Thin Layer Chromatography Analysis Research Articles

Thin Layer Chromatographic Analysis of Flavonoid in Mulberry Leaf Extract

Abstract: Morus alba L., also known as white mulberry, has long been used in traditional medicine to treat a variety of health conditions. The leaves of the plant are rich in polyphenols such as quercetin 3-(- malonylglucoside), rutin, isoquercitin, and cyaniding-3-rutinoside apigenin, luteolin, caffeic acid, gallic acid and umbelliferone, chlorogenic acid, kaempferol, and flavonoids. In the present study, qualitative tests of leaf extracts showed significant indication of the presence of Flavonoids were found to be present in the leaf extracts, with a maximum concentration by using methanol and ethanol solvents. Using thin layer chromatography (TLC) profiling of plant extracts gives an impressive result that directing towards the existence of phytoconstituents. In addition, the alcoholic extracts for mulberry leaves contain a higher content of bioactive compounds, which can be used for further researches on this plant.

Exploration and Evaluation of Secondary Metabolites from Trichoderma harzianum: GC-MS Analysis, Phytochemical Profiling, Antifungal and Antioxidant Activity Assessment

In this study, we investigated in vitro the potential of Trichoderma harzianum to produce bioactive secondary metabolites that can be used as alternatives to synthetic compounds. The study focused on analyzing two extracts of T. harzianum using ethyl acetate and n-butanol solvents with different polarities. The extracts were examined using phytochemical analysis to determine the content of polyphenols, flavonoids, tannins, and alkaloids. Thin-layer chromatography (TLC) and Gas chromatography-mass spectroscopy (GC-MS) analysis were used to profile volatile organic metabolites (VOCs) present in the extracts. Furthermore, the extracts were tested for their antifungal ability using the poison food technique. For measuring antioxidant activity, the 1,1-diphenyl-2-picryl-hydrazyl (DPPH) test was used. Trichoderma harzianum was shown to have a significantly high content of tannins and alkaloids, with a noticeable difference between the two extracts. GC-MS analysis identified 33 potential compounds with numerous benefits that could be used in agriculture and the medicinal industry. Moreover, strong antifungal activity was identified against Sclerotinia sclerotiorum by 94.44%, Alternaria sp. by 77.04%, and Fusarium solani by 51.48; similarly, the IC50 of antioxidant activity was estimated for ethyl acetate extract by 71.47% and n-butanol extract by 56.01%. This leads to the conclusion that Trichoderma harzianum VOCs play a significant role as an antifungal and antioxidant agent when taking into account the advantageous bioactive chemicals noted in the extracts. However, to our knowledge, this is the first study in Algeria presenting detailed phytochemical analysis and GC-MS profiling of Trichoderma harzianum for two extracts, ethyl acetate and n-butanol.

Phytochemical Screening, Chromatographic and Quantitative Study of Phenols and Flavonoids in Leaves of Oroxylum indicum and Pongamia pinnata

Number of secondary compounds is produced by plants as natural antioxidants. The current study's goal was to identify the phytochemical components, thin-layer chromatographic profile, total phenol, and total flavonoid content in the leaves of Pongamia pinnata and Oroxylum indicum (O. indicum) (P. pinnata). The well-known test protocol described in the literature was used to determine the qualitative analysis of different phytochemical constituents, thin-layer chromatography, and quantitative analysis of total phenolics and flavonoids. By using the Folins Ciocalteau reagent method and the aluminium chloride method, respectively, phenolic and flavonoid quantities were analysed quantitatively. Both plants contained glycosides, flavonoids, diterpenes, phenol, proteins, saponins, and carbohydrates, according to a phytochemical analysis. Studies using thin-layer chromatography and various solvent systems revealed homogeneous spots with various Rf values. According to the findings of the current study, O. indicum and P. pinnata crude extracts are a rich source of secondary phytoconstituents that have significant antioxidant potential. The pharmaceutical industries, pharmacologists, and phytochemists will all benefit from the study's findings.

