Glutamine, valine, isoleucine, and leucine are baseline resolved in a dietary supplement with a mobile phase of acetonitrile, water, and sulfuric acid with ultraviolet (UV) detection at 210 nm on a Primesep 100 column. Increasing the sulfuric acid concentration allows elution of arginine. Primesep 100 separates underivatized amino acids and amines by combining reversed-phase and ion-exchange mechanisms. The HPLC separation uses a mobile phase of water, acetonitrile (MeCN, ACN), and sulfuric acid with ultraviolet (UV) detection at 210 nm.
| Column | Primesep 100, 4.6×250 mm, 5 µm, 100 Å |
|---|---|
| Mobile Phase | MeCN/H2O – 30/70% |
| Buffer | H2SO4 |
| Flow Rate | 1.0 mL/min |
| Detection | UV, 210 nm |