Choosing sample volume to achieve maximum detection sensitivity and resolution with high-performance liquid...

The relationship between sample volume and the chromatographic performance factors of detection sensitivity and resolution were explored for high-performance liquid chromatography (HPLC) columns of 1.0, 2.1 and 4.6 mm I.D. Performance was evaluated for isocratic and gradient elution in systems having low, intermediate and high extracolumn dispersion. Sample volumes ranged from 0.4 microliter to 2 ml. This paper provides guideline for selecting sample injection parameters so that the performance of microbore columns is not needlessly sacrificed by using too small or too large a sample volume. The guidelines are extracted from the works of others and are modified by our own experimental results. Comments are also made on the relative merits of various injection techniques.