Chloroform-methanol Extraction Research Articles

In vitro α-amylase and α-glucosidase inhibitory effects and antioxidant potential of new dihydrochalcones from Baphia massaiensis Taub

Two previously undescribed dihydrochalcones, 4,2′-dihydroxy- [2″, 2″-dimethylpyrano (3″, 4″: 4′, 5′]) dihydrochalcone (1) and 4, 2′, 4′, 2″-tetrahydroxy-5′-(3″-methylbut-3″-enyl) dihydrochalcone (2), along with six known compounds (3-8) were isolated from chloroform-methanol (1:1) extract of Baphia Massaiensis twigs. Their chemical structures were determined using NMR analysis, mass spectrometry, electronic circular dichroism (ECD), density functional theory (DFT) calculations, and a thorough comparison with reported data from the literature. All the isolated compounds (1−8) were assessed for their antioxidant activities, α-amylase and α-glucosidase inhibition effects. Dihydrochalcone 1 showed significant antioxidant activity with IC50 value of 118.8 ± 2.95 µM, and a more potent antioxidant activity was exhibited by compounds 3 and 6 with IC50 values of 84.3 ± 0.07 µM and 62.1 ± 1.59 µM, respectively, compared to the positive control, gallic acid with IC50 value of 92.9 µM. Dihydrochalcone 2 showed moderate inhibition against α-amylase and α-glucosidase with IC50 values of 44.0 ± 4.83 µM and 51.7 ± 9.72 µM compared to the standard acarbose with IC50 values of 22.9 ± 0.92 µM and 31.5 ± 0.63 µM, respectively. Molecular docking studies of the new dihydrochalcones 1 and 2, revealed the active binding sites which support the observed α-amylase and α-glucosidase inhibitory effects.

Isolation of two steroidal saponins with antileishmanial activity from Allium giganteum L.

Alliums are rich sources of steroidal saponins, flavonoids, and sulphoric compounds of which steroidal saponins have recently received more attention due to their important pharmacological activities. Allium giganteum (giant onion) which is named locally "Couria" in the Northeast of Iran, is grown widely in "Kouh-Sorkh" mountains in Khorasan province. Phytochemical investigation of chloroform-methanol and aqueous extract of the plant resulted in the isolation and identification of two steroidal saponins, using comprehensive spectroscopic methods including 1D and 2D NMR and MS. The chemical structures of the isolated saponins were determined as (22S)-cholesta-1b,3b,16b,22b-tetraol 5-en, and 3-O-β-D-glucopyranosyl26-O-β-D-glucopyranosside and (25R)-26-O-β-D-glucopyranosyl-5α-furostan-1α,3β,22α,26-tetraol3-O-{β-D-galactopyranosyl-(1→2)-O-[β-D-xylopyranosyl- (1→3)]-O-β-D-glucopyranosyl-(1→4)-β-D-galactopyranoside}. Investigation of in vitro antileishmanial activity of the isolated compounds at 10, 50, and 100 μg/mL exhibited significant leishmanicidal against the promastigotes of Leishmania major. The results established a valuable basis for further studies about A. giganteum and the anti-parasitic activity of steroidal saponins.

Pengujian Toksisitas dan Antioksidan Ekstrak Etilasetat dan n-Butanol Hipokotil Sarang Semut, Myrmecodia pendens Merr. & Perry

Sarang semut, Myrmecodia pendens Merr. & Perry (Rubiaceae) is an epiphytic plant from Papua which is used to cure various diseases. The purpose of this study is to see the chromatogram profile [Thin Layer Chromatography (TLC) and High Performance Liquid Chromatography (HPLC)] analysis, and biological test of ethyl acetate and n-butanol of Sarang semut’s hypocotyl extracts from methanol extracts. Meanwhile, the biological tests carried out were toxicity tests on Artemia salina L shrimp larvae and antioxidant assay with free radical scavenging methods (DPPH). The results of the antioxidant activity test showed that the antioxidant activity of the ethyl acetate extract was higher (IC50=18.23μg/ml) than the n-butanol extract (IC50=29.58μg/ml). The results of the toxicity test on the ethyl acetate extract were higher (LC50=101.84 g/ml) than the n-butanol extract (LC50=154.59 g/ml). TLC analysis results showed that the mobile phase for the best separation of chemical compounds in ethyl acetate extract was chloroform-methanol (10:1) and n-butanol extract was the mobile phase chloroform-methanol-water (5:5:1), while HPLC analysis showed that the mobile phase for the separation of chemical compounds in the ethyl acetate extract was n-hexane-ethyl acetate (10:1) and the n-butanol extract was methanol-water (10:1).

