UHPLC analysis and in vitro biological activities of Areca triandra Roxb. ex Buch.-Ham.: a substitute of areca nut

2021 ◽  
Vol 16 (12) ◽  
pp. 109-118
Author(s):  
Ashutosh Pathak ◽  
Rahul Das ◽  
Deepu Vijayan ◽  
Nripemo Odyuo ◽  
David L. Baite ◽  
...  
Keyword(s):  
Phenolic Acids ◽  
Clinical Laboratory ◽  
Biological Activities ◽  
Antifungal Susceptibility ◽  
Reducing Power ◽  
Total Phenolic ◽  
Areca Nut ◽  
Chromatography Method ◽  
Minimum Effective Concentration

Areca triandra Roxb. ex Buch.-Ham. (Arecaceae) distributed in Assam, North Bengal plains, Meghalaya andaman and Nicobar Islands is used as a substitute for A. catechu L. (Areca Nut). A. catechu is widely used as masticator in South Asia. The present study was conducted to find out the phenolic acids present in A. triandra and potential of it in inhibiting the reactive oxygen species (ROS). Plant extracts obtained from leaf and nut were analyzed qualitatively, quantitatively and phenolic acids were identified and quantified using reverse phase chromatography method developed on through Hypersil Gold aQ C18 column in Thermo Scientific Dionex Ultimate 3000 UHPLC. Three antioxidant assays viz. DPPH, ABTS and reducing power assay were performed to assess the potential of A. triandra as an antioxidant. Antifungal susceptibility test was performed by Clinical Laboratory Standard Institute (CLSI) recommended broth microdilution assay and disk diffusion susceptibility assay using Tinidazole as standard. Total catechin content was 19.51% of total phenolic content in NME. The antioxidant property was observed in NME only for DPPH, ABTS and RPA assays. In antifungal activity, minimum effective concentration (MEC) was observed in NME at the concentration of 1.0mg/ml and above whereas in LME, no MEC was observed up to tested concentrations. A. triandra possesses high catechin content and has promising potential as an antioxidant agent.

Zawartość związków fenolowych w ekstrakcie z propolisu oraz ocena jego aktywności przeciwutleniającej i cytoochronnej względem erytrocytów ludzkich w warunkach stresu oksydacyjnego in vitro

2019 ◽  
Vol 20 (1) ◽  
Author(s):  
Magdalena Woźniak ◽  
Lucyna Mrówczyńska ◽  
Agnieszka Waśkiewicz ◽  
Marta Babicka ◽  
Elżbieta Hołderna-Kędzia ◽  
...  
Keyword(s):  
Free Radicals ◽  
Phenolic Acids ◽  
Biological Activities ◽  
Radical Scavenging ◽  
Reducing Power ◽  
Human Erythrocytes ◽  
Antioxidant Potential ◽  
Natural Material ◽  
Propolis Extract

Introduction. Propolis (bee glue) is a natural product collected by honeybees from buds of various trees, shrubs and other plant species. Extracts of propolis possess numerous biological activities, including antioxidant, antibacterial, antifungal and anticancer. For this reason, propolis is currently used in many applications, such as preparations for cold syndrome, dermatological preparations or as a constituent of nutritional supplements and health food. The chemical composition of this natural material is very complex and depending on many factors, including method of extraction and selection of the solvent for the extraction process. Aim. The aim of the study was to determine concentration of selected phenolic compounds (flavonoids and phenolic acids) in extract of Polish propolis and estimate its antioxidant activity and effect on human red blood cells. Material and methods. In the propolis extract was determined concentration of 14 flavonoids and 9 phenolic acids using ultra-performance liquid chromatography equipped with a photodiode detector and a triple quadrupole mass spectrometer. The antioxidant potential of propolis extract was evaluated applying DPPH˙ free radical scavenging activity assay and Fe3+ reducing power assay. Moreover, the cytotoxicity and cytoprotective potential of propolis extract was estimated using human erythrocytes in vitro. Results. The propolis extract contained high concentration of pinocembrin, galangin, chrysin, apigenin, kaempferol, coumaric acid and cinnamic acid. It exhibited also high antioxidant potential. The antiradical activity of examined propolis extract was equal to 75% approx. activity of both standard antioxidants used in the study, namely Trolox and BHT. The reducing power of extract was equal to 65% approx. of Trolox and 80% of BHT, respectively. The propolis extract had no hemolytic activity, moreover, effectively protected human erythrocytes against free radicals-induced damage in vitro. Conclusions. The results of this study indicate that the propolis extract of national origin is a rich source of flavonoids and phenolic acids. Therefore, the propolis extract possesses a high antioxidant potential and can protect erythrocytes against free radicals-induced oxidative hemolysis.

