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The document details a quantitative amino acid analysis method combining automated online derivatization with o-phthalaldehyde (OPA) for primary amino acids and 9-fluorenylmethyl chloroformate (FMOC) for secondary amino acids. This procedure is integrated with rugged HPLC analysis using the Agilent 1100 system to achieve rapid, accurate, and reproducible results.
| Analysis Time (75-mm column) | 14 min |
|---|---|
| Analysis Time (150-cm column) | 26 min |
| Column Dimensions (High-Throughput) | 4.6 x 75 mm, 3.5 um |
| Column Dimensions (High-Resolution) | 4.6 x 150 mm |
| Detection Wavelength (OPA) | 338 nm |
| Detection Wavelength (FMOC) | 262 nm |
Primary amino acids are reacted first with OPA using 3-mercaptopropionic acid (3-MPA).
Secondary amino acids are derivatized using FMOC after they do not react with OPA.
The total analysis time on a 75-mm column is 14 minutes, including re-equilibration.