability for producing pectinase and cellulase. The anti-oxidant activity of isolates showed that A. alternata extract (IC50 = 471 ± 29 µg/mL) had the highest anti-oxidant properties, followed by A. tenuissima extract (IC50 = 512 ± 19 µg/mL). Also, the extract of A. alternata had the greatest amount of total phenols and flavonoids contents (8.2 ± 0.4 mg GAL/g and 2.3 ± 0.3 mg QE/g, respectively). The quantification analysis of phenolic acid showed that rosmarinic acid, para-coumaric acid, and meta-coumaric acid (42.02 ± 1.31, 7.53 ± 0.19, 5.41 ± 0.21 mg/g, respectively) were the main phenolic acids in the studied fungi. The analysis of fatty acids confirmed that, in all fungi, the main fatty acids were stearic acid (27.9–35.2%), oleic acid (11.3–17.3%), palmitic acid (16.9–23.2%), linoleic acid (5.8–11.6%), and caprylic acid (6.3–10.9%). Our finding showed that endophytic fungi are a source of bioactive compounds, which could be used in various industries. This is the first report of endophytic fungi associated with G. tournefortii, which provides knowledge on their future use on biotechnological processes.

Anticancer effect of sequential extracts from Curcuma caesia rhizomes on human cancer cell lines and characterization of selected polyphenols in active extracts by LC-MS/MS

The objective of the present study is to quantify and compare in vitro cytotoxic properties of crude extracts sequentially extracted in solvents from the least polar hexane to the most polar water (hexane, petroleum ether, benzene, chloroform, ethyl acetate, methanol and water) from fresh and dry rhizomes of black turmeric on human cancer cell lines and further identification of phenolic acids and flavonoids by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in the active extracts. Of the extracts analyzed, chloroform and ethyl acetate extracts exhibited significant cytotoxicity on the tested cells. LC-MS/MS analysis of the active extracts (chloroform and ethyl acetate) revealed the presence of 14 phenolic acids and 9 flavonoids for the first time in the rhizomes of C. caesia, phenolic acids present in high concentration being gallic and vanillic acids in chloroform extracts, vanillic and protocatechuic acids in ethyl acetate extracts and catechin being the most abundant flavonoid. The active ingredients gallic acid, vanillic acid, protocatechuic acid and catechin present in these extracts may act as lead compounds for the development of new drugs against cancer.

QSAR model of pancreatic lipase inhibition by phenolic acids and their derivatives based on machine learning and multi-descriptor strategy

Inhibition of pancreatic lipase can be effective in controlling or treating obesity. Phenolic acids have good pancreatic lipase inhibitory effects and are widely found in plants. The identification of phenolic acids and their derivatives that can efficiently inhibit pancreatic lipase has attracted widespread attention. Therefore, it is crucial to screen natural pancreatic lipase inhibitors efficiently by establishing quantitative structure-activity relationships (QSAR) models. In this study, machine learning was used to perform multiple computational analyses of 28 phenolic acids and their derivatives with pancreatic lipase inhibitory activity, and multiple QSAR models were established. Molecular parameters were analyzed, IC50 was predicted, and molecular docking and molecular dynamics simulations analysis were done for the screened natural compounds. The results showed that the model with the highest level of accuracy is the 2D-3D mix model with a numerator parameter value of 8 (R2fitting of 0.9537 and MAE of 0.2230), and MATS6c is a key descriptor. Further screened 5 compounds that have not yet been reported and have the potential to inhibit lipase activity (IC50pre ≤ 16.69 μM), these all bound to key residues (Tyr114 and Ser152) of pancreatic lipase, of which 3–4,1,5-carboxylic acid bound the most tightly (affinity of −9.8 kJ/mol). And the compounds-pancreatic lipase complexes (2-3-2-enoate) had good stability. The three compounds with high bioavailability have the potential to become novel pancreatic lipase inhibitors. This study provides a novel approach for the prediction of phenolic acids and their derivatives capable of inhibiting pancreatic lipase.

