(Bio)inert RP/HILIC (U)HPLC Columns: YMC Accura Triart

LC-MS has been the standard choice for the analysis of signalling lipids, but the method poses specific challenges, such as peak tailing and loss of recovery for phosphate groups containing lipids. Explore an optimized LC-MS/MS method from YMC Europe that covers 388 signalling lipids from 17 lipid classes. Discover how the bioinert coated YMC Accura Triart C18 column combines high resolution with high sensitivity for phosphate groups containing lipids.

The lipid signature of biological samples, or lipidome, is remarkably different between health and disease states. Therefore, lipids are good candidates to produce potent biomarkers. Analysis of lipidomes deals with a large number of isomeric compounds, making comprehensive separation essential to generate a biologically informative datasets. Explore how YMC uses human plasma samples to demonstrate that separations under 10 minutes are possible with real samples using a less hydrophobic YMC Accura Triart C8 column with bioinert coating to facilitate higher levels of sensitivity and recovery.

Traditional methods such as ELISA and SDS-PAGE are labor-intensive and imprecise, whereas reversed phase liquid chromatography (RP-LC) coupled with mass spectrometry (MS) offers better accuracy for quantifying viral proteins VP1, VP2, and VP3. YMC Europe presents the YMC-Accura Triart Bio C4 column as the optimal choice for analyzing adeno-associated virus (AAV) capsid proteins, which are crucial for gene therapy applications. Further, YMC Europe highlights a study that compares the bioinert YMC-Accura Triart Bio C4 column to standard columns, demonstrating that the bioinert coating significantly improves resolution, peak shape, and retention times by preventing protein adsorption to the column hardware.

A particular challenge for chromatography is to find the balance between sensitivity and covering a broad spectrum of substances. Hydrophilic liquid chromatography (HILIC) is therefore especially suitable for such analyses containing polar compounds. It works with mass spectrometry (MS) and has very good chromatographic performance. With this combination, non-targeted analysis is well suited to the screening of polar metabolites in human plasma. YMC Europe highlights the non-targeted screening of polar metabolites in human plasma using a bioinert YMC-Accura Triart Diol-HILIC column. The bioinert coated stainless-steel hardware is essential to achieve the highest sensitivity and chromatographic resolution of endogenous isomers.

The YMC-Accura column hardware is designed to eliminate any interaction between sample and stainless steel due to a strict coating of column body and frit. It allows sharp peaks, stable recoveries and eliminates consequential carry-over effects. YMC Europe highlights results obtained with standard column hardware in comparison with the bioinert coated YMC-Accura Triart C18 column. All peptides were detected with much higher recovery rate than with the standard column hardware. Even challenging peptides can be analysed reproducibly. The use of bioinert YMC-Accura Triart columns has several benefits: higher recovery, better peak shapes, greater reproducibility, and little or no conditioning required.

Phospholipids are key components of cell membranes and have applications in nanotechnology. However, their phosphate groups pose challenges in chromatographic analyses by adsorbing onto metallic surfaces, especially at low to neutral pH. Bioinert systems like YMC-Accura Triart columns, which have a bioinert coating to prevent these interactions, are beneficial. YMC Europe highlights a study to demonstrate that the YMC-Accura Triart C18 UHPLC column provides excellent peak shapes for LPC and PC compared to stainless steel columns, which show significant peak tailing.

Oligonucleotides are crucial in genetic testing, research, and forensics, requiring robust and sensitive analytical methods. Ion pair reversed phase liquid chromatography is the gold standard, but stainless steel tubing and columns pose challenges due to corrosion and undesired ionic interactions with oligonucleotides. These interactions can negatively impact recovery and peak shape, especially at low to neutral pH. Traditional passivation methods are time-consuming and temporary. A more robust solution is using bioinert systems and columns. YMC Europe presents the YMC-Accura Triart series, with bioinert coatings and multi-stage endcapping, offering an ideal solution for oligonucleotide analysis. For short PS RNA oligonucleotides, YMC reports that the YMC-Accura Triart Bio C18 provides the optimum separation results compared to other columns specifically designed for oligonucleotide separations, and from a chromatographic point of view, it provided the best peak shapes and resolution for both buffer concentrations.

The use of stainless steel column hardware can negatively affect the separation of certain small molecules, especially metal-coordinating compounds like chelating agents, leading to issues such as loss of recovery, poor peak shapes, and sample carryover. This problem is more pronounced with new columns, requiring extensive pre-conditioning. YMC Europe presents the YMC-Accura column, with a strict bioinert coating, that prevents unwanted interactions with metal surfaces. YMC compares the analysis of two chelating agents, 8-hydroxyquinoline and hinokitiol, using the bioinert YMC-Accura Triart C18 column versus a conventional stainless steel column, and reports that the bioinert coated YMC-Accura Triart C18 column led to higher peak intensities and superior peak shapes, and virtually no tailing could be observed.

In this application note, YMC Europe provides a selection of studies, use case scenarios, and specifications for its range of oligonucleotide columns. Oligonucleotides are a powerful tool with vast potential in biotechnology and medicine. These short DNA or RNA sequences can target specific genes for therapeutic purposes, enhance diagnostics, and fuel advancements in synthetic biology, offering exciting opportunities for precision medicine and scientific innovation.

In this application note, YMC Europe addresses challenges in analyzing sensitive molecules using standard liquid chromatography methods. Sensitive compounds like peptides, proteins, and oligonucleotides often experience issues like peak tailing, loss of recovery, and retention time shifts due to unwanted interactions with metal surfaces in the chromatographic system. These interactions are more pronounced with acidic molecules containing phosphate or multiple carboxylate groups. The extent of adsorption and peak deformations depend on factors such as pH and ionic strength, with higher ionic strength reducing adsorption and a low to neutral pH increasing it. The number of such moieties also affects adsorption, as seen with adenosine triphosphate (ATP), which shows higher adsorption and does not elute compared to adenosine diphosphate (ADP) or adenosine monophosphate (AMP), with ADP still exhibiting more deformed peaks than AMP.

New strategies pair high-performance chromatography with digital tools to tackle persistent pollutants

From sample limitations to isomeric separations, see how the right column choice transformed a challenging lipidomics project