ZNF143 inhibits hepatocyte mitophagy and promotes non-alcoholic fatty liver disease by targeting increased lncRNA NEAT1 expression to activate ROCK2 pathway

ABSTRACT Background: Nonalcoholic fatty liver disease (NAFLD) is the most common hepatic disorders worldwide. The mitophagy is suggested to be repressed in NAFLD, but the mechanism remains to be elucidated. Methods: NAFLD cell and mouse models were established by treating with free fatty acid (FFA) and feeding a high fat diet (HFD), respectively. QRT-PCR, Western blotting, or IHC measured the expression of ZNF143, lncRNA NEAT1, ROCK2, and lipid formation/mitophagy-related proteins. Cell viability and mitophagy were evaluated by MTT and immunofluorescence. The chloroform-methanol extraction method measured triglyceride and total cholesterol levels. ELISA detected ALT and AST levels. The interactions among ZNF143, lncRNA NEAT1 and SND1 were analysed by ChIP, dual-luciferase reporter, pull-down, and RIP. The lipid droplets were determined by Oil-red O and HE staining. Results: ZNF143 and lncRNA NEAT1 were upregulated in hepatic cells treated with FFA (p < 0.01 and p < 0.001). Knockdown of ZNF143 or lncRNA NEAT1 inhibited lipid droplets formation, while promoting mitophagy (p < 0.01 and p < 0.001). ZNF143 promoted lncRNA NEAT1 transcriptional expression through binding to its promoter. LncRNA NEAT1 increased ROCK2 mRNA stability by targeting SND1. LncRNA NEAT1 or ROCK2 overexpression reversed the effect of ZNF143 or lncRNA NEAT1 knockdown on hepatic steatosis and mitophagy (p < 0.01 and p < 0.001). ZNF143 or lncRNA NEAT1 knockdown inhibited HFD-induced steatosis and promoted mitophagy in vivo (p < 0.01 and p < 0.001). Conclusion: The upregulation of lncRNA NEAT1 caused by ZNF143 promoted NAFLD through inhibiting mitophagy via activating ROCK2 pathway by targeting SND1, providing potential targets for NAFLD therapy.

Evaluation of the Total Content of Phenols and Flavonoids in Two Different Extracts of Orbea wissmannii O. Schwart and their Toxic Effect on WI38 and HepG2 Cell Lines.

This study aimed at estimating the total phenolic content, total flavonoid content and evaluation of toxic effect of chloroform and aqueous methanol extracts of Orbea wissmannii against WI38 and HepG2 cell lines. Orbea wissmannii was collected from Qaren region near Amran governorate, Yemen. Chloroform and methanol extracts of whole parts of Orbea wissmannii were prepared by maceration and decantation. The total polyphenol content was estimated using Folin–Ciocalteau method. Aluminum chloride colorimetric method was applied for the determination of total flavonoid content. The cytotoxic activity was measured using the MTT assay. The total phenolic content of chloroform and methanol extracts was 70.21 mg GAE/g, 122 mg GAE/g respectively (milligrams of gallic acid equivalents per gram), while the flavonoid content was 21.80 mg QE/g and 54.24 mg QE/g respectively (milligrams of quercetin dihydrate equivalents per gram). Dependently, the two extracts significantly and concentratedly reduced the viability of lung tissue-derived fibroblasts (wi38) and human hepatoma cells (HepG2). The two extracts showed effective effect on HepG2 at 250, 500 and 1000 μg/ml, while the same extracts showed their effect on the WI38 at 500 and 1000 μg/ml. The toxicological properties shown by the extracts of this plant require further study to isolate and purify the active substances responsible for this high anti-cancer efficacy. Phytochemical investigation on chloroform/methanol extracts and their structures is recommended.

