IMPROVED HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY OF AMINO-ACIDS DERIVATIZED WITH 9-FLUORENYLMETHYL CHLOROFORMATE

An improved high-performance liquid chromatography method for the sepa ration of amino acids derivatised with 9-fluorenylmethyl chloroformate (Fmoc) is described. This method, in conjunction with a multi-tasking program, not only allows high automatic throughput but also offers ba seline resolution of the common Fmoc-amino acids from a linear acetoni trile gradient, greater chromatographic reproducibility over the life of the column (800 runs) and easy column regeneration. The methods des cribed are shown to be suitable for analysis of hydrolysates of protei ns from two-dimensional gels for protein identification purposes, and will also be useful for routine quality control and screening of biolo gical samples.