A Test Mixture for Performance Verification of Multi-User UHPLC–MS Instruments
March 1st 2017Chromatographic techniques with mass spectrometric detection are important enablers in modern drug discovery. With the development of robust instrumentation and implementation of user-friendly software (or software packages), non-expert users can now walk up to easily accessible advanced chromatographic systems and perform experiments at their own convenience. Although remarkable improvements in robustness and ease-of-use have happened since the introduction of the first high performance liquid chromatography–mass spectrometry (HPLC–MS) systems, the instrument performance still needs to be qualified and monitored to ensure consistent high-quality results. This article will demonstrate how a simple test mixture of carefully selected compounds can facilitate both the development of generic ultrahigh-pressure liquid chromatography–mass spectrometry (UHPLC–MS) methods and automated performance monitoring of multiple instruments located in separate laboratories and buildings.
Count the Cost, Part 1: Increasing Resolution by Increasing Column Efficiency
March 1st 2017When considering column efficiency, more is not always better. We look at some ways to quickly estimate the effects of changes in column length and particle diameter rather than trying the experiments in the laboratory.
Improving GC Performance Systematically
March 1st 2017Gas chromatographers can control several variables that affect their separations: carrier-gas flow, column temperature, column dimensions, and stationary-phase chemistry. When faced with less than optimum resolution or separation speed, a strategy of changing just one variable at a time can be more productive than trying to hit the goal in one attempt. This month’s “GC Connections” examines how to use such a plan to obtain better gas chromatography results.
Characterizing Monoclonal Antibodies and Antibody–Drug Conjugates Using 2D-LC–MS
March 1st 2017Two-dimensional liquid chromatography (2D-LC) has in recent years seen an enormous evolution, and with the introduction of commercial instrumentation, the technique is no longer considered a specialist tool. One of the fields where 2D-LC is being widely adopted is in the analysis of biopharmaceuticals, including monoclonal antibodies (mAbs) and antibody–drug conjugates (ADCs). These molecules come with a structural complexity that drives state-of-the-art chromatography and mass spectrometry (MS) to its limits. Using practical examples from the authors’ laboratory complemented with background literature, the possibilities of on-line 2D-LC for the characterization of mAbs and ADCs are presented and discussed.