scholarly journals Correlation between Antifungal Susceptibilities ofCoccidioides immitis In Vitro and Antifungal Treatment with Caspofungin in a Mouse Model

2001 ◽  
Vol 45 (6) ◽  
pp. 1854-1859 ◽  
Author(s):  
Gloria M. González ◽  
Rolando Tijerina ◽  
Laura K. Najvar ◽  
Rosie Bocanegra ◽  
Michael Luther ◽  
...  
Keyword(s):  
Susceptibility Testing ◽  
Clinical Laboratory ◽  
Antifungal Susceptibility ◽  
National Committee ◽  
In Vitro Susceptibility ◽  
Minimum Effective Concentration ◽  
Study Results ◽  
Significant Difference

ABSTRACT Caspofungin (Merck Pharmaceuticals) was tested in vitro against 25 clinical isolates of Coccidoides immitis. In vitro susceptibility testing was performed in accordance with the National Committee for Clinical Laboratory Standards document M38-P guidelines. Two C. immitis isolates for which the caspofungin MICs were different were selected for determination of the minimum effective concentration (MEC), and these same strains were used for animal studies. Survival and tissue burdens of the spleens, livers, and lungs were used as antifungal response markers. Mice infected with strain 98-449 (48-h MIC, 8 μg/ml; 48-h MEC, 0.125 μg/ml) showed 100% survival to day 50 when treated with caspofungin at ≥1 mg/kg. Mice infected with strain 98-571 (48-h MIC, 64 μg/ml; 48-h MEC, 0.125 μg/ml) displayed ≥80% survival when the treatment was caspofungin at ≥5 mg/kg. Treatment with caspofungin at 0.5, 1, 5, or 10 mg/kg was effective in reducing the tissue fungal burdens of mice infected with either isolate. When tissue fungal burden study results were compared between strains, caspofungin showed no statistically significant difference in efficacy in the organs of the mice treated with both strains. A better in vitro-in vivo correlation was noted when we used the MEC instead of the MIC as the endpoint for antifungal susceptibility testing. Caspofungin may have a role in the treatment of coccidioidomycosis.

scholarly journals Phenylalanine Increases the Production of Antioxidant Phenolic Acids in Ginkgo biloba Cell Cultures

Molecules ◽  
2021 ◽  
Vol 26 (16) ◽  
pp. 4965
Author(s):  
Agnieszka Szewczyk ◽  
Inga Kwiecień ◽  
Mariusz Grabowski ◽  
Karolina Rajek ◽  
Emilia Cavò ◽  
...  
Keyword(s):  
Phenolic Acids ◽  
Cell Cultures ◽  
Ginkgo Biloba ◽  
Condensed Tannin ◽  
Antioxidant Properties ◽  
Reducing Power ◽  
Total Phenolic ◽  
Moderate Activity ◽  
High Concentration