Biological activity of extracts and hydrolysates from early- and adult-stage edible grasshopper Sphenarium purpurascens.

Edible insects have become a promising food source because they are rich in protein, fatty acids, minerals, among others. In recent years, edible insects have been proposed to be used as innovative functional ingredients in terms of biological activity. The present study aimed to determine and compare biological activities of the extracts and hydrolysates obtained from early- and adult-stage edible grasshoppers Sphenarium purpurascens to evaluate their potential as a source of bioactive compounds. Proximal analyses showed that in adult grasshoppers (AGs), the percentage of protein (48.9% ± 1.2), crude fat (13.1% ± 0.09), and chitin (15.6% ± 0.81) was significantly higher than early grasshoppers (EGs) (42.2% ± 0.55, 9.35% ± 0.08, and 10.5% ± 0.15, respectively). Total phenolic compounds, 2,2-diphenyl-1-picrylhydrazyl (DPPH•), and 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS•+) free radical scavenging were analyzed and reported. Enzymatic hydrolysis increased the concentration of total phenolic compounds and higher antioxidant capacity (up to 252.78 mM trolox). Once fractionated by ultrafiltration, the fraction that presented the highest antioxidant activity against DPPH• and ABTS•+ was that with molecules ≤ 10 kDa. Furthermore, the bioaccessibility of the samples was analyzed by in vitro protein digestion using a multienzymatic method, and a recovery index (RI) was reported. Extracts and hydrolysates were analyzed by UPLC-MS, and this allowed the identification of phenolic acids and flavonoids. The results obtained in this work suggest that the grasshopper can be used as a possible source of bioactive compounds that can be used in the food or pharmaceutical industry.

Hydroxypropyl methyl cellulose-Satureja hortensis L. ethanol extract mixtures as antimicrobial coating for sutures, identification of phenolic acids by using LC-MS-MS and TLC techniques

In this study, phenolic acid content of Satureja hortensis L. ethanol extract was investigated using TLC and LC-MS-MS methods and thymol, vanillic acid and caffeic acid were identified. Antimicrobial activity of prepared Hydroxypropyl methylcellulose (HPMC)-plant ethanol extract mixture was screened against E. coli (ATCC 25922), S. aureus (ATCC 25923) and B. cereus (ATCC 14579) species by using TLC-bioautography, disk and well diffusion methods. Both plant extract and the extract-biopolymer mixture showed inhibition activity on Gram-positive species according to TLC-bioautography results. Considering the disk diffusion test results, it was found that the inhibition activity increased with biopolymer plant extract mixtures for Gram-positive bacteria. Finally, the combination of plant extract with HPMC was used as coating for surgical sutures and antimicrobial activities were performed by using agar diffusion method. All coated sutures showed antimicrobial activity against S. aureus. Absorbable coated suture also exhibited antimicrobial activity against Bacillus cereus.

Identification of phenolic acids in rhizosphere soil of continuous cropping of Salvia miltiorrhiza. Bge

Identification of phenolic acids in rhizosphere soil of continuous cropping of Salvia miltiorrhiza. Bge

Separation and Identification of Phenolic Acids from Red Grape Seeds by HPLC

Red grape seed extract (GSE) was examined for its phenolic acids content. Grape seed powder was defatted twice with hexane for 1h and the defatted residues were kept in methanol for 2h with gentle agitation then centrifuged at 5000rpm for 10 minutes, and the precipitates were extracted using 2M NaOH for 4h with gentle agitation, acidified with HCl to pH 2.2 and centrifuged at 5000 rpm for 10 minutes. The pooled supernatants were extracted with a mixture of diethyl ether and ethyl acetate (1:1 v/v) twice. The organic layer (upper layer) was extracted using a separating funnel. The pooled extracts were collected, the solvent was evaporated, and the extracted residues were dissolved in methanol: water (4:1 v/v) and subjected to analysis by high-performance liquid chromatography (HPLC). Ferulic and coumaric acids were abundant among other phenolic acids detected in GSE, as their concentrations reached 103.16±6.18 and 93.66±3.42 mg/100g dry weight (DW) respectively. Caffeic, gallic, hydroxybenzoic, and vanillic acids were also identified and quantified in GSE but were at relatively low concentrations of 0.05±0.01; 2.37±0.07; 1.19±0.03; and 1.96±0.04 mg/100g DW respectively. Moreover, chlorogenic and syringic acids were also identified in GSE however; they were not quantified. The results of the present study indicated that red grape seeds contain considerable concentrations of ferulic and coumaric acids. These findings support the postulation that grape seeds have a potential for application in the pharmaceuticals and functional food industry.