GC-MS Analysis of Bioactive Compounds Extracted from Plant Rhazya stricta Using Various Solvents.

Worldwide, human beings have traditionally employed many folkloric herbal resources as complementary and alternative remedies, and these remedies have played a pivotal role in modern medicines for many decades, as scientists have used them to develop drugs. We studied the effects of employing solvents with varying polarity on the yields of phytochemical components extracted from the plant Rhazya stricta. We used chloroform-methanol (1:1), methanol, ethanol, diethyl ether, and ethyl acetate as extraction solvents. The results showed that the efficiencies of the solvents at extracting phytochemical compounds were in this order: chloroform-methanol < ethanol < methanol < diethyl ether < ethyl acetate extract. The chloroform-methanol extract produced the highest concentration of phenolic and flavonoid contents among the five solvents tested (13.3 mg GAE/g DM and 5.43 CE/g DM). The yields of the extracted phytochemical compounds ranged from 47.55 to 6.05%. The results revealed that the properties of the extraction solvents considerably impacted the extraction yield and the phytochemical components of the R. stricta extract. Furthermore, compared with the other solvents, the chloroform-methanol extraction led to the highest yield (47.55%) and to more phytochemical substances being extracted. The aim of this study is to investigate the phytochemical compounds extracted from R. stricta with different solvents that have different polarities.

Low-cost rapid workflow for honey adulteration detection by UV–Vis spectroscopy in combination with factorial design, response surface methodology and supervised machine learning classifiers

Honey is often adulterated and heat treated for higher profits. This study aims to develop a low-cost, easy to perform, and less time-consuming method for screening the honey status. Known (12: adulterated 4; unadulterated 8) and unknown (7) honey samples of different geographical and biological sources were used to simulate adulteration and heat treatment effects by factorial design. Crude methanol-chloroform extracts of simulated samples were used for generating UV–Vis spectra (200-700 nm) and the effects were assessed by response surface methodology. Four machine learning classifiers were applied to predict the sample status, and among them Neural network and Random-forest were found to be satisfactory. The selected classifiers successfully detected 2 adulterated, and 4 unadulterated honeys among 7 unknown samples. Present workflow is proved to be an effective technique in detecting honey adulteration, and can be applied in honey and other food industries for optimization and quality control of their products.

&lt;i&gt;Lentinus squarrosulus&lt;/i&gt; Mont. Mushroom: Molecular Identification, In vitro Anti-Diabetic, Anti-Obesity, and Cytotoxicity Assessment

Mushrooms are fungi with nutritional and health benefits. Lentinus squarrosulus Mont., an edible fungus, has traditional usage and relevance in local therapy for managing metabolic diseases. In that view, this study aimed to evaluate the in vitro anti-obesity, anti-diabetic, and cytotoxic potential of the chloroform/methanol extract (CME) and aqueous extract (AE) of the mushroom. L. squarrosulus was identified using molecular biology tools. The CME and AE were obtained sequentially and, then, subjected to α-amylase, α-glucosidase, and lipase inhibitory enzyme assays as well as total phenolic content (TPC) and flavonoid content (TFC) estimations. The cytotoxic potential of extract fractions of L. squarrosulus was assessed using the brine shrimp lethality assay. The molecular identification of the mushroom displayed that the internal transcribed spacer sequence was an equivalent match to that of L. squarrosulus with a high percentage similarity, and thus assigned a unique accession number (KT120043.1). The CME of L. squarrosulus had higher TPC, TFC, and α-glucosidase inhibitory activity than AE. Furthermore, AE of the mushroom showed a higher lipase inhibitory potential with an IC50 value of 22.28 ± 0.65 μg/mL than the CME, while that of the reference, i.e. orlistat was 2.28 ± 0.34 μg/mL. However, these extracts exhibited very low or no α-amylase inhibitory and cytotoxic activity at the tested concentrations. This study reveals that CME of L. squarrosulus, rich in polyphenols and flavonoids, possesses considerable α-glucosidase and lipase inhibitory activities.

