Concentrations Of Methanolic Extracts Research Articles

Does Erodium trifolium (Cav.) Guitt exhibit medicinal properties? Response elements from phytochemical profiling, enzyme-inhibiting, and antioxidant and antimicrobial activities

Abstract Geraniaceae are typically used as diuretic, anti-diarrhoeal, stomachic, and anti-hemorrhagic drugs. This study examined the phytochemicals and bioactivities in methanolic extract (ME) and petroleum ether extract (PEE) of the Erodium trifolium aerial part. Inductively coupled plasma mass spectrometry was used to assess the mineral profiles, high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS) and high-performance liquid chromatography with diode-array detection (HPLC-DAD) were used to assess the phenolic content of ME, and gas chromatography-mass spectrometry (GC-MS) was used to assess the fatty acid and volatile composition of the PEEs. In addition, the bioactivities of extracts were evaluated by using the 1,1-diphenyl-2-picrylhydrazyl, ABTS, and cupric reducing antioxidant capacity assays, including enzyme inhibition against acetylcholinesterase (AChE), butyrylcholinesterase (BChE), α-amylase, α-glucosidase activities, and antibacterial properties. HPLC-MS identified eight compounds in ME: rutin, catechin, and caffeine were the major phenolic compounds detected. HPLC-DAD analysis showed that 11 compounds detected among epicatechin, catechin, chlorogenic acid, and gallic acid were predominant. GC-MS analysis revealed 30 fatty acids in PEE, with palmitic acid and oleic acid being predominant. The mineral content showed that Fe was the abundant microelement, and Ca and K were the abundant macroelements. The ME exhibited the highest activity compared to the PEE in all tests regarding antioxidant and antibacterial activities. Furthermore, all the extracts showed moderate inhibition against AChE, BChE, α-amylase, and α-glucosidase. In conclusion, E. trifolium may be employed to separate novel bioactive metabolites with potential pharmaceutical activities.

The Impact of Growth Years on the Medicinal Material Characteristics and Metabolites of Stellaria dichotoma L. var. lanceolata Bge. Reveals the Optimal Harvest Age.

The original plant of Chinese medicine Stellariae Radix (Yin Chai Hu) is Stellaria dichotoma L. var. lanceolata Bge (abbreviated as SDL). SDL is a perennial herbaceous plant and a characteristic crop in Ningxia. Growth years are vital factors that affect the quality of perennial medicinal materials. This study aims to investigate the impact of growth years on SDL and screen for the optimal harvest age by comparing the medicinal material characteristics of SDL with different growth years. Additionally, metabolomics analysis using UHPLC-Q-TOF MS was employed to investigate the impact of growth years on the accumulation of metabolites in SDL. The results show that the characteristics of medicinal materials and the drying rate of SDL gradually increase with the increase in growth years. The fastest development period of SDL occurred during the first 3 years, after which the development slowed down. Medicinal materials characteristics of 3-year-old SDL exhibited mature qualities with a high drying rate, methanol extract content, and the highest content of total sterols and total flavonoids. A total of 1586 metabolites were identified, which were classified into 13 major classes with more than 50 sub-classes. Multivariate statistical analysis indicated significant differences in the diversity of metabolites of SDL in different growth years, with greater differences observed in metabolites as the growth years increased. Moreover, different highly expressed metabolites in SDL at different growth years were observed: 1-2 years old was beneficial to the accumulation of more lipids, while 3-5 years old was conducive to accumulating more alkaloids, benzenoids, etc. Furthermore, 12 metabolites accumulating with growth years and 20 metabolites decreasing with growth years were screened, and 17 significantly different metabolites were noted in 3-year-old SDL. In conclusion, growth years not only influenced medicinal material characteristics, drying rate, content of methanol extract, and total sterol and flavonoid contents, but also had a considerable effect on SDL metabolites and metabolic pathways. SDL planted for 3 years presented the optimum harvest time. The screened significantly different metabolites with biological activity, such as rutin, cucurbitacin e, isorhamnetin-3-o-glucoside, etc., can be utilized as potential quality markers of SDL. This research provides references for studying the growth and development of SDL medicinal materials, the accumulation of metabolites, and the selection of optimal harvest time.

Profil Fitokimia dan Penentuan Fenolik Total, Flavonoid Total, dan Uji Aktivitas Antioksidan Ekstrak Daun Sungkai (Peronema canescens Jack) dari Daerah Kota Padang

Sungkai (Peronema canescens Jack) is a plant that is easy to find in Indonesia. Sungkai leaves are used by the community as herbal medicine to boost immunity. The purpose of this study was to determine the phytochemical profile, total phenolic content and total flavonoids and test the antioxidant activity of Sungkai leaf extract. Sungkai leaf extract was obtained by maceration method in stages. The results of the phytochemical screening showed that ethyl acetate extract and methanol extract contained phenolic and flavonoid compounds. Total phenolic test using the method Folin-Ciocalteu with the highest total phenolic content in methanol extract namely 172.75 mg GAE/g sample. Total flavonoid test using the AlCl3 method with the highest total flavonoid content in the ethyl acetate extract, namely 825.714 mg QE/g sample. Antioxidant activity test using the ABTS method with a very strong antioxidant activity in methanol extract with IC50 of 13.589 mg/L.

