Advanced Peak Processing to Reduce Efforts in Method Optimization
December 10th 2019Partial coelution of chromatographic peaks is an often-encountered issue in high performance liquid chromatography (HPLC) analysis, despite best efforts in method development and optimization. Even though there are several ways of integration of overlapping signals, accurate quantification of single compounds using conventional photodiode array (PDA) detection is almost impossible without baseline separation. While extensive signal processing is well established in spectroscopic analyses such as infrared (IR) or nuclear magnetic resonance (NMR), it has not yet been commonly adapted to improve chromatographic data evaluation. This article introduces the theory and application of a novel data analysis technique for PDA detection to accurately determine and quantify single compounds, even from overlapping peaks, without the need for mass spectrometry (MS) detection.
Tips & Tricks GPC/SEC: “Green” GPC/SEC and Solvent Alternatives
December 10th 2019The “greenest solution” is certainly using no solvent but gel permeation chromatography/size-exclusion chromatography (GPC/SEC) as a liquid chromatography (LC) technique requires the use of a mobile phase. The growing awareness of the need for more sustainable (greener) solutions has focused attention on environmentally- and health-friendly solvents and solutions.
The Basics of HPLC Peptide Analysis
December 10th 2019To fully characterize a protein biopharmaceutical, it must be broken down into smaller segments (peptides). Several high performance liquid chromatography (HPLC) techniques can be used to provide a wealth of information on everything from post-translational modifications (PTMs) to the glycoprofile to information on similarity when characterizing biosimilars.