Qualitative Phytochemical Screening and Antimicrobial Activities of n-Hexane, Chloroform, Ethyl Acetate and Methanol Extracts of Ficus capensis Stem Bark

Ficus capensis (farin baure) has wide application in traditional medicine especially in the Northan part of Nigeria in the treatment and management of diarrhoea and digestive distress. This is attributed to the presence of many biologically active compounds in their different parts. In this study, stem bark of F. capensis was extracted with four (4) solvents of different polarities ranging from non-polar, moderately polar to polar. Qualitative phytochemical screening of these extracts was conducted using standard laboratory procedures. The results revealed that only alkaloids, glycosides and carbohydrates were detected from the n-hexane (non-polar) extract of F. capensis while the chloroform and ethyl acetate (moderately polar solvents) and methanol (polar solvent) extracts showed the presence of tannins, saponins, alkaloids, carbohydrates, glycosides, flavonoids and proteins. But anthraquinones were not detected in all the extracts.&#x0D; Antimicrobial properties of the extracts were evaluated using clinical isolates of four different bacteria and fungi each. Different concentration of extracts was prepared and activity was measured using zone of inhibition in millimetres (mm). Methanol extracts showed activities against E. coli with the zone of inhibition 10 mm, 11 mm, 12 mm and 14 mm; B. subtilis with the zone of inhibition 10 mm, 11 mm, 13 mm and 15 mm all for 10, 20, 25 and 30% concentration of extracts respectively. Chloroform and ethyl acetate extracts showed antimicrobial activities against S. aureus (12 mm, 14 mm), E. coli (12 mm, 14 mm), P. aeruginosa (14 mm, 12 mm) and B. subtilis (12 mm, 16 mm) respectively for 20% concentration of the extracts. The methanol and ethyl acetate extracts showed considerable antifungal activities against Aspergillus flavus and Aspergillus fungatus. But the n-hexane extract did not show any reasonable antibacterial and no antifungal activities.

Cytotoxic effects of Chloroform-methanol extract, protein extract and various fractions of ethanolic extract of Curcuma amada rhizome on HCT116 colon cancer cell line

Cytotoxic effects of chloroform-methanol extract, protein extract and various fractions of ethanolic extracts of Curcuma amada rhizome were investigated on Human colorectal cancer cell line (HCT116). The ethanolic extract showed excellent cytotoxicity of all extracts with an IC50 value of 14.46µg followed by the Chloroform-methanol extract with an IC50 value of 18.31µg. The various fractions of ethanolic extracts fractionated with petroleum ether, ethyl acetate, n-butanol were also tested for cytotoxicity independently. Ethyl acetate fraction gave an IC50 value of 1012µg and n-butanol IC50 value was 43.19µg, with petroleum ether fraction giving no cytotoxicity. The crude proteins isolated from the rhizome showed good cytotoxicity and gave an IC50 of 43.1µg.

Ecological and metabolomic responses of plants to deer exclosure in a suburban forest.