Integrated methods for fingerprinting of monofloral honeys

Honey is an important functional food due to its content of polyphenolic compounds with beneficial effects on human body. The analysis of polyphenolic compound in honey is very promising in determination of floral origin and so for quality control. This study approaches a new methodology using integrated methods for the determination of honey authenticity. Polyphenolic compounds from seven different types of monofloral honey from two different sources were extracted and analyzed by thin layer chromatography (TLC) and UV-Vis spectroscopy. Chromatographic analysis was performed on TLC Silica gel 60 RP-18 F254 plates using a mobile phase consisting in a mixture of methanol: water: formic acid (5: 5: 0.1, v/v/v). The images from the TLC analysis were processed and the obtained chromatograms together with the UV-Vis deconvoluted spectra of honey were subjected to visual and statistical analysis. The obtained results showed notable differences between the honey samples, which can be used to establish their floral origin and for their authentication. The proposed methodology is easy to perform, cost-effective and offers complementary information, therefore it has the potential of being used for the honey authentication.

Migration of Eastern North American monarch butterflies via the South-east and the Atlantic: evidence from stable isotopes, thin layer chromatography, DNA and phenotype

Abstract Monarch butterflies, Danaus plexippus (L.), from eastern North America are well known for their incredible autumn migration to Mexico; however, not all monarchs follow this route. There is evidence of monarchs overwintering and reproducing in Florida, arriving to insular and continental Caribbean, and roosting by the thousands in the Yucatán, Mexico. This work aims to present evidence that these monarchs are part of two current migratory routes, that we consider as the south-eastern and the Atlantic routes, routes that were probably more prominent in the past. Monarchs were collected for 12 months in south Florida, for 4 years in Cuba in November and once in March, in the Yucatán and Guatemala at different times, and once in northern Venezuela. We used two independent techniques, stable hydrogen isotope (δ2H) analyses of the wings and/or thin layer chromatography (TLC), to trace the monarch’s natal grounds. We analysed the DNA of monarchs collected in St. Marks in the Florida Panhandle, Cuba and Guatemala, and compared those data against previously-analysed DNA data from monarchs in the Americas to characterize their genetic structure and to assess the possible movement and presence of North American monarchs and/or their alleles outside the USA. Our results support the existence of south-eastern and Atlantic migratory routes. TLC, isotope and DNA analyses showed the arrival of likely North American monarchs in Cuba, Yucatán, Guatemala, Venezuela and other areas of the Americas. North American monarchs found in these areas have different natal grounds, phenotypic traits and DNA signature than Mexican migrants. Monarchs from the south-eastern route mostly originated in the south-east USA and fed on local Asclepias spp., such as Asclepias viridis, Asclepias humistrata, Asclepias perennis and Asclepias asperula. Butterflies from this migratory route move east, enter the Florida Peninsula, pass to Cuba, fly to the Yucatán and then to Guatemala where they appear to overwinter in the high mountains of Guatemala where Abies guatemalensis occurs. Monarchs that are part of the Atlantic route move east of the Appalachians, enter the Florida Peninsula, and from there pass to the insular and continental Caribbean. The main host plants for the Atlantic monarchs are A. perennis and A. humistrata; in contrast, Mexican monarchs mainly feed on Asclepias syriaca. Some monarchs from these two proposed migratory routes, south-eastern and Atlantic, will stay in the places where they travelled and others will return via Florida and Mexico. We propose a scenario for how the different migratory routes evolved.

Cross-linked phytase aggregates for improved phytate degradation at low pH in animal feed