Qualitative and Quantitative Analysis of Phytochemical Constituents of Alternanthera brasiliana (L.) Kuntze and Cassia alata (L.) using Different Organic Solvents

The therapeutic qualities of Alternanthera brasiliana and Cassia alata are due to the phytochemicals contained in these plants. This research was conducted to study the phytochemical analysis of leaf extracts of these plants using methanol, hexane, and chloroform solvents. The leaves were collected, cleaned, and dried for eight hours at 60°C in a cabinet dryer. Using the solvent extraction method, the extract was made. Qualitative and quantitative analysis was done to determine the occurrence of flavonoids, terpenoids, tannins, phenols, amino acids, alkaloids, phlobatannins, glycosides, and saponins. The extraction yields in methanol, hexane, and chloroform were 15.77%, 3.01%, and 3.16% for A. brasiliana and 26.91%, 19.5%, and 18.32% for C. alata. Alkaloids, terpenoids, tannins, phenols, glycosides, flavonoids, carbohydrates, phlorotannins, and saponins were all found in A. brasiliana. Alkaloids, tannins, phenols, glycosides, flavonoids, carbohydrates, and saponins were found in C. alata. Total phenolic, flavonoid, and tannin content in methanol extract for A. brasiliana were found to be 127.76 mg GAE/g, 136.48 mg GAE/g, and 58.88 mg GAE/g. The total phenolic, flavonoid and tannin content found were 41.76 mg GAE/g, 103.21 mg GAE/g, and 58.67 mg GAE in hexane extract. Similarly, 102.4 mg GAE/g, 112.49 mg GAE/g, and 41.76 mg GAE/g in the chloroform extract. Total phenolic, flavonoid, and tannin content of C. alata in the methanol extract were 36.3 mg GAE/g, 22.43 mg GAE/g, and 10.89 mg GAE/g, in the hexane extract, 25.98 mg GAE/g, 16.68 mg GAE/g, and 3.25 mg GAE/g, in the chloroform extract, 28.52 mg GAE/g, 20.33 mg GAE/g, and 4.56 mg GAE/g. From extraction yields and qualitative and quantitative analysis, methanol was the best solvent.

In vitro investigation of antimicrobial, enzyme inhibitory and free radical scavenging activities of Inula salicina L.

In this study, in vitro biological activities and total phenol/flavonoid contents of methanol extract (ISM) and its hexane (ISH), chloroform (ISC), ethyl acetate (ISEA) and aqueous methanol (ISAM) fractions obtained from aerial parts of Inula salicina were investigated. ISEA showed the highest antioxidant activity against DPPH and ABTS radicals with an IC50 value of 0.014 mg ml-1for both assays. ISEA exhibited a good anti-inflammatory activity with an IC50 value of 0.060 mg ml-1. ISEA was found to exhibit a moderate level of antidiabetic activity against α amylase enzyme with an IC50 value of 0.290 mg ml-1. ISEA and ISM presented low and moderate inhibitory activity against acetylcholinesterase and butyrylcholinesterase enzymes with IC50 values of 0.577 and 0.279 mg ml-1, respectively. ISC with MIC values of 78 and 156 µg ml-1 displayed a significant antimicrobial activity against Staphylococcus aureus and S. epidermidis, respectively. Almost all extracts had moderate effect against Candida species. The highest total phenolic and flavonoid contents were determined in ISEA with 574.8 mg GAE (gallic acid equivalent) g-1 extract and 30.48 mg QE (quercetin equivalent) g-1 extract, respectively. These results showed that ISEA had a good antioxidant and anti-inflammatory activity with moderate α-amylase and butyrylcholinesterase inhibitory activity. Also, ISC exhibited a significant antimicrobial activity against Staphylococcus species.

