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Separations
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New Trends in Thin-Layer Chromatography
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New Trends in Thin-Layer Chromatography

A special issue of Separations (ISSN 2297-8739).

Deadline for manuscript submissions: closed (15 February 2015) | Viewed by 59629

Special Issue Editor

Dr. Mark Devlin Maloney

E-Mail Website
Guest Editor
Department of Biology, Spelman College, Atlanta, GA 30314, USA
Interests: immune system; B cell development; thin-layer chromatography; glycolipid-protein interaction

Special Issue Information

Dear Colleagues,

For many years, Thin-Layer Chromatography (TLC) has served science and industry well in the efficient and cost-effective separation, purification and identification of many chemical substances. Advancements in manufacturing have improved separation and isolation efficiencies of high-performance TLC (HPTLC) plates, and the number of substances incorporated into the stationary phase of thin-layer plates has allowed for diverse and specialized applications. TLC remains an economical and expedient method for the isolation, purification and identification of many different types of molecules. TLC overlay and TLC blotting techniques exploit the specificity of protein probes such as antibodies and toxins in aqueous solutions for the identification of glycolipids and other complex molecules. Coupling HPTLC with the analytical capabilities of techniques such as mass spectrometry and Raman scattering also has added new dimensions to the usefulness of TLC. This issue is dedicated to demonstrating the utility of the many improvements in TLC sorbent and binder technology and the ability to couple TLC separation capabilities with other analytical techniques that have allowed for such a multitude of applications.

Prof. Dr. Mark Devlin Maloney
Guest Editor

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Keywords

  • chromatography
  • stationary phase
  • HPTLC
  • TLC overlay
  • TLC blot
  • TLC IR-MALDI-o-TOF MS
  • TLC-SERS

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Published Papers (6 papers)