Phytases are widely used food and feed enzymes to improve phosphate availability and reduce anti-nutritional factors. Despite the benefits, enzyme usage is restricted by the harsh conditions in a gastrointestinal tract (pH 2–6) and feed pelleting conditions at high temperatures (60–90 °C). The commercially available phytase Quantum® Blue has been immobilized as CLEAs using glutardialdehyde and soy protein resulting in a residual activity of 33%. The influence of the precipitating agent, precipitant concentration, cross-linker concentration and cross-linking time, sodium borohydride as well as the proteic feeders gluten, soy protein and bovine serum albumin (BSA) has been optimized. The best conditions were 90% (v/v) ethyl lactate as precipitating reagent, 100 mM glutardialdehyde and a soy protein concentration of 227 mg/L with a cross-linking time of 1 h. The intrinsically stable phytase remained its high thermal stability and temperature optimum. The phytase-CLEA achieved a 425% increase of residual activity under harsh acidic conditions between pH 2.2 and 3.5 compared to the free enzyme. The free and immobilized phytase were deployed in an in vitro assay simulating the acidic conditions in the gizzard of poultry at pH 2. The hydrolysis of phytate was monitored using a novel high-performance thin-layer chromatography (HPTLC) analysis and DAD scanner to study the InsPx fingerprint. All lower inositol phosphate pools (InsP1–InsP6) and free phosphate were separated and analyzed. The phytase-CLEA efficiently released 80% of the total phosphate within 180 min, whereas the free enzyme only released 6% in the same time under the same conditions.

Isolation, development and validation of HPTLC method for the estimation of β-carotene from Gymnosporia senegalensis (Lam.) Loes.

The present study is aimed to isolate terpenoids from Gymnosporia senegalensis through analytical and preparative thin-layer chromatography (TLC) and to determine their antioxidant activity using the 2, 2-diphenyl-1- picrylhydrazyl (DPPH) assay and to find out the presence of β-carotene through high-performance thin-layer chromatography (HPTLC). The validation included linearity, limit of detection (LOD), limit of quantification (LOQ), specificity, precision, recovery, and robustness. All the isolated compounds from TLC exhibited significant antioxidant activity. Among all, isolated compounds from leaf showed highest IC50 values. The highest total terpenoid content (TTC) was found 51.6 ± 0.06 in stem, then 49.02 ± 0.01 in bark, and 46.27 ± 0.01 in leaf. DPPH results indicated that leaf-isolated compound 1 (LIC1) showed the highest IC50 at 7.55 ± 0.02 and stem-isolated compound 2 (SIC2) showed the lowest IC50 at 0.616 ± 0.01 among all the isolated compounds of G. senegalensis. HPTLC separation was carried out on aluminium plates pre-coated with silica gel 60 F254 as the stationary phase and n-hexane: ethyl acetate (6:4, v/v) as the mobile phase. Quantification was achieved based on a densitometric analysis of β-carotene in the concentration range of 100–500 ng/band at 254 nm. For the calibration plots, linear regression produced r2 = 0.96450 and Rf = 0.27. The LOD and LOQ were 10.15 and 30.76 ng/mL for HPTLC and relative standard deviation were 137.26 ± 2.03 and 160.43 ± 2.95 (intra-day) and 127.88 ± 2.14 and 157.27 ± 1.90 (inter-day) for 200 and 400 ng/band, respectively. The present study shows the presence of various types of terpenoids through TLC whereas the HPTLC results indicated that the developed methods were accurate and precise. It also shows that the approach is appropriate for its intended use in routine quality control testing of commercially available tablet formulations and drug assay to assist both industries and researchers in making important decisions at a reasonable cost. Moreover, due to the use of a safer and more environmentally friendly mobile phase in comparison to the toxic mobile phases used in recent analytical techniques to estimate β-carotene, this methodology is also secure and sustainable.

Solar light assisted degradation of Rhodamine B dye by chemically synthesized silver nanoparticles

Utilization of dyes and dyestuff is increasing day by day, which affects environmental health and sustainability. Rhodamine B (RhB) is a synthetic xanthene class of dye that has a major application in cosmetic, painting, printing, and textile industries. The toxicity of RhB has been reported in animals, insects as well as humans in parts per billion (ppb) concentration. Exposure to this dye causes skin irritation, nervous system issues, and developmental changes. Therefore, the remediation of the dye is a crucial factor. The aim of this study was solar-assisted photocatalytic degradation of RhB. Synthesis of silver nanoparticles (AgNPs) was carried out with chemical reduction method, and characterization with the X-ray diffraction (XRD). The photocatalytic activity was checked with 1000 mL wastewater in which 10 ppm dye was added. Optimization of physicochemical parameters was performed by one-factor-at-a-time (OFAT) method. Maximum 99.32 % decolourization was observed at 2 g/L catalyst concentration, pH = 6, and 10 ppm dye concentration in 50 min irradiation time at 0.19 mg/L/min rate of decolourization. The initial degradation was confirmed based on chemical oxygen demand (COD) reduction; 89.25 % COD removal at a 0.17 mg/L/min rate was observed. The confirmation of degradation was carried out with UV-visible spectrophotometer and high-performance thin-layer chromatography (HPTLC) analysis. AgNPs were the efficient catalyst for the degradation of RhB. The toxicity study proved that, after degradation of RhB with AgNPs, the degraded products were less toxic than the original ones.