Effect of methanolic extract of Mimosa malacophylla A.Gray in vero and HEK-293 cell lines, and in the morphology of kidney and bladder of rats with induced urolithiasis

Ethnopharmacological relevanceUrolithiasis is the presence of stones in the kidney, ureters, bladder and/or urethra; it is the third most frequent disease of the urinary tract. Mimosa malacophylla A. Gray, is a species distributed in northern Mexico, where people traditionally use it for its diuretic effect, and to treat kidney diseases; however, no scientific reports have been found in relation to its antiurolithic properties. Aim of the studyThis study aimed to obtain a qualitative phytochemical profile of the methanolic extract (ME) of M. malacophylla, and to evaluate its potential cytotoxic effect in vitro and its antiurolithic activity in vivo. Material and methodsPhytochemical screening was performed to demonstrate the presence of secondary metabolite groups in the methanolic extract of M. malacophylla. In vitro cytotoxicity assays (MTT and nucleotide labeling with DAPI) were performed to evaluate the effect of the extract on kidney cell lines. Urolithiasis was induced in the bladder of Wistar rats introducing zinc disks for the calculus formation and exposed to three concentrations of ME. ResultsPhytochemical screening showed phenols, steroids, terpenoids and carbohydrates. In vitro analysis demonstrated that concentrations below 300 μg/mL of ME did not produce a cytotoxic effect on renal Vero and HEK-293 cells. In vivo analysis of 15 days of exposition, revealed that the extract at concentrations of 50 mg/kg to 150 mg/kg were effective as an antiurolithic treatment, and did not produce morphological alterations in kidney or bladder in murine model of induced urolithiasis. ConclusionsThe antiurolithic activity may be attributed to the presence of flavonoids, steroids and terpenes detected in the phytochemical screening which have been reported to possess this activity. These results could be useful to evaluate new alternatives and their potential therapeutic effect to treat renal or urinary affections.

Cytotoxic Effects of Some Nepeta Species against Breast Cancer Cell Lines and Their Associated Phytochemical Properties.

Nepeta is one of the largest genera of the Lamiaceae family. Nepeta species are commonly employed in traditional medicine for a variety of ailments, as well as food additives. In addition, they also come to the fore with their rich phytochemical content. In the present study, the quantitative phytochemical content of methanolic extracts and infusions prepared from the aerial parts of 14 Nepeta taxa collected from Turkey and their cytotoxic effects on two breast cancer cell lines, MCF-7 and MDA-MB-231, were investigated by using the MTT (3-(4,5-dimethylthiazol-2-yl))-2,5-diphenyltetrazolium-bromide) test. According to HPLC-PDA analysis, N. racemosa methanolic extract had the highest ursolic acid content with 165.9 mg/g extract. Total sterol, total iridoid, and total triterpenoid content were determined to be greatest in the methanolic extracts of N. meyeri, N. trichocalyx and N. phyllochlamys. The MTT experiment demonstrated that certain Nepeta species suppressed the growth of MCF-7 and MDA-MB-231 cells in a dose-dependent manner. Statistical analysis revealed a strong correlation between the cytotoxic effects of the extracts and their triterpene content. In conclusion, the data obtained from this study are important in terms of forming a basis for advanced anticancer activity studies on breast cancer with Nepeta sp.

Flavonoid components, chemotypes, and candidate chemical markers of Teucrium (Lamiaceae) species using HPLC-MQ-API-MS/MS

Belonging to Lamiaceae family, the genusTeucriumL. is a rich source of secondary metabolites used in pharmacology and industrial medicine. This study aimed to investigate the flavonoid contents of methanol extract and the chemical markers ofTeucriumat specific and infraspecific levels and identify the chemotypes of its accessions. Nineteen accessions, eight species, and five subspecies were collected from three sections (Teucris, Polium, andScordium) belonging to the natural habitats in the west, southwest, centre, and south of Iran. Total flavonoid extraction was attained by the air-dried leaf ofTeucriumspecies and 90% methanol. HPLC coupled with triple quadrupole mass analyser was applied to screen and identify the chemical constituents. Multivariate analyses were implemented by cluster and principal component analysis. Chemical investigation specified 148 chemical compounds, of which 135 flavonoids were tentatively identified. The most abundant derivatives belonged to flavones (41 derivatives), flavonols (37 derivatives), and isoflavonoids (22 derivatives). Based on the cluster analysis, 13 groups of chemical markers were formed in sectionsTeucrisandScordiumand five groups in sectionPoliumas chemotaxonomic indicators. The specified chemotypes were represented by ten in the sectionsTeucrisandScordiumand by six in the sectionPolium. A relationship was suggested between type and flavonoid quantity (number), altitude, and geographical location. The present study provided valuable information for pharmacological research, bioactivity assessment, and chemotaxonomic purposes. This is the first report on the presence of chemotypes, chemical markers, and quality assessments for the genusTeucrium.

Suppression of Oxidative Stress and Proinflammatory Cytokines Is a Potential Therapeutic Action of Ficus lepicarpa B. (Moraceae) against Carbon Tetrachloride (CCl4)-Induced Hepatotoxicity in Rats.