The aims of this study were to evaluate the antioxidant properties, to investigate the content of major secondary metabolites in Ginkgo biloba cell cultures, and to determine the change in the production of phenolic acids by adding phenylalanine to the culture medium. Three in vitro methods, which depend on different mechanisms, were used for assessing the antioxidant activity of the extract: 1,1-diphenyl-2-picrylhydrazil (DPPH), reducing power and Fe2+ chelating activity assays. The extract showed moderate activity both in the DPPH and in the reducing power assays (IC50 = 1.966 ± 0.058 mg/mL; ASE/mL = 16.31 ± 1.20); instead, it was found to possess good chelating properties reaching approximately 70% activity at the highest tested dose. The total phenolic, total flavonoid, and condensed tannin content of G. biloba cell culture extract was spectrophotometrically determined. The phenolic acid content was investigated by RP-HPLC, and the major metabolites—protocatechuic and p-hydroxybenzoic acids—were isolated and investigated by 1H NMR. The results showed that phenylalanine added to G. biloba cell cultures at concentrations of 100, 150, and 200 mg/150 mL increased the production of phenolic acids. Cultures that were grown for 3 weeks and collected after 4 days of phenylalanine supplementation at high concentration showed maximal content of phenolic acids (73.76 mg/100 g DW).

scholarly journals A 20-Year Antifungal Susceptibility Surveillance (From 1999 to 2019) for Aspergillus spp. and Proposed Epidemiological Cutoff Values for Aspergillus fumigatus and Aspergillus flavus: A Study in a Tertiary Hospital in China

2021 ◽  
Vol 12 ◽  
Author(s):  
Xinyu Yang ◽  
Wei Chen ◽  
Tianyu Liang ◽  
JingWen Tan ◽  
Weixia Liu ◽  
...  
Keyword(s):  
Aspergillus Fumigatus ◽  
Aspergillus Flavus ◽  
Clinical Laboratory ◽  
Dynamic Change ◽  
Antifungal Susceptibility ◽  
Surveillance Period ◽  
Minimum Effective Concentration ◽  
Cutoff Values ◽  
Broth Microdilution Method

The emergence of resistant Aspergillus spp. is increasing worldwide. Long-term susceptibility surveillance for clinically isolated Aspergillus spp. strains is warranted for understanding the dynamic change in susceptibility and monitoring the emergence of resistance. Additionally, neither clinical breakpoints (CBPs) nor epidemiological cutoff values (ECVs) for Aspergillus spp. in China have been established. In this study, we performed a 20-year antifungal susceptibility surveillance for 706 isolates of Aspergillus spp. in a clinical laboratory at Peking University First Hospital from 1999 to 2019; and in vitro antifungal susceptibility to triazoles, caspofungin, and amphotericin B was determined by the Clinical and Laboratory Standards Institute (CLSI) broth microdilution method. It was observed that Aspergillus fumigatus was the most common species, followed by Aspergillus flavus and Aspergillus terreus. Forty isolates (5.7%), including A. fumigatus, A. flavus, A. terreus, Aspergillus niger, and Aspergillus nidulans, were classified as non-wild type (non-WT). Importantly, multidrug resistance was observed among A. flavus, A. terreus, and A. niger isolates. Cyp51A mutations were characterized for 19 non-WT A. fumigatus isolates, and TR34/L98H/S297T/F495I was the most prevalent mutation during the 20-year surveillance period. The overall resistance trend of A. fumigatus increased over 20 years in China. Furthermore, based on ECV establishment principles, proposed ECVs for A. fumigatus and A. flavus were established using gathered minimum inhibitory concentration (MIC)/minimum effective concentration (MEC) data. Consequently, all the proposed ECVs were identical to the CLSI ECVs, with the exception of itraconazole against A. flavus, resulting in a decrease in the non-WT rate from 6.0 to 0.6%.

scholarly journals Potential Investigation of In Vitro Antioxidant, Anti-Inflammatory and Anti-Haemolytic Activities from Polar Solvent Extracts of Pterocarpus marsupium

2017 ◽  
Vol 9 (1) ◽  
Author(s):  
Ramesh L. Londonkar ◽  
Aruna L. H. ◽  
Amarvani P. Kanjikar
Keyword(s):  
Methanol Extract ◽  
Protein Denaturation ◽  
Biological Activities ◽  
Reducing Power ◽  
Total Phenolic ◽  
Pterocarpus Marsupium ◽  
Anti Inflammatory ◽  
Phenolic And Flavonoid ◽  
In Vitro Antioxidant Activity

