Sample preparation layer

Ng (1991) has provided a review on TLC and HPTLC analysis of the more popular pharmaceuticals and illicit drugs covering the literature from about 1975 to 1988. This reference includes information on sample preparation, layers, mobile phases, detection reagents, and structures of popular human and animal drugs. Numerous examples are provided for qualitative and quantitative TLC analysis of pharmaceutical drugs in various dosage forms. The review of Ng (1991) was extended and updated by Szepesi and Nyiredy (1996) to include references through 1995. 

Touchstone (1992) has provided information on methods of sample preparation, layers, mobile phases, and detection reagents for pharmaceuticals, particularly drugs of abuse such as amphetamines, barbiturates, benzodiazepines, canna-binoids, cocaine, methaqualone, opiates, and anorexicants. 

One of trends of development of thin-layer chromatography implies that replacement of aqueous-organic eluents by micellar surfactants solution. This is reduces the toxicity, flammability, environmental contamination and cost of the mobile phases, reduce sample prepar ation in some cases. 

In the early days of TEM, sample preparation was divided into two categories, one for thin films and one for bulk materials. Thin-films, particularly metal layers, were often deposited on substrates and later removed by some sort of technique involving dissolution of the substrate. Bulk materials were cut and polished into thin slabs, which were then either electropolished (metals) or ion-milled (ceramics). The latter technique uses a focused ion beam (typically Ar+) of high-energy, which sputters the surface of the thinned slab. These techniques produce so-called plan-view thin foils. 

Static SIMS is also capable of analyzing liquids and fine particles or powders. A liquid is ofren prepared by putting down an extremely thin layer on a flat substrate, such as a silicon wafer. Particles are easily prepared by pressing them onto doublesided tape. No further sample preparation, such as gold- or carbon-coating, is required. 

Even though the mechanical profiler provides somewhat limited two dimensional information, no sample preparation is necessary, and results can be obtained in seconds. Also, no restriction is imposed by the need to measure craters through several layers of different composition or material type. 

The STM uses this eflFect to obtain a measurement of the surface by raster scanning over the sample in a manner similar to AFM while measuring the tunneling current. The probe tip is typically a few tenths of a nanometer from the sample. Individual atoms and atomic-scale surface structure can be measured in a field size that is usually less than 1 pm x 1 pm, but field sizes of 10 pm x 10 pm can also be imaged. STM can provide better resolution than AFM. Conductive samples are required, but insulators can be analyzed if coated with a conductive layer. No other sample preparation is required. 

These reactions can be carried out during sample preparation or directly on the layer at the start after application of the sample. Reactions have also been described in the capillaries employed for application. 

HPTLC plates Sihca gel 60 (Merck). Before apphcation of the samples the layer was prewashed once with the mobile phase and dried at 110°C for 20 min. Before it was placed in the developing chamber the prepared HPTLC plate was preconditioned for 30 min at 0% relative humidity (over cone, sulfuric acid). 

Thus, for the investigation of buried polymer interfaces, several techniques with molecular resolution are also available. Recently NMR spin diffusion experiments [92] have also been applied to the analysis of a transition zone in polymer blends or crystals and even the diffusion and mobility of chains within this layer may be analyzed. There are still several other techniques used, such as radioactive tracer detection, forced Rayleigh scattering or fluorescence quenching, which also yield valuable information on specific aspects of buried interfaces. They all depend very critically on sample preparation and quality, and we will discuss this important aspect in the next section. 

Experiments at present are concentrated on sd-metals and Pt-group metals. The sp-metals, on which theories of the double layer have been based, are somewhat disregarded. In some cases the most recent results date back more than 10 years. It would be welcome if double-layer studies could be repeated for some sp-metals, with samples prepared using actual surface procedures. For instance, in the case of Pb, the existing data manifest a discrepancy between the crystalline system and the crystal face sequence of other cases (e.g., Sn and Zn) the determination of EgaQ is still doubtful. For most of sp-metals, there are no recent data on the electron work function. 

State-of-the-art TOF-SIMS instruments feature surface sensitivities well below one ppm of a mono layer, mass resolutions well above 10,000, mass accuracies in the ppm range, and lateral and depth resolutions below 100 nm and 1 nm, respectively. They can be applied to a wide variety of materials, all kinds of sample geometries, and to both conductors and insulators without requiring any sample preparation or pretreatment. TOF-SIMS combines high lateral and depth resolution with the extreme sensitivity and variety of information supplied by mass spectrometry (all elements, isotopes, molecules). This combination makes TOF-SIMS a unique technique for surface and thin film analysis, supplying information which is inaccessible by any other surface analytical technique, for example EDX, AES, or XPS. 

In this case, three particles are shown, a 40 u, 20 p and a 10 p particle. The most important step is sample preparation on the microscope slide, since only a pinch of materied is used, one must be sure that the sample is uniform and representative of the material. Also, since most materials tend to agglomerate due to accumulated surface charge in a dry state, one adds a few drops of alcohol and works it with a spatula, spreading it out into a thin layer which dries. Too much working breaks down the original peirticles. 

In a study of dental silicate cements, Kent, Fletcher Wilson (1970) used electron probe analysis to study the fully set material. Their method of sample preparation varied slightly from the general one described above, in that they embedded their set cement in epoxy resin, polished the surface to flatness, and then coated it with a 2-nm carbon layer to provide electrical conductivity. They analysed the various areas of the cement for calcium, silicon, aluminium and phosphorus, and found that the cement comprised a matrix containing phosphorus, aluminium and calcium, but not silicon. The aluminosilicate glass was assumed to develop into a gel which was relatively depleted in calcium. 

Manufacturers of TLC materials and accessories are well prepared to satisfy the needs for professionally performed PLC. High-quality precoated preparative plates are available from a number of eommercial sources. Alternatively, less expensive or specialty preparative plates ean be homemade in the laboratory, and loose sorbents and coating devices ean be purehased for this purpose. More-or-less-automated devices can also be purehased for band application of higher quantities of sample solutions to preparative layers. At least for some users, sophisticated densitometric and other instrumental techniques are available as nondestructive tools for preliminary detention and identification of separated compounds in order to enhance the effieiency of their isolation. The only aid still missing, and maybe the most important of all, is a comprehensive monograph on PLC that might encourage and instruct many potential users on how to fully benefit from this very versatile, efficient, relatively inexpensive, and rather easy to use isolation and purification technique. This book was planned to fill that void. 

Chapter 4 discusses the selection and optimization of mobile phases for successful separations in PLC. Chapter 5 details procedures for sample application and development of layers, and Chapter 6 complements Chapter 5 by dealing specifically with the use of horizontal chambers for the development of preparative layers, including linear, continuous, two-dimensional, gradient, circular, and anticircular modes. 

An alternative to spray-on in the form of a rectangular area, the solid phase sample apphcation (SPSA) is suited for apphcation of especially large, nonvolatile sample volumes on preparative layers [2]. Therefore, the sample is dissolved in a suitable... 

Sample recovery tubes are available commercially for the removal and recovery of adsorbent from a preparative layer. An example is shown in Figure 8.2. A similar sample recovery tube is available from Bodman (Aston, PA Catalog No. GSRT-2). With these tubes, the scrapings are direcdy recovered from the plate using suction, and elution is accomphshed with an appropriate solvent under vacuum while the adsorbent is retained on the disc. 

---