Local tribes use the leaves of Ficus lepicarpa B. (Moraceae), a traditional Malaysian medicine, as a vegetable dish, a tonic, and to treat ailments including fever, jaundice and ringworm. The purpose of this study was to look into the possible therapeutic effects of F. lepicarpa leaf extract against carbon tetrachloride (CCl4)-induced liver damage in rats. The DPPH test was used to measure the antioxidant activity of plants. Gas chromatography-mass spectrometry was used for the phytochemical analysis (GCMS). Six groups of male Sprague-Dawley rats were subjected to the following treatment regimens: control group, CCl4 alone, F. lepicarpa 400 mg/kg alone, CCl4 + F. lepicarpa 100 mg/kg, CCl4 + F. lepicarpa 200 mg/kg and CCl4 + F. lepicarpa 400 mg/kg. The rats were euthanized after two weeks, and biomarkers of liver function and antioxidant enzyme status were assessed. To assess the extent of liver damage and fibrosis, histopathological and immunohistochemical examinations of liver tissue were undertaken. The total phenolic content and the total flavonoid content in methanol extract of F. lepicarpa leaves were 58.86 ± 0.04 mg GAE/g and 44.31 ± 0.10 mg CAE/g, respectively. F. lepicarpa’s inhibitory concentration (IC50) for free radical scavenging activity was reported to be 3.73 mg/mL. In a dose-related manner, F. lepicarpa was effective in preventing an increase in serum ALT, serum AST and liver MDA. Histopathological alterations revealed that F. lepicarpa protects against the oxidative stress caused by CCl4. The immunohistochemistry results showed that proinflammatory cytokines (tumour necrosis factor-α, interleukin-6, prostaglandin E2) were suppressed. The antioxidative, anti-inflammatory, and free-radical scavenging activities of F. lepicarpa can be related to its hepatoprotective benefits.

PHYTOCHEMICAL CONSTITUENT, ACUTE AND SUBACUTE TOXICITY STUDIES OF METHANOL EXTRACT OF LAPORTEA AESTUANS LEAF

Laportea aestuans is an African herbaceous weed. This work assessed the bioactive compound content of methanol extract of Laportea aestuans leaf (LALME) and its acute and subacute toxicity effects in Male Wistar rats. According to qualitative and quantitative phytochemical analysis, LALME possesses a significant amount of phenolics, flavonoids, alkaloids, tannin, carotenoids, and anthraquinones. LALME is regarded as safe because it did not cause death in rats when given doses of up to 2,000 mg/kg body weight. LALME at 200 and 400 mg/kg markedly increased (p<0.05) serum activity of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and gamma-glutamyl transferase (GGT). There was a decrease in liver LDH activity in rats administered LALME, but no significant alteration was noticed in the serum of the rats. The total bilirubin concentration in the serum of rats given 200 mg/kg LALME and in the liver of rats administered 200 and 400 mg/kg LALME was significantly higher (p<0.05) than that in the serum and liver of control rats. The serum of rats given LALME had a nearly threefold increase in direct bilirubin compared to control. However, rats administered 200 mg/kg of LALME had an increased serum urea concentration compared to control. When comparing photomicrographs of the liver and kidney of rats given LALME to those of control rats, the liver and kidney of rats given LALME showed minor congestion and renal dissemination, respectively. In conclusion, LALME was found to be safe in acute toxicity; nevertheless, subacute administration may cause variations in

Crinum jagus (J. Thomps. Dandy): Antioxidant and protective properties as a medicinal plant on toluene-induced oxidative stress damages in liver and kidney of rats