Co-Solvent Selection for Supercritical Fluid Extraction (SFE) of Phenolic Compounds from Labisia pumila.

A preliminary study was conducted to study the effects of different types and concentrations of co-solvents based on yield, composition and antioxidants capacity of extract prior to optimization studies of supercritical fluid extraction (SFE) of Labisia pumila (locally referred to as ‘kacip fatimah’). The following co-solvents were studied prior to the optimization of supercritical carbon dioxide (SC–CO2) technique: ethanol, water, methanol, as well as aqueous solutions of ethanol–water and methanol–water (50% and 70% v/v). By using the selected co-solvents, identification of phenolic acids (gallic acid, methyl gallate and caffeic acid) was determined by using High-Performance Liquid Chromatography (HPLC). Then, the antioxidant capacity was evaluated by using three different assays: total phenolic content (TPC), ferric reducing/antioxidant power (FRAP) and free radical-scavenging capacity of 2,2-diphenyl-1-picrylhydrazyl (DPPH). SC–CO2 with 70% ethanol–water co-solvent was superior in terms of a higher combination of phenolic compounds extracted and antioxidants capacity. Overall, SC–CO2 with co-solvent 70% ethanol–water technique was efficient in extracting phenolic compounds from L. pumila, and thus the usage of this solvent system should be considered for further optimization studies.

Effects of phenolic and protein extracts from Melipona beecheii honey on pathogenic strains of Escherichia coli and Staphylococcus aureus.

Antimicrobial effects of Melipona beecheii honey have been attributed to diverse factors, in this sense, certain components such as proteins and phenolics could explain relevant aspects of its antimicrobial activity. The aim of this study was to evaluate the antibacterial activity of phenolic and protein extracts from M. beecheii honey against two bacterial pathogens: Staphylococcus aureus and Escherichia coli. With respect to phenolic content, HPLC analysis allowed the identification of phenolic acids like chlorogenic acid, caffeic acid, and flavonoids like catechin, myricetin, quercetin and apigenin. On the other hand, seven bands with molecular weight from 7.6 to 95kDa were detected in protein extract by SDS-PAGE system. It was determined the antibacterial activity of both extracts, with MICs lower than 145µg/mL and 60µg/mL for the phenolic and protein extracts respectively. These results indicate that phenolic and protein components of M. beecheii honey contribute significantly to the antibacterial activity.

Exploring the Synergy Between HPTLC and HPLC-DAD for the Investigation of Wine-Making By-Products.

Both environmental and economic issues are increasingly pushing for the revalorization of agri-food by-products, including those arising from wine industry. Wastes produced from wine-making processes are important sources of biologically active compounds, mainly phenolic acids and flavonoids, which could be re-used for several applications, for example as additive surrogates or new ingredients in foodstuffs and/or pharmaceuticals. Therefore, the development of methods aimed at isolating, characterizing and quantifying molecules present in winery by-products acquires considerable importance in view of their re-utilization on a large scale. In this connection, this study demonstrated that high-performance thin-layer chromatography (HPTLC) and high-performance liquid chromatography with diode array detection (HPLC-DAD) can operate in synergy for the investigation of pomace and seed materials arising from both white and red cultivars of Vitis Vinifera. By virtue of fingerprint profiling, mass spectrometry (MS) interfacing and band comparison method, HPTLC enabled detection and identification of phenolic acids, non-anthocyanic flavonoids and anthocyanins. On the contrary, only anthocyanins could be identified by HPLC-DAD, and their subsequent quantification showed that malvidin-3-O-glucoside (oenin) was the most abundant one. In parallel, HPTLC has allowed to detect and quantify proanthocyanidins (PAC), showing that only catechin was present in the test samples. Both quantitative analytical methods were validated in terms of linearity, detection and quantification limits and precision.

