Thin-layer chromatography specificity

Analytical methods iaclude thin-layer chromatography (69), gas chromatography (70), and specific methods for determining amine oxides ia detergeats (71) and foods (72). Nuclear magnetic resonance (73—75) and mass spectrometry (76) have also been used. A frequentiy used procedure for iadustrial amine oxides (77) iavolves titratioa with hydrochloric acid before and after conversion of the amine to the quaternary ammonium salt by reaction with methyl iodide. A simple, rapid quaHty control procedure has been developed for the deterrniaation of amine oxide and unreacted tertiary amine (78). 

Thin-Layer Chromatography (tic). Tic (126) is used widely for quahtative analysis and micro-quantity separation of amino acid mixtures. The amino acids detected are developed by ninhydrin coloring, except for proline and hydroxyproline. Isatia has been recommended for specific coloring of pToline (127). 

Commercially available pre-coated plates with a variety of adsorbents are generally very good for quantitative work because they are of a standard quality. Plates of a standardised silica gel 60 (as medium porosity silica gel with a mean porosity of 6mm) released by Merck have a specific surface of 500 m /g and a specific pore volume of 0.75 mL/g. They are so efficient that they have been called high performance thin layer chromatography (HPTLC) plates (Ropphahn and Halpap J Chromatogr 112 81 1975). In another variant of thin layer chromatography the... 

The Determination of 6 Specific Polynuclear Aromatic Hydrocarbons in Waters [Using High-Performance Liquid Chromatography,Thin-layer Chromatography], 1985... 

The lipophilicity (7 m value) and specific hydrophobic surface area of 1 ]/f-pyrido[2,]-fi]quinazolin-] 1-one and its isomeric 6//-pyrido[l, 2-u]qui-nazolin-6-one were determined by reversed-phase thin-layer chromatography (98MI4). 

Recent developments in the practice of thin-layer chromatography have resulted in a breakthrough in performance which has led to the expression high performance thin-layer chromatography . These developments have not been the result of any specific advance in instrumentation (as with HPLC), but rather the culmination of improvements in the various operations involved in TLC. The three chief features of HPTLC are summarised below, but for a comprehensive account of the subject the reader is recommended to consult a more specialised text.59... 

The dinitrophenylhydrazones were separated from the reaction mixture by thin-layer chromatography (silica gel G developed with benzene) and further purified by thin-layer chromatography on aluminum oxide G (petroleum ether-diethyl ether (96 to 4), silica gel G (chloroform), and silica gel G (diethyl ether)). In all cases, the specific activities of the dinitrophenylhydrazones remained constant over the course of the last two purifications. 

This volume is the second of a series of practice-orientated TLC/HPTLC books published in excellent quality by VCH Publishers. As in the first volume, a series of reagents and detection methods have been reviewed with the intention of helping the practical analyst increase the detection specificity of routine samples separated by thin-layer chromatography. 

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). 

It is very often advantageous in thin-layer chromatography to be able to obtain a preliminary impression of a substance separation by first exposing the plate to a rapidly carried out, economically priced universal reaction before passing on to final characterization using group-specific or even better substance-specific reactions. 

Mendoza CE, Shields JB. 1971. Esterase specificity and sensitivity to organophosphorus and carbamate pesticides Factors affecting determination by thin layer chromatography. J Assoc Off Anal Chem 54 507-512. 

Patil VB, Sevaikar MT, Padalikar SV. 1987. Specific spray reagent for the detection of endosulfan by thin-layer chromatography. J Chromatogr 396 441-443. 

In conclusion, synthetic dyes can be determined in solid foods and in nonalcoholic beverages and from their concentrated formulas by spectrometric methods or by several separation techniques such as TEC, HPLC, HPLC coupled with diode array or UV-Vis spectrometry, MECK, MEECK, voltammetry, and CE. ° Many analytical approaches have been used for simultaneous determinations of synthetic food additives thin layer chromatography, " " derivative spectrophotometry, adsorptive voltammetry, differential pulse polarography, and flow-through sensors for the specific determination of Sunset Yellow and its Sudan 1 subsidiary in food, " but they are generally suitable only for analyzing few-component mixtures. 

Table 1 provides the urinalysis test results for the 4,847 arrestees, While PCP was tested for by an EMIT test only, cocaine, opiates and methadone were tested for by both EMIT and thin layer chromatography (TLC). (The EMIT test for opiates is not specific to morphine, the metabolite of heroin, and can detect the recent use of a variety of opiates. A specimen positive for opiates is most likely to indicate the use of heroin in this population, however.) Our analyses will use only the results from the EMIT tests, because we have learned that the TLC general drug screen is less sensitive for detecting recent use of these illicit street drugs (Wish et al. 1983 Wish et al. 1984). 

Some studies have evaluated the quantity of a specific constituent in various herbal products by a thin-layer chromatography spectrophotometric method. Of 44 feverfew products that were evaluated, 14 (32%) did not contain the minimum of 0.2% parthenolide content (active ingredient) and 10 (22%) did not contain any detectable levels of parthenolide [30]. 

Chemicals. Purified, P Cj-labelled alachlor (specific activity = 17 mCi/mM), butylate (specific activity = 2.54 mCi/mM) and metolachlor (specific activity = 4.5 mCi/mM) were used in the leaching, adsorption, and diffusion studies. The radiopurity of these compounds was greater than 95% as determined by thin-layer chromatography. All other studies were conducted using analytical grade, non-radioactive material (purity 5 95%). 

The theoretical work that exploited the advantages of the multidimensional separation format appears to have been developed much later than the original experimental work. One of the earliest studies was conducted by Connors (1974), who assumed that the distribution of spots on a two-dimensional thin-layer chromatography (2DTLC) plate could be modeled using a Poisson distribution of data on each retention axis. He then constructed equations that related the number of chromatographic systems needed to resolve a specific number of compounds. One... 

Lipophilicity and specific hydrophobic surface area were determined by using reversed-phase thin-layer chromatography for fused heterocyclic ring systems including five pyrido[2,iy][l,2,4]triazine derivatives <1998MI64>. 

Another cultured cell line of Catharanthus roseus (EU4A), which does not produce detectable amounts of vinblastine and other bisindole alkaloids, was also examined for its ability to transform 78 (183). Cell-free extracts of the culture line were prepared, and the 35,000 X g supernatant solution was used. Incubations with 2r-tritioanhydiovinblastine yielded a mixture from which radioactive vinblastine (52) was isolated. The labeled vinblastine was reisolated after unlabeled carrier was added and rigorously purified by successive thin-layer chromatography, reversed-phase HPLC, and crystallization to constant specific activity. Boiled extracts could not produce labeled 52, thus supporting the involvement of enzymes in the conversion process. 

The technique of thin-layer chromatography (TLC) has been used extensively in the domain of pharmaceutical analysis for a variety of specific and useful applications, for example ... 

If the compound shows no convenient intrinsic feature, it may be possible to develop a detectable characteristic by chemical modification and, particularly in thin-layer chromatography and electrophoresis, many locating reagents which are specific for various groups of compounds are available. 

Aromatic amines that have been used include o-toluidine, p-aminosali-cylic acid, p-aminobenzoic acid, diphenylamine and p-aminophenol. Their ability to react preferentially with a particular carbohydrate or class of carbohydrate is often useful, e.g. p-aminophenol, which shows some specificity for ketoses compared with aldoses and is useful for measuring fructose. These reagents have proved particularly useful for the visualization and identification of carbohydrates after separation of mixtures by paper or thin-layer chromatography, when colour variations and the presence or absence of a reaction aid the interpretation of the chromatogram. 

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