Pterocarpus marsupium is a well known plant in ayurvedic system of medicine. The plant has been found to possess diverse number of biological activities and is thus commercially exploited. In the present study, methanol and aqueous extracts of P.marsupium bark were used to evaluate in vitro antioxidant, anti-inflammatory and anti-haemolytic activities. In vitro antioxidant activity was carried out by DPPH, ABTS, phosphomolybdenum, reducing power assays, anti- inflammatory activity by inhibition of protein denaturation and anti-haemolytic by hyposaline induced haemolysis. Total phenolic and flavonoid content was also estimated. Methanol extract showed an effective pharmacological activity in all assays when compared with their respective standards. The maximum phenolic and flavonoid contents were found to be (280 ±0.47mg/g) and (620±0.81mg/g) respectively in methanol extract of Pterocarpus marsupium. This study indicates the presence of active constituents which can be exploited for the treatment of various diseases and also could be used in pharmaceutical industry.

scholarly journals Hepatoprotective studies on methanolic extract fractions of Lindernia ciliata and development of qualitative analytical profile for the bioactive extract

2019 ◽  
Vol 5 (1) ◽  
Author(s):  
Praneetha Pallerla ◽  
Narsimha Reddy Yellu ◽  
Ravi Kumar Bobbala
Keyword(s):  
Methanolic Extract ◽  
Antioxidant Properties ◽  
Radical Scavenging ◽  
Liver Homogenate ◽  
Reducing Power ◽  
Hepatoprotective Activity ◽  
Total Phenolic ◽  
Effective Fraction

Abstract Background The objective of the study is to evaluate the hepatoprotective activity of methanolic extract fractions of Lindernia ciliata (LC) and development of qualitative analytical profile of the bioactive fraction using HPLC fingerprinting analysis. All the fractions of methanolic extract of Lindernia ciliata (LCME) are assessed for their total phenolic, flavonoid contents and in vitro antioxidant properties by using DPPH, superoxide, nitric oxide, hydroxyl radical scavenging activities and reducing power assay. Acute toxicity study was conducted for all the fractions and the two test doses 50 and 100 mg/kg were selected for the hepatoprotective study. Liver damage was induced in different groups of rats by administering 3 g/kg.b.w.p.o. paracetamol and the effect of fractions were tested for hepatoprotective potential by evaluating serum biochemical parameters and histology of liver of rats. The effective fraction was evaluated for its antihepatotoxic activity against D-Galactosamine (400 mg/kg b.w. i.p.) and in vivo antioxidant parameters viz., Glutathione (GSH), Melondialdehyde (MDA) and Catalase (CAT) levels are estimated using liver homogenate. Results Among all the fractions, butanone fraction of LCME, (BNF-LCME) has shown better hepatoprotective activity and hence it is selected to evaluate the antihepatotoxicity against D-GaIN. The activity of BNF-LCME is well supported in in vitro and in vivo antioxidant studies and may be attributed to flavonoidal, phenolic compounds present in the fraction. Hence, BNF-LCME was subjected to the development of qualitative analytical profile using HPLC finger printing analysis. Conclusions All the fractions of LCME exhibited significant hepatoprotective activity and BNF-LCME (50 mg/kg) was identified as the most effective fraction.

scholarly journals Characterization, Stability, and Bioaccessibility of Betalain and Phenolic Compounds from Opuntia stricta var. Dillenii Fruits and Products of Their Industrialization

Foods ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1593
Author(s):  
Iván Gómez-López ◽  
Gloria Lobo-Rodrigo ◽  
María P. Portillo ◽  
M. Pilar Cano
Keyword(s):  
Phenolic Compounds ◽  
Phenolic Acids ◽  
Total Phenolic ◽  
Antioxidant Compounds ◽  
Gastrointestinal Digestion ◽  
Fruit Peel ◽  
Full Characterization ◽  
In Vitro Gastrointestinal Digestion ◽  
Quercetin Glycoside

