This collection of technical articles, interviews, and news pieces delves into the latest innovations in LC methods, including advance in high performance liquid chromatography (HPLC), ultrahigh-pressure liquid chromatography (UHPLC), liquid chromatography–mass spectrometry (LC–MS), and multidimensional LC.
Liquid chromatography (LC) is a key element of modern analytical science, offering precision and versatility for separating, identifying, and quantifying compounds in complex mixtures. This collection of technical articles, interviews, and news pieces delves into the latest innovations in LC methods, including advance in high performance liquid chromatography (HPLC), ultrahigh-pressure liquid chromatography (UHPLC), liquid chromatography–mass spectrometry (LC–MS), and multidimensional LC. These articles explore pharmaceutical analysis, biopharmaceutical analysis, environmental monitoring, food analysis, sustainability, and method development amongst others.
In-Loop Analyte Degradation in Two-Dimensional Liquid Chromatography: Example and Solutions
By Ziqing Lin, Qinggang Wang, and Dwight R. Stoll
The use of two-dimensional liquid chromatography (2D-LC) to assess peak purity is becoming common in the pharmaceutical industry. In this application space, it is critically valuable to demonstrate that all compounds eluted from a column during an LC assay are accounted for, and that no impurities are “hiding” under the peaks of known compounds. Although the addition of the second dimension (2D) separation makes 2D-LC an exquisitely powerful tool, it also warrants additional care when interpreting the results to avoid misidentification of apparently new peaks as coeluted impurities, when in fact they are analytical artifacts. In this installment of “LC Troubleshooting,” we describe one such possible artifact that can arise because of the degradation of compounds during the transfer of fractions of the first dimension (1D) column effluent to the 2D separation. The authors suggest simple experiments to determine whether new peaks observed in 2D chromatograms result from degradation, ultimately increasing our confidence in the interpretation of 2D-LC results.
By Alasdair Matheson
As part of our RAFA 2024 coverage, we interviewed Rudolf Krska of BOKU University about his research on food safety, specifically regarding using analytical methods to mycotoxins and other secondary metabolites in plants and foods.
Applying Sustainability Concepts to Modern Liquid Chromatography
By Matt James, Tony Edge, and Arianne Soliven
Governments are striving to implement policy changes towards a greater use of green technology, specifically around the generation of energy. Individuals and industrial organizations have also taken up the challenge, and now many companies are driving to significantly reduce their environmental footprint, or indeed become carbon negative within very short time frames.
Detailed Glycosylation Analysis of Therapeutic Enzymes Using Comprehensive 2D-LC–MS
By Koen Sandra, Kathleen Piens, Debby Bracke, Pat Sandra, and Wouter Vervecken
The use of comprehensive two-dimensional liquid chromatography (LC×LC) coupled to mass spectrometry (MS) for characterizing glycosylation of therapeutic enzymes is presented. Recombinant human acid α-glucosidase (rhGAA) was digested, and the resulting peptides were separated by reversed-phase LC (RPLC) at high and low pH in, respectively, the first and second dimension. Glycopeptide peaks were then selectively detected and identified by MS operated in all-ion fragmentation mode. The study of first generation rhGAA (myozyme), expressed in Chinese hamster ovary (CHO) cells, and next-generation glyco-engineered rhGAA, produced in yeast cells to fine-tune the mannose-6-phosphate (M6P) content, is described.
Exploring The Chemical Subspace of RPLC: A Data-driven Approach
By Alasdair Matheson
Saer Samanipour from the Van ‘t Hoff Institute for Molecular Sciences (HIMS) at the University of Amsterdam spoke to LCGC International about the benefits of a data-driven reversed-phase liquid chromatography (RPLC) approach his team developed to enhance RPLC method development, including increased efficiency for non-targeted analysis and suspect screening, a reduction in the amount of false positives produced, and a predictive way to determine if a chemical can be separated using RPLC.
Investigating PFAS in Plastic Food Storage Bags Using LC–MS/MS
By Alasdair Matheson
Yelena Sapozhnikova from the United States Department of Agriculture (USDA) spoke to The Column about her innovative research investigating per- and polyfluoroalkyl substances (PFAS) in plastic food storage bags using targeted and non-targeted liquid chromatography–tandem mass spectrometry (LC–MS/MS). Rigorous QA–QC procedures were implemented to overcome challenges associated with background contamination and interferences.
By Ann Marie Rojahn, Mathias Hehn, and Daniel Eber
This article describes a comprehensive study on optimizing ion-pair reversed-phase liquid chromatography for analyzing protected and unprotected single-stranded DNA oligonucleotides. The study identifies optimal conditions for improved chromatographic resolution and discusses the advantages of using bioinert column hardware and specific temperature settings to enhance the recovery and peak shapes of oligonucleotides, providing a detailed guide for similar analytical setups in the field.
Saliva Sampling Methods Optimized Using UHPLC-MS
By Aaron Acevedo
Scientists from the Institute of Analytical Chemistry of the Czech Academy of Sciences and Masaryk University recently tested various saliva sampling methods to optimize bile acid analysis.
By Pascal Marillier, Neal Adams, Steven W. Baertschi, John M. Campbell, Chris Foti, Juçara Ribeiro Franca, Simon Hicks, Dorina Kotoni, Christian Laue, Stacey Marden, Liping Meng, Ana Claudia de Oliveira Santos, Mariah Ultramari, An Van Cleempoe, Chloe Wang, Todd Zelesky, and Zongyun Huang
This review article discusses scientific rationales and current best practices in the pharmaceutical industry for performing chromatographic peak purity assessments (PPA). These activities are associated with the development and validation of liquid chromatographic (LC) stability-indicating analytical methods applicable to regulatory submissions of small-molecule drug candidates. The discussion includes a comprehensive overview of the PPA-related regulatory and scientific landscape and common industry approaches to obtain PPA results, as well as the strengths and weaknesses of PDA-facilitated ultraviolet (UV) PPA and other PPA techniques.
A Career in Liquid Chromatography: An Interview with Wolfgang Lindner
By Patrick Lavery
At Pittcon 2024, LCGC International sat down with Wolfgang Lindner of the University of Vienna to talk about his career in liquid chromatography (LC). Lindner is the 2024 recipient of LCGC’s Lifetime Achievement in Chromatography Award.
Next Generation Peak Fitting for Separations
December 11th 2024Separation scientists frequently encounter critical pairs that are difficult to separate in a complex mixture. To save time and expensive solvents, an effective alternative to conventional screening protocols or mathematical peak width reduction is called iterative curve fitting.
Mobile Phase Buffers in Liquid Chromatography: A Review of Essential Ideas
December 11th 2024In this installment of "LC Troubleshooting," Dwight Stoll discusses several essential principles related to when and why buffers are important, as well as practical factors, such as commonly used buffering agents, that are recommended for use with different types of detectors.
The Chromatographic Society 2025 Martin and Jubilee Award Winners
December 6th 2024The Chromatographic Society (ChromSoc) has announced the winners of the Martin Medal and the Silver Jubilee Medal for 2025. Professor Bogusław Buszewski of Nicolaus Copernicus University in Torun, Poland, has been awarded the prestigious Martin Medal, and the 2025 Silver Jubilee Medal has been awarded to Elia Psillakis of the Technical University of Crete in Greece.