Electrokinetic instruments

The presence of surface conductance behind the slip plane alters the relationships between the various electrokinetic phenomena [83, 84] further complications arise in solvent mixtures [85]. Surface conductance can have a profound effect on the streaming current and electrophoretic mobility of polymer latices [86, 87]. In order to obtain an accurate interpretation of the electrostatic properties of a suspension, one must perform more than one type of electrokinetic experiment. One novel approach is to measure electrophoretic mobility and dielectric spectroscopy in a single instrument [88]. 

Recently a decreased level of CE activity has been noticed with a shift of attention towards other separation techniques such as electrochromatography. CE is apparently not more frequently used partly because of early instrumental problems associated with lower sensitivity, sample injection, and lack of precision and reliability compared with HPLC. CE has slumped in many application areas with relatively few accepted routine methods and few manufacturers in the market place. While the slow acceptance of electrokinetic separations in polymer analysis has been attributed to conservatism [905], it is more likely that as yet no unique information has been generated in this area or eventually only the same information has been gathered in a more efficient manner than by conventional means. The applications of CE have recently been reviewed [949,950] metal ion determination by CE was specifically addressed by Pacakova et al. [951]. 

Suspensions are generally evaluated with respect to their particle size, electrokinetic properties (zeta potential), and rheological characteristics. A detailed discussion on the methods/techniques and relevant instrumentation is given in Sec. VII. A number of evaluating methods done specifically with suspension dosage forms, such as sedimentation volume, redispersibility, and specific gravity measurements, will be treated in this section. 

PANFILI G, MANZI P, COMPAGNONE D, SCARCIGLIA L and PALLESCHI G (2000), Rapid assay of chohne in foods using microwave hydrolysis and a choline biosensor , J Agric Food Chem, 48, 3403-7. pant I and trennery v c (1995), The determination of sorbic acid and benzoic acid in a variety of beverages and foods by micellar electrokinetic capillary chromatography , Food Chem, 53(2), 219-26. pare j r j and Belanger j m r (1997), Instrumental Methods in Food Analysis. Series Techniques and instrumentation in analytical chemistry - Vol. 18, Amsterdam, Elsevier. 

One of the major advantages of CE as a separation technique is the wide variety of separation modes available. Analytes can be separated on the basis of charge, molecular size or shape, pi, or hydrophobicity. The same CE instrument can be used for zone electrophoresis, IEF, sieving separations, isotachophoresis, and chromatographic techniques such as MEKC and capillary electrokinetic chromatography. This section provides a brief description of each separation mode. Zone electrophoresis, IEF, and sieving are the primary modes used for protein separations, and these will be discussed in detail in the following sections. 

The injection volumes in CE are extremely small because of the use of capillaries with very small diameters. Typical injection volumes are in the order of 10—50nE (a fog droplet is +10 nL). Injection of such small volumes of sample into the capillary is very challenging and requires specific approaches including use of rotary-, split- and micro-injectors, electrokinetic and hydrodynamic injection. Although all these injection techniques have shown to be quite appropriate, electrokinetic and hydrodynamic injection methods are mostly applied. Recent commercial instruments are usually equipped with these two injection modes as standard methods.Chapter 3 provides more details on the different injection modes. 

Subsequently four different CE modes are described in the sections Capillary Zone Electrophoresis, Capillary Gel Electrophoresis, Capillary Isoelectric Focussing, and Micellar Electrokinetic Chromatography (MEKC), respectively. The fundamental principles of the specific separation modes are briefly explained, using appropriate equations where required. In Table 3 all equations are listed. In addition, the influence of both instrumental parameters and electrolytic solution parameters on the optimization of separations is described. 

Samples are introduced into the capillary by either electrokinetic or hydrodynamic or hydrostatic means. Electrokinetic injection is preferentially employed with packed or monolithic capillaries whereas hydrostatic injection systems are limited to open capillary columns and are primarily used in homemade instruments. Optical detection directly through the capillary at the opposite end of sample injection is the most employed detection mode, using either a photodiode array or fluorescence or a laser-induced fluorescence (LIF) detector. Less common detection modes include conductivity [1], amperometric [2], chemiluminescence [3], and mass spectrometric [4] detection. 

Sample injection in NCE is very important for reproducible results with low limits of detection. In spite of some development in NCE very little effort has been made to develop sample injection devices in this technique. Of course sample injection in NCE is a challenging job due to small volume requirement [87], The controlled injection of small amounts of sample is a prerequisite for successful analysis in NCE. Electrokinetic injection (based on electroosmotic flow) is the preferred method and Jacobson et al. [88] optimized sample injection using this approach. Pinched injection allowing injection in minute quantities [89,90] and double-T shaped fluidic channels [91] have also been used for this purpose. Furthermore, Jacobson et al. [92] used a single high voltage source to simplify instrumentation. Similarly Zhang and Manz [93] developed a narrow sample channel injector to improve... 

One key feature of CE is the overall simplicity of the instrumentation. Briefly, the ends of a narrow-bore, fused silica capillary (25-75 pm i.d., 350-400 pm o.d., and 10-100 cm in length) are placed in buffer reservoirs. The content of the reservoirs is identical to that within the capillary. The reservoirs also contain the electrodes used to make electrical contact between the high voltage power supply and capillary. The sample is loaded into the capillary as follows one of the reservoirs (usually at the anode) is replaced by the sample reservoir and either an electric field (electrokinetic... 

