***Thursday and Friday May 13-14 | May 20-21, 2021*** In the relaxed and unconventional talks in this symposium, highly experienced speakers from industry and academia will impart key learnings from their long experience with chromatography—from heroic failures that have taught them valuable lessons, to moments of inspirational serendipity, and everything in between. CHROMTalks will deliver lifetimes of learning in condensed form, covering topics across implementation, troubleshooting, and method development of analytical techniques. Time spent at CHROMTalks will pay back as soon as you enter your laboratory! ***Available On Demand Until May 21, 2022***
In biotherapeutics manufacturing, controlling size variants of therapeutic monoclonal antibodies (mAbs) is critical for ensuring product quality.
This concise yet comprehensive overview of sample preparation for bioanalysis looks at sample preparation fundamentals, best practices, and modern trends—all illustrated with a case study.
Gas chromatography–mass spectrometry (GC–MS) with cold electron ionization (EI) is based on interfacing the GC and MS instruments with supersonic molecular beams (SMB) along with electron ionization of vibrationally cold sample compounds in SMB in a fly-through ion source (hence the name cold EI). GC–MS with cold EI improves all the central performance aspects of GC–MS. These aspects include enhanced molecular ions, improved sample identification, an extended range of compounds amenable for analysis, uniform response to all analytes, faster analysis, greater selectivity, and lower detection limits. In GC–MS with cold EI, the GC elution temperatures can be significantly lowered by reducing the column length and increasing the carrier gas flow rate. Furthermore, the injector temperature can be reduced using a high column flow rate, and sample degradation at the cold EI fly-through ion source is eliminated. Thus, a greater range of thermally labile and low volatility compounds can be analyzed. The extension of the range of compounds and applications amenable for analysis is the most important benefit of cold EI that bridges the gap with LC–MS. Several examples of GC–MS with cold EI applications are discussed including cannabinoids analysis, synthetic organic compounds analysis, and lipids in blood analysis for medical diagnostics.
High-resolution mass spectrometry (HRMS) is an increasingly critical tool for identifying, characterizing, and monitoring attributes of protein-based therapeutics.
In this installment of “LC Troubleshooting,” we discuss strategies that can be used to minimize the likelihood of compound degradation in the isolation process.
An introduction to this special issue by our guest editor.
Liquid chromatography (LC) is a platform technology amenable to portable and “at-site” or deployable applications. This has awoken end-users to new possibilities and potential cost savings and process improvements.
When can analyte retention deviate from what is expected or normal? We explain three subtle causes.
Thursday, March 17th, 2022 at 10am EDT | 7am PDT | 3pm CET | 2pm GMT Learn how fast and efficient HPLC methods using BIA Separations, now a Sartorius company, PATfix™ system allow process optimization in both USP and DSP part of AAV manufacturing, by assessing the purity from the in-process sample up to the final product.
Per- and polyfluoralkyl substances (PFAS) are found in our food. Sensitive, precise, and accurate analytical methods are needed to estimate human exposure to these chemicals. A comparative study was performed between two extraction and cleanup methods (solid-phase extraction [SPE] and dispersive SPE) for the analysis of PFAS in apples. Both methods showed excellent sensitivity, precision, and accuracy. dSPE has some benefits over conventional SPE, and vice versa. The advantages and disadvantages of both methods are discussed.
Pyrolysis–gas chromatography–mass spectrometry has advantages for the analysis of environmental microplastic samples compared to other leading analytical methods, including spectroscopic techniques.
A simple LC–MS method has been developed and validated for the simultaneous determination of 18 synthetic food additives and caffeine in soft and energy drinks, and in various alcoholic beverages. Nine food colours (tartrazine, sunset yellow FCF, azorubine, ponceau 4R, allura red AC, patent blue V, brilliant blue FCF, green S, brilliant black BN), two preservatives (sorbic and benzoic acid) and seven sweeteners (acesulfame K, aspartame, cyclamic acid, saccharin, sucralose, neohesperidin DC, neotame) were targeted food additives. The method employs reversed-phase ultra-high performance liquid chromatography (UHPLC) for analyte separation and a single quadrupole mass spectrometer for their detection. The limits of quantification were low enough to enable a reliable control of maximum limits set for some additives (Regulation [EC] No. 1333/2008). The method was applied for analysis of a wide range of samples collected at a typical supermarket: 14 soft drinks, 19 energy drinks, and 43 alcoholic beverages.
Crucial phosphorylated compounds can be analyzed by a polymeric iHILIC-(P) Classic HILIC column and MS. The method is applicable for targeted and non-targeted metabolomics.
Reversed-phase LC–MS has limitations in numerous analytical applications. Alternative MS-compatible chromatographic techniques separate analytes in the liquid phase based on different retention mechanisms compared with reversed-phase LC. This article describes these alternative chromatographic approaches, relevant applications, and the future of these techniques.
Innovative native separation strategies coupled with native mass spectrometry techniques to characterize biopharmaceuticals close to their natural state are described.
The article discusses the application of a hybrid organic/inorganic surface technology that forms a barrier between the sample and the metal surfaces of both the HPLC system and chromatographic column. Formed by a vapor deposition of an ethylene-bridged siloxane polymer on metal substrates), this technology effectively addresses common chromatographic challenges such as analyte loss, carryover, and peak tailing due to metal-analyte interactions. It improves peak symmetry and areas, as well as reproducibility, thereby not only benefiting challenging analytes but also increasing confidence in analytical results. We demonstrate the benefits of this technology through the analysis of B-group vitamins, steviol glycosides, and dextran oligosaccharides typically found in food, beverages, and dietary supplements, showcasing its critical role in improving chromatographic performance.
One of the fundamental tenets of chromatography is the interaction between an analyte and stationary phase.
Improved analysis of pharmaceutical and natural medicine products requires advances in reversed-phase LC stationary phases. We examine two synthesized stationary phases with applicability in quality control and chiral separation for analysis of natural products.
Microextraction techniques have evolved since their appearance three decades ago, moving toward the use of greener and sustainable materials and solvents.
A column with chemically modified column hardware showed improvements in analytical performance for siRNA compared to a conventional stainless-steel column.
The efficiencies of microbore-2 columns, which are prepared from blanks that have a wide variety of inner surface roughness, drop sharply when the size of individual surface roughness features approaches the particle size of the packing material. The results suggest that two categories of packed column structure relate to the surface features and yield high and low efficiency columns. This installment of "Column Watch" discusses this conclusion in terms of the stability of an agglomerated layer of packing particles on the blank wall when subjected to shear forces during column packing.
The UHPLC–MS/MS method can accurately determine the presence of these illegal feed additives in swine tissues.
A highly sensitive method for quantitative determination of contaminants in surface waters, with and without particulate matter, was tested using stir-bar sorptive extraction (SBSE).
Measuring volatile phenols in wine is essential in ensuring superior wine quality. A new analytical technique, called solid-phase mesh-enhanced sorption from headspace (SPMESH), was modified with direct immersion (DI) conditions and coupled to direct analysis in real time–mass spectrometry (DART–MS) to be used to detect smoke taint in winemaking.
The challenges that arise during cannabis metabolomics analysis using ultrahigh-performance reversed-phase liquid chromatography coupled with high-resolution tandem mass spectrometry (UHPLC–reversed-phase–HRMS/MS) are presented.