Crinum jagus (C. jagus; J. Thomps.) Dandy (Liliaceae) is a pantropical plant known for its medicinal values and pharmacological properties. The study assessed the protective effects and changes in oxidative stress indices due to C. jagus leaf extracts on the toluene-induced liver and kidney injuries in rats. The study was conducted on 8-week-old male Wistar rats (n = 80), weighing 243.3 ± 1.42 g. Group I, 1 ml/kg distilled water for 7 days; Group II, 4.5 ml/kg toluene once, 1 ml/kg distilled water for 7 days; Group III, 4.5 ml/kg toluene once, 500 mg/kg methanolic extract for 7 days; Group IV, 4.5 ml/kg toluene once, 500 mg/kg aqueous extract for 7 days; Group V, 500 mg/kg methanolic extract for 7 days; Group VI, 500 mg/kg aqueous extract for 7 days; Group VII, 500 mg/kg of vitamin C for 7 days; Group, VIII, 4.5 ml/kg toluene once, 500 mg/kg vitamin C for 7 days, all administrations were given by oral gavage. The phytochemical contents, absolute and relative organ weights of liver and kidneys, liver and kidney function tests, antioxidant status, as well as histological tests were analyzed using standard protocols. The tannins, flavonoids, and polyphenols were in highest concentration in both extracts, content in methanol extract (57.04 ± 1.51 mgg−1, 35.43 ± 1.03 mgg−1, 28.2 ± 0.34 mgg−1 respectively) > aqueous extract (18.74 ± 1.01 mgg−1, 13.43 ± 0.47 mgg−1, 19.65 ± 0.21 mgg−1 respectively). In the negative control group (II), bodyweights significantly (P < 0.05) reduced by 22%, liver weight and kidney weight significantly (P < 0.05) increased by 42% and 83% respectively, liver-to-bodyweight and kidney-to-bodyweight ratios increased significantly (P < 0.05); serum liver function tests (LFTs) i.e., bilirubin, alkaline phosphatase (ALP), Alanine aminotransferase (ALT), Aspartate aminotransferase (AST), Gamma-glutamyl transferase (GGT), and serum kidney function tests (creatinine and urea) were significantly (P < 0.05) elevated; oxidant status (tissue malondialdehyde; MDA) was significantly (P < 0.05) elevated, antioxidant status i.e., tissue superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH) levels was significantly (P < 0.05) reduced; with markedly visible renal and hepatic histopathological findings, compared to the normal control group. In C. jagus extract test groups (III and IV), the parameters were significantly (P < 0.05) alleviated and reversed to normal/near normal compared to the negative control. The LFTs, kidney function tests, and antioxidant status were significantly (P < 0.05) more improved with the methanol extract test and standard control groups compared to the aqueous extract test group; Also, the methanol extract test group showed better histological features than the aqueous extract test and standard control groups. The methanolic extract shows better antioxidant potential due to the availability of more nonenzymatic antioxidants (tannins, flavonoids, and polyphenols). The findings showed that toluene is a very aggressive xenobiotic due to the promotion of oxidative stress and peroxidation of cellular lipids, but C. jagus leaves provide significant protection through the reducing power of nonenzymatic antioxidants and their ability to induce endogenous antioxidant enzymes (SOD, CAT, and glutathione reductase or GR) causing reduced cellular lipid peroxidation and tissue damages, quickened tissue repair, and improved cell biology of liver and kidneys during toluene toxicity. The methanol leaf extract provides better protection and should be advanced for more experimental and clinical studies to confirm its efficacy in alleviating oxidative stress tissue injuries, specifically due to toluene.

Evaluation of Antileishmania Effect of Methanolic Extract of Dandelion Root (Taraxacum Officinale) on Leishmanaia Major Promastigotes in Vitro Techniques

Background and Objectives: Cutaneous leishmaniasis is one of the common diseases between humans and animals that is considered a health problem in Iran and many countries around the world. Currently, glucantime is used to treat cutaneous leishmaniasis. Due to its high side effects and resistance, the use of alternative therapies, especially plants and natural compounds, has been highly recommended by researchers. The aim of this study is to investigate the effect of the methanolic extract of dandelion root (Taraxacum officinale) on Leishmania major in vitro techniques. Methods: In this experimental study, the extract was prepared by the Soxhlet method and to evaluate the effect of methanolic extract of dandelion root on L. major parasite at concentrations of 300-1200 mg/ml. Amphotericin B and distilled water were considered as positive and negative controls, respectively. Then, 106 live parasites were added to all tubes, and all groups were kept at 25 ±1 °C. At 24, 48, and 72 hours, the number of live parasites was counted by Trypan Blue using a neobar slide and light microscope (Hemocytometer method). Then Half-maximal inhibitory concentration (IC50) value for the above extract was calculated using SigmaPlot™ 13 software. All steps of the experiment were done in triplicate and the results were considered as average. Results: The IC50 content of methanolic extract of dandelion roots after 24, 48, and 72 hours on L. major were calculated to be 1.04, 0.9, and 0.68 µg/ml. The highest growth inhibition (100%) was observed at a concentration of 1200 μg/ml after 72 hours of exposure. There was a significant difference between the IC50 extract and amphotericin B drug after 24, 48 and 72 hours (P&lt;0.05). Conclusion: Methanolic extract of dandelion root in different concentrations has an inhibitory effect on the growth of L. major and it is suggested that more and more complete studies be performed on the components of this plant and the lethal effect of the parasite in vivo.

An insight of anopheline larvicidal mechanism of Trichoderma asperellum (TaspSKGN2)

