Chromatography open-column

Open-column chromatography on alumina or silica has been extensively used in surfactant analysis. It is inexpensive but tedious and is now being superseded by HPLC. 

Hydrocarbons, secondary alcohols, primary alcohols, waxes and fatty esters, lanolin, ethoxylates, alkanolamides, anionics. 

Separations on silica are mostly in the nonionics field and include the following general separation [17]  

Hydrocarbons + fatty acid methyl esters, glycerides 4- fatty alcohols + fatty acids, fatty acid ethoxylates + alkylphenol ethoxylates, other ethoxylates, glycerol + high-molecular-weight polyethylene glycols. 

Separations usually take a long time, use large volumes of solvents and often do not give clean separations. A description of the practical procedure is given in chapter 3. Some specific uses are described under the appropriate surfactants. 

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Spectrophotometric deterrnination at 550 nm is relatively insensitive and is useful for the deterrnination of vitamin B 2 in high potency products such as premixes. Thin-layer chromatography and open-column chromatography have been appHed to both the direct assay of cobalamins and to the fractionation and removal of interfering substances from sample extracts prior to microbiological or radioassay. Atomic absorption spectrophotometry of cobalt has been proposed for the deterrnination of vitamin B 2 in dry feeds. Chemical methods based on the estimation of cyanide or the presence of 5,6-dimethylben2irnida2ole in the vitamin B 2 molecule have not been widely used. 

High performance liquid chromatography (HPLC) has been by far the most important method for separating chlorophylls. Open column chromatography and thin layer chromatography are still used for clean-up procedures to isolate and separate carotenoids and other lipids from chlorophylls and for preparative applications, but both are losing importance for analytical purposes due to their low resolution and have been replaced by more effective techniques like solid phase, supercritical fluid extraction and counter current chromatography. The whole analysis should be as brief as possible, since each additional step is a potential source of epimers and allomers. 

Although saponification was found to be unnecessary for the separation and quantification of carotenoids from leafy vegetables by high performance liquid chromatography (HPLC) or open column chromatography (OCC), saponification is usually employed to clean the extract when subsequent purification steps are required such as for nuclear magnetic resonance (NMR) spectroscopy and production of standards from natural sources. 

In order to reduce the time-consuming open-column chromatographic processes, conventional methods of hydrocarbon-group-type separation have been replaced by MPLC and HPLC. Flash column chromatography is a technique less commonly applied than open-column version, but several applications have been described [2,24—27]. The common technique version is to use a silica-gel-filled column for example, 230 to 400 mesh 20 X 1 cm column size with a back pressure of 1.5 X 10 Pa of an ambient gas such as nitrogen. Solvents are similar to the ones apphed in the case of open-column chromatography fractionations. 

Although SPE can be done in a batch equilibration similar to that used in LLE, it is much more common to use a small tube (minicolumn) or cartridge packed with the solid particles. SPE is often referred to as LSE, bonded phase or sorbent extraction SPE is a refinement of open-column chromatography. The mechanisms of retention include reversed phase, normal phase, and ion exchange. 

Applications Open-column chromatography was used for polymer/additive analysis mainly in the 1950-1970 period (cf. Vimalasiri et al. [160]). Examples are the application of CC to styrene-butadiene copoly-mer/(additives, low-MW compounds) [530] and rubbers accelerators, antioxidants) [531]. Column chromatography of nine plasticisers in PVC with various elution solvents has been reported [44], as well as the separation of CHCI3 solvent extracts of PE/(BHT, Santonox R) on an alumina column [532]. Similarly, Santonox R and Ionol CP were easily separated using benzene and Topanol CA and dilaurylthiodipropionate using cyclohexane ethyl acetate (9 1 v/v) [533]. CC on neutral alumina has been used for the separation of antioxidants, accelerators and plasticisers in rubber extracts [534]. Column chromatography of polymer additives has been reviewed [160,375,376]. 

FIGURE 11.14 The classic open-column chromatography configuration with fraction collector. 

The urine of people who are heavy smokers contains mutagenic chemicals, chemicals that cause mutations in biological cells. Bioanalytical laboratories can analyze urine samples for these chemicals, but the samples must be cleaned up first prior to extraction with methylene chloride. The procedure for this cleanup utilizes an open column chromatography. Columns several inches tall and about an inch wide are prepared by packing them with an adsorbing resin that has been treated with methyl alcohol. The urine samples are passed through these columns as part of the sample preparation scheme. 

Compare high-performance liquid chromatography with open-column chromatography in terms of... 

Partition chromatography Ion exchange chromatography Paper chromatography Open-column chromatography High-performance liquid chromatography... 

This is invariably referred to in various literature as open-column chromatography drop chromatography strip-chromatography spread-layer chromatography surface chromatography . 

Cleanup or fractionation procedures that have been used in the more recent fat-soluble vitamin assays include sterol precipitation, open-column chromatography, solid-phase extraction, and high-pressure gel permeation chromatography. High-performance LC has been used on a semipreparative scale in vitamin D and vitamin K assays to obtain purified fractions of sample extracts. This technique is discussed in Sec. V.B.3. 

The more recent applications of open-column chromatography in fat-soluble vitamin assays utilize liquid-solid (adsorption) chromatography using gravity-flow glass columns dry-packed with magnesia, alumina, or silica gel. Such columns enable separations directly comparable with those obtained by thin-layer chromatography to be carried out rapidly on a preparative scale. 

Three recent reviews specifically cover HPLC methods for quantitating riboflavin in foods. In addition to HPLC methods, Nielsen (81) summarized paper chromatography, TLC, and open-column chromatography procedures for quantitating total riboflavin and the individual vitamers in foods, pharmaceuticals, and biological samples. Russell (44) included a brief discussion of the standard methods, along with HPLC and flow injection analyses published between 1990 and 1994 for total riboflavin and the individual vitamers in foods. Ball (45) reviewed HPLC methods for quantitation of riboflavin, as well as chemical and microbiological riboflavin assays for foods. 

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