Extraction of phenolic compounds with antioxidant potential from coconut (Cocos nucifera L.) testa and identification of phenolic acids and flavonoids using UPLC coupled with TQD-MS/MS

Coconut testa, a brown skin covering of a coconut endosperm, is a rich source of phenolics. It is one of the by-products obtained in the coconut processing industries and currently underutilized despite being rich in phenolic compounds. A study was conducted to identify the suitable solvent system for maximum extractability of total phenolic content (TPC) and total flavonoid content (TFC) with antioxidant potential from the testa. Individual phenolic acids and flavonoids of different solvent extracts were also determined using UPLC-H class coupled with TQD-MS/MS. The TPC and TFC ranged from 4.9 to 167 mg GAE/g and 8.84–115 mg QE/g defatted testa, respectively; significant differences were observed for various solvent systems studied. Acidification of solvents significantly increased the extractability of TPC and reduced the extractability of TFC. A total of 28 phenolics comprising 16 phenolic acids and 12 flavonoids were identified. Protocatechuic acid, p-coumaric acid and ferulic acid are the major phenolic acids identified whereas, catechin, apigenin and kaempferol are the major flavonoids identified. In summary, this study proved that coconut testa is a natural source of multiple phenolics acids and flavonoids with potent antioxidant capacity, and it can be used as a natural source of antioxidants.

Determination of polyphenolic content, HPLC analyses and DNA cleavage activity of Malaysian Averrhoa carambola L. fruit extracts

In developing countries, the increasing gap between population growth and food supply has created renewed interest in finding reliable and cheap natural resources of nutraceutical value and health promoting properties. Therefore, the present study deals with the phytochemical analyses and DNA cleavage activity of Averrhoa carambola L. fruit (starfruit) extracts. The phytochemical studies involve colour tests and quantification of phenolics and flavonoids of the prepared ethanolic and aqueous extracts. Identification of phenolic acids and flavonoids present in the extracts were conducted by high performance liquid chromatography (HPLC) equipped with diode array detector (DAD). DNA cleavage activity of the extracts was evaluated through gel electrophoresis against plasmid Escherichia coli DNA at different concentrations (0.125–0.60μg/μl). The results of the study exhibited that the starfruit is a rich source of polyphenols and all the extracts exhibited a dose dependent DNA cleavage activity, whereas ethanolic extract induced more cleavage as compared to the aqueous extract. In conclusion, the present study provides preliminary evidence with regard to nutraceutical value of the fruit. So, further extensive study is a prerequisite to exploit DNA cleaving properties of the fruit extracts for therapeutic application.

HPLC/PDA–ESI/MS Identification of Phenolic Acids, Flavonol Glycosides and Antioxidant Potential in Blueberry, Blackberry, Raspberries and Cranberries

Berry fruits are known to be a rich source of natural compounds which provide them many health benefits. The most common compounds that occur in berries are vitamins, flavonoids, anthocyanins and phenolic acids. The objective of this work was to identify and compare the phenolic acids, flavonol glycosides content and antioxidant potential in blueberry, blackberry, raspberries and cranberries. Berries methanolic extracts were analyzed by HPLC/PDA–ESI/MS. Antioxidant activity was determinate using FRAP assay. Antioxidant activity of analyzed berries varied considerably. Blueberry extract demonstrated the highest ferric reducing antioxidant potential (48.3 μM Fe<SUP>2</SUP>/g) while lower values were obtained for cranberries extract (19.6 μM Fe<SUP>2</SUP>/g). Berries extracts were characterized by the presence of 16 compounds in different ratios. Blueberry extract was characterized by the presence of a large amount of phenolic acids such as chlorogenic and caffeic acid while raspberries extract was found to have high amount of ellagic acid. All analyzed berries contain higher levels of bioactive compounds which are responsible for their antioxidant potential. Based on quantitative analysis of phenolics these fruits can be highly recommended for daily consumption<i>.</i>

