Chromatographic separation thin layer chromatography

Solute identification means qualitative analysis. Various methods are used today to identify a separated substance on a thin-layer chromatographic plate. Of all chromatographic methods, thin-layer chromatography (TLC) provides a unique simultaneous separation of up to 70 samples on the same plate therefore the reproducibility of the experimental conditions is not an issue because the experimental conditions are the same for all samples. This, together with the advantage of separating compounds with very different polarity and the possibility of using different detection methods for the same spot or for adjacent spots on the same plate, is the power of TLC. 

Residues are determined in the purified extracts by chromatographic or immunochemical techniques. In the chromatographic systems, thin-layer chromatography (TEC), liquid chromatography (LC), and GC, the analytes are separated on plates or columns and determined by colorimetry, by spectrophotometry (ultraviolet (UV), infrared (IR, Fourier transform infrared (FTIR)), by fluorescence, by selective detectors (in GC analysis ECD, flame photometric (FPD), nitrogen/phosphorus (NPD, TSD), etc.), or by MS. Separations may also be achieved by... 

An important publication by Kost et al. (63JGU525) on thin-layer chromatography (TLC) of pyrazoles contains a large collection of Rf values for 1 1 mixtures of petroleum ether-chloroform or benzene-chloroform as eluents and alumina as stationary phase. 1,3- and 1,5-disubstituted pyrazoles can be separated and identified by TLC (Rf l,3>i y 1,5). For another publication by the same authors on the chromatographic separation of the aminopyrazoles, see (63JGU2519). A-Unsubstituted pyrazoles move with difficulty and it is necessary to add acetone or methanol to the eluent mixture. Other convenient conditions for AH pyrazoles utilize silica gel and ethyl acetate saturated with water (a pentacyanoamine ferroate ammonium disodium salt solution can be used to visualize the pyrazoles). 

There is no other facet where thin-layer chromatography reveals its paper-chromatographic ancestry more clearly than in the question of development chambers (Fig. 56). Scaled-down paper-chromatographic chambers are still used for development to this day. From the beginning these possessed a vapor space, to allow an equilibration of the whole system for partition-chromatographic separations. The organic mobile phase was placed in the upper trough after the internal space of the chamber and, hence, the paper had been saturated, via the vapor phase, with the hydrophilic lower phase on the base of the chamber. 

The development and adaptation of procedures for the separation, isolation, purification, identification, and analysis of the components of the pyrethrum mixture have been studied and evaluated. Results of studies to determine the molar extinction coefficient of pyrethrin I as well as a gas chromatographic procedure for the determination of pyrethrins are reported. The use of chromatographic separation procedures (including partition, adsorption, gas, and thin-layer chromatography), colorimetry, and infrared spectrophotometry are discussed. 

Besides the calculation of the different sulfonated species, it is also possible to determine them directly by chromatographic methods. Separation of the ester sulfonate and the disodium salt is achieved by thin-layer chromatography on silica gel plates. With a solvent mixture of acetone and tetrahydrofuran (90 10 v/v) the disodium salt stays at the start whereas the ester sulfonate has an R value of 0.2. With the more polar solvent 0.1 N H2S04 + methanol + chloroform the ester sulfonate and the disalt have Rf values of 0.36 and 0.14. For visualization, the plate is sprayed with pinacryptol yellow. In UV light (254 and... 

Reactions can be exploited more speciHcally if it is known that particular functional groups are present [cf. Chapter 2]. They still do not allow direct identification, but they increase the specificity of the evidence. The chromatographic separation carried out before detection also contributes to this. This reduces the number of potential components. However, this does not exclude the possibility that there might be several substances in the particular part of the chromatogram involved. This not only applies to thin-layer chromatography but also applies with equal force to other microanalytical separation methods (GC, HPLC). 

Thin-layer chromatography usually involves the adsorption chromatographic separation of substance mixtures into polarity groups. It is well known that clean looking chromatographic peaks can hide several substances. For instance, primary, secondary and tertiary alcohols are to be found at very nearly the same hRf. 

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