The aim of the present study was the full characterization, quantification, and determination of the digestive stability and bioaccessibility of individual betalain and phenolic compounds of Opuntia stricta, var. Dillenii fresh fruits (peel, pulp, and whole fruit) and of the products of the industrialization to obtain jam (raw pressed juice (product used for jam formulation), by-product (bagasse), and frozen whole fruit (starting material for jam production)). Opuntia stricta var. Dillenii fruits and products profile showed 60 betalain and phenolic compounds that were identified and quantified by HPLC-DAD-ESI/MS and HPLC-DAD-MS/QTOF, being 25 phenolic acids (including isomers and derivatives), 12 flavonoids (including glycosides), 3 ellagic acids (including glycosides and derivative), and 20 betanins (including degradation compounds). In vitro gastrointestinal digestion was performed by INFOGEST® protocol. Fruit pulp showed the greater content of total betalains (444.77 mg/100 g f.w.), and jam only showed very low amounts of two betanin degradation compounds, Cyclo-dopa-5-O-β-glucoside (and its isomer) (0.63 mg/100 f.w.), and two Phyllocactin derivatives (1.04 mg/100 g f.w.). Meanwhile, fruit peel was the richer tissue in total phenolic acids (273.42 mg/100 g f.w.), mainly in piscidic acid content and total flavonoids (7.39 mg/100 g f.w.), isorhamnetin glucoxyl-rhamnosyl-pentoside (IG2) being the most abundant of these compounds. The stability of betalains and phenolic compounds during in vitro gastrointestinal digestion is reported in the present study. In Opuntia stricta var. Dillenii pulp (the edible fraction of the fresh fruit), the betanin bioaccessibility was only 22.9%, and the flavonoid bioaccessibility ranged from 53.7% to 30.6%, depending on the compound. In non-edible samples, such as peel sample (PE), the betanin bioaccessibility was 42.5% and the greater bioaccessibility in flavonoids was observed for quercetin glycoside (QG1) 53.7%, the fruit peel being the most interesting material to obtain antioxidant extracts, attending to its composition on antioxidant compounds and their bioaccessibilities.

scholarly journals Triterpene Acid and Phenolics from Ancient Apples of Friuli Venezia Giulia as Nutraceutical Ingredients: LC-MS Study and In Vitro Activities

Molecules ◽  
2019 ◽  
Vol 24 (6) ◽  
pp. 1109 ◽  
Author(s):  
Stefania Sut ◽  
Gokhan Zengin ◽  
Filippo Maggi ◽  
Mario Malagoli ◽  
Stefano Dall’Acqua
Keyword(s):  
Radical Scavenging ◽  
Reducing Power ◽  
Total Phenolic ◽  
Inhibitory Effects ◽  
Triterpene Acid ◽  
Beneficial Effects ◽  
Triterpene Acids ◽  
Phenolic Constituents ◽  
Friuli Venezia Giulia

Triterpene acid and phenolic constituents from nine ancient varieties of apple (Malus domestica) fruits cultivated in Fanna, Friuli Venezia Giulia region, northeast Italy, were analyzed and compared with four commercial apples (‘Golden Delicious’, ‘Red Delicious’, ‘Granny Smith’ and ‘Royal Gala’). Total phenolic and flavonoid contents were measured by spectrophotometric assays. The quali-quantitative fingerprint of secondary metabolites including triterpene acid was obtained by LC-DAD-(ESI)-MS and LC-(APCI)-MS, respectively. Based on the two LC-MS datasets, multivariate analysis was used to compare the composition of ancient fruit varieties with those of four commercial apples. Significant differences related mainly to the pattern of triterpene acids were found. Pomolic, euscaphyc, maslinic and ursolic acids are the most abundant triterpene in ancient varieties pulps and peels, while ursolic and oleanolic acids were prevalent in the commercial fruits. Also, the content of the phenolic compounds phloretin-2-O-xyloglucoside and quercetin-3-O-arabinoside was greater in ancient apple varieties. The antioxidant (radical scavenging, reducing power, metal chelating and phosphomolybdenum assays) and enzyme inhibitory effects (against cholinesterase, tyrosinase, amylase and glucosidase) of the samples were investigated in vitro. Antioxidant assays showed that the peels were more active than pulps. However, all the samples exhibited similar enzyme inhibitory effects. Ancient Friuli Venezia Giulia apple cultivars can be a source of chlorogenic acid and various triterpene acids, which are known for their potential anti-inflammatory activity and beneficial effects on lipid and glucose metabolism. Our results make these ancient varieties suitable for the development of new nutraceutical ingredients.

