Component Mobile Phase

Nonpolar organic mobile phases, such as hexane with ethanol or 2-propanol as typical polar modifiers, are most commonly used with these types of phases. Under these conditions, retention seems to foUow normal phase-type behavior (eg, increased mobile phase polarity produces decreased retention). The normal mobile-phase components only weakly interact with the stationary phase and are easily displaced by the chiral analytes thereby promoting enantiospecific interactions. Some of the Pirkle-types of phases have also been used, to a lesser extent, in the reversed phase mode. 

But there can be no question of chamber saturation if the TLC plate is then placed directly in the chamber. But at least there is a reduction in the evaporation of mobile phase components from the layer. Mobile phase components are simultaneously transported onto the layer (Fig. 57). In the case of multicomponent mobile phases this reduces the formation of / -fronts. 

Note The background color depends on the pH of the layer, it is, therefore, affected by the efficiency of removal of acidic mobile phase components before staining. 

Many chromatographic techniques have been named and are practiced in various regions of the fluid continuum. These regions are identified in Figures 7.3-7.8. We have not specified the mobile-phase components, and not all of these techniques are necessarily practical with the same mobile-phase component choices. However, the general view is valid. 

Figure 7.4 The Subcritical Fluid Cliromatography range. This occupies the volume in the phase diagram below the locus of critical temperatures, above and below the locus of critical pressures, and is composed mostly of the more volatile mobile-phase component. Reproduced by peimission of the American Chemical Society.

The mobile phase is always freshly made up. This is done by mk-ing the three mobile phase components in a separating funnel and shaking vigorously several times the top phase is used as mobile phase. 

If an adequate resolution is not obtained, the mobile phase components should be changed for a better selectivity, preserving the same strength. 

The adsorption mechanism in chromatography on alumina differs from that on silica gel because of the structural differences between these adsorbents. Relationships between the values of solutes and the adsorption data for the mobile phase components on sihca gel G and alumina G have been investigated by Rozylo [64,65]. The theoretical and experimental results obtained by the relation 2 = /( 1) show a good agreement for the two adsorbents. 

Snyder and Soczewinski created and published, at the same time, another model called the S-S model describing the adsorption chromatographic process [19,61]. This model takes into account the role of the mobile phase in the chromatographic separation of the mixture. It assumes that in the chromatographic system the whole surface of the adsorbent is covered by a monolayer of adsorbed molecules of the mobile phase and of the solute and that the molecules of the mobile phase components occupy sites of identical size. It is supposed that under chromatographic process conditions the solute concentrations are very low, and the adsorption layer consists mainly of molecules of the mobile phase solvents. According to the S-S model, intermolecular interactions are reduced in the mobile phase but only for the... 

A detailed description of sources used in atmospheric pressure ionization by electrospray or chemical ionization has been compiled.2 Atmospheric pressure has been used in a wide array of applications with electron impact, chemical ionization, pressure spray ionization (ionization when the electrode is below the threshold for corona discharge), electrospray ionization, and sonic spray ionization.3 Interferences potentially include overlap of ions of about the same mass-charge ratio, mobile-phase components, formation of adducts such as alkali metal ions, and suppression of ionization by substances more easily ionized than the analyte.4 A number of applications of mass spectroscopy are given in subsequent chapters. However, this section will serve as a brief synopsis, focusing on key techniques. 

Since selectivity in HPLC involves both the stationary and mobile phases [5-9,58-60], it is important to note that the solvent strength of the mobile phase, as compared to the stationary phase, (composed of mobile-phase components reversibly retained by the bonded phase and silica support) determines the elution order or k of the retained components. Unfortunately, the columns with the same stationary phase can exhibit significant variabilities from one manufacturer to another and even from the same manufacturer [5-8]. Based on discussions heard at various scientific meetings, this situation has not changed much. Variabilities can occur in the packing process even where all other conditions are supposedly constant. These factors have to be considered prior to developing an understanding as to how separations occur in HPLC. 

Normal-phase HPLC has also found application in the analysis of pigments in marine sediments and water-column particulate matter. Sediments were extracted twice with methanol and twice with dichloromethane. The combined extracts were washed with water, concentrated under vacuum and redissolved in acetone. Nomal-phase separation was performed with gradient elution solvents A and B being hexane-N,N-disopropylethylamine (99.5 0.5, v/v) and hexane-2-propanol (60 40, v/v), respectively. Gradient conditions were 100 per cent A, in 0 min 50 per cent A, in 10 min 0 per cent A in 15 min isocratic, 20 min. Preparative RP-HPLC was carried out in an ODS column (100 X 4.6 mm i.d. particle size 3 jum). Solvent A was methanol-aqueous 0.5 N ammonium acetate (75 25, v/v), solvent B methanol-acetone (20 80, v/v). The gradient was as follows 0 min, 60 per cent A 40 per cent A over 2 min 0 per cent A over 28 min isocratic, 30 min. The same column and mobile phase components were applied for the analytical separation of solutes. The chemical structure and retention time of the major pigments are compiled in Table 2.96. 

The swabs present several problems. With swabs there are many steps that need to be validated to insure an accnrate result. The analyst must be able to quantitatively remove the analyte from the swab. The analyte is added to the swab as a solution, dried, and quantitatively extracted off for analysis. The preferable extraction solntion is the swabbing solvent. The HPLC mobile phase or a mobile phase component could be used, but would necessitate a dilution of the swabbing solution. Standard addition and recovery data of clarithromycin added to different lots of polyester fiber are present in Table 3. ... 

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