Biochemical analyses of a novel acidophilic GH5 β-mannanase from Trichoderma asperellum ND-1 and its application in mannooligosaccharides production from galactomannans.

In this study, an acidophilic GH5 β-mannanase (TaMan5) from Trichoderma asperellum ND-1 was efficiently expressed in Pichia pastoris (a 2.0-fold increase, 67.5 ± 1.95 U/mL). TaMan5 displayed the highest specificity toward locust bean gum (Km = 1.34 mg/mL, Vmax = 749.14 μmol/min/mg) at pH 4.0 and 65°C. Furthermore, TaMan5 displayed remarkable tolerance to acidic environments, retaining over 80% of its original activity at pH 3.0-5.0. The activity of TaMan5 was remarkably decreased by Cu2+, Mn2+, and SDS, while Fe2+/Fe3+ improved the enzyme activity. A thin-layer chromatography (TLC) analysis of the action model showed that TaMan5 could rapidly degrade mannan/MOS into mannobiose without mannose via hydrolysis action as well as transglycosylation. Site-directed mutagenesis results suggested that Glu205, Glu313, and Asp357 of TaMan5 are crucial catalytic residues, with Asp152 playing an auxiliary function. Additionally, TaMan5 and commercial α-galactosidase displayed a remarkable synergistic effect on the degradation of galactomannans. This study provided a novel β-mannanase with ideal characteristics and can be considered a potential candidate for the production of bioactive polysaccharide mannobiose.

THE PROBLEMS TO IMPROVE THE STANDARDISATION OF YARROW HERB

Relevance. It is known the method of quantitative determination of the total flavonoids in the herb of common yarrow (Achillea millefolium L.) includ-ed in the State Pharmacopoeia of the Russian Federation XIV edition, according to which at the stage of extraction of these raw materials should be ac-id hydrolysis with ethyl alcohol 96% containing 1% hydrochloric acid concentrated, followed by determination of the amount of flavonoids in recalcula-tion on luteolin. The flavonoid glycosides, including 7-O-glucosides luteolin (cynaroside) and apigenin (cosmosiin), difficult to acid hydrolysis, which does not allow for complete hydrolysis and exhaustive extraction of the target substances. Determination of the main groups of biologically active substances by thin-layer chromatography (TLC) involves the use of a standard sample of sudan III, not contained in the raw material. The purpose of this study is the improvement of methods of qualitative and quantitative analysis of herb of Achillea millefolium L. Material and methods. Yarrow herb harvested in 2021 was used as study materials. Extraction of individual substances from yarrow herbs was car-ried out using column chromatography on silica gel KSC 50/100 under conditions of gradient elution with chloroform-ethanol solvent mixture in different ratios. Along with the isolated individual flavonoids, we used working standard samples of flavonoids obtained by the authors in previous studies of herbal plants containing flavonoids, characterized using UV and NMR spectroscopy: rutin, quercetin, kaempferol, cynaroside, luteolin. Flavonoid content was determined by thin-layer chromatography and by differential spectrophotometry. Results. TLC analysis revealed the presence of three dominant and diagnostically significant flavonoids: cosmosiin, cynaroside and apigenin. Analysis of UV spectra also confirmed that the spectral characteristics of the water-alcoholic extracts from common yarrow herbs, especially under the condi-tions of differential spectrophotometry, are mainly determined by the dominant flavones (cosmosiin, cynaroside and apigenin). Conclusions. The obtained results of studies on the flavonoid composition and spectral characteristics of the isolated dominant flavonoids suggest the feasibility of conducting of the qualitative and quantitative analysis of common yarrow herbs using cynaroside as a standard sample.

