By Joseph Sherma, Bernard Fried
Written via over forty across the world acclaimed specialists on thin-layer chromatography (TLC), this accomplished moment version provides the newest strategies, instrumentation, and functions of overpressurized, rotational, and high-performance quantitative TLC.
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Extra info for Handbook of Thin-Layer Chromatography
The plate is placed layer down over a tray with various compartments, which can hold different solvents, humidity-control liquids, and volatile acids and bases whose vapors will impregnate and condition or preload the layer. Developing solvent is in a separate tray and is transferred to the layer by a wick. The Vario chamber can be used to test six mobile phases side-by-side on one plate for solvent optimization, to determine if layer preequilibration (preloading) is advantageous, to ascertain if S- or N-chamber configuration is best, and to test different humidity conditions.
New detection reagents are updated biennially in the Analytical Chemistry review of TLC written by Sherma (25). A book on physical and chemical detection methods in TLC was published (157). Compounds that are naturally colored are detected on layers directly, while compounds with native fluorescence are viewed under UV light (Fig. 8). In some cases, fluorescence is visible on a wetted plate only, so the layer is impregnated with a nonvolatile liquid. , quinine at low pH. Compounds that absorb UV light can be detected on layers containing an indicator (phosphor) that fluoresces upon excitation with 254 nm or 366 nm UV light.
When laned or channeled plates are used, the length of the band is confined within the channel. Sample application is fast and simple for relatively large volumes (up to ca. 50 ul for TLC and 25 |il for HPTLC). High efficiency can be obtained for HPTLC by spotting larger volumes of dilute solutions rather than nanoliter volumes of highly concentrated solutions. Crude samples can be directly spotted, and salts and other impurities may be retained in the preadsorbent and not interfere with sample resolution or detection.