The AminoPac® PA10 strong anion-exchange column is a microbore column used for high-resolution separations of free amino acids by AAA-DirectTM.
Carbohydrates and amino acids monitored simultaneously
>50× more sensitive than ninhydrin-based analyzers; mid-femtomole to low-picomole detection limits
Fast and simple analyses of difficult protein samples (e.g., collagen)
Compatible with all commonly used hydrolysis procedures
Can be configured for on-line analysis
The AminoPac PA10 column is used for high-resolution separations of free amino acids by AAA-Direct. AAA-Direct is a technique that eliminates the need for any form of derivatization, and is therefore compatible with on-line monitoring of cell cultures and fermentation broths. Amino sugars and carbohydrates are detected simultaneously with amino acids if they are present in the sample. Additional capabilities include separation and detection of a wide range of sugars, phosphorylated amino acids, and common oxidation products of sulfur-containing amino acids, such as cysteic acid, methionine sulfone, and methionine sulfoxide.
Detection in AAA-Direct is by integrated pulsed amperometric detection (IPAD) and detection limits compare favorably with precolumn derivatization methods. The MDLs range from 110 femtomoles for histidine up to 4.42 pmole for leucine under standard conditions. AAA-Direct is directly compatible with all four of the commonly used protein hydrolysis procedures. The only sample preparation required with AAA-Direct following hydrolysis is filtration and dilution. This preparation is particularly advantageous when using the alkaline hydrolysis procedure since the hydrolysate must be neutralized when using other techniques. Even difficult samples are straightforward by AAA-Direct. Collagen is considered a difficult sample using classical ninhydrin amino acid analysis, because two runs are required at different dilutions as a consequence of the limited dynamic range of the ninhydrin postcolumn detection, and the disparate concentration levels of the amino acids present in collagen hydrolysates. This application is performed in a single run by AAA-Direct.
The AminoPac PA10 column is packed with a hydrophobic, polymeric, pellicular, anion-exchange resin that is stable over the range of pH 0–14. The unique pH stability of the packing material allows the use of eluent compositions that are conducive to anodic oxidation of amino acids at gold electrodes. The AminoPac PA10 column is recommended for use with the AAA-Direct Amino Acid Analyzer and Dionex Sodium Acetate Reagent, allowing direct detection of primary and secondary amino acids by IPAD, with no need for pre- or postcolumn derivatization.