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Research

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3747 KiB  
Article
The Dramatic Modulatory Role of the 2'N Substitution of the Terminal Amino Hexose of Globotetraosylceramide in Determining Binding by Members of the Verotoxin Family
by Murugesapillai Mylvaganam, Beth Binnington, Monique Budani, Anna M. Soltyk and Clifford A. Lingwood
Chromatography 2015, 2(3), 529-544; https://doi.org/10.3390/chromatography2030529 - 14 Aug 2015
Cited by 1 | Viewed by 5508
Abstract
Although globotetraosylceramide (Gb4) is only recognized by a single member of the verotoxin family namely, the pig edema disease toxin (VT2e), removal of the acetyl group from the terminal N-acetyl hexosamine of Gb4 to generate the free amino sugar containing [...] Read more.
Although globotetraosylceramide (Gb4) is only recognized by a single member of the verotoxin family namely, the pig edema disease toxin (VT2e), removal of the acetyl group from the terminal N-acetyl hexosamine of Gb4 to generate the free amino sugar containing species (aminoGb4) results in the generation of a glycolipid preferentially recognized by all members of the verotoxin family (i.e., VT1, VT2, VT2c, and VT2e). GT3, a site-specific mutant of VT2e, in which Gb4 recognition is lost but Gb3 binding is retained, also binds aminoGb4. We have now compared the binding of VT1, VT2, VT2e, and GT3 to a series of aminoGb4 derivatives using a TLC overlay technique. DimethylaminoGb4 is bound by VT1 and VT2 but not VT2e or GT3; formylaminoGb4 binds all toxins but poorly to VT2 and preferentially VT2e; trifluoroacetylaminoGb4 binds only VT2e and GT3; isopropylaminoGb4 binds VT1 and poorly to VT2; benzylaminoGb4 binds all four toxins. Thus, there is a marked distinction between the permissible amino substitutions for VT1 and VT2e binding. GT3 is a hybrid between these in that, according to the substitution, it behaves similarly either to VT1 or to VT2e. For each species, GT3 does not however, show a hybrid binding between that of VT1 and VT2e. Analysis of the binding as a function of pH shows opposite effects for VT1 and VT2e: decreased pH increases VT1, but decreases VT2e receptor glycolipid binding. Full article
(This article belongs to the Special Issue New Trends in Thin-Layer Chromatography)
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664 KiB  
Article
Effects of Different Levels of Echinostoma caproni Miracidial Dose on Glucose and Maltose Composition of Biomphalaria glabrata Snails as Determined by High Performance Thin-Layer Chromatography-Densitometry
by Dolcie DeGrandchamp, Sage Hartlaub, Bernard Fried and Joseph Sherma
Chromatography 2015, 2(2), 188-194; https://doi.org/10.3390/chromatography2020188 - 9 Apr 2015
Viewed by 5490
Abstract
The effects of 5, 25, and 40 Echinostoma caproni miracidia on the sugar content of young adult and mature adult Biomphalaria glabrata were studied using high performance thin layer chromatography (HPTLC)-densitometry. Analysis was done on the snail’s digestive gland gonad complex (DGG) at [...] Read more.
The effects of 5, 25, and 40 Echinostoma caproni miracidia on the sugar content of young adult and mature adult Biomphalaria glabrata were studied using high performance thin layer chromatography (HPTLC)-densitometry. Analysis was done on the snail’s digestive gland gonad complex (DGG) at two and four weeks postmiracidial exposure. The sugars were extracted from the DGG using 70% ethanol and analyzed on silica gel HPTLC plates with a preadsorbent zone using 1-butanol-glacial acetic acid-diethyl ether-deionized water (27:18:5:3) mobile phase. The separated bands were then detected using alpha-naphthol-sulfuric reagent and quantified by densitometry at 515 nm. Significant differences were found in the maltose content between two and four weeks post exposure for both age groups. Additionally, significantly lower maltose and glucose levels were observed in the high exposure groups of both ages. Full article
(This article belongs to the Special Issue New Trends in Thin-Layer Chromatography)
700 KiB  
Article
A Hyphenated Technique based on High-Performance Thin Layer Chromatography for Determining Neutral Sphingolipids: A Proof of Concept
by Andrés Domínguez, Carmen Jarne, Vicente L. Cebolla, Javier Galbán, María Savirón, Jesús Orduna, Luis Membrado, María-Pilar Lapieza, Elena Romero, Isabel Sanz Vicente, Susana De Marcos and Rosa Garriga
Chromatography 2015, 2(2), 167-187; https://doi.org/10.3390/chromatography2020167 - 8 Apr 2015
Cited by 11 | Viewed by 7605
Abstract
Hyphenated HPTLC has been used to analyze several neutral sphingolipids acting as lysosomal storage disease (LSD) biomarkers. Automated multiple development (AMD) provides separation of lipid peaks, which are detected and quantified using fluorescence detection by intensity changes (FDIC) after primuline post-impregnation. A final [...] Read more.
Hyphenated HPTLC has been used to analyze several neutral sphingolipids acting as lysosomal storage disease (LSD) biomarkers. Automated multiple development (AMD) provides separation of lipid peaks, which are detected and quantified using fluorescence detection by intensity changes (FDIC) after primuline post-impregnation. A final online transfer to a mass spectrometer by means of an elution-based interface allows their identification using electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI).Given that the increases in fluorescent emission detected by FDIC are produced by non-specific, electrostatic interactions between the primuline and hydrocarbon chains in the ceramide backbones of sphingolipids, it is a non-destructive detection technique, allowing the precise location and transfer of biomarker peaks to a mass spectrometer using an elution interface. By using primuline as a fluorophore, the technique is also compatible with ESI-APCI and does not interfere with the MS of sphingolipids. APCI provides useful and complementary structural information to the ESI for sphingolipid identification. Moreover, FDIC emission can be used for quantitative purposes. Results include the determination of sphingomyelin (SM) in human-plasma samples (RSD < 6%) by means of a standard addition method with non-linear calibration, and the identification of globotriaosylceramide (Gb3) in the plasma of a Fabry patient. Only one HPTLC plate is needed to perform the analysis. Full article
(This article belongs to the Special Issue New Trends in Thin-Layer Chromatography)
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2435 KiB  
Article
Analysis of Bioactive Components of Oilseed Cakes by High-Performance Thin-Layer Chromatography-(Bio)assay Combined with Mass Spectrometry
by Sue-Siang Teh and Gertrud E. Morlock
Chromatography 2015, 2(1), 125-140; https://doi.org/10.3390/chromatography2010125 - 17 Mar 2015
Cited by 16 | Viewed by 10194
Abstract
Hemp, flax and canola seed cakes are byproducts of the plant oil extraction industry that have not received much attention in terms of their potential use for human food instead of animal feed. Thus, the bioactivity profiling of these oilseed cakes is of [...] Read more.
Hemp, flax and canola seed cakes are byproducts of the plant oil extraction industry that have not received much attention in terms of their potential use for human food instead of animal feed. Thus, the bioactivity profiling of these oilseed cakes is of interest. For their effect-directed analysis, planar chromatography was combined with several (bio)assays, namely 2,2-diphenyl-1-picrylhydrazyl scavenging, acetylcholine esterase inhibition, planar yeast estrogen screen, antimicrobial Bacillus subtilis and Aliivibrio fischeri assays. The streamlined high-performance thin-layer chromatography (HPTLC)-bioassay method allowed the discovery of previously unknown bioactive compounds present in these oilseed cake extracts. In contrast to target analysis, the direct link to the effective compounds allowed comprehensive information with regard to selected effects. HPTLC-electrospray ionization-mass spectrometry via the elution-head based TLC-MS Interface was used for a first characterization of the unknown effective compounds. The demonstrated bioactivity profiling on the feed/food intake side may guide the isolation of active compounds for production of functional food or for justified motivation of functional feed/food supplements. Full article
(This article belongs to the Special Issue New Trends in Thin-Layer Chromatography)
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Review

