Collector chromatography

Column chromatography apparatus, including a suitable low-pressure pump, on-line UV monitoring at 280 nm and a fraction collector. Chromatography media Sephacryl S-200 (SF), Sephadex G-25 (SF) (Pharmacia). 

The conditions for gas chromatography are given in Note 9. The product was found mainly in fractions 2-20 by the submitters. The first 25-ml. fraction contained considerable amounts of byproducts, while fractions 21 and higher contained 1-adamantanol. The checkers collected 10-ml. fractions with an automatic fraction collector. 

Figure 4-1. Components of a simple liquid chromatography apparatus. R Reservoir of mobile phase liquid, delivered either by gravity or using a pump. C Glass or plastic column containing stationary phase. F Fraction collector for collecting portions, called fractions, of the eluant liquid in separate test tubes.

Figure 2, Block diagram of a liquid chromatograph. A, solvent reservoir B, filter C, pump D, pulse dampener (optional) E, pre-column (used only in liquid-liquid chromatography) F, pressure gauge G, infector H, column I, detector J, fraction collector K, recorder or oth readout device.

FIGURE 10.5 Elution profile on OH-B12 treated by microwave heating for 6 min during silica gel 60 column chromatography. Fifty milliliters of the treated OH-B12 solution (5 mmol/1) was evaporated to dryness and dissolved in a small amount of w-butanol/2-pro-panol/water (10 7 10, v/v) as a solvent. The concentrated solution was put on a column (1.4 X 15.0 cm) of silica gel 60 equilibrated with the same solvent and eluted with the same solvent in the dark. The eluate was collected at 4.0 ml with a fraction collector. Fractions I to V were pooled, evaporated to dryness, dissolved with a small amount of distilled water, and analyzed with silica gel TLC. Inset represents the mobile pattern of the OH-B12 degradation products of fractions I to V on the TLC plate. Data are typical, taken from one of five experiments. (Reprinted with permission from Watanabe, F. et al., J. Agric. Food Chem., 46, 5177-5180, 1998. Copyright (1998) American Chemical Society.)... 

In many cases, data can be obtained offline and analyzed in a comprehensive manner from a fraction collector. For example, LC and gas chromatography (GC) can be utilized to form a 2D experiment — LC/GC. This is quite useful as the retention... 

Collect fractions starting from the bottom of the gradient, while recording an absorbance profile at 254 nm (which is dominated by the very abundant ribosomal RNA). We use a setup consisting of a peristaltic pump, ultraviolet (UV) detector, and fraction collector commonly used for chromatography experiments (GE Healthcare Life Sciences). 

A. Rottenbach, T. Uhl, A. Hain, A. Scharf, K. Kritzler and W. Kretschmer, Development of a fraction collector for coupling gas chromatography with an AMS facility, Nucl. Instrum. Methods B 266, 2238 2241 (2008). 

In the analysis of clinical, biological and environmental samples it is often important to have information on the speciation of the analyte, e.g. metal atoms. Thus an initial sample solution may be subjected to a separation stage using chromatography or electrophoresis. Measurements may, of course, be made on fractions from a fraction collector, but with plasma sources, interfacing in order to provide a continuous monitoring of the column effluent can be possible. This relies upon the ability of the high-temperature plasma to break down the matrix and produce free ions. 

Nonsolvent bath, polymer precipitation by immersion in, 15 808-811 Nonspecific elution, in affinity chromatography, 6 398, 399 Nonstationary Poisson process, in reliability modeling, 26 989 Non-steady-state conduction, 9 105 Nonsteroidal antiinflamatory agents/drugs (NSAIDs) 21 231 for Alzheimer s disease, 2 820 for cancer prevention, 2 826 Nonsulfide collectors, 16 649 Nonsulfide flotation, 16 649-650 Nonsulfide mineral flotation collectors used in, 16 648-649t modifiers used in, 16 650, 651t Nonsulfide ores, 16 598, 624... 

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

Chromatography and Electrophoresis of Antigens. Cotton dust (20-30 mg) was dissolved in water and passed through a small column (30 x 1.5 cm id) packed with 3 g of Sephadex G-75 that had been preswelled in water. Chromatograms were obtained with an LKB Uvicord ultraviolet monitor and a chopper bar recorder. Volumes of 5 ml each were collected in test tubes with an automatic fraction collector. Two components were detected at 250 nm. Samples of 10 mg were analyzed by disc gel electrophoresis according to procedures described by Davis (36), and Zacharius (37). 

Fig. A.S Flow Chart for column Chromatography. The Central part Is the column from the sample and/or solvent is loaded at a controlled flow rate with a pump. The eluates from the column are collected in tubes of a fraction collector.

Since the development of HPLC as a separation technique, considerable effort has been spent on the design and improvement of suitable detectors. The detector is perhaps the second-most important component of an HPLC system, after the column that performs the actual separation it would be pointless to perform any separation without some means of identifying the separated components. To this end, a number of analytical techniques have been employed to examine either samples taken from a fraction collector or the column effluent itself. Although many different physical principles have been examined for their potential as chromatography detectors, only four main types of detectors have obtained almost universal application, namely, ultraviolet (UV) absorbance, refractive index (RI), fluorescence, and conductivity detectors. Today, these detectors are used in about 80% of all separations. Newer varieties of detector such as the laser-induced fluorescence (LIE), electrochemical (EC), evaporative light scattering (ELS), and mass spectrometer (MS) detectors have been developed to meet the demands set by either specialized analyses or by miniaturization. 

To avoid re-mixing of separated components, make the distance between column outlet, detector, and fraction collector as short as possible. For normal pressure chromatography use polypropylene or Teflon tubing with an inner diameter of about 1 mm. 

Sephadex G-25-150 slurry in buffer Chromatography columns 1.5 X 15 cm Fraction collector... 

Experimental studies to construct a binding curve will be described at pH 4.5. To prepare the gel filtration column, obtain a chromatography column (about 1.5 X 15 cm). It should be equipped with a porous glass disk and a stopcock at the bottom to control flow rate. Clamp the column to a ring stand and connect the bottom tubing to a fraction collector containing 50 test tubes. If... 

Column chromatography apparatus, including a suitable low-pressure pump, online UV monitoring at 280 nm and a fraction collector. 

We shall not endeavor to describe in this chapter the various types of fraction collectors that have been used in column chromatography. For urine analysis 1-ml fractions are necessary ( °/o error). In Moore and Stein s 1958 procedure (see above (c) Amberlite IR 120-X8 columns), 2-ml fractions can also be collected. 

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