Anopheline larvicidal property of T. asperellum has been found recently in medical science. The mechanism of actions exhibited by T. asperellum to infect mosquito larvae is the pivotal context of our present study. To infect an insect, entomopathogens must undergo some events of pathogenesis. We performed some experiments to find out the mechanisms of action of T. asperellum against anopheline larvae and compared its actions with other two well recognized entomopathogens like Metarhizium anisopliae and Beauveria bassiana. The methodology adopted for this includes Compound light and SE Microscopic study of host–pathogen interaction, detection of fungal spore adhesion on larval surface (Mucilage assay), detection of cuticle degrading enzymes (Spore bound pr1, chitinase and protease) by spectro-photometric method, Quantitative estimation of chitinase and protease enzymes, and determination of nuclear degeneration of hemocyte cells of ME (methanolic extract) treated larvae by T. asperellum under fluorescence microscope. Compound light microscopic studies showed spore attachment, appressorium and germ tube formation, invasion and proliferated hyphal growth of T. asperellum on epicuticle and inside of dead larvae. SEM study also supported them. After 3 h of interaction, spores were found to be attached on larval surface exhibiting pink colored outer layer at the site of attachment indicating the presence of mucilage surrounding the attached spores. The enzymatic cleavage of the 4-nitroanilide substrate yields 4-nitroaniline which indicates the presence of spore-bound PR1 protein (Pathogenecity Related 1 Protein) and it was highest (absorbance 1.298 ± 0.002) for T. asperellum in comparison with control and other two entomopathogens. T. asperellum exhibited highest enzymatic index values for both chitinase (5.20) and protease (2.77) among three entomopathogens. Quantitative experiment showed that chitinase enzyme concentration of T. asperellum (245 µg mL−1) was better than other two M. anisopliae (134.59 µg mL−1) and B. bassiana (128.65 µg mL−1). Similarly protease enzyme concentration of this fungus was best (298.652 µg mL−1) among three entomopathogens. Here we have detected and estimated fragmentized nuclei of hemocyte cells by fluorescence microscopy in treated larvae with different ME doses of T. asperellum, and also observed that mosquito larvae exposed to 0.1 mg mL−1 dose of ME showed maximum (100%) nuclear fragmentations of hemocytes and while 20, 45, 70 and 85% of nuclear deformities were recorded at 0.02, 0.04, 0.06 and 0.08 mg mL−1 concentrations of ME. The knowledge of this work certainly will help in understanding of mechanism of action of T. asperellum for anopheline larval killing and consequently in eradication of malaria vector.

Quality control parameters of prickles of Bombax ceiba Linn.

Bombax ceiba Linn. is a deciduous tree known as Red silk cotton tree. Bombax ceiba stem, stem bark and prickle are used for the treatment of rheumatism, swellings, asthma and inflammation of the legs. The stem bark of Bombax ceiba is official in the ayurvedic Pharmacopoeia of India recommends its use in acne. Prickles are also used and recommended in anti-acne marketed formulation. However, limited work was carried out on the prickle toward establishing quality control parameters. Studies were therefore carried out to determine the phytochemical and pharmacognostic profile of prickle of Bombax ceiba Linn. The study, including determination of macroscopic, microscopic characters, ash values, extractive values, loss on drying, phytochemical screening, total phenolic content, TLC and HPTLC fingerprinting were carried out. Successive extracts were prepared using soxhlet extraction. The phytochemical analysis of extracts revealed the presence of tannin, saponin, phenol, carbohydrate, steroids, triterpenoids and flavonoids. Total phenolic and tannin content of methanol extract was found 327.78±1.09mg GAE/gm equivalent and 4.98±0.22%w/w, respectively. The mobile phase showing distinct spots in TLC was found in Toluene: Ethyl acetate (9.5:0.5) and (9:1) for petroleum ether and methanol extracts, respectively. HPTLC fingerprinting shows 8 & 3 peaks at 366nm for petroleum ether and methanol extract, respectively.

Improvement of endothelial function by Gunnera tinctoria extract with antioxidant properties

BackgroundGunnera tinctoria has been collected by Mapuche-Pewenche people for food and medicinal purposes. The high polyphenol content of methanolic extract from G. tinctoria leaves with chemical constituents such as ellagic acid and quercetin derivatives suggests its application to prevent endothelial dysfunction and oxidative stress. The aim of this study was to provide evidence of the protective effect of this extract on endothelial function by reducing oxidative stress induced by high d-glucose and H2O2, as well as by stimulating nitric oxide (NO) levels in human umbilical vein endothelial cells (HUVECs).ResultsA methanolic extract with a high content of polyphenols (520 ± 30 mg gallic acid equivalents/g dry extract) was obtained from G. tinctoria leaves. Its main constituent was ellagic acid. The results of Ferric reducing antioxidant power and 2,2-diphenyl-1-picrylhydrazyl radical scavenging assays of the extract confirmed its antioxidant activity by inhibition pathway of radical species. The incubation of HUVECs with the extract decreased the apoptosis and reactive oxygen species (ROS) synthesis induced by high extracellular concentration of d-glucose or hydrogen peroxide. The extract increased endothelial NO levels and reduced vasoconstriction in human placental vessels.ConclusionsThis study provides evidence about the antioxidant and endothelial protective properties of methanolic G. tinctoria leaf extract. The extract improves the availability of NO in HUVECs, inhibiting the production of ROS and vasoconstriction.