Use of spectrophotometric reading for in vitro antifungal susceptibility testing ofAspergillusspp.

1999 ◽  
Vol 45 (10) ◽  
pp. 871-874 ◽  
Author(s):  
Eric Dannaoui ◽  
Florence Persat ◽  
Marie-France Monier ◽  
Elisabeth Borel ◽  
Marie-Antoinette Piens ◽  
...  
Keyword(s):  
Amphotericin B ◽  
Susceptibility Testing ◽  
Clinical Laboratory ◽  
Antifungal Susceptibility ◽  
Reference Method ◽  
Antifungal Susceptibility Testing ◽  
Aspergillus Species ◽  
National Committee ◽  
Broth Microdilution Method

A comparative study of visual and spectrophotometric MIC endpoint determinations for antifungal susceptibility testing of Aspergillus species was performed. A broth microdilution method adapted from the National Committee for Clinical Laboratory Standards (NCCLS) was used for susceptibility testing of 180 clinical isolates of Aspergillus species against amphotericin B and itraconazole. MICs were determined visually and spectrophotometrically at 490 nm after 24, 48, and 72h of incubation, and MIC pairs were compared. The agreement between the two methods was 99% for amphotericin B and ranged from 95 to 98% for itraconazole. It is concluded that spectrophotometric MIC endpoint determination is a valuable alternative to the visual reference method for susceptibility testing of Aspergillus species.Key words: antifungal, susceptibility testing, Aspergillus, spectrophotometric reading.

scholarly journals Quantitation of Candida albicans Ergosterol Content Improves the Correlation between In Vitro Antifungal Susceptibility Test Results and In Vivo Outcome after Fluconazole Treatment in a Murine Model of Invasive Candidiasis

2000 ◽  
Vol 44 (8) ◽  
pp. 2081-2085 ◽  
Author(s):  
Beth A. Arthington-Skaggs ◽  
David W. Warnock ◽  
Christine J. Morrison
Keyword(s):  
Murine Model ◽  
Invasive Candidiasis ◽  
Clinical Laboratory ◽  
Antifungal Susceptibility ◽  
Resistant Isolate ◽  
National Committee ◽  
Ergosterol Content ◽  
In Vitro Antifungal Susceptibility

ABSTRACT MIC end point determination for the most commonly prescribed azole antifungal drug, fluconazole, can be complicated by “trailing” growth of the organism during susceptibility testing by the National Committee for Clinical Laboratory Standards approved M27-A broth macrodilution method and its modified broth microdilution format. To address this problem, we previously developed the sterol quantitation method (SQM) for in vitro determination of fluconazole susceptibility, which measures cellular ergosterol content rather than growth inhibition after exposure to fluconazole. To determine if SQM MICs of fluconazole correlated better with in vivo outcome than M27-A MICs, we used a murine model of invasive candidiasis and analyzed the capacity of fluconazole to treat infections caused by C. albicansisolates which were trailers (M27-A MICs at 24 and 48 h, ≤1.0 and ≥64 μg/ml, respectively; SQM MIC, ≤1.0 μg/ml), as well as those which were fluconazole sensitive (M27-A and SQM MIC, ≤1.0 μg/ml) and fluconazole resistant (M27-A MIC, ≥64 μg/ml; SQM MIC, 54 μg/ml). Compared with the untreated controls, fluconazole therapy increased the survival of mice infected with a sensitive isolate and both trailing isolates but did not increase the survival of mice infected with a resistant isolate. These results indicate that the SQM is more predictive of in vivo outcome than the M27-A method for isolates that give unclear MIC end points due to trailing growth in fluconazole.

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