Theory and Mechanism of Thin-Layer Chromatography

Kaczmarski, Krzysztof; Kowalska, Teresa; Prus, Wojciech

doi:10.1201/9780203912430.ch2

Cited by 5 publications

References 26 publications

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Capillary action liquid chromatography

Zhang

Bergström

Goodall³

et al. 2009

J of Separation Science

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Capillary action LC (caLC) is introduced as a technique using capillary action as the driving force to perform LC in capillary columns packed with HPLC type microparticulate materials. A dry packing method with centrifugal force was developed to prepare capillary columns in parallel (10 columns per 3 min) to support their disposable use in caLC. Using a digital microscope for real-time imaging and recording separations of components in a dye mixture, caLC was found to have flow characteristics similar to TLC. Based on the investigation of microparticulate HPLC silica gels of different size (1.5-10 microm) and a typical TLC grade irregular medium, Merck 60G silica, the van Deemter curves suggested molecular diffusion as the major contribution to band broadening in caLC. With Waters Xbridge 2.6 microm silica, plate heights down to 8.8 microm were obtained, comparable to those achievable in HPLC. Assisted by an image-processing method, the visual caLC separation was converted to a classical chromatogram for further data analysis and such a facility confirmed the observation of highly efficient bands.

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Capillary action liquid chromatography

Zhang

Bergström

Goodall³

et al. 2009

J of Separation Science

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High Performance Thin-Layer Chromatography

Milojković‐Opsenica

Andrić

2013

Green Chromatographic Techniques

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A sustainable HPTLC approach for green assessment of Tyrosine to phenylalanine ratio in chronic kidney disease

Kamel,

Abdel-aal,

Osman

et al. 2024

Sci Rep

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The kidney is considered the main site for the net release of Tyrosine (Tyr) to the circulation through hydroxylation of Phenylalanine (Phe) using phenylalanine hydroxylase enzyme. In chronic kidney disease (CKD) patients the enzyme activity is impaired, therefore the serum concentration ratio of Tyr/Phe is reduced compared to healthy individuals. This motivates us to develop a cost effective, green, simple, sensitive, and selective HPTLC method to measure the levels of Tyr and Phe in serum samples. The analysis was carried out using acetonitrile: ethanol: 25% ammonia solution: ethyl acetate (6.5: 1.5: 1: 0.5) as a mobile phase. Rf values were 0.55 ± 0.05 for Phe and 0.39 ± 0.05 for Tyr. Densitometry scanning was performed using UV detector and dual wavelengths of 210 and 225 nm were obtained. A linear correlation was observed between the levels of Phe and Tyr, ranging from 50 to 700 ng band− 1 and 50 to 600 ng band− 1, respectively, under the optimum conditions. The method selectivity, linearity, precision, accuracy, and robustness were all confirmed in accordance with ICH recommendations. Calculations of the separation and resolution factors, number of theoretical plates, and height equivalent to theoretical plates prove to the chromatographic system accuracy and high separation efficiency. The developed method exhibits an acceptable eco-scale when measuring the method greenness using AGREE and GAPI softwares. It was applied for the determination of Phe and Tyr concentrations in human serum samples.

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Theory and Mechanism of Thin-Layer Chromatography

Cited by 5 publications

References 26 publications

Capillary action liquid chromatography

Capillary action liquid chromatography

High Performance Thin-Layer Chromatography

A sustainable HPTLC approach for green assessment of Tyrosine to phenylalanine ratio in chronic kidney disease

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