Genotypic characterization and steviol glycoside quantification in a population of Stevia rebaudiana Bertoni from Paraguay

Stevia rebaudiana (Bertoni) Bertoni is a widely grown species in various regions of the world, mainly due to its sweetening properties attributed to steviol glycosides. Despite the fact that Paraguay is the center of origin of S. rebaudiana, genetic characterization of local genotypes has not been studied. Considering that accumulation of steviol glycosides is a complex trait, molecular breeding could be an effective method for its improvement. Therefore, the identification of molecular markers associated with steviol glycoside accumulation is important. The aim of this study was to assess the genetic relationship among eight advanced lines of S. rebaudiana and two varieties selected from local germoplasm and quantify their steviol glycoside content. The stevioside and rebaudioside A content in methanolic extract was determined by high-performance liquid chromatography (HPLC), and the molecular characterization of the ten genotypes of S. rebaudiana was performed using SSR and ISSR markers. A wide range of variability was observed for the metabolites of interest under greenhouse conditions. The variety Eirete presented highest stevioside and rebausiode A content, followed by the variety Katupyry. One of the advanced line presented both metabolite content similar to that of Katupyry. Genotyping of these lines showed two main clusters. Cluster I contained the both varieties and advanced lines with higher stevioside and rebaudioside A content. Our results suggest that S. rebaudiana lines with genetic profile similar to that of cluster I, determined mainly by genotype at the markers SUGMS 35 and ISSR 47, might present higher stevioside and rebaudioside A content.

Total Phenolic and Flavonoid Content, Antioxidant Activity of Talaromyces funiculosus Strains

Microscopic fungi synthesize a wide range of phenolic compounds that play an important role in cell wall formation, as well as perform signaling and protective functions. One of the important functions of phenolic compounds, primarily polyphenols, is antioxidant activity. The most active polyphenols are flavonoids, their synthesis in microscopic fungi has not been studied enough. The aim of the study was to determine the total phenols and flavonoids content and antioxidant activity of Talaromyces funiculosus strains isolated from different econiches. Methods. 20 strains of the microscopic fungus T. funiculosus – 10 endophytes and 10 saprotrophs were used in the experiments. Cultivation of filamentous fungi was performed under submerged conditions with shaking at 210–230 rpm in Erlenmeyer flasks containing 200 ml of Czapek liquid medium at a temperature 26±2°C for 6 days. The biomass of the strains was filtered, dried at 70°C and extracted with 95% methanol (1:50 w/v). The total phenols content was determined using Folin-Ciocalteu reagent by the method of Arora, Chandra (2010) with some modifications and expressed as mg of gallic acid equivalents (GAE) per g of dry weight mycelium. Flavonoids in each extract were determined spectrophotometrically with AlCl3 and expressed as mg of quercetin equivalents (QE) per g of dry weight mycelium. Antioxidant activity of methanolic extracts was measured as radical scavenging activity by the percentage inhibition of the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical. Results. The total phenolic content of methanolic extracts of T. funiculosus strains ranged from 0.12±0.007 to 1.97±0.150 mg GAE/g. The highest level of phenolic compounds was found for saprotroph UCM F-16790 (1.97±0.150) and endophyte UCM F-16795 (1.71±0.137 mg GAE/g). The flavonoid content in methanolic extracts of the studied strains varied in the range of 0.02±0.001–0.39±0.041 mg QE/g. All studied strains of T. funiculosus showed antioxidant activity against DPPH radical, which was 12.19±2.34–60.37±7.24% inhibition. Statistical analysis revealed a strong positive correlation between the total phenols and flavonoids content (Pearson’s correlation coefficient r=0.92); between antioxidant activity, phenols (r=0.82) and flavonoids (r=0.81) content in methanol extracts of T. funiculosus strains. The total content of phenols and flavonoids and antioxidant activity did not depend on the trophic group of the studied strains. The moderate correlation between flavonoids content and antioxidant activity of methanolic extracts of endophytic T. funiculosus strains (r=0.46) may be related to their ability to synthesize a wider range of polyphenolic compounds with antioxidant activity. Conclusions. The studied T. funiculosus strains have the ability to synthesize phenolic compounds and flavonoids in different concentrations. Correlation analysis revealed strong correlations between total phenolic and flavonoid contents and antioxidant activity. The reason for phenomenon of moderate correlation (r=0.46) between flavonoid content and antioxidant activity in endophytic strains may be the synthesis of other compounds by individual strains. The determination of individual components synthesized by the most active strains is of scientific interest for further study. T. funiculosus UСM F-16795 and UСM F-16790 strains had a quite high level of phenols and flavonoids. Obtained data may be promising for the search for phenolic compounds with antioxidant properties to increase plant resistance to negative impacts of environmental stress factors.