Trees and shrubs in suburban forests can be subject to chronic herbivory from abundant white-tailed deer, influencing survival, growth, secondary metabolites, and ecological success in the community. We investigated how deer affect the size, cover, and metabolomes of four species in the understory of a suburban forest in central New Jersey, USA: the woody shrubs Euonymus alatus and Lindera benzoin, the tree Nyssa sylvatica, and the semi-woody shrub Rosa multiflora. For each species, we compared plants in 38 16 m2 plots with or without deer exclosure, measuring proportion cover and mean height after 6.5 years of fencing. We scored each species in all plots for deer browsing over 8 years and assessed selection by deer among the species. We did untargeted metabolomics by sampling leaves from three plants of each species in an equal number of fenced and unfenced plots, conducting chloroform-methanol extractions followed by LC-MS/MS, and conducting statistical analysis on MetaboAnalyst. The proportion of a species browsed ranged from 0.24 to 0.35. Nyssa sylvatica appeared most selected by and susceptible to deer; in unfenced plots, both its cover and mean height were significantly lower. Only cover or height was lower for E.alatus and L.benzoin in unfenced plots, while R.multiflora height was greater. The metabolomic analysis identified 2333 metabolites, which clustered by species but not fencing treatment. However, targeted analysis of the top metabolites grouped by fencing for all samples and for each species alone and was especially clear in N.sylvatica, which also grouped by fencing using all metabolites. The most significant metabolites that were upregulated in fenced plants include some involved in defense-related metabolic pathways, e.g., monoterpenoid biosynthesis. In overbrowsed suburban forests, variation of deer impact on species' ecological success, potentially mediated by metabolome-wide chemical responses to deer, may contribute to changes in community structure.

Phenolic Profile, Antioxidant, Anti-Enzymatic and Cytotoxic Activity of the Fruits and Roots of Eleutherococcus senticosus (Rupr. et Maxim.) Maxim

Eleutherococcus senticosus (Rupr. et Maxim.) Maxim. is well-known for its adaptogenic properties in traditional Eastern medicine. It has been categorized as an endangered species due to the over-exploitation of the roots. As a result, alternatives must be found, including the usage of renewable aerial parts such as fruits. The goal of this research was to determine the phenolic compounds and the enzymatic, antioxidant, and cytotoxic activities of the intractum gained from the E. senticosus fruits and the mixture of chloroform-methanol roots extract with naringenin (3:7:5). The obtained results showed, that the intractum contained 1.02 mg/g ext. of polyphenols, 0.30 mg/g ext. of flavonoids, and 0.19 mg/g ext. of phenolic acids. In turn, the mixture of chloroform-methanol roots extract with naringenin (3:7:5) contained 159.27 mg/g ext. of polyphenols, 137.47 mg/g ext. of flavonoids, and 79.99 mg/g ext. of phenolic acids. Regarding the anti-enzymatic assay, the IC50 values for tyrosinase and hyaluronidase were equal to 586.83 and 217.44 [μg/mL] for the intractum, and 162.56 and 44.80 [μg/mL] for the mixture, respectively. Both preparations have possessed significant antioxidant activity in the ABTS, DPPH, and ferrozine tests. No cytotoxic effect on the FaDu and HEP G2 cancer cell lines was observed. Our findings support the traditional use of fruits and roots. Moreover, the results indicate also that adaptogens are rather nontoxic for normal and cancer cells, which corresponds with some hypotheses on adaptogens activity.

Antileishmanial Activity of New Steroidal Saponin Isolated from the Flowers of Allium Austroiranicum

Background &amp; Objective: Plants are reservoirs of bioactive compounds, which are known to be chemically balanced, effective and least injurious as compared to synthetic medicines. The current resistance and the toxic effects of the available drugs have brought the trend to assess the antileishmanial effect of various plant extracts and their purified compound/s. Alliums are rich sources of steroidal saponins, flavonoids, and sulphuric compounds of which steroidal saponins have recently received more attention due to their important pharmacological activities. Allium austroiranicum is a common edible vegetable in western regions of Iran, especially in “Chaharmahal and Bakhtiari” province, where it is named “Lopo” and is considerably used as a raw vegetable, flavoring agent, and as a medicinal plant.&#x0D; Materials &amp; Methods: The chloroform-methanolic extract was fractionated using MPLC, and the appropriate fractions were then subjected to isolation and purification of the constituents by HPLC. Structure elucidation was done using comprehensive spectroscopic methods including 1D and 2D NMR. Antileishmanial effects of the isolated compound against the promastigotes of Leishmania major were evaluated using MTT method.&#x0D; Results: Phytochemical investigation of chloroform-methanol extract of the plant resulted in the isolation and identification of a Nicotianoside C related steroidal saponin and its chemical structure was determined as (25S)-5α-Spirostan-1β,3βdiol-3-O-{α-L-rhamnopyranosyl-(1→2)-{α-L-rhamnopyranosyl-(1→4)}-[β-D-glucopyranoside}. Investigation of in vitro antileishmanial activity of the isolated compound, in 10 and 50 and 100 μg/mL concentrations, exhibited significant leishmanicidal effects against the promastigotes of Leishmania major.&#x0D; Conclusion: The results established a valuable basis for further studies about A. austroiranicum and anti-parasitic activity of steroidal saponins.

