Guest editors Valentina D’Atri from the University of Geneva in Switzerland and Jelle De Vos from the Research Institute for Chromatography (RIC) in Belgium, interviewed leading luminary of biopharmaceutical analysis, Reed Harris on critical quality attributes (CQAs), contemporary trends in biopharmaceutical characterization, and how he earned the moniker “Dr. Doom.”
The 51st International Symposium on High Performance Liquid Phase Separations and Related Techniques (HPLC 2023), chaired by Michael Lammerhofer and Oliver J. Schmitz, was held from 18-22 June in Dusseldorf, Germany. This instalment of "Column Watch" presents many of the highlighted topics and trends observed at this exceptionally well-run symposium.
A novel liquid chromatography-tandem mass spectrometry (LC-MS/MS) method with a dual electrospray ionization (ESI) and atmospheric-pressure chemical ionization (APCI) source was developed for analyzing 102 pesticides and five mycotoxins that are regulated by the state of Colorado in hemp. The limit of quantitation (LOQ) of the 102 pesticides and 5 mycotoxins met Colorado state action limit requirements for these analytes in a hemp matrix. For this study, 88 out of 102 pesticides and all 5 mycotoxins were analyzed using LC-MS/MS with an ESI source, whereas the remaining 14 pesticides were determined using LC-MS/MS with an APCI source. A simple, fast, green, and cheap acetonitrile solvent extraction method was used to extract the pesticides and mycotoxins from the hemp matrix with good extraction efficiency in the range of 80-120%. A hemp matrix is challenging and causes matrix effects such as ion suppression or enhancement. The LC method was optimized and 30 internal standards were added to reduce and compensate for these matrix effects to obtain method accuracy in the range of 70-120%. The ionization mechanism of nonpolar pesticides (normally analyzed by gas chromatography-tandem mass spectrometry [GC-MS/MS]) with an APCI ion source was elucidated.
“Questions of Quality” is 30 years old! What, if anything, has changed in chromatography laboratories over that time? What, if anything, remains the same?
Therapeutic mRNA is receiving growing interest in various therapeutic applications such as genome editing, cancer immunotherapy and prophylactic vaccines. As with other drugs, it is essential to guarantee product quality. Among the critical quality attributes of therapeutic mRNA, characterization of the capping and poly(A) tail are of the greatest importance because of their involvement in mRNA stability and in the efficiency of protein synthesis. This article presents a method for the simultaneous characterization of both attributes in a single sample preparation workflow. The method involves lipid extraction, various RNAse enzymes, purification steps and LC–MS to analyze the capping and poly(A) tailing.
