Summary.Transfer of four rapid thin-layer chromatography (TLC) screening methods used to detect substandard and counterfeit pharmaceutical products to quantitative high-performance TLC (HPTLC)-densitometry methods is demonstrated. These methods for acetaminophen, acetylsalicylic acid, ibuprofen, and chlorpheniramine maleate are contained in a Compendium of methods developed by Kenyon and Layloff for use in countries with limited resources. The new quantitative methods use Merck HPTLC silica gel 60 F 254 glass plates, automated standard and sample application, and automated densitometry for detection, identification, and quantification. Standard and sample solution preparation and application procedures for obtaining calibration curves and bracketed samples are described. The HPTLC plates give better efficiency, selectivity, and resolution than TLC, and the new methods overcome the deficiencies in technology related to manual application and visual zone comparison that do not allow the Compendium TLC procedures to support regulatory compliance actions. These transferred methods can be fully validated according to International Conference on Harmonization (ICH) guidelines or by interlaboratory studies if their applications require. The approach described can be used to transfer the remaining Compendium methods as well as the GPHF [Global (formerly German) Pharma Health Fund E.V.] Minilab kit TLC screening methods.
This review examines metabolic profiling of Schistosoma mansoni and Echinostoma caproni in their definitive and intermediate hosts. The earlier coverage of the literature on metabolic profiling was reviewed by Wang et al. 2010, Advances in Parasitology, 73, 373-404 and covered mainly studies using proton nuclear magnetic resonance spectroscopy. The methods focused upon in our review are mainly chromatographic. In the studies reviewed, various metabolites were analyzed in hosts infected with either E. caproni or S. mansoni and compared to the uninfected controls.
Seven different thin-layer chromatography stationary phases, one additional stationary layer pretreatment, eight mobile phases, two spotting techniques, and three detection reagents were evaluated for the determination of glucose in mouse fecal samples. Quantitative analysis was performed by slit-scanning densitometry. The optimal system was found to be Merck silica gel HPTLC plates with a concentrating zone developed with 1-butanol-glacial acetic acid-diethyl ether-deionized water 27:18:5:3. α-Naphthol-sulphuric acid detection reagent was found to give the best quantitative results, while the naphthoresorcinol reagent was the most useful for qualitative analysis. Semiautomatic application of samples with a CAMAG Linomat was found to give more compact bands and better separations than manual application. Using this system, quantification of glucose was achieved in mouse fecal samples. The amounts of glucose in the fecal samples of BALB/c mice infected with the intestinal trematode E. caproni were compared to control samples of uninfected mice. On the third and tenth days of postinfection, it was determined that the amount of glucose in the infected fecal samples was significantly greater than in the control samples. This indicates that metabolic profiling of glucose using TLC is possible in the mouse model and that TLC may potentially be used to test for the presence of E. caproni in humans.
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