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Phase separation mobile

Cjg, C8, or Cj siloxane stationary phases may be used for reversed-phase separations. Mobile phases of high pH values should be avoided as hydrolysis of the siloxane stationary phase occurs and so the packing material degrades over time. Phosphate or acetate buffers may be used to control the pH of the mobile phase, although the concentration of these salts should be kept to a minimum so that clogging of the torch injector and sampler cone does not occur. [Pg.381]

In reverse-phase chromatography, which is the more commonly encountered form of HPLC, the stationary phase is nonpolar and the mobile phase is polar. The most common nonpolar stationary phases use an organochlorosilane for which the R group is an -octyl (Cg) or -octyldecyl (Cig) hydrocarbon chain. Most reverse-phase separations are carried out using a buffered aqueous solution as a polar mobile phase. Because the silica substrate is subject to hydrolysis in basic solutions, the pH of the mobile phase must be less than 7.5. [Pg.580]

A useful guide when using the polarity index is that a change in its value of 2 units corresponds to an approximate tenfold change in a solute s capacity factor. Thus, if k is 22 for the reverse-phase separation of a solute when using a mobile phase of water (P = 10.2), then switching to a 60 40 water-methanol mobile phase (P = 8.2) will decrease k to approximately 2.2. Note that the capacity factor decreases because we are switching from a more polar to a less polar mobile phase in a reverse-phase separation. [Pg.581]

Thus far all the separations we have considered involve a mobile phase and a stationary phase. Separation of a complex mixture of analytes occurs because each analyte has a different ability to partition between the two phases. An analyte whose distribution ratio favors the stationary phase is retained on the column for a longer time, thereby eluting with a longer retention time. Although the methods described in the preceding sections involve different types of stationary and mobile phases, all are forms of chromatography. [Pg.597]

Capillary Electrochromatography Another approach to separating neutral species is capillary electrochromatography (CEC). In this technique the capillary tubing is packed with 1.5-3-pm silica particles coated with a bonded, nonpolar stationary phase. Neutral species separate based on their ability to partition between the stationary phase and the buffer solution (which, due to electroosmotic flow, is the mobile phase). Separations are similar to the analogous HPLC separation, but without the need for high-pressure pumps, furthermore, efficiency in CEC is better than in HPLC, with shorter analysis times. [Pg.607]

Otto and Wegscheider report the following capacity factors for the reverse phase separation of 2-aminobenzoic acid on a C18 column when using 10% v/v methanol as a mobile phase. ... [Pg.617]

Another example is the purification of a P-lactam antibiotic, where process-scale reversed-phase separations began to be used around 1983 when suitable, high pressure process-scale equipment became available. A reversed-phase microparticulate (55—105 p.m particle size) C g siUca column, with a mobile phase of aqueous methanol having 0.1 Af ammonium phosphate at pH 5.3, was able to fractionate out impurities not readily removed by hquid—hquid extraction (37). Optimization of the separation resulted in recovery of product at 93% purity and 95% yield. This type of separation differs markedly from protein purification in feed concentration ( i 50 200 g/L for cefonicid vs 1 to 10 g/L for protein), molecular weight of impurities (<5000 compared to 10,000—100,000 for proteins), and throughputs ( i l-2 mg/(g stationary phasemin) compared to 0.01—0.1 mg/(gmin) for proteins). [Pg.55]

At lower frequencies, orientational polarization may occur if the glass contains permanent ionic or molecular dipoles, such as H2O or an Si—OH group, that can rotate or oscillate in the presence of an appHed electric field. Another source of orientational polarization at even lower frequencies is the oscillatory movement of mobile ions such as Na". The higher the amount of alkaH oxide in the glass, the higher the dielectric constant. When the movement of mobile charge carriers is obstmcted by a barrier, the accumulation of carriers at the interface leads to interfacial polarization. Interfacial polarization can occur in phase-separated glasses if the phases have different dielectric constants. [Pg.333]

In reeent years, tire use of elevated temperatures has been reeognised as a potential variable in method development. Witlr inereased temperature, aqueous-organie mobile phases separations ean improve, viseosity deereases and diffusion inereases so baek pressures are redueed. At higher temperatures (usually with superheated water > 100 °C under modest pressures) water alone ean be used as the mobile phase and eair provide unique separation opportunities. The absenee of an organie solvent enables the use in HPLC of alternative deteetors sueh as FID or on-line LC-NMR using deuterium oxide as the eluent. [Pg.16]

Silica gel, per se, is not so frequently used in LC as the reversed phases or the bonded phases, because silica separates substances largely by polar interactions with the silanol groups on the silica surface. In contrast, the reversed and bonded phases separate material largely by interactions with the dispersive components of the solute. As the dispersive character of substances, in general, vary more subtly than does their polar character, the reversed and bonded phases are usually preferred. In addition, silica has a significant solubility in many solvents, particularly aqueous solvents and, thus, silica columns can be less stable than those packed with bonded phases. The analytical procedure can be a little more complex and costly with silica gel columns as, in general, a wider variety of more expensive solvents are required. Reversed and bonded phases utilize blended solvents such as hexane/ethanol, methanol/water or acetonitrile/water mixtures as the mobile phase and, consequently, are considerably more economical. Nevertheless, silica gel has certain areas of application for which it is particularly useful and is very effective for separating polarizable substances such as the polynuclear aromatic hydrocarbons and substances... [Pg.93]

The interactions between solute and the pha.ses are exactly the same as those present in LC separations, namely, dispersive, polar and ionic interactions. At one extreme, the plate coating might be silica gel, which would offer predominately polar and induced polar interactions with the solute and, con.sequently, the separation order would follow that of the solute polarity. To confine the polar selectivity to the stationai y phase, the mobile phase might be -hexane which would offer only dispersive interactions to the solute. The separation of aromatic hydrocarbons by induced polar selectivity could be achieved, for example, with such a system. [Pg.443]

The observed reversal in the thermal stability of the copolymer at a critical composition, which appears to be between 30 and 40 mol% of ethylene, may be explained on the basis of the emergence of phase-separation between the nonpolar ethylene and polar vinyl chloride blocks. Although crystallization of the ethylene blocks in the copolymer is only observed when more than 70 mol% ethylene units are present, the possibility of phase-separation occurring at lower contents of ethylene units cannot be excluded. Also, round about the critical copolymer composition, the Tg of the copolymer may be reduced to a level that would facilitate separation between the unlike phases by increased molecular mobility within the polymer matrix. As has been discussed earlier, occurrence of phase-separation in the copolymer would not only make the mechanism of stabilization due... [Pg.334]

The separation was carried out on a column 25 cm long, 4.6 mm in diameter and packed with a Cl8 reversed phase. The mobile phase was programmed from a 93% acetonitrile 7% water to 99% acetonitrile 1% water over a period of 30 minutes. The gradient was linear and the flow... [Pg.182]

Reverse phase chromatography is finding increasing use in modern LC. For example, steroids (42) and fat soluble vitamins (43) are appropriately separated by this mode. Reverse phase with a chemically bonded stationary phase is popular because mobile phase conditions can be quickly found which produce reasonable retention. (In reverse phase LC the mobile phase is typically a water-organic solvent mixture.) Rapid solvent changeover also allows easy operation in gradient elution. Many examples of reverse phase separations can be found in the literature of the various instrument companies. [Pg.240]


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See also in sourсe #XX -- [ Pg.516 ]




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