Phenolic substances in Ailanthus glandulosa Desf

Abstract The aim of this work was the isolation and identification of phenolic acids from fruit and leaves of Ailanthus glandulosa Desf. The methods used in the isolation and identification of the compounds were: isolation of phenolic acids modified by Ibrahim and Towers, acidic and alkaline hydrolysis by Schmidtlein and Herrmann and identification of phenolic acids in the isolated fraction of two-dimensional thin layer chromatography (TLC) on cellulose plates. In the course of the study, the presence of phenolic acids in leaves and fruit of Ailanthus glandulosa Desf was confirmed. Overall, proportions of 15 phenolic acids were found and identified in the analyzed material. These are: gallic acid, ellagic acid, caffeic acid, gentisic acid, protocatechuic acid, p-hydroxybenzoic acid, m-hydroxybenzoic acid, ferulic acid, p-coumaric acid, syryngic acid, vanillic acid, salicylic acid, 2-hydroxy-4- methoxybenzoic acid, 2,5-dimethoxycinnamic, p-hydroxyphenylacetic acid, and 3 other compounds named A, B, C, whose identification was not possible due to the lack of suitable reference patterns. Studies have shown that leaves and fruit of the plant Ailanthus glandulosa Desf. contain a large number of phenolic acids which possess many important pharmacological activities.

Chromatographical analysis of phenolic acids in some species of Polygonum L. genus. Part 1 Qualitative analysis by two-dimensional thin layer chromatography (TLC)

The Two-Dimensional Thin Layer Chromatography method has been used for the separation and identification of phenolic acids from six taxons of &lt;em&gt;Pohygonum&lt;/em&gt; L. genus. The following acids were found: caffeic, p-coumaric, ferulic, p-hydroxybenzoic, m-hydroxybenzoic, vanillic, syringic, p-hydroxyphenylacetic, o-hydroxyphenylacetic, synapic, melillotic, salicylic, gentisic, elagic, gallic, chlorogenic, protocatechuic and homoprotocatechuic. Gallic, ferulic, vanillic, p-coumaric and p-hydroxybenzoic acids were isolated from herb &lt;em&gt;Polygonum convolvulus&lt;/em&gt; L. using column chromatography.

Identification of phenolic acids in wheat plants after fumigation with SO&lt;sub&gt;2&lt;/sub&gt; in open-top chambers (OTCs)

Identification of phenolic acids in wheat plants after fumigation with SO&lt;sub&gt;2&lt;/sub&gt; in open-top chambers (OTCs)

Clitocybe alexandri extract induces cell cycle arrest and apoptosis in a lung cancer cell line: Identification of phenolic acids with cytotoxic potential

Mushrooms are a possible rich source of biologically active compounds with the potential for drug discovery. The aim of this work was to gain further insight into the cytotoxicity mechanism of action of Clitocybe alexandri ethanolic extract against a lung cancer cell line (NCI-H460 cells). The effects on cell cycle profile and levels of apoptosis were evaluated by flow cytometry, and the effect on the expression levels of proteins related to cellular apoptosis was also investigated by Western blot. The extract was characterised regarding its phenolic composition by HPLC-DAD, and the identified compounds were studied regarding their growth inhibitory activity, by sulforhodamine B (SRB) assay. The effect of individual or combined compounds on viable cell number was also evaluated using the Trypan blue exclusion assay. It was observed that the C. alexandri extract induced an S-phase cell cycle arrest and increased the percentage of apoptotic cells. In addition, treatment with the GI50 concentration (concentration that was able to cause 50% of cell growth inhibition; 24.8μg/ml) for 48h caused an increase in the levels of wt. p53, cleaved caspase-3 and cleaved poly (ADP–ribose) polymerase (PARP). The main components identified in this extract were protocatechuic, p-hydroxybenzoic and cinnamic acids. Cinnamic acid was found to be the most potent compound regarding cell growth inhibition. Nevertheless, it was verified that the concomitant use of the individual compounds provided the strongest decrease in viable cell number. Overall, evidence was found for alterations in cell cycle and apoptosis, involving p53 and caspase-3. Furthermore, our data suggests that the phenolic acids identified in the extract are at least partially responsible for the cytotoxicity induced by this mushroom extract.