Demonstration of the Antioxidant Potential of Three Newly Isolated Carotenoid-Producing Microscopic Algae from KwaZulu-Natal (South Africa)

Natural carotenoids from green microalgae exhibit beneficial effects in treating health-related diseases, primarily due to their antioxidant capacity. Therefore, carotenoid-producing microalgae were isolated and characterized from aqueous samples in KwaZulu-Natal (South Africa) under photoautotrophic conditions. Three isolates were characterized comprehensively using light and electron microscopy. In addition, the growth of the three selected microalgae was evaluated under photoautotrophic and photoheterotrophic conditions. Based on the cytological characteristics, the three strains matched the genera Haematococcus, Parachlorella, and Scenedesmus, the latter two additionally confirmed via analysis of the 18S rRNA gene sequence at the genus level. Light and electron microscopy and UV-Vis spectroscopy confirmed pigment production by all three microalgal strains. Both isolate Parachlorella sp. AA1 and Scenedesmus sp. AA2 showed the highest growth rate when cultured with acetate (25 mM) photoheterotrophically, while the isolate Haematococcus sp. AA3 grew best with glucose (50 mM). In addition, all three isolates utilized salicylate as a carbon source under photoheterotrophic conditions, evidently resulting in increased carotenoid production in strains AA1 and AA3. The 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) antioxidant assay confirmed that methanol extracts of all three isolates contained carotenoids that can scavenge radicals, and thin layer chromatography (TLC) analysis showed that β-carotene and astaxanthin were formed by strain AA3 as main carotenoids.

Extraction, HPTLC Analysis and Antiobesity Activity of Jatropha tanjorensis and Fraxinus micrantha on High-Fat Diet Model in Rats.

The accumulation of body fat due to an imbalance between calorie intake and energy expenditure is called obesity. Metabolic syndrome increases the risk of heart disease, type 2 diabetes, and stroke. The purpose of this study was to determine the effect of Jatropha tanjorensis (J.T.) and Fraxinus micrantha (F.M.) leaf extracts on high-fat diet-induced obesity in rats. Normal control, high-fat diet (HFD) control, orlistat standard, and test groups were created using male Albino Wistar rats (n = 6 per group) weighing 190 ± 15 g. Except for the control group, all regimens were administered orally and continued for 6 weeks while on HFD. Evaluation criteria included body weight, food intake, blood glucose, lipid profile, oxidative stress, and liver histology. High-Performance Thin Layer Chromatography (HPTLC) analysis was performed using a solvent system (7:3 hexane: ethyl acetate for sitosterol solution and Jatropha tanjorensis extracts and 6:4 hexane: ethyl acetate: 1 drop of acetic acid for esculetin and Fraxinus micrantha extracts). There were no deaths during the 14 days before the acute toxicity test, indicating that aqueous and ethanolic extracts of both J.T. and F.M. did not produce acute toxicity at any dose (5, 50, 300, and 2000 mg/kg). The ethanolic and aqueous extracts of J.T. and F.M. leaves at 200 and 400 mg/kg/orally showed a reduction in weight gain, feed intake, and significant decreases in serum glucose and lipid profile. As compared to inducer HFD animals, co-treatment of aqueous and ethanolic extract of both J.T. and F.M. and orlistat increased the levels of antioxidant enzymes and decreased lipid peroxidation. The liver's histological findings showed that the sample had some degree of protection. These results indicate that ethanolic samples of J.T. have antidiabetic potential in diabetic rats fed an HFD. The strong antioxidant potential and restoration of serum lipid levels may be related to this. Co-treatment of samples JTE, JTAQ, FME, FMAQ and orlistat resulted in an increase in antioxidant enzymes and reduction in lipid peroxidation as compared to inducer HFD animals. We report, for the first time, on using these leaves to combat obesity.

