A comprehensive study on the phenomenon of total breakthrough in liquid chromatography

Expediting the chromatographic analysis of COVID-19 antibody therapeutics with ultra-short columns, retention modeling and automated method development

“…Flow rate was set as 0.6 mL/min and 0.5 µL of 0.8 mg/mL protein material (dissolved in 20% of MPA) was injected. After the optimal conditions were found, an initial short peak compression step was added - prior to the analytical gradient - in which the %B was rapidly increased (in 0.5 min) from 20% to starting conditions of the gradient (to avoid break-through effects and peak distortion [27] ).…”

An approach to high throughput measurement of accurate retention data in liquid chromatography

“…Increasing complexity of challenges faced by separation scientists along with the ever-increasing drive for more efficient method development is fueling continuing interest in modeling and simulation of a variety of aspects of liquid phase separations [1][2][3][4][5][6][7]. For example, recent studies by different research groups have focused on aspects including the effect of the volume and composition of the injected sample on separation quality [2,4,5,8], the effect of temperature on analyte retention in reversed-phase liquid chromatography (RPLC) [9], the effect of pump nonidealities on the prediction of retention time when using gradient elution conditions [10], and resolution of difficult-to-separate mixtures by serially coupling columns with different selectivities [11,12]. Currently, these efforts depend on experimental data to build models that are accurate enough to be useful in method development.…”

Enhanced Sensitivity in Comprehensive Liquid Chromatography: Overcoming the Dilution Problem in LC × LC via Temperature-Responsive Liquid Chromatography

“…If the injection solvent is of lower elution strength than the 2 D mobile phase, the injection plug and the volumes of the transferred peaks are reduced by on-column focusing . In this way, the combination of orthogonal separation modes can lead to severe injection effects in the 2 D due to the ensuing solvent strength mismatch . This is often observed for, e.g., HILIC × RPLC or NPLC × RPLC.…”

“…An RPLC × RPLC method applying two 2.1 mm columns in both dimensions revealed that when refocusing is obtained in the 2 D, large effluent volumes could be transferred in LC × LC, reducing the dilution factor and its detrimental effects on sensitivity while benefitting from additional gains in peak capacity in comparison to 1D-LC. 24 It was further shown that 2D-LC can also be promising for the determination of low abundant compounds such as impurities at a 0.05% level in pharmaceutical quality control. 25 Stoll et al 25 found that quantitation limits are, for example, dependent on the peak location in the gradient window, as baseline disturbances will be higher near the dead time.…”