Identification and quantification of flavonoids and phenolic acids in burr parsley (Caucalis platycarpos L.), using high-performance liquid chromatography with diode array detection and electrospray ionization mass spectrometry.

A sensitive method coupling high-performance liquid chromatography (HPLC) with diode-array detector (DAD) and electrospray ionization mass spectrometry (MS) was optimized for the separation and identification of phenolic acids, flavonoid glycosides and flavonoid aglycones in the extract of burr parsley (Caucalis platycarpos L.). Fragmentation behavior of flavonoid glycosides and phenolic acids were investigated using ion trap mass spectrometry in negative electrospray ionization. The MS, MSn and UV data together with HPLC retention time (TR) of phenolic acids and flavonoids allowed structural characterization of these compounds. Caffeoylquinic acid (CQA) isomers, p-coumaroyl-quinic acids (p-CoQA), feruloylquinic acids (FQA), dicaffeoylquinic acids (diCQA), luteolin-7-O-rutinoside, apigenin-7-O-rutinoside as well as isolated chrysoeriol-7-O-rutinoside have been identified as constituents of C. platycarpos for the first time. An accurate, precise and sensitive LC-DAD method for quantification of four phenolic acids (3-O-caffeoylquinic, caffeic, p-coumaric, o-coumaric acid), four flavonoid glycosides (luteolin-7-O-glucoside, apigenin-7-O-glucoside, quercetin-3-O-galactoside, quercetin-3-O-rhamnoside), and three flavonoid aglycones (luteolin, apigenin, chrysoeriol) in C. platycarpos extract was validated in terms of linearity, limit of detection, limit of quantification, precision and accuracy. 3-O-caffeoylquinic acid was the predominant phenolic acid and luteolin-7-O-glucoside was the predominant flavonoid glycoside.

Identification of phenolic acids in herbs and fruits of Myrrhis odorata (L.) Scop

Identification of phenolic acids in herbs and fruits of Myrrhis odorata (L.) Scop

THE CONTENT AND ANTIOXIDANT ACTIVITY OF PHENOLIC COMPOUNDS IN COLD‐PRESSED PLANT OILS

ABSTRACT Many nontraditional vegetable oils have been introduced to the market relatively recently, and therefore data on their antioxidant potential have not been reported. Such data are of importance for the evaluation of the nutritional and health impact of these oils. Antioxidant properties of the methanolic extracts of cold‐pressed oils such as soybean, sunflower, rapeseed, corn, grapeseed, hemp, flax, rice bran and pumpkin were studied. The methanolic extracts were obtained by solid phase extraction and separation, and identification of phenolic acids was conducted by high‐performance liquid chromatography. The obtained extracts were characterized by different scavenging activities of 2,2‐diphenyl‐1‐picrylhydrazyl radicals. The best antioxidant properties were displayed by the extract from hemp, pumpkin and rapeseed oils. The highest content of total phenolic compounds was determined for the pumpkin and hemp oils – about 2.4 mg/100 g. Rapeseed oil was characterized by the highest content of phenolic acids, especially sinapic acid. To better understand the beneficial effects of antioxidant compounds in vegetable oils, it is important to investigate whether these bioactive compounds in oils differ in their reactions with free radicals. PRACTICAL APPLICATIONSPhenolic compounds have been reported to be present in all vegetable oils, which is very important for the oxidative stability of the polyunsaturated fatty acids of these oils. Additionally, edible oils rich in natural antioxidants may play a role in reducing the risk of chronic diseases. Thus, the oils examined may be used in different food applications to provide nutrition and health benefits.