Repeatability and reproducibility of retention data and band profiles on reversed-phase liquid chromatography columns I. Experimental protocol

A procedure is described for the determination of the short-term and long-t erm repeatabilities and of the column-to-column and lot-to-lot reproducibil ities of the retention and profile characteristics of the peaks obtained wi th a number of different reversed-phase liquid chromatography (RPLC) C-18-b onded silica columns of several commercial brands. Data characterizing the retention, the steric selectivity, the hydrogen bonding capacity, the hydro phobic interaction selectivity, the column efficiency and the peak asymmetr y will be acquired for all the probe compounds. These include 30 neutral, a cidic and basic compounds distributed into five groups to be eluted under a s many different sets of chromatographic conditions. The data will be obtai ned using an HP 1100 liquid chromatograph. The compounds and experimental c onditions selected were similar to those previously used by different autho rs for comparison purposes. Careful attention will be paid to minimization of external error contributions by adhering to a strict operational procedu re. The precision expected will be high because preliminary results gave st andard deviations of around 0.04% for the separation factors, below 0.15% f or the retention times and around 1% for the column efficiency in short-ter m repeatability experiments performed with one column. (C) 1949 Elsevier Sc ience B.V. All rights reserved.