title: Effect of pH additive and column temperature on kinetic performance of two different sub-2 μm stationary phases for ultrafast separation of charged analytes.
abstract: The performance characteristics of separation were studied for small pharmaceuticals and larger charged molecules (peptides) in various mobile phase conditions on two 5-cm long narrow bore columns packed with 1.7 μm core-shell and totally porous particles respectively. The effect of temperature and pH additives (formic acid, trifluoroacetic acid, ammonium formate, ammonium acetate and ammonia) on column efficiency was investigated through a kinetic study based upon data obtained under gradient elution conditions. Sample peak capacities were calculated and compared in all studied conditions for a sample of ten representative peptides having masses ranging from 500 to 2000 Da. The elevation of temperature was found to be significantly beneficial. The effect of flow-rate on peak shape was also investigated. Ammonium acetate at neutral pH led to the best results in terms of both efficiency and peak capacity. It was found that column performance was strongly dependent on the type of stationary phase, especially in acidic medium.
authors: Laboratoire des Sciences Analytiques, UMR CNRS 5280, Université de Lyon, 43 Boulevard du 11 Novembre 1918, 69622 Villeurbanne Cedex, France. sabine.heinisch@univ-lyon1.fr
authors: Sabine
authors: S
authors: Heinisch
authors: Amélie
authors: A
authors: D'Attoma
authors: Candice
authors: C
authors: Grivel
mesh_terms: D000085:Acetates / Q000737:chemistry; D000097:Acetonitriles / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation* / Q000379:methods*; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D010455:Peptides / Q000302:isolation & purification; D004364:Pharmaceutical Preparations / Q000302:isolation & purification
chemical_list: D000085:Acetates; D000097:Acetonitriles; D010455:Peptides; D004364:Pharmaceutical Preparations; C018824:ammonium acetate; C032159:acetonitrile