Hepatoprotective effect of Physalis divaricata in paracetamol induced hepatotoxicity: In vitro, in silico and in vivo analysis

Ethnopharmacological relevancePhysalis divaricata D. Don. is an erect weed of family Solanaceae. The root extract of this plant is used by the indigenous communities of Sub-Himalayan region of Uttarakhand, India for the treatment of liver disorders. Aim of the studyTo evaluate hepatoprotective potential of P. divaricata in paracetamol (PCM) induced hepatotoxicity in rats. Materials and methodsThe dried roots of P. divaricata were subjected to extraction using different solvents. The chloroform extract, methanol extract and bioactive aqueous fraction of methanol extract were evaluated for hepatoprotective effect. After initial in vitro screening, all extracts were screened for hepatoprotective potential in PCM (3 g/kg p.o) induced hepatotoxicity. Following PCM administration, extracts were administered orally for 7 days in increasing dose concentrations. All the animals were euthanized on eighth day, serum and liver tissues were collected and subjected to various biochemical and histopathological analysis. Aqueous fraction of methanol extract was further analyzed using LC- MS analysis. ResultsMethanol extract and its bioactive aqueous fraction exhibited significant and better in vitro antioxidant and antiproliferative activity as compared to chloroform extract. PCM treatment caused hepatotoxicity as assessed by altered levels of various hepatic biomarkers (increase in the levels of ALT, AST, ALP, albumin, triglycerides, cholesterol, TBARS, and AOPPs as well as decrease in GSH and TrxR levels) along with histopathological changes (portal to portal bridging, necrosis, and inflammation). Methanolic extract (200, 400 and 800 mg/kg) and its aqueous fraction treatment (25, 50 and 100 mg/kg) significantly restored elevated hepatic biomarkers, oxidative stress, and protected normal hepato-architecture. LC-MS analysis of aqueous fraction showed presence of rutin and kaempferol. In silico analysis further showed the capability of rutin to make complex with TNF-α and block its interaction with the target site. ConclusionAqueous fraction showed maximum hepatoprotective potential as conceived through in vitro and in vivo studies. Presence of rutin may explain hepatoprotective potential of P. divaricata.

In Vitro Investigation of the Antioxidant and Cytotoxic Potential of Tabernaemontana ventricosa Hochst. ex A. DC. Leaf, Stem, and Latex Extracts