Triterpenes, steroids and lichexantone from Macrolobium acaciifolium leaves and branches hexane extracts and antibacterial activity

Macrolobium acaciifolium belongs to Fabaceae family and only few phytochemical studies were carried out in the scientific literature consulted. The objective of this work was to isolate and identify the secondary metabolites from M. acaciifolium leaves and branches hexane extracts, in addition to evaluate the antibacterial potential of the extracts. The plant materials were dried and extracted with hexane, then methanol and water. The hexane extracts showed the presence of terpenoids (including steroids) by thin layer chromatography and nuclear magnetic resonance analysis. The phytochemical fractionation allowed to isolate the triterpenes: lupeol, β-amyrin and 24-methylenecycloartanol from hexane leaves extract. The mixture of the triterpene friedelin and the xanthone 1-hydroxy-3,6-dimethoxy-8-methyl-9h-xanthen-9-one (lichexanthone) were isolated from branches hexane extract. In both extracts, the mixture of the steroids β-sitosterol and stigmasterol were also isolated. The leaves hexane extract showed to be toxic to Aeromonas hydrophila with 87% growth inhibition at a concentration of 1000 µg/mL. This is the first description of the isolation of these substances in Macrolobium acaciifolium and the antibacterial activity.

A Comparative Analysis of Abhavitha and Bhavitha Choorna of Dried Whole Plant of Mandukaparni- Centella Asiatica (Linn.) Urban. Through High Performance Thin Layer Chromatography

HPTLC (High Performance Thin Layer Chromatography) is a powerful analytical technique in the process of drug discovery and evaluation. Due to its dependability and simplicity, it is used as a tool for the identification, authentication, and quality control of herbal drugs. Mandukaparni- Centella asiatica (Linn.) Urban. is a significant and well-known drug expound discussed in the Ayurvedic classics. It is a prostrate, stoloniferous herb with enriched therapeutic potential. The herb is rich in terpinoids, flavonoids, iso-flavonoids, alkanes tannins, saponins, aminoacids and phenylpropanoids, which accounts for its multiple beneficial action in the therapeutics. It has strong anti-inflammatory and antioxidant properties. The Abhavitha choorna (dried powder) and Bhavitha choorna (powder soaked in Mandukaparni swarasa) of dried whole plant of Mandukaparni- Centella asiatica (Linn.) Urban. was subjected to High performance thin layer chromatography and comparative analysis of different peaks in both the Choornas were obtained. The results suggest that change in concentration of phytochemical constituents occur on Abhavitha choorna (powder) to Bhavitha (soaked in Mandukaparni swarasa).

Bioactive compounds from four Indian medicinal plants have different potency to induce sex reversal in Nile tilapia: A chromatographic, molecular docking and in silico analysis.

