After stabilizing the column with the mobile phase, samples (50 ml) were injected onto the HPLC system.
ME from mobile phases, sample solvents, pre-treatment methods, sample origins and instruments was evaluated, and its impact on the results of IDMS was assessed using the IDMS correction factor.
In principle, the procedure of SIC is based on a sequential injection, i.e. a discontinuous-flow approach, of a mobile phase and samples.
The mobile phase used for sample application, elution and sample preparation was pH 7.4, 0.067 M phosphate buffer.
In this case study, we systematically examined operating parameters such as mobile phase flow rate, sample injection volume and feed protein concentration on the characteristics of flow-through and bound-and-eluted protein peaks.
The developed and validated method is rapid, reproducible with simple mobile phase, trouble-free sample preparation steps, improved sensitivity and a short chromatographic run time, which therefore serves as a tool for the quality control of pharmaceutical dosage forms.
Resolution between differently sized molecules is a function of resin pore size, bed height, flow rate, mobile phase composition, and sample size.
A mixture of methanol (80%) and formic acid (0.1%) was used as a mobile phase and the sample flow rate was set to 300 μL/min.
The mobile phase for urine samples will be 10 mM ammonium formate, adjusted to pH 3.75 with formic acid and 2% acetonitrile.
The mobile phase for urine samples consisted of acetonitrile: 25 mM di-potassium hydrogen phosphate (39 61, v/v) adjusted to pH 8.9 with 85% orthophosphoric acid at a flow rate of 1.0 ml/min, and the mobile phase for plasma samples consisted of acetonitrile: 25 mM di-potassium hydrogen phosphate (15:75, v/v) adjusted to pH 3.0 with 85% orthophosphoric acid at a flow rate of 1.5 ml/min.
