Separation of o-phthalaldehyde-mercaptoethanol derivatives of amino acids from blood plasma on reversed-phase Nova-Pak C18 cartridges.

A separation of 25 o-phthalaldehyde-mercaptoethanol derivatives of primary amino acids in plasma prepared from human blood has been developed for Waters 10 cm x 0.8 cm I.D., 4-microns Nova-Pak C18 Radial-Pak cartridges. A binary gradient system with solvent-switching capability for the A pump is required. Computer methodologies have been utilized to develop mobile phase mixtures of phosphate buffer (pH 6.9), water, methanol and tetrahydrofuran. Advantages of the method include simple sample preparation, fast turnover time (67 min including the pre-column Autotag derivatization procedure) and exceptional column durability (several hundred analyses).