Tabernaemontana ventricosa (Apocynaceae) a latex-bearing plant is used in traditional medicine for its therapeutic benefits in reducing fever and hypertension and wound healing. Due to limited information on the plant’s pharmacological activities, this study aimed to investigate the antioxidant potential of the leaf, stem, and latex extracts of T. ventricosa, using the Folin-Ciocalteu (total phenolics), aluminum chloride colorimetric (total flavonoids), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and ferric reducing antioxidant power (FRAP) assays. The cytotoxic activity was evaluated in the human HEK293 (embryonic kidney), HeLa (cervical carcinoma), and MCF-7 (breast adenocarcinoma) cell lines using the MTT assay. The latex extracts possessed the highest total phenolic content (115.36 ± 2.89 mg GAE/g), followed by the stem hexane extracts (21.33 ± 0.42 mg GAE/g), the chloroform leaf (7.89 ± 0.87 mg GAE/g), and the chloroform stem (4.69 ± 0.21 mg GAE/g) extracts. The flavonoid content was substantially high ranging from 946.92 ± 6.29 mg QE/g in the stem hexane, 768.96 ± 5.43 mg QE/g in the latex, 693.24 ± 4.12 mg QE/g in the stem chloroform, and 662.20 ± 1.00 mg QE/g in the leaf hexane extracts. The DPPH assays showed the highest percentage of inhibition at 240 µg/mL, for the stem hexane (70.10%), stem methanol (65.24%), and stem chloroform (60.26%) extracts, with their respective IC50 values of 19.26 µg/mL (stem hexane), 6.19 µg/mL (stem methanol), and 22.56 µg/mL (stem chloroform). The FRAP assays displayed minimal inhibition ranging from 4.73% to 14.40%, except for the latex extracts which displayed moderate inhibition at 15 µg/mL (21.82%) and substantial inhibition at 240 µg/mL (98.48%). The HeLa and MCF-7 cell lines were the most sensitive to the extracts, with the hexane, chloroform, and methanol leaf and stem, and latex extracts significantly affecting the percentage cell survival. Overall, the various parts of T. ventricosa exhibited strong antioxidant activity correlating to its cytotoxicity. Further studies should focus on the isolation of specific antioxidant compounds that could be investigated for their anticancer potential.

Optimization of Extraction Process and Composition Identification of Polar Lipids in Milk

In order to develop and utilize the milk polar lipids, the chloroform-methanol extraction method was used to extract the polar lipids from buttermilk. The phosphorus content as an index, the condition of chloroform-methanol volume ratio, liquid-to-methanol ratio, extraction temperature and time were optimized by single-factor experiment and the analysis of response surface methodology determined the optimum extraction conditions. The obtained samples were analyzed and identified by thin-layer chromatography(TLC). The results indicated that the optimal extraction conditions for polar lipids were: Chloroform-methanol volume ratio 2:1(v/v, %), 1:4(v/v, %) for the ratio of material to liquid, 20 ℃ for extraction temperature, and 10 min for extraction time. The phospholipid contents in the sample under this extracted condition was (1.647±0.010) mg/g, and there were polar lipids such as phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, and sphingomyelin. This process aims to provide a research basis for the extraction and development of milk polar lipids.

Isolation and identification of 6-methoxy parillin and coniferin from the bulbs of Allium affine L.

Phytochemically, Allium species are a rich source of important secondary metabolites especially steroidal saponin and sapogenins, flavonoids, and sulfur compounds. As a member of this genus, Allium affine, which is locally known as "tareh kouhi", is an endemic plant of middle Asian countries. Bulbs of A. affine were collected and air-dried in the shade. The chloroform-methanol (9:1) extract of the sample was subjected to purification by MPLC and HPLC. Structure elucidation of isolated compounds was done using comprehensive spectroscopic methods including 1D-NMR, 2D-NMR, and MS. A steroidal saponin structurally related to parillin and a phenylpropanoid glycoside (coniferin) were isolated and identified from the plant chloroform-methanol extract. To the best of our knowledge isolation of these potentially medicinal compounds from A. affine was reported for the first time in this study.

Phytochemical profile, in vitro antioxidant, and anti-protein denaturation activities of Curcuma longa L. rhizome and leaves

Abstract Curcuma longa L. is a famous spice cultivated in many countries with significant variations reported in its phytochemical contents and biological potential. For the first time, the present work is aimed to identify the major phytochemicals present in methanol:chloroform (MC) and petroleum ether (PE) extracts of Curcuma longa rhizome and leaves (by determining polyphenols and GC/MS analysis), and their in-vitro antioxidant and anti-protein denaturation potential. Results showed that the highest value (P &lt; 0.05) of polyphenolic content was in MC extract of rhizome (51.46 ± 0.46 mg GAE/g) followed by 31.20 ± 0.53 mg GAE/g in MC leaves extract. The strong antiradical activity was evaluated in MC extract of rhizome with IC50 value of 92 ± 0.02 µg/mL. MC extracts of both the rhizome and leaves exerted a potent inhibitory effect against protein denaturation with IC50 values of 106.21 ± 0.53 and 108.06 ± 4.67 μg/mL (P &gt; 0.5), respectively. GC/MS analysis showed that α-tumerone was the main component in the rhizome oil (32.44%), whereas in the leaf oil, palmitic acid was the prominent constituent (28.33%) and α-phellandrene recorded a comparable percentage (7.29). In conclusion, C. longa is a valuable source of natural antioxidants and anti-inflammatory constituents, as indicated by its high polyphenolic content and by its considerable in vitro antiradical and anti-protein denaturation potential.

