Regeneration, column

An additional problem exists in which impurities in the displacer itself complicate separation.54 Also, the displacer itself must be removed from the column, which lengthens regeneration time and can adversely affect throughput. Ironically, while the difficulties involved in identifying displacers and in column regeneration have retarded use of displacement as a preparative method, there has been renewed interest in using displacement chromatography in analytical and semi-preparative applications for enrichment of trace compounds.55 56... 

The vast majority of purification procedures employ at least one ion-exchange step it represents the single most popular chromatographic technique in the context of protein purification. Its popularity is based upon the high level of resolution achievable, its straightforward scale-up (for industrial application), together with its ease of use and ease of column regeneration. In addition,... 

The dwell time was 200 msec for the analytes and 100 msec for the IS. At least 500 extracted samples were injected onto each column without any column regeneration. No solvent evaporation and reconstitution steps were involved. Ethyl acetate was preferred over methyl t-butyl ether (MTBE) because MTBE caused pulp-up of the mat. Six blank plasma lots were tested for matrix interference and none was detected in the analyte or IS region. When 100 ng/mL of the analytes were spiked into the blank plasma samples, the relative standard deviations were 1.0 and 1.5% for omeprazole and its metabolite, respectively. Precision and accuracy figures are given in Table 1.9. 

FIGURE 2.1 Flow diagram for dual-stream HPLC with guard column regeneration. 

FIGURE 2.3 Autosampler set-up for performing dual-stream HPLC with guard column regeneration (A) overall view, (B) zoomed view of the valving configuration. 

FIGURE 4.11 Timeshare of MS without MS modification. Flow diagram also shows guard column regeneration set-up. (Source Chang, M. et al., presentation at Pittsburgh Conference and Exposition, 2000. With permission.)... 

The following operating guides are recommended for maintaining reversed-phase columns. Consult the vendor s column instructions for precautions and column regeneration guides. 

Step 3 Detect excess label and/or elute label for detection (column regeneration)... 

On-line size exclusion chromatographic (SEC) analyses were performed with a Waters Model 401 differential refractometer (DR), a Waters Model 480 ultraviolet (UV) variable wavelength spectrophotometer and a Foxboro Miran lA infrared (IR) photometer, equipped with a zinc selenide ultramicro flowcell of 1.5 mm nominal pathlength and 4.5 /xl volume, purchased from the same supplier. A set of ten Mycrostyra-gel (Waters Associates) columns, regenerated by Analytical Sciences Inc. (ASI) and of nominal porosities 100, 500 (two) 10 (two), 10 (three), 10 and lO X, in the order given and a mobile phase flow rate of 1 ml/min was used. The column set had a specific resolution of 19.7 in 1,4-dioxane, as determined by the method of Yau(2). 

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