Herbal plants such as Basella alba (Family: Basellaceae), Tribulus terrestris (Family: Zygophyllaceae), Asparagus racemosus (Family: Asparagaceae) and Mucuna pruriens (Family: Fabaceae) are mentioned in Indian traditional system of medicine Ayurveda to possess androgenic activity and increase male virility. The plants have been reported to improve testosterone level and sperm production in experimental male rodents as well. Male Nile tilapias grow more quickly than females and hence are preferred for monosex Nile tilapia culture. Ethanol extracts of B. alba leaves (EB) and T. terrestris seeds (ET), and methanol extract of A. racemosus roots (MA) and M. pruriens seeds (MM) were found effective to induce masculinization in Nile tilapia. The present study intends to evaluate the anti-aromatase activity of EB, ET, MA and MM, to identify the androgenic bioactive compounds in the extracts, and to determine their pharmacokinetics. The study may validate the use of those plant extracts and their major bioactive phytoconstituents in the field of aquaculture and pharmaceuticals. The four crude plant extracts were first fractioned through column and thin layer chromatography (TLC). Three days old Nile tilapia juveniles (mean weight 0.025±0.009g; mean length 12.50±0.12mm; n=50 fish/replicate, 3 replicates/treatment) were then fed diets fortified with the obtained fractions for 30 days. After 30 days, fish were sacrificed and gonad aromatase mRNA expression, and 11-ketotestosterone (11-KT) and estradiol (E2) levels were measured. Fractions yielding the highest male percentage for each plant were subject to gas chromatography-mass spectrometry (GC-MS) analysis. The in silico docking and SwissADME study were conducted with the components showing higher peak percentage in chromatogram. After column chromatography and TLC analysis, EB, ET, MM and MA yielded 6 (EB1 - EB6), 8 (ET1- ET8), 14 (MM1-MM14) and 5 (MA1- MA5) fractions, respectively. Fish fed EB2, ET2, MA2 and MM13 fraction fortified diets showed significantly (p<0.05) higher male percentage (92.32%-98.39%) compared to other treatment groups. EB2, ET2, MA2 and MM13 fed fish showed significantly (p<0.05) higher 11-KT level compared to control male (+247.52 - +397.76%) and lower E2 level compared to control female (-95.92% to -90.65%). Aromatase mRNA expression was significantly (p<0.05) down-regulated by all these four fractions (-1.32 to -5.65 fold) with respect to control female. GC-MS analysis revealed the presence of 1-Octadecene (OD) in EB2, Phenol, 2,4-bis(1,1-dimethylethyl) (PD) in ET2 and MA2, 9,12-Octadecadienoic acid (Z,Z)- (ODDA) in MM13. In silico molecular docking indicated that PD is more effective than ODDA and OD to inhibit aromatase. In addition, PD showed better pharmacokinetics and more drug-likeness compared to OD and ODDA in SwissADME analysis. The present results indicate that ET and MA are more potent to produce all-male tilapia by means of aromatase inhibition. PD can be an ideal compound to achieve masculinization in Nile tilapia through dietary administration, but further investigation is required.

In situ monitoring of galactolipid digestion by infrared spectroscopy in both model micelles and spinach chloroplasts

Galactolipids are the main lipids from plant photosynthetic membranes and they can be digested by pancreatic lipase related protein 2 (PLRP2), an enzyme found in the pancreatic secretion in many animal species. Here, we used transmission Fourier-transform infrared spectroscopy (FTIR) to monitor continuously the hydrolysis of galactolipids by PLRP2, in situ and in real time. The method was first developed with a model substrate, a synthetic monogalactosyl diacylglycerol with 8-carbon acyl chains (C8-MGDG), in the form of mixed micelles with a bile salt, sodium taurodeoxycholate (NaTDC). The concentrations of the residual substrate and reaction products (monogalactosylmonoglyceride, MGMG; monogalactosylglycerol, MGG; octanoic acid) were estimated from the carbonyl and carboxylate vibration bands after calibration with reference standards. The results were confirmed by thin layer chromatography analysis (TLC) and specific staining of galactosylated compounds with thymol and sulfuric acid. The method was then applied to the lipolysis of more complex substrates, a natural extract of MGDG with long acyl chains, micellized with NaTDC, and intact chloroplasts isolated from spinach leaves. After a calibration performed with α-linolenic acid, the main fatty acid (FA) found in plant galactolipids, FTIR allowed quantitative measurement of chloroplast lipolysis by PLRP2. A full release of FA from membrane galactolipids was observed, that was not dependent on the presence of bile salts. Nevertheless, the evolution of amide vibration band in FTIR spectra suggested the interaction of membrane proteins with NaTDC and lipolysis products.

Development of a bis-pyrene phospholipid probe for fluorometric detection of phospholipase A2 inhibition

In this work, we report the design, synthesis, and application of a bis-pyrene phospholipid probe for detection of phospholipase A2 action through changes in pyrene monomer and excimer fluorescence intensities. Continuous fluorometric assays enabled detection of the activities of multiple PLA2 enzymes as well as the decrease in catalysis by PLA2 from honey bee venom caused by the inhibitor p-bromo phenacylbromide. Thin-layer chromatography and mass spectrometry analysis were also used to validate probe hydrolysis by PLA2. Mass spectrometry data also supported cleavage of the probe by phospholipase C and D enzymes, although changes in fluorescence were not observed in these cases. Nevertheless, the bis-pyrene phospholipid probe developed in this work is effective for detection of PLA2 enzyme activity through an assay that enables screening for inhibitor development.

Lipid Peroxidation Inhibitory and Cytotoxic Activities of Two Camellia Species Growing Wild in Vietnam

Background Leaves, flowers and young shoots of yellow Camellia species are often used as a substitute for green tea in Vietnam. Yellow Camellia species contain high levels of polyphenol compounds, and have shown activities such as antioxidant, anticancer, and cytotoxic. Objectives We qualitatively screened phytochemicals, quantified total polyphenols of 70% ethanol (EtOH) extracts of leaves, young shoots, and flowers of Camellia vuquangensis and Camellia hatinhensis in Ha Tinh Province, Vietnam. Furthermore, these extracts have been evaluated for lipid peroxidation inhibitory activities at the in vivo level in the BALB/c mouse model, and in vitro cytotoxic activities. Materials and Methods Chemical methods and thin-layer chromatographic (TLC) analysis were used for the qualitative screening of phytochemicals. High-performance liquid chromatography (HPLC) was used to determine total polyphenols. Cell survival was determined through optical density (OD) measured when the protein composition of the cells was stained with sulforhodamine B (SRB). Inhibition of lipid peroxidation was evaluated by determining the content of malondialdehyde (MDA), which is a product of membrane lipid peroxidation. Results The total polyphenol contents of all parts of the two species were comparable to that of green tea with values between 319.3 and 342.6 mg gallic acid equivalents (GAE/g) dry weight. Extracts of leaves, flowers of C. vuquangensis and C. hatinhensis showed strong lipid peroxidation inhibitory activities (IC50: 7.92–17.45 µg/mL) and moderate cytotoxic activities against cell lines HepG2, A549, MCF7, SK-Mel-2, HT-29, and AGS (IC50: 34.73–80.58 µg/mL). Conclusion These two tea species may be considered as herbal teas with various health benefits.

The Use of Chemical Bleaching and Thin-layer Chromatographic Methods for the Detection and Identification of Sudan-III Dye in Adulterated Palm Oil

The deleterious effect of consuming azo dye adulterants in palm oil is well documented including cancer. The presence of Sudan-III dye in palm oil cannot be detected by mere visual inspection. This study was aimed at developing a simple, cheap and convenient protocol for detection and identification of Sudan-III and other azo dyes in adulterated palm oil. The results revealed that the refractive index could be used to screen for azo dye adulteration in palm oil samples as the parameter increases with increasing concentration of Sudan-III dye in palm oil and were statistically different ((p&lt;0.05) from crude unadulterated palm oil samples. Analytical thin layer chromatography and chemical bleaching using 20% v/v each of concentrated sulphuric acid and 30% hydrogen peroxide in palm oil was effective in detecting Sudan III dye adulteration in concentrations of 10mg/L and above.

Phenotypic and genomic analysis of inulin consumption byLactiplantibacillus plantarumstrains from Sichuan pickle

To investigate the capability, properties, and molecular mechanism of inulin fermentation by lactic acid bacteria (LAB) from Sichuan pickle. A total of 79 LAB strains were purified from 30 aged Sichuan pickle brine samples, and only 21 Lactiplantibacillus plantarum strains (26.58%, 21/79) derived from 15 samples grew well through utilizing inulin as a carbon source. The fermentation tests through using long-chain inulin (lc-inulin) as a carbon source showed that only 6 L. plantarum strains grew well, while other 15 strains could only utilize short-chain oligofructose (FOS), and thin-layer chromatography analysis evidenced a strain specificity of inulin consumption patterns. Lactiplantibacillus plantarum YT041 is a vigorous inulin fermenter, and whole genome sequencing data revealed that sacPTS1 and fosRABCDXE operons might be associated with the fermentation of FOS and lc-inulin, respectively. The phenotype of inulin consumption is commonly present in LAB from Sichuan pickle, which is strain-specific and largely depends on their specific ecological niche and degree of polymerization.