Advances of 2 mm I.D. Monolithic Silica Columns for the Rapid, Sensitive and Highly Efficient Separation of Small and Large Molecules in UHPLC

Petra Lewits, Merck KGaA, Darmstadt, Germany (petra.lewits@merckgroup.com)
Cory Muraco, Milliporesigma, Bellefonte, Usa
Benjamin Peters, Merck Kgaa, Darmstadt, Germany

The performance of HPLC columns has improved dramatically in terms of separation power by a reduction in particle size. However, not all samples benefit equally from this development: clean samples with a relatively simple matrix are ideally suited for sub-2 µm UHPLC columns. But for samples with a more difficult and complex matrix, the benefits are often accompanied by significant limitations which results in increased need for sample preparation.

Monolithic HPLC columns have significant advantages over particle packed columns in that they can be run at higher flow rates with minimal backpressure and that they are resistant to matrix effects from samples.  These columns have been used for rapid separations of small molecules, and more recently, in separating larger, biomacromolecules. In addition to expanding the utility of these columns into large molecule separations, advances in the overall architecture of the monolithic silica skeleton and geometry of the monolithic column have led to increases in the efficiency of this type of column.

Monolithic 2 mm I.D. columns overcome the limitations of typical UHPLC columns packed with small particles due to their robust bi-modal silica structure.  This unique feature allows for cost and time efficient HPLC analysis of matrix-rich samples with simplified sample preparation and excellent separation power for small and large molecule analysis as well as rapid, UHPLC-like separations at significantly lower column backpressure.

This seminar will discuss these advances in monolith design from both a fundamental level and from an application standpoint.  Utilizing these modified monolithic 2 mm I.D. HPLC columns, application examples for rapid, sensitive, and highly efficient separation for both small and large molecules will be showcased, demonstrating the ability of these columns to separate a broad range of compounds.

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Short Biography of Presenting Author

Petra works with Chromatography for more than 35 years. She started her career in separation science in 1983 using several chromatographic methods, including HPLC, TLC, