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Showing papers on "Thin-layer chromatography published in 2015"


Patent
16 Jun 2015
TL;DR: In this article, a method for manufacturing a chromatography apparatus such as a thin layer chromatography (TLC) plate is disclosed, which includes forming a layer of elongated nanostructures (e.g., carbon nanotubes), and at least partially coating the oxidized elongated nano-structures with a coating.
Abstract: In an embodiment, a method for manufacturing a chromatography apparatus such as a thin layer chromatography (“TLC”) plate is disclosed. The method includes forming a layer of elongated nanostructures (e.g., carbon nanotubes), and at least partially coating the oxidized elongated nanostructures with a coating. The coating includes a stationary phase and/or precursor of a stationary phase and at least one photoluminescent material for use in chromatography. Embodiments for TLC plates and related methods are also disclosed.

105 citations


Journal ArticleDOI
TL;DR: The study gives new insights into goat casein hydrolysates with identified DPP-IV-inhibitory peptides efficiently isolated by 2D-TLC, which provides a simple and cost-efficient separation process and is compatible with liquid chromatography-tandem mass spectrometry (LC-MS/MS) identification.
Abstract: New dipeptidyl peptidase IV (DPP-IV)-inhibitory peptides from trypsin/chymotrypsin-treated goat milk casein hydrolysates were isolated and identified by two-dimensional silica thin-layer chromatography (2D-TLC) combined to nano LC-MS/MS 2D-TLC with chloroform/methanol/25% ammonia (2:2:1) and n-butanol/acetic acid/water (4:1:1) as the first- and second-dimension eluents, respectively, in analytical and semipreparative scales, was set up and verified by reversed-phase high-performance liquid chromatography (RP-HPLC) to be feasible and efficient to separate the hydrolysates Five new DPP-IV-inhibitory peptides, four relatively large oligopeptides (MHQPPQPL, SPTVMFPPQSVL, VMFPPQSVL, and INNQFLPYPY), and AWPQYL were identified, and INNQFLPYPY showed a notable IC50 value of 4008 μM as an uncompetitive inhibitor Interactive effects on DPP-IV inhibition were also observed among separated fractions and pure synthetic peptide mixtures with concentration-dependent activity The study gives new insights into goat casein hydrolysates with identified DPP-IV-inhibitory peptides efficiently isolated by 2D-TLC, which provides a simple and cost-efficient separation process and is compatible with liquid chromatography-tandem mass spectrometry (LC-MS/MS) identification

89 citations


Journal ArticleDOI
TL;DR: A sample preparation method was developed for the analysis of chicken eggs to determine 97 GC and 81 LC amenable residues, including organophosphates, organochlorines, pyrethroids, triazoles, carboxyl-containing compounds, and the indicator PCBs, which offered a higher sample throughput and a lower solvent consumption than the currently established EN 1528 method.

57 citations


Journal Article
TL;DR: The methods adopted and active metabolites extracted and purified can be fruitfully employed for obtaining novel antibiotic compounds to treat human pathogenic bacterial and fungal infections.
Abstract: Objective: The objective was to study the antimicrobial activity of active eluent of thin layer chromatography (TLC) from Streptomyces isolated from marine sample. Methods: To determine the antimicrobial activity by agar well diffusion method. Active compounds were extracted by solvent extraction method and purified using TLC. The presence of active compounds was confirmed by gas chromatography-mass spectrometry (GC-MS) analysis. Identification of active Streptomyces by 16S rRNA partial gene sequencing method. Results: The active strain was identified as Streptomyces cacaoi strain SU2 (JF730119). The GC-MS analysis revealed that the presence of dodecane, eicosane, cetene, diethyl phthalate, phthalic acid isobutyl nonyl ester, thieno[3,2-e] benzofuran and bis (2-ethylhexyl) phthalate. Conclusion: The methods adopted and active metabolites extracted and purified can be fruitfully employed for obtaining novel antibiotic compounds to treat human pathogenic bacterial and fungal infections. Keywords: Streptomyces, GC-MS, Antimicrobial, Thin layer chromatography, Marine soil.

42 citations


Journal ArticleDOI
06 Feb 2015-PLOS ONE
TL;DR: A combined method using Sepbox chromatography and thin-layer chromatography (TLC) bioautography was developed to probe α-glucosidase inhibitors further, and this is the first report of separation of α- glucosodase inhibitors from P. tuberosa.
Abstract: Alpha-glucosidase inhibitors currently form an important basis for developing novel drugs for diabetes treatment. In our preliminary tests, the ethyl acetate fraction of Phlomis tuberosa extracts showed significant α-glucosidase inhibitory activity (IC₅₀ = 100 μg/mL). In the present study, a combined method using Sepbox chromatography and thin-layer chromatography (TLC) bioautography was developed to probe α-glucosidase inhibitors further. The ethyl acetate fraction of P. tuberosa extracts was separated into 150 individual subfractions within 20 h using Sepbox chromatography. Then, under the guidance of TLC bioautography, 20 compounds were successfully isolated from these fractions, including four new diterpenoids [14-hydroxyabieta-8,11,13-triene-11-carbaldehyde-18-oic-12-carboxy-13-(1-hydroxy-1-methylethyl)-lactone (1), 14-hydroxyabieta-8,11,13-triene-17-oic-12-carboxy-13-(1-hydroxy-1-methylethyl)-lactone (2), 14,16-dihydroxyabieta-8,11,13-triene-15,17-dioic acid (3), and phlomisol (15,16-eposy-8,13(16),14-labdatrien-19-ol) (4)], and 16 known compounds. Activity estimation indicated that 15 compounds showed more potent α-glucosidase inhibitory effects (with IC50 values in the range 0.067–1.203 mM) than the positive control, acarbose (IC50 = 3.72 ± 0.113 mM). This is the first report of separation of α-glucosidase inhibitors from P. tuberosa.

35 citations


Journal ArticleDOI
TL;DR: An indigenous mixed bacterial culture was screened and acclimated to decolorize a sulfonated azo dye; acid red 88 (AR88) under changing growth conditions as mentioned in this paper.
Abstract: An indigenous mixed bacterial culture was screened and acclimated to decolorize a sulfonated azo dye; Acid Red 88 (AR88) under changing growth conditions. The process parameters influencing the average decolorization rate, ADRAR88 (μg min−1) was optimized using Taguchi's orthogonal array (OA-L25), under design of experiments. The value of ADRAR88 is 888.86 (μg min−1) when the operational parameters like pH, 8.0, temperature, 35°C and AR88 concentration, 500 (mg L−1) were maintained for 6 h. The mixed culture exhibited 87.89 and 90.58% reduction in the level of chemical oxygen demand (COD) and aromatic amines, respectively. It was found that increasing AR88 concentration, were found to be insignificant on the basis of analysis of variance (ANOVA) and signal-to-noise ratio (SNR) variation for the obtained responses from Taguchi's OA experiments. The decolorization by the mixed culture was confirmed by UV–vis spectrophotometry, whereas biotransformation was evidenced by spectroscopic studies like Fourier transform infrared (FTIR), proton nuclear magnetic resonance (1H-NMR), and chromatographic techniques like high performance liquid chromatography (HPLC), thin layer chromatography (TLC), and gas chromatography coupled with mass spectrometry (GC-MS) analyses. The induction and activities of various AR88 degrading oxido-reductive enzymes was elucidated. The phytotoxicity test using plant seeds revealed the extent of detoxification of parent dye compound by the mixed culture. © 2015 American Institute of Chemical Engineers Environ Prog, 34: 1455–1466, 2015

31 citations


Journal ArticleDOI
TL;DR: The results indicated that HEO‑A may serve as an effective healthcare food and source of natural antioxidant compounds.
Abstract: Oligosaccharide are carbohydrate molecules, comprising repeating units joined together by glycosidic bonds. In recent years, an increasing number of oligosaccharides have been reported to exhibit various biological activities, including antitumor, immune-stimulation and antioxidation effects. In the present study, crude water‑soluble oligosaccharides were extracted from the fruiting bodies of Hericium erinaceus with water and then successively purified by diethylaminoethyl‑cellulose 52 and Sephadex G‑100 column chromatography, yielding one major oligosaccharide fraction: Hericium erinaceus oligosaccharide (HEO‑A). The structural features of HEO‑A were investigated by a combination of monosaccharide component analysis by thin layer chromatography, infrared spectroscopy, nuclear magnetic resonance spectroscopy, scanning electron microscopy and high‑performance gel permeation chromatography. The results indicated that HEO‑A was composed of D‑xylose and D‑glucose, and the average molecular size was ~1,877 Da. The antioxidant activity of HEO‑A was evaluated using three biochemical methods to determine the scavenging activity of HEO‑A on 1,1‑diphenyl‑2‑picrylhydrazyl, hydrogen peroxide and 2,2'‑azino‑bis(3‑ethylbenzthiazoline‑6‑sufonic acid) diammonium radicals. The results indicated that HEO‑A may serve as an effective healthcare food and source of natural antioxidant compounds.

26 citations


Journal ArticleDOI
TL;DR: The poly(L-lactic acid)-silica column was found to work in both modes, and the retention of test compounds depending on acetonitrile content exhibited "U-shaped" curves, which was an indicator of reversed-phase liquid chromatography/hydrophilic interaction liquid Chromatography mixed-mode retention behavior.
Abstract: Poly(L-lactic acid) is a linear aliphatic thermoplastic polyester that can be produced from renewable resources. A poly(L-lactic acid)-modified silica stationary phase was newly prepared by amide bond reaction between amino groups on aminopropyl silica and carboxylic acid groups at the end of the poly(L-lactic acid) chain. The poly(L-lactic acid)-silica column was characterized in reversed-phase liquid chromatography and hydrophilic interaction liquid chromatography with the use of different mobile phase compositions. The poly(L-lactic acid)-silica column was found to work in both modes, and the retention of test compounds depending on acetonitrile content exhibited "U-shaped" curves, which was an indicator of reversed-phase liquid chromatography/hydrophilic interaction liquid chromatography mixed-mode retention behavior. In addition, carbonyl groups included into the poly(L-lactic acid) backbone work as an electron-accepting group toward a polycyclic aromatic hydrocarbon and provide π-π interactions.

18 citations


Journal ArticleDOI
TL;DR: The adiponectin receptors (AdIPoR1 and AdipoR2) are membrane proteins with seven transmembrane helices that regulate glucose and fatty acid metabolism, thereby ameliorating type 2 diabetes.
Abstract: The adiponectin receptors (AdipoR1 and AdipoR2) are membrane proteins with seven transmembrane helices. These receptors regulate glucose and fatty acid metabolism, thereby ameliorating type 2 diabetes. The full-length human AdipoR1 and a series of N-terminally truncated mutants of human AdipoR1 and AdipoR2 were expressed in insect cells. In small-scale size exclusion chromatography, the truncated mutants AdipoR1Δ88 (residues 89–375) and AdipoR2Δ99 (residues 100–386) eluted mostly in the intact monodisperse state, while the others eluted primarily as aggregates. However, gel filtration chromatography of the large-scale preparation of the tag-affinity-purified AdipoR1Δ88 revealed the presence of an excessive amount of the aggregated state over the intact state. Since aggregation due to contaminating nucleic acids may have occurred during the sample concentration step, anion-exchange column chromatography was performed immediately after affinity chromatography, to separate the intact AdipoR1Δ88 from the aggregating species. The separated intact AdipoR1Δ88 did not undergo further aggregation, and was successfully purified to homogeneity by gel filtration chromatography. The purified AdipoR1Δ88 and AdipoR2Δ99 proteins were characterized by thermostability assays with 7-diethylamino-3-(4-maleimidophenyl)-4-methyl coumarin, thin layer chromatography of bound lipids, and surface plasmon resonance analysis of ligand binding, demonstrating their structural integrities. The AdipoR1Δ88 and AdipoR2Δ99 proteins were crystallized with the anti-AdipoR1 monoclonal antibody Fv fragment, by the lipidic mesophase method. X-ray diffraction data sets were obtained at resolutions of 2.8 and 2.4 A, respectively.

18 citations


Journal ArticleDOI
Joseph Sherma1
TL;DR: In addition to references on residue analysis, studies such as pesticide structure – retention relationships, identification and characterization of natural and synthesized pesticides, metabolism, bioactivity, degradation, soil mobility, and lipophilicity are covered.
Abstract: Publications reporting techniques and applications of thin layer chromatography (planar chromatography) for the separation, detection, qualitative, and quantitative determination, and preparative isolation of pesticides and their metabolites are reviewed for the period from November 1, 2012 to November 1, 2014. Analyses are described for a variety of sample types and pesticide classes. In addition to references on residue analysis, studies such as pesticide structure - retention relationships, identification and characterization of natural and synthesized pesticides, metabolism, bioactivity, degradation, soil mobility, and lipophilicity are covered.

17 citations


Journal ArticleDOI
TL;DR: Normal and reversed-phase chromatography can be easily illustrated using thin layer chromatography for the separation of green leaf extracts within a short time and at a low cost.
Abstract: Normal and reversed-phase chromatography can be easily illustrated using thin layer chromatography for the separation of green leaf extracts within a short time and at a low cost.

Journal ArticleDOI
TL;DR: Investigation by thin-layer chromatography (TLC) demonstrated its ability to hydrolyze raffinose and stachyose, suggesting that PMG can be exploited in degradation of non-digestible oligosaccharides from food and feed industries.
Abstract: An alpha-galactosidase was purified from Pseudobalsamia microspora (PMG) to 1224.1-fold with a specific activity of 11,274.5 units/mg by ion-exchange chromatography and gel filtration. PMG is a monomeric protein with a molecular mass of 62 kDa as determined by SDS-PAGE and by gel filtration. Chemical modification using N-bromosuccinimide (NBS) resulted in a complete abrogation of the activity of PMG, suggesting that Trp is an amino acid essential to its activity. The activity was strongly inhibited by Hg(2+), Cd(2+), Cu(2+), and Fe(3+) ions. Three inner peptide sequences for PMG were obtained by liquid chromatography-tandem mass spectrometry (LC-MS-MS) analysis. When 4-nitrophenyl α-D-glucopyranoside (pNPGal) was used as substrate, the optimum pH and temperature of PMG were 5.0 and 55 °C, respectively. The Michaelis constant (K m) value of the alpha-galactosidase on pNPGal was 0.29 mM, and the maximal velocity (V max) was 0.97 μmol ml(-1) min(-1). Investigation by thin-layer chromatography (TLC) demonstrated its ability to hydrolyze raffinose and stachyose. Hence, it can be exploited in degradation of non-digestible oligosaccharides from food and feed industries.

Journal ArticleDOI
10 Nov 2015-PeerJ
TL;DR: This is the first report of chemotaxis towards organophosphate compound by a thermo-halo tolerant bacterium.
Abstract: An organophosphate (OP) degrading chemotactic bacterial strain BUR11 isolated from an agricultural field was identified as a member of Pseudomonas genus on the basis of its 16S rRNA gene sequence. The strain could utilize parathion, chlorpyrifos and their major hydrolytic intermediates as sole source of carbon for its growth and exhibited positive chemotactic response towards most of them. Optimum concentration of parathion for its growth was recorded to be 200 ppm and 62% of which was degraded within 96 h at 37 °C. Growth studies indicated the strain to be moderately thermo-halo tolerant in nature. Investigation based on identification of intermediates of parathion degradation by thin layer chromatography (TLC), high performance liquid chromatography (HPLC), gas chromatography (GC) and liquid chromatography mass spectrometry (LC-MS/MS) provided evidence for possible existence of two pathways. The first pathway proceeds via 4-nitrophenol (4-NP) while the second proceeds through formation of 4-aminoparathion (4-APar), 4-aminophenol (4-AP) and parabenzoquinone (PBQ). This is the first report of chemotaxis towards organophosphate compound by a thermo-halo tolerant bacterium.

Journal ArticleDOI
TL;DR: A new thin layer chromatographic layer using gold nanoparticles grafted 3-triethoxysilyl propylamine modified silica gel (Au NPs-APTS modifiedsilica gel) was developed as a stationary phase for separation and determination of two steroid hormones, namely progesterone and testosterone, and it is applicable to separation and determined in biological matrices such as urine samples.

Journal ArticleDOI
TL;DR: The results indicated that the developed HPLC method is simple, sensitive and reliable and can be readily utilized as a quality control method for horse chestnut and other related medicinal plants.
Abstract: urpose: To separate and quantify four major saponins in the extracts of the skin and the endosperm of seeds of horse chestnut (Aesculus hippocastanum L.) using ultrasonic solvent extraction followed by a high performance liquid chromatography-diode array detector (HPLC-DAD) with positive confirmation by thin layer chromatography (TLC). Methods: The saponins: escin Ia, escin Ib, isoescin Ia and isoescin Ib were extracted using ultrasonic extraction method. The optimized extraction conditions were: 70% methanol as extraction solvent, 80 C as extraction temperature, and the extraction time was achieved in 4 hours. The HPLC conditions used: Zorbax SB-ODS-(150 mm × 2.1 mm, 3 m) column, acetonitrile and 0.10% phosphoric acid solution (39:61 v/v) as mobile phase, flow rate was 0.5 mL min-1 at 210 nm and 230 nm detection. The injection volume was 10 L, and the separation was carried out isothermally at 30 C in a heated chamber. Results: The results indicated that the developed HPLC method is simple, sensitive and reliable. Moreover, the content of escins in seeds decreased by more than 30% in endosperm and by more than 40% in skin upon storage for two years. Conclusion: This assay can be readily utilized as a quality control method for horse chestnut and other related medicinal plants.

Journal ArticleDOI
TL;DR: The received results demonstrate that the RP-TLC method may be the only reliable technique for fluoroquinolones in describing their lipophilic nature as well as the activity.
Abstract: The lipophilicity of fifteen fluoroquinolones has been investigated. Reversed-phase thin layer chromatography with densitometric detection was applied to determine the RM0 factor. The RP-TLC investigations were performed in mixtures of organic modifier–water. The partition coefficients of examined fluoroquinolones were also calculated with computational programs. All the obtained data, from experimental methods and theoretical calculations, were compared and a suitable conclusion was reached. The received results demonstrate that the RP-TLC method may be the only reliable technique for fluoroquinolones in describing their lipophilic nature as well as the activity.

Journal ArticleDOI
TL;DR: It is believed that Ch147 is a potential candidate for the conversion of waste materials into simple sugar for productions of biofuels and also can be used as an alternative option for biological control of plant pathogens being unfriendly to chemicals.
Abstract: Streptomyces sp. CS147 grown on chitin liquid medium incorporating 0.5 % colloidal chitin as a sole carbon source produced an extracellular chitinase (Ch147). The enzyme (Ch147) was purified using Sepharose CL-6B column chromatography and biochemically characterized. The enzymatic reaction products, analyzed using high-performance liquid chromatography and thin layer chromatography clearly indicates the production of N-acetyl-d-glucosamine (GlcNAc) as principal product which can be further hydrolyzed for the production of alcohol, a second generation biofuel. Ch147 hydrolyzed colloidal chitin to 0.278, 0.817, and 1.058 mg/mL of (GlcNAc) as a major product, at retention time of 4.3, respectively, when incubated for 8, 16, and 24 h at 50 °C. GlcNAc is a monosaccharide that usually polymerize linearly through (1, 4) β linkage. The 41 kDa molecular mass chitinase, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), has the amino acid sequences DINGGGATLPQKLYL significantly different from other chitinase. Ch147 had K m and V max values of 2.05 ± 5.3 mg/mL and 467.2 ± 2.4 mmol/min, respectively. Further, the purified enzyme (5 U) inhibits the fungal phytopathogens belonging to the genera Fusarium and Aspergillus. We believe that Ch147 is a potential candidate for the conversion of waste materials into simple sugar for productions of biofuels and also can be used as an alternative option for biological control of plant pathogens being unfriendly to chemicals.

Journal ArticleDOI
Naila Hassan1, Javed Ahamad1, Saima Amin1, Mohd Mujeeb1, Showkat R. Mir1 
TL;DR: Estimation of erythrocentaurin in extracts and fractions based on high-pressure thin-layer chromatography was carried out on silica gel 60 F(254) plates with toluene/ethyl acetate/formic acid as the mobile phase and showed good linearity in the concentration range of 200-1500 ng/band.
Abstract: Erythrocentaurin is a relatively simple natural product present among the members of Gentianaceae. A preparative method for the isolation of erythrocentaurin from the ethyl acetate fraction of Enicostemma littorale using medium-pressure liquid chromatography has been reported. The method consisted of a simple step gradient from 10 to 20% ethyl acetate in n-hexane. Using a 70 × 460 mm Si60 column, this method is capable of processing 20 g of material in <3 h (purity ≈ 97%). The recovery of erythrocentaurin was 87.77%. Estimation of erythrocentaurin in extracts and fractions based on high-pressure thin-layer chromatography was carried out on silica gel 60 F(254) plates with toluene/ethyl acetate/formic acid (80:18:2 v/v/v) as the mobile phase. The densitometric analysis was performed at 230 nm. A well-separated compact band of erythrocentaurin appeared at R(f )0.54 ± 0.04. The analytical method showed good linearity in the concentration range of 200-1500 ng/band with a correlation coefficient of 0.99417. The limits of detection and quantification were found to be ≈60 and ≈180 ng/band, respectively. Erythrocentaurin exhibited a concentration-dependent α-amylase inhibition (IC(50) 1.67 ± 0.28 mg/mL). The outcome of the study should be considered for pharmacokinetic and biotransformation studies involving E. littorale.

Journal ArticleDOI
TL;DR: The developed HPLC and HPTLC fingerprinting method can be used for the quality control, and standardization of WG and its extracts used as nutritional supplement.
Abstract: Aim: Wheatgrass (WG) is the shoot of Triticum aestivum Linn. belongs to the family Gramineae, and possess high chlorophyll content and essential vitamins, minerals, vital enzymes, amino acids, dietary fibers etc., It has been shown to possess anti-cancer, anti-ulcer, antioxidant, and anti-arthritic activity due to the presence of biologically active compounds, and minerals. Therefore, in the present study, high-performance thin layer chromatography (HPTLC), and high-performance liquid chromatography (HPLC) methods for qualitative and quantitative analysis have been proposed, which will help in quality evaluation of wheat grass extract. Materials and Methods: Samples for analysis were prepared in methanol and water simply by sonication. These were applied on pre-coated silica plate and chromatograms were developed using toluene: Ethyl acetate: Formic acid. HPLC analysis was done on Waters HPLC system using water, methanol, and acetonitrile as mobile phase. Merck C18 column has been used. Results: HPTLC finger printing of alcoholic extracts of WG was carried out and found 10–11 spots at different wavelengths 254, 366, and 435 nm. HPLC fingerprinting produced 22 peaks at 256 nm. Quantitative HPTLC analysis was done to determine the gallic acid content, and was found to be 0.077% w/w in aqueous extract. By HPLC, the content of gallic acid and rutin was found to be 0.07%, and 0.04% w/w in aqueous extract of WG. Conclusion: The developed HPLC and HPTLC fingerprinting method can be used for the quality control, and standardization of WG and its extracts used as nutritional supplement.

Journal ArticleDOI
TL;DR: The developed assay is able to detect acetylcholinesterase inhibitors present in complex matrixes that were chromatographed in normal phase or reversed-phase thin-layer chromatography.
Abstract: A dual readout autographic assay to detect acetylcholinesterase inhibitors present in complex matrices adsorbed on reversed-phase or normal-phase thin-layer chromatography plates is described. Enzyme gel entrapment with an amphiphilic copolymer was used for assay development. The effects of substrate and enzyme concentrations, pH, incubation time, and incubation temperature on the sensitivity and the detection limit of the assay were evaluated. Experimental design and response surface methodology were used to optimize conditions with a minimum number of experiments. The assay allowed the detection of 0.01% w/w of physostigmine in both a spiked Sonchus oleraceus L. extract chromatographed on normal phase and a spiked Pimenta racemosa (Mill.) J.W. Moore leaf essential oil chromatographed on reversed phase. Finally, the reversed-phase thin-layer chromatography assay was applied to reveal the presence of an inhibitor in the Cymbopogon citratus (DC.) Stapf essential oil. The developed assay is able to detect acetylcholinesterase inhibitors present in complex matrixes that were chromatographed in normal phase or reversed-phase thin-layer chromatography. The detection limit for physostigmine on both normal and reversed phase was of 1×10(-4) μg. The results can be read by a change in color and/or a change in fluorescence.

Journal ArticleDOI
TL;DR: This is the first report of achyrobichalcone isolation on a semi-preparative scale by high-speed countercurrent chromatography, which afforded achyrocline satureioides in good yield and purity for further biopharmaceutical studies.
Abstract: Achyrobichalcone is a biflavonoid recently found in Achyrocline satureioides. This substance has unprecedented chemical structure and occurrence, but resembles other bioactive bichalcones, which have important pharmacological properties, such as anticancer activity. The major challenge for evaluation of the physicochemical and biological properties of this new molecule is the isolation step, which affects the purity and yield of the isolated product. Thus, the objective of this work was to develop a semi-preparative method for achyrobichalcone isolation from Achyrocline satureioides by high-speed countercurrent chromatography. The high-speed countercurrent chromatography separation was achieved in two steps. In the first step, an enriched fraction of achyrobichalcone from the freeze-dried extract was obtained, using the solvent system hexane-ethyl acetate-methanol-water 0.8:1:0.8:1, v/v. The purification of achyrobichalcone from the enriched fractions was achieved by further high-speed countercurrent chromatography fractionation with hexane-ethyl acetate-methanol-water 0.9:0.9:0.8:1, v/v. The final isolated product was obtained using preparative thin layer chromatography and crystallization procedure. A yellow semi-crystalline solid with purity close to 90% was obtained as the final product. The mass recovery of achyrobichalcone isolation was near 67%. The structural identification from spectroscopic and chromatographic techniques confirmed the achyrobichalcone structure. This is the first report of achyrobichalcone isolation on a semi-preparative scale by high-speed countercurrent chromatography. This method afforded achyrobichalcone in good yield and purity for further biopharmaceutical studies.

Journal ArticleDOI
TL;DR: Optimum performance was obtained after spotting samples onto thin-layer chromatography plates as sample substrates for a custom-built solvent-assisted desorption/ionization mass spectrometry (DI-MS) interface.
Abstract: In this study, we demonstrate the application of ambient mass spectrometry for measuring fatty acids from various biological sample matrices such as olive oil, fish oil, salmon, and human serum. Optimum performance was obtained after spotting samples onto thin-layer chromatography (TLC) plates as sample substrates for a custom-built solvent-assisted desorption/ionization mass spectrometry (DI-MS) interface. Good to excellent linearities (coefficients of determination, 0.9856 to 0.9977) and reproducibilities (average 6 % relative standard deviation (RSD) using syringe deposition) were obtained after application of an internal standard. Signal suppression phenomena were minimized by separating the analytes by TLC to some extent prior to DI-MS, leading to a fourfold increase of signal-to-noise ratios as compared to single spot mixture analysis without TLC separation.

Journal ArticleDOI
TL;DR: In this paper, the influence of the following variables such as adsorbent type, type and concentration of organic modifier in mobile phase and pH of the mobile phase buffer on retention of some synthetic peptides in reversed-phase high-performance thin-layer chromatography systems has been investigated.
Abstract: Influence of the following variables such as adsorbent type, type and concentration of organic modifier in mobile phase, type and concentration of ion-pairing reagent, or pH of the mobile phase buffer on retention of some synthetic peptides in reversed-phase high-performance thin-layer chromatography systems has been investigated. The investigations have been also focused on influence of the variables mentioned on solute zone shape regarding optimization of their separation. Remarks about solute retention mechanism have been also provided.

Journal ArticleDOI
TL;DR: The proposed HPTLC method proved to be more sensitive, while the HPLC gave more reproducible results besides saving time; indicating the ability of the proposed methods to be reliable and suitable for routine analysis of drug product.
Abstract: Validated sensitive and highly selective methods were developed for the quantitative determination of cefoperazone sodium (CEF) in the presence of its reported impurities; 7-aminocephalosporanic acid (7-ACA) and 5-mercapto-1-methyl-tetrazole (5-MER). Method A is high-performance liquid chromatography (HPLC), where the mixture of CEF and the reported impurities; 7-ACA and 5-MER were separated on a C8 column (5 µm ps, 250 mm × 4.6 i.d.) using methanol:0.05 M KH2PO4 buffer (22.5:77.5 v/v, pH 7.5) as a mobile phase. The three components were detected at 254 nm with a concentration range of 10-90 µg mL(-1) and the mean percentage recovery 99.67% (SD 1.465). Method B is high-performance thin layer chromatography (HPTLC), where the mixture of CEF and the reported impurities were separated on silica gel HPTLC F254 plates using (acetone:methanol:ethyl acetate:2% sodium lauryl sulfate:glacial acetic acid) (3:2:3:0.8:0.2, by volume) as a developing system and scanning at 254 nm over a concentration range of 1-10 µg per band with the mean percentage recovery 99.95% (SD 1.335). The proposed methods were statistically compared with a reported HPLC method with no significant difference regarding accuracy and precision; indicating the ability of the proposed methods to be reliable and suitable for routine analysis of drug product. The proposed HPTLC method proved to be more sensitive, while the HPLC gave more reproducible results besides saving time.

Journal ArticleDOI
Xiang Ding1, Yiling Hou1, Wanru Hou1, Yuanxiu Zhu1, Lei Fu1, Hongqing Zhu1 
TL;DR: The LDGO-A might serve as an effective healthcare food and source of natural anti-tumor compounds and also effected the expression of some housekeeping genes mRNA in S180 tumor.
Abstract: Background: Oligosaccharides are composed of a variable number of monosaccharide units and very important in the biologically diverse of biological systems. Materials and Methods: Crude water-soluble oligosaccharide was extracted from the fruiting bodies with water and then successively purified by DEAE-cellulose 52 and Sephadex G-100 column chromatography, yielding one major oligosaccharides fractions: LES-A. Structural features of Lactarius deliciosus (L. ex Fr.) Gray oligosaccharide (LDGO-A) were investigated by a combination of monosaccharide component analysis by thin layer chromatography, infrared spectra, nuclear magnetic resonance spectroscopy, scanning electron microscopy, and high-performance gel permeation chromatography analysis. Result: The results indicated that LDGO-A was composed of D-glucose and D-xylose, and the average molecular sizes was approximately 945 Da. The anti-tumor activity of LDGO-A was evaluated in vivo. The inhibitory rate in mice treated with 40 mg/kg LDGO-A can reach 40.02%, being the highest in the three doses, which may be comparable to mannatide. Histology of immune organs shows that the tissues arranged more regular and firmer, but the tumor tissue arranged looser in LDGO-A group than those in the control group. Meanwhile, there is no obvious damage to other organs, such as heart. The anti-tumor activity of the LDGO-A was usually believed to be a consequence of the stimulation of the cell-mediated immune response because it can significantly promote the lymphocyte and macrophage cells in the dose range of 100-400 μg/mL in vitro. LDGO-A also effected the expression of some housekeeping genes mRNA in S180 tumor. Conclusion: Accordingly, the LDGO-A might serve as an effective healthcare food and source of natural anti-tumor compounds.

01 Jan 2015
TL;DR: In this article, the authors used thin layer chromatography (TLC) to measure the number of compounds in hexane:ethyl acetate: methanol solvent system for cocoa pod and chloroform:methanol solvent for cocoa leaf under UV illuminator.
Abstract: The cocoa leaf (CL), seeds (CS) and pod (CP) were studied for the estimation of medicinal properties that they contain and the focus was screening for anticancer ability. Extraction of the samples in Soxhlet apparatus was done with methanol as the solvent followed by phytochemical analysis, antimicrobial activity, antioxidant assay and anticancer activity. Thin Layer Chromatography (TLC) showed the number of compounds in hexane:ethyl acetate: methanol solvent system for cocoa pod and chloroform:methanol solvent for cocoa leaf under UV illuminator. The antimicrobial assay by agar well diffusion method of cocoa seeds showed zone of inhibition against pathogens Serratia marcescens, Staphylococcus aureus, Salmonella sp. and Shigella dysenteriae. Further, antioxidant assay was performed using DPPH radical scavenging assay. The sample was then tested for cytotoxicity assay against MG63 osteosarcoma cell lines and active compounds were identified by doing Gas chromatography and Mass spectroscopy (GCMS).

Journal ArticleDOI
TL;DR: Methods had been validated as per the International Conference of Harmonization guideline and the results indicated that the suggested methods were reproducible, reliable and applicable for rapid routine analysis.
Abstract: This work was concerned with development, optimization, application and validation of reversed phase high performance liquid chromatography (RP-HPLC) and thin layer chromatography (TLC)-densitometric methods for analysis of cetylpyridinium chloride, chlorocresol and lidocaine in Canyon(®) gel. The first developed RP-HPLC method depended on chromatographic separation on a ZORBAX Eclipse Plus C8 column, with elution with a mobile phase consisting of 0.05% phosphoric acid solution : acetonitrile : methanol (15 : 24 : 61, by volume), pumping the mobile phase at a flow rate of 1.00 mL min(-1), with ultraviolet detection at 220 nm. While in the subsequently developed method, the TLC-densitometric method, complete separation of the studied mixture was achieved using methanol : acetone : acetic acid (7 : 3 : 0.2, by volume) as a mobile phase, aluminum plates precoated with silica gel 60 F254 as a stationary phase and 215 nm as the scanning wavelength. Factors affecting the developed methods were studied and optimized; moreover, methods had been validated as per the International Conference of Harmonization guideline and the results indicated that the suggested methods were reproducible, reliable and applicable for rapid routine analysis. Statistical comparison of the two developed methods with the reported HPLC ones using F- and Student's t tests showed no significant difference.

Journal ArticleDOI
Xiang Ma1, Erpei Wang1, Yuyun Lu1, Yong Wang1, Shiyi Ou1, Rian Yan1 
22 Jun 2015-PLOS ONE
TL;DR: This study selectively acylated the primary hydroxyl groups on flavonoids in antioxidant of bamboo leaves (AOB) using lauric acid with Candida antarctica lipase B in tert-amyl-alcohol to reduce acrylamide formation in fried potato crisps.
Abstract: This study selectively acylated the primary hydroxyl groups on flavonoids in antioxidant of bamboo leaves (AOB) using lauric acid with Candida antarctica lipase B in tert-amyl-alcohol. The separation and isolation of acylated derivatives were performed using silica gel column chromatography with a mixture of dichloromethane/diethyl ether/methanol as eluents. Both thin layer chromatography and high-performance liquid chromatography analyses confirmed the high efficiency of the isolation process with the purified orientin-6″-laurate, isoorientin-6″-laurate, vitexin-6″-laurate, and isovitexin-6″-laurate that were obtained. The addition of AOB and acylated AOB reduced acrylamide formation in fried potato crisps. Results showed that 0.05% AOB and 0.05% and 0.1% acylated AOB groups significantly (p < 0.05) reduced the content of acrylamide in potato crisps by 30.7%, 44.5%, and 46.9%, respectively.


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TL;DR: Soil and water samples were collected from various regions of SIPCOT and nearby Vanappadi Lake, Ranipet, Tamilnadu, India and a new compound is proposed to be 6-propionyl pterin (2-amino-6- Propionyl-3H-pteridin-4-one).
Abstract: Soil and water samples were collected from various regions of SIPCOT and nearby Vanappadi Lake, Ranipet, Tamilnadu, India. Based on their colony morphology and their stability during subculturing, 72 bacteria were isolated, of which 14 isolates were actinomycetes. Preliminary selection was carried out to exploit the ability of the microorganisms to utilize sodium cyanate as nitrogen source. Those organisms that were able to utilize cyanate were subjected to secondary screening viz., utilization of sodium cyanide as the nitrogen source. The oxygenolytic cleavage of cyanide is dependent on cyanide monooxygenase which obligately requires pterin cofactor for its activity. Based on this, the organisms capable of utilizing sodium cyanide were tested for the presence of pterin. Thin layer chromatography (TLC) of the cell extracts using n-butanol: 5 N glacial acetic acid (4:1) revealed that 10 out of 12 organisms that were able to utilize cyanide had the pterin-related blue fluorescent compound in the cell extract. The cell extracts of these 10 organisms were subjected to high performance thin layer chromatography (HPTLC) for further confirmation using a pterin standard. Based on the incubation period, cell biomass yield, peak height and area, strain VPW3 was selected and was identified as Bacillus subtilis. The Rf value of the cell extract was 0.73 which was consistent with the 0.74 Rf value of the pterin standard when scanned at 254 nm. The compound was extracted and purified by preparative High Performance Liquid Chromatography (HPLC). Characterization of the compound was performed by ultraviolet spectrum, fluorescence spectrum, Electrospray Ionization-Mass Spectrometry (ESI-MS), and Nuclear Magnetic Resonance spectroscopy (NMR). The compound is proposed to be 6-propionyl pterin (2-amino-6-propionyl-3H-pteridin-4-one).