Sample loading capacity

Fast chromatography involves the use of narrow-bore columns (typically 0.1-mm i.d.) that will require higher inlet pressures compared with the conventional wide-bore capillary columns. These columns require detectors and computing systems capable of fast data acquisition. The main disadvantage is a much-reduced sample loading capacity. Advances in GC column technology, along with many of the GC-related techniques discussed below, were recently reviewed by Eiceman et... 

For the analysis of nonvolatile compounds, on-line coupled microcolumn SEC-PyGC has been described [979]. Alternatively, on-line p,SEC coupled to a conventional-size LC system can be used for separation and quantitative determination of compounds, in which volatility may not allow analysis via capillary GC [976]. An automated SEC-gradient HPLC flow system for polymer analysis has been developed [980]. The high sample loading capacity available in SEC makes it an attractive technique for intermediate sample cleanup [981] prior to a more sensitive RPLC technique. Hence, this intermediate step is especially interesting for experimental purposes whenever polymer matrix interference cannot be separated from the peak of interest. Coupling of SEC to RPLC is expected to benefit from the miniaturised approach in the first dimension (no broadening). Development of the first separation step in SEC-HPLC is usually quite short, unless problems are encountered with sample/column compatibility. 

Fused silica capillaries are almost universally used in capillary electrophoresis. The inner diameter of fused silica capillaries varies from 20 to 200 pm, and the outer diameter varies from 150 to 360 pm. Selection of the capillary inner diameter is a compromise between resolution, sensitivity, and capacity. Best resolution is achieved by reducing the capillary diameter to maximize heat dissipation. Best sensitivity and sample load capacity are achieved with large internal diameters. A capillary internal diameter of 50 pm is optimal for most applications, but diameters of 75 to 100 pm may be needed for high sensitivity or for micropreparative applications. However, capillary diameters above 75 pm exhibit poor heat dissipation and may require use of low-conductivity buffers and low field strengths to avoid excessive Joule heating. 

Chiral SFC can be performed in open tubular [41,42], and packed column [43,44] modes. Packed column SFC can be further categorized into analytical, semipreparative, and preparative SFC [7, 8], Packed column SFC is more suitable for fast separations than open tubular column SFC, since a packed column generally provides low mass transfer resistance and high selectivity [45, 46], Packed column SFC also provides high sample loading capacity [27,47], which can increase sensitivity. Only packed column SFC is suitable for preparative-scale enantioseparation. This chapter will focus on chiral separation using packed column SFC in the analytical scale. 

However, issues remain with sample loading capacity, and CapLC-NMR may be best suited to mass-limited samples where the component of interest is present at a sufficient concentration such that column loading does not become an issue [88]. Capillary probes have therefore been used most effectively where... 

Sellergren B. Molecular imprinting by noncovalent interactions tailor-made chiral stationary phases of high selectivity and sample load capacity. Chirality 1989 1 63-68. 

The same experimental setup was used in the separation of metabolites of paracetamol from a human urine extract [73] and Thomapyrin, containing acetaminophen, caffeine, and acetylsalicylic acid [74], Compared to isocratic CEC—NMR, gradient CEC—NMR offers increased sample loading capacity because of preconcentration at the front of the column and higher separation efficiency together with a reduction in analysis time [75],... 

Using SEC, most accomplishments of the commonly used RP-LC technique can be employed, such as various detector options, high sample loading capacity, variability in stationary phases and up-scaling option. Often in SEC, non-size-exclusion effects such as electrostatic and hydrophobic interactions between the analyte and stationary phase may be observed. The separation efficiency can be improved by optimizing the mobile phase, flow rate, column length, and sample volume. Practical guidelines for SEC method development have been described [42]. 

Pellicular or controlled surface porosity particles were introduced in the late 1960s these have a solid inert impervious spherical core with a thin outer layer of interactive stationary phase, 1-2 pm thick [13]. Originally, the inner sphere was a glass bead, 35-50 pm i.d., with a thin active polymer film or a layer of sintered modified silica particles on its surface. Such particles were not very stable, had very low sample load capacities because of low surface areas and are not used any more. Nowadays, this type of material is available as micropellicular silica or polymer-based particles of size 1.5 to 2.5 pm [14]. Micropellicular particles are usually packed in short columns and because of fast mass-transfer kinetics have outstanding efficiency for the separation of macromolecules. Because the solutes are eluted as very sharp narrow peaks, such columns require a chromatograph designed to minimise the extra-column contributions to band broadening. 

A curing step (120°C) applied to the L-PA imprinted polymers led to an apparent increase in sample load capacity [7]. As seen in Table 5.9 this treatment does not result in any significant differences in the dry state porosity and swelling of the material. However, it is likely that the number of unreacted double bonds is lower in these materials in view of the higher conversions observed in thermally versus photochemically polymerised materials (see Chapter 2). 

The attainable enrichment and clean-up in SPE depend primarily on the selectivity and affinity of the sorbent for the selected target analyte or analytes, the sample load capacity for the analytes and the rate of mass transfer to and from the binding sites, the latter affecting the minimum desorption volume and thus the enrichment that can be obtained. Other factors of importance are the reproducibility of the recovery yields and the stability and reusability of the sorbent when online procedures are desired. For hydrophobic analytes satisfactory results are usually obtained using standard reversed phase sorbents. Thus hydrophobised silica (C8, Cl8), styrene-divinylbenzene copolymers (PS-DVB) and graphitised carbon black (GCB) are the conventional sorbent materials used in SPE (Fig. 15.2)... 

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