Chromatographic techniques fractionation

Chemically cross-linked dextrans Water-insoluble chromatographic techniques (fractionation and purification... 

Linking TLC with a tandem instrument differs from combining GC or LC with an appropriate spectrometer. Hyphenation of planar chromatographic techniques represents a niche application compared to HPLC-based methods. Due to the nature of the development process in TLC, the combination is often considered as an off-line in situ procedure rather than a truly hyphenated system. True in-line TLC tandem systems are not actually possible, as the TLC separation must be developed before the spots can be monitored. It follows that all TLC tandem instruments operate as either fraction collectors or off-line monitoring devices. Various elaborate plate extraction procedures have been developed. In all cases, TLC serves as a cleanup method. 

Applications If an extract needs further cleanup, it is possible to couple it with multidimensional chromatographic techniques such as LC-LC or LC-GC. The first chromatographic step can then be used for the on-line cleanup and concentration of the extract, and the second one for the final separation. Large-volume, on-column injection (LVI-COC) is particularly useful for coupled LC-GC in which 100-350 xL fractions of eluent from the NPLC cleanup separation step are transferred on-line to the GC column. For example, on-line removal of high-MW interfering material, such as polymers from a polymer/additive dissolution, can be achieved easily by using SEC before the fraction containing additives is transferred to the GC. 

Purification. The commercial LDPE was dissolved in hot toluene and precipitated in methanol. The resulting small chips were dried in a vacuum oven overnight at 80° C. HDPE was purified in the same manner, except that in this case much of the commercial material was insoluble in toluene, and so significant fractionation apparently occurs during t.he purification. The model compounds were purified by the flash chromatographic technique of Still et al. (10). 

Figure 2.2. Fractionation of protein extracts before 2D gel electrophoresis. Crude lysates can be fractionated by affinity purification or by a number of chromatographic techniques. In addition, organelles or other cellular structures can be purified and lysates from these organelles can be fractionated or separated directly on 2D gels. By repeating this procedure using a number of conditions it may be possible to visualize a large fraction of a cell s proteome.

In addition to GC/MS, high performance liquid chromatography (HPLC/MS) has been used to analyse natural resins in ancient samples, particularly for paint varnishes containing mastic and dammar resins [34]. A partial limitation of chromatographic techniques is that they do not permit the analysis of the polymeric fraction or insoluble fraction that may be present in the native resins or formed in the course of ageing. Techniques based on the direct introduction of the sample in the mass spectrometer such as direct temperature resolved mass spectrometry (DTMS), direct exposure mass spectrometry (DE-MS) and direct inlet mass spectrometry (DI-MS), and on analytical pyrolysis (Py-GC/MS), have been employed as complementary techniques to obtain preliminary information on the... 

Removal of viruses from the product stream can be achieved in a number of ways. The physicochemical properties of viral particles differ greatly from most proteins, ensuring that effective fractionation is automatically achieved by most chromatographic techniques. Gel-filtration chromatography, for example, effectively separates viral particles from most proteins on the basis of differences in size. 

A novel pigment has been isolated from the petals of Rosa hybrida with complex chromatographic techniques and the structure was elucidated with spectroscopic methods and high resolution fast-atom bombardement mass spectrometry, lH NMR, and FTIR. Anthocyanins were extracted from 7.9 kg of petals of Rosa hybrida cv. M me Violet with 80 per cent aqueous ACN containing 0.1 per cent TFA. The extract was purified in a Sephadex LH-20 column, and the fraction eluted with 80 per cent ACN was further fractionated in a HP-20 column using water, 15, 20 and 30 per cent ACN as mobile phases. The last fraction was lyophilized and separated by preparative RP-HPLC using an ODS column (50 X 5 cm i.d.). Solvents were 0.5 per cent aqueous TFA (A) and water-methanol... 

The following chromatographic techniques are representative of those used by a number of investigators for the further enrichment and fractionation of analytes. The concentrated dialysate is subjected to SEC to remove co-dialyzed lipid... 

Thus, Freudenberg concluded that the cyclodextrins are not preformed in starch, but that their formation is made possible by the helicity of the starch chain. Freudenberg s hypotheses concerning the starch structure (that is, the amylose fraction) and the Bacillus macerans amylase mechanism have been confirmed by X-ray crystallography and chromatographic techniques. ... 

The detection, identification and estimation of impurities by various chromatographic techniques is so well documented that few comments are required here. It must be remembered, however, that since we are concerned with extremely low concentrations, one cannot be sure of finding an impurity whose retention time on a column is close to that of the main compound, and also that a very small fraction of the main compound may undergo transformations on the column, especially if it is at an elevated temperature, so that spurious impurities may be produced in this way. If the main compound is sensitive to any of the components of air, especial precautions must be taken in transferring the sample from its evacuated container to the inlet of the chromatograph. 

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