Adapting an Ergosterol Extraction Method with Marine Yeasts for the Quantification of Oceanic Fungal Biomass.

Ergosterol has traditionally been used as a proxy to estimate fungal biomass as it is almost exclusively found in fungal lipid membranes. Ergosterol determination has been mostly used for fungal samples from terrestrial, freshwater, salt marsh- and mangrove-dominated environments or to describe fungal degradation of plant matter. In the open ocean, however, the expected concentrations of ergosterol are orders of magnitude lower than in terrestrial or macrophyte-dominated coastal systems. Consequently, the fungal biomass in the open ocean remains largely unknown. Recent evidence based on microscopy and -omics techniques suggests, however, that fungi contribute substantially to the microbial biomass in the oceanic water column, highlighting the need to accurately determine fungal biomass in the open ocean. We performed ergosterol extractions of an oceanic fungal isolate (Rhodotorula sphaerocarpa) with biomass concentrations varying over nine orders of magnitude. While after the initial chloroform-methanol extraction ~87% of the ergosterol was recovered, a second extraction recovered an additional ~10%. Testing this extraction method on samples collected from the open Atlantic Ocean, we successfully determined ergosterol concentrations as low as 0.12 pM. Thus, this highly sensitive method is well suited for measuring fungal biomass from open ocean waters, including deep-sea environments.

Relative Contributions of All-Trans and 11-Cis Retinal to Formation of Lipofuscin and A2E Accumulating in Mouse Retinal Pigment Epithelium.

Purpose Bis-retinoids are a major component of lipofuscin that accumulates in the retinal pigment epithelium (RPE) in aging and age-related macular degeneration (AMD). Although bis-retinoids are known to originate from retinaldehydes required for the light response of photoreceptor cells, the relative contributions of the chromophore, 11-cis retinal, and photoisomerization product, all-trans retinal, are unknown. In photoreceptor outer segments, all-trans retinal, but not 11-cis retinal, is reduced by retinol dehydrogenase 8 (RDH8). Using Rdh8−/− mice, we evaluated the contribution of increased all-trans retinal to the formation and stability of RPE lipofuscin.Methods Rdh8−/− mice were reared in cyclic-light or darkness for up to 6 months, with selected light-reared cohorts switched to dark-rearing for the final 1 to 8 weeks. The bis-retinoid A2E was measured from chloroform-methanol extracts of RPE-choroid using HPLC-UV/VIS spectroscopy. Lipofuscin fluorescence was measured from whole flattened eyecups (excitation, 488 nm; emission, 565–725 nm).ResultsCyclic-light-reared Rdh8−/− mice accumulated A2E and RPE lipofuscin approximately 1.5 times and approximately 2 times faster, respectively, than dark-reared mice. Moving Rdh8−/− mice from cyclic-light to darkness resulted in A2E levels less than expected to have accumulated before the move.ConclusionsOur findings establish that elevated levels of all-trans retinal present in cyclic-light-reared Rdh8−/− mice, which remain low in wild-type mice, contribute only modestly to RPE lipofuscin formation and accumulation. Furthermore, decreases in A2E levels occurring after moving cyclic-light-reared Rdh8−/− mice to darkness are consistent with processing of A2E within the RPE and the existence of a mechanism that could be a therapeutic target for controlling A2E cytotoxicity.