HPLC Column Tests

Since it is not possible to use test solutes which would be of interest to all users of HPLC columns, test mixtures should be composed of simple, stable, pure, and readily available organic compounds. 

I 4 Comparison and Selection of Modem HPLC Columns Test 2... 

However, the use of a HPLC separation step enabled a remarkable acceleration of the deconvolution process. Instead of preparing all of the sublibraries, the c(Arg-Lys-O-Pro-O-P-Ala) library was fractionated on a semipreparative HPLC column and three fractions as shown in Fig. 3-2 were collected and subjected to amino acid analysis. According to the analysis, the least hydrophobic fraction, which eluted first, did not contain peptides that included valine, methionine, isoleucine, leucine, tyrosine, and phenylalanine residues and also did not exhibit any separation ability for the tested racemic amino acid derivatives (Table 3-1). 

Techniques for packing hplc columns and for testing the packed column are described. 

A suitable, globally available HPLC column should be used in accordance with the manufacturer s specifications and its durability should be tested using the specified test conditions. An alternative column to be used in the method should be considered and appropriately validated. The initial pressure for a typical new column of 15-cm length and 5- um particles, using typical HPLC conditions like a flow rate of 0.5-1.0mL and ordinary buffer systems should be <200 bars. 

Significant time is saved in this calibration procedure by minimizing sample preparation and the number of calibrated test apparatus wherein only a balance, a 5-mL volumetric flask, a stopwatch, a thermal probe, two standards, one HPLC column, and one mobile phase are used. The use of MS Word template forms (see Figure 10) saves considerable time by eliminating any notebook entries and also improves the consistency of calibration records. Two important system parameters, dwell volume, and... 

Toxin Separations. A number of columns have been evaluated for suitability using the HPLC method. Of all columns tested to date, the Hamilton PRP-1 column (polystyrene divinylbenzene resin) has proven to be the most useful. Toxin retention on this column is controlled by 1) methanol concentration, 2) mobile phase ionic strength, 3) chain length of the ion-pair reagent, and M) mobile phase pH. 

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