The small dimensions associated with CE preclude the injection of large volumes. The sample may be introduced to the capillary either by a diplacement technique (i.e., pressure, vacuum, or siphoning) or via electrokinetic injection. The majority of commercial instruments apply a pressure differ-... 

Terabe, S., Micellar Electrokinetic Chromatography. Beckman Instruments, Fullerton, California, 1993. 

For isocratic mode of CEC separations, standard CE instrumentation is sufficient. This applies particularly for equipment that has the provision of column pressurization. In practice this is achieved by applying a gas under a pressure of 2-12 bar to both inlet and outlet vials. Column thermostating in CEC is regarded mandatory to avoid excessive radial temperature gradients within the capillary. In such instruments, sample is typically injected electrokinetically and alternatively by applying the external gas pressure to the sample vial. Detection occurs on-column i.e. directly through a non-packed section of the capillary following immediately the end of the bed. 

Kappes et al. evaluated the potentiometric detection of acetylcholine and other neurotransmitters through capillary electrophoresis [209]. Experiments were performed on an in-house capillary electrophoresis instrument that made use of detection at a platinum wire, dip-coated in 3.4% potassium tetrakis (4-chlorophenyl) borate/64.4% o-nitrohenyl octyl ether/32.2% PVC in THF. The results were compared to those obtained using capillary electrophoresis with amperometric detection at a graphite electrode. Samples prepared in the capillary electrophoresis buffer were electrokinetically injected (7 s at 5 kV) into an untreated fused silica capillary (88 cm x 25 pm i.d.) and separated with 20mM tartaric acid adjusted to pH 3 with MgO as the running buffer. The system used an applied potential of 30 kV, and detection versus the capillary electrophoresis ground electrode. 

Another well documented, but less widely used method for formulation development is the measurement of electrokinetic properties [14]. These tests require more disdnc-tive/elaborate sample preparation and are mosdy restricted to use during development. Also widely used and relied on are rheological measurements. The scope of rheological measurements ranges from viscosity measurements to the determination of yield points or oscillatory properties, such as the G and G -modules [15]. Since suspoemulsions are not ideal viscous but mostly viscoelastic or dilatant, a wide range of characterization techniques exists. Instrumentation required for this are simple rotary viscosimeters (Brookfield) or more sophisticated stress or shear controlled rotational viscosimeters. 

CRM for road dust (BCR-723) containing 81.3 2.5 Jg/kg Pt, 6.1 1.9 ig/ kg Pd, and 12.8 1.3 Jg/kg Rh, was introduced [49, 228]. It is widely used for quality control of results obtained in the analysis of environmental materials (e.g., airborne particulate matters, dusts, soils, and sediments). Comparison of results obtained using different analytical procedures and interlaboratory studies are recommended when there is a lack of suitable CRM (e.g., in examination of clinical samples). The use of standards based on real matrices (e.g., saliva, plasma, ultrafiltrates, and lung fluids) instead of synthetic solutions is recommended in such analyses. Difficulties with the identification and quantification of different metal species in examined samples make the reliability of results of great importance. The use of various instrumental techniques for examination of particular samples can be helpful. The application of chromatography, mass spectrometry, and electrochemistry [199] HPLC ICP MS and HPLC MS/MS [156] ESI MS and MALDI [162] micellar electrokinetic chromatography, NMR, and MS [167] AAS, ESI MS, and CD spectroscopy [179] SEC IC ICP MS and EC ESI MS [180] and NMR and HPLC [229] are examples of such approaches. 

Fig. 16.1. Schematic of a CEC instrument. The capillary column is typically 20-50 cm long with an inner diameter of 50-100 pm. The high voltage supply normally delivers 0-30 kV. About 1-10 nL of sample is usually injected electrokinetically.

A CEC instrument basically consists of a system for injection (pressure driven or electrokinetic), a column in which the separation takes place, a detector and a high voltage supply (Fig. 16.1). The most commonly used detector so far has been UV with transmission through the capillary outside of the packed bed. Laser induced fluorescence detection has been employed in several studies. Also, mass-spectrometry has been used. Normally, isocratic CEC is performed, but approaches to gradient CEC have been reported [29]. However, special equipment must be employed in most cases. 

After eliminating the effects of the pressure measurement instrumentation, electrokinetic and roughness effects were studied. 

A wide variety of electrokinetic measurements or instruments can be used to quantify the electrostatic stability of the dispersed phase. These measurements will only be summarized here. 

Capillary electrophoresis is an exciting, new, high resolution separation technique useful for the determination of drugs and their metabolites in body fluids. The first commercial capillary electrophoresis instruments began to emerge on the market in 1988. Today approximately a dozen companies manufacture electrokinetic capillary instrumentation, with many of these fully automated, that comprise auto samplers with computerized data evaluation.f Capillary electrophoresis involves the electrophoretic separations of minute quantities of molecules in solution according to their different velocities in an applied electrical field. The velocity of these molecules... 

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