The Effect of Methanolic Extract and Ethyl Acetate Fraction of Kepel Fruit (Stelechocarpus burahol) to α-Glutathione S-Transferase Enzyme Concentration of Rat Liver and Blood Induced by CCl4

Degenerative diseases are mostly caused by high free radical concentrations in the body. Kepel fruit(Stelechocarpus burahol) is known to contain flavonoid compounds, a class of compounds that has free radicalscavenging activity in the body and could affect the activity of antioxidant enzymes. There have been reportsthat methanolic extract and ethyl acetate fractions of Kepel fruit have the lowest IC50 values, compared toextracts or fractions which are made using other organic solvents, on in vitro assays so that flavonoids aresuspected as responsible compounds to the free radical scavenging activity. This study aimed to identify thepresence of flavonoid content in methanolic extract and the fraction of ethyl acetate of Kepel fruit and also itseffect on rat alpha Glutathione S-Transferase (α-GST) enzyme concentration for the function to prevent hepaticcell damage due to carbon tetrachloride exposure. The identification of flavonoids content used a series ofcolor chemical reaction tests and thin layer chromatography that used silica gel 60F254 as the solid phase andmixture of ethyl acetate, formic acid, glacial acetic acid, and water (100: 11 : 11: 20 v/v) as the mobile phase.This study used 30 Sprague-Dawley female rats (age 2-3 months) that were grouped into 5 groups as solventcontrol (CMC-Na) group, negative control (CCl4) group, positive control (vitamin C) group, methanolic extractgroup, and ethyl acetate fraction group. All groups were induced with carbon tetrachloride (CCl4) compoundsexcept for the CMC-Na solvent control group. Blood sampling at 0 h, 24 h, and 72 h, while liver sampling at 24h and 72 h, with 3 samples each group. The analysis of rat α-GST enzyme concentrations used theEnzyme-linked Immunosorbent Assay (ELISA) method. Based on the phytochemical test, the methanolicextract and the ethyl acetate fraction of Kepel fruit have been identified contains flavonoids compound. Basedon the in vivo study, the positive control group's α-GST enzyme concentration was decreased, while thenegative control group's α-GST enzyme concentration was increased. Administration of methanolic extractand ethyl acetate fraction also decreased both liver and blood rat α-GST enzyme concentrations, althoughwithout significant correlation, and still could prevent the hepatic cell damage due to carbon tetrachlorideexposure.

Total Flavonoid Content and Antioxidant Activity of Temu Mangga (Curcuma mangga Val. & Zijp) and its Classification with Chemometrics

Indonesia is a rich country in terms of its biodiversity. Herbs that are widely used tomaintain a healthy body comes from the Zingiberaceae family, including temu mangga. Temu mangga(Curcuma mangga Val. &amp; Zijp) is effective for relieving gastric pain, diarrhea, menstrual pain, acneand ulcers, and increasing appetite. Besides, temu mangga is considered to have antioxidant activitydue to the presence of the phenolic and flavonoid compounds. This study aims to determine thecorrelation and classification of temu mangga from various places based on total flavonoid contentand antioxidant activities. The samples were extracted with methanol, then tested for totalflavonoid content by AlCl3 method and antioxidant activity by DPPH free radical capture method.Analysis was performed using correlation coefficient value and chemometrics PCA (PrincipalComponent Analysis) and CA (Cluster Analysis) to see the relationship between total flavonoidcontent with their antioxidant activity and to see their classification. Total flavonoid content ofmethanolic extract of temu mangga gives a moderate correlation to its antioxidant activity with r =-0.6085 and clustering samples successfully determined by PCA and CA which resulted in 4 groups.Keywords: Curcuma mangga Val. &amp; Zijp, Methanolic Extract, Flavonoid, Antioxidant, PCA, CA

Polyphenols/Flavonoids Analysis and Antimicrobial Activity in Pomegranate Peel Extracts

The use of plants still plays a vital role to establish the basic health requirements in developing countries. The extracts of P. granatum and its compounds have shown antimicrobial activity. This study showed that the acetone (125.36±0.01 and 70.5±0.01) extract contained the highest amount of total polyphenols (TPPC) and total flavonoids content (TFC) respectively compared to methanol (105.6±0.005 and 20.99±0.004) extract while methanol extract content was higher than ethanol (94.14±0.02 and14.48 ±0.005). The aqueous extracts content was the less one (83.17 ± 0.003 and 10.65±0.003). When study the antimicrobial activity of four different prepared concentrations of four pomegranate peel extracts against microorganisms including two Gram-positive bacteria [P. aeruginosa and S. aureus], one Gram-negative bacteria [A. baumannii] and Candida parapsilosis. The result shown the antimicrobial activity of acetone extract on all microorganisms the highest value followed by methanol, ethanol, and aqueous extract activity. Besides, the activity of all extracts was good excepted aqueous extract has low antimicrobial activity against S. aureus.