Analysis of amino acids using on-line pre-column derivatization with OPA with rapid separation HPLC
Author(s)
Jasco

The high selectivity and high sensitivity measurement of amino acids requires the derivatization as amino acids do not absorb in UV-Vis region or fluorescence. Pre-column and post-column derivatization methods can be used for the analysis of amino acids. In the case of pre-column derivatization, the sample is derivatized in the autosampler and then separated in a C18 or similar column and detected. On the other hand, in the case of post-column derivatization, the sample is separated using an ion-exchange column or similar first, then mixed with reaction solution for derivatization and then detection. 
 

Jasco offers several analysis systems:

  • Pre-column derivatization system using orthophtalaldehyde (OPA) for fluorescence detection
  • Pre-column derivatization system using Dabsyl chloride for absorbance detection
  • Post-column derivatization system using OPA for fluorescence detection
  • Post-column derivatization system using ninhydrin for absorbance detection. 

The pre-column derivatization method has many advantages such as a simple system configuration, wide selectivity of derivatization reagents, and offers a high sensitive measurement.

In this application, 19 amino acids were measured using OPA pre-column derivatization through the automated pre-column derivatization function of autosampler. The RHPLC conditions drastically reduced the measurement time.