Presenting a new method for oxysterol separations at CHA25

Berlin: – The MOLNÁR-INSTITUTE for Applied Chromatography will be a prominent presence at the upcoming CHanalysis meeting of Swiss Analytical Scientists, as an official partner of the event.

The 9th meeting of the Swiss Analytical Scientists (CHA25) is being hosted at a resort in the Swiss Alpine village of Beatenberg, with its famous views over the Bernese Oberland mountains.

The MOLNÁR-INSTITUTE is also contributing to the scientific program with a DryLab use case on the second day of the conference as part of the session on Data, Quality and Metrology, chaired by Hanspeter Andres.

Simplifying Oxysterol Separations

Dr. Andrea Castellaneta from the Department of Chemistry at the University of Bari will present his research titled “Expanding the Analytical Design Space for Oxysterol Separation by HPLC”. This work was carried out in collaboration with Dr. Imre Molnár, President of the Molnár-Institute, and Dr. Hans-Jürgen Rieger, Vice President of the Molnár-Institute.

The presented study demonstrates the successful application of separation modeling to enhance the robustness of hyphenated mass spectrometric liquid chromatography (LC-MS) for the analysis of oxysterol-compounds. Oxysterols are valuable biological markers that require precise qualitative and quantitative determination. However, conventional GC-MS and LC-MS methods have several limitations. In LC-MS, for example, prior derivatization is often necessary, which can compromise the primary analytical objective of accurately characterizing oxysterol composition. Additionally, challenges related to separation—such as ionization efficiency and sensitivity issues—are common. A further complication is that many oxysterols undergo extensive in-source fragmentation, resulting in the formation of isomeric ions from non-isomeric species, which further complicates method optimization efforts.

In this study, promising results were achieved through the application of DryLab modeling to optimize the chromatographic separation, specifically utilizing C18 and cyanopropyl stationary phases. The systematic method development strategy enabled effective separation without the need for derivatization agents, representing a significant advancement over conventional approaches. These findings may further contribute to the development of more reliable RPLC-FTMS methods in future studies, offering a robust and efficient platform with minimal sample preparation requirements.

Multidimensional Modeling Support

The team conducted multidimensional Design-of-Experiments (DoEs) to evaluate the impact of key separation parameters, including gradient steepness (tG), column temperature (T) and mobile phase ternary composition (tC). Dr. Castellaneta will demonstrate how the resulting visualized modeling maps captured distinct separation patterns for each stationary phase, revealing specific selectivity opportunities and enabling the clear identification of optimal separation conditions. This approach not only offers a practical strategy for method optimization on a single stationary phase but also facilitates subsequent column interchangeability studies. Such studies can be particularly valuable for identifying general operational conditions across different columns, enhancing flexibility in both method development and routine analysis.

About The MOLNÁR-INSTITUTE

Founded in 1981, The MOLNÁR-INSTITUTE develops DryLab®4, a software for UHPLC modelling for a world-wide market. Its powerful modules gradient editor, peak tracking, automation, robustness and Design Space Comparison allow for the most sophisticated method development as required across modern pharma industries. Analytical scientists use DryLab®4 to understand chromatographic interactions, to reduce analysis time, to increase robustness, and to conform to Analytical Quality by Design (AQbD) principles, according to the recently published ICH Q14 regulatory framework.

The MOLNÁR-INSTITUTEis a registered partner of the US-FDA, CDC and many other regulatory bodies. DryLab®4 pioneered AQbD long before regulatory agencies across the world encouraged such submissions. Widely implemented by thought leaders, the software contributes substantially to the paradigm shift towards a science and risk driven perspective on HPLC Quality Control and Assurance.

Further information at: http://www.molnar-institute.com/

About CHA 25

The CHanalysis series is an annual forum for the Swiss analytical science community, hosted by the Analytical Science Division (DAS) of the Swiss Chemical Society (SCS).

The 9th meeting of the Swiss Analytical Scientists (CHA25) is a two-day meeting opening March 19, 2025, at the Dorint Ressort Blüelisalp in Beatenberg, with its famous views over the Swiss Alpine peaks of the Bernese Oberland including the ‘Big Three’ mountains: Eiger, Mönch and Jungfrau.

The conference will feature sessions on Materials, Pollutants, Data, Quality & Metrology, and Forensic & Cultural Heritage Science.

The event is organised by DAS/SCS with further information at: https://cha25.scg.ch

Resources

Click on Molnár-Institute Method Development Services for other information.