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3090 KiB  
Review
TLC-Direct Bioautography as a High Throughput Method for Detection of Antimicrobials in Plants
by Irena M. Choma and Wioleta Jesionek
Chromatography 2015, 2(2), 225-238; https://doi.org/10.3390/chromatography2020225 - 18 May 2015
Cited by 56 | Viewed by 24373
Abstract
The richness of bioactive compounds in plant materials encourages continuous development of separation methods and bioassays for their isolation and identification. Thin-layer chromatography-direct bioautography links separation on the adsorbent layer with biological tests performed directly on it. Therefore, the method is very convenient [...] Read more.
The richness of bioactive compounds in plant materials encourages continuous development of separation methods and bioassays for their isolation and identification. Thin-layer chromatography-direct bioautography links separation on the adsorbent layer with biological tests performed directly on it. Therefore, the method is very convenient for searching plant constituents with biological activity, such as antibiotics. Test bacteria grow directly on a plate surface excluding places where antibacterials are located. They can be detected with reagents converted by living bacteria. TLC-DB is a high throughput method enabling analyses of many samples in parallel and the comparison of their activity. Both screening and semi-quantitative analysis is possible. The targeted compounds can be identified using spectroscopic methods, mostly mass spectrometry, that can be performed directly on a TLC plate. This paper discusses all above mentioned aspects of TLC-DB, illustrating them with literature, schemes and our own results. Full article
(This article belongs to the Special Issue New Trends in Thin-Layer Chromatography)
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633 KiB  
Review
Recent High Performance Thin Layer Chromatographic Studies on Biomphalaria glabrata (Gastropoda)
by Bernard Fried and Aditya Reddy
Chromatography 2015, 2(1), 118-124; https://doi.org/10.3390/chromatography2010118 - 10 Mar 2015
Cited by 2 | Viewed by 4367
Abstract
This review examines the recent high performance thin layer chromatography (HPTLC) literature on the effects of biotic and abiotic factors on certain analytes in the medically important freshwater snail, Biomphalaria glabrata. The analytes studied were lipids, lipophilic pigments, amino acids, and carbohydrates. [...] Read more.
This review examines the recent high performance thin layer chromatography (HPTLC) literature on the effects of biotic and abiotic factors on certain analytes in the medically important freshwater snail, Biomphalaria glabrata. The analytes studied were lipids, lipophilic pigments, amino acids, and carbohydrates. As determined by HPTLC, various factors, such as larval parasitism, estivation, temperature changes, and others, alter the metabolism of the snail and cause significant changes in the chemical contents of the analytes under study. Full article
(This article belongs to the Special Issue New Trends in Thin-Layer Chromatography)
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