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Improved Amino Acid Methods using Agilent ZORBAX Eclipse Plus C18 Columns for a Variety of Agilent LC Instrumentation and Separation Goals

Abstract: The continuous improvement in HPLC columns and instrumentation presents an opportunity to improve HPLC methods. A proven ortho-phthalaldehyde/9-fluorenylmethyl chloroformate (OPA/FMOC) derivatized amino acid analysis method developed on the HP 1090 Series HPLC Systems, and later updated for the Agilent 1100 Series HPLC Systems, has now evolved further taking advantage of the Agilent 1200 Series SL and Agilent ZORBAX Eclipse Plus C18 stationary phase columns. [1-5] Benefits of this protocol over previous iterations include: • Better retention of the first two eluting amino acids, aspartic and glutamic acid. • Higher resolution of several closely eluting amino acid pairs depending on the column configuration used. • Ten column configurations, including three particle sizes and several column lengths and diameters, allowing the analyst to customize the separation to his specifications and constraints (for example, available Agilent HPLC model, desired throughput, or desired resolution). • A quaternary pump option. Of course, the previous protocols' benefits are still retained, especially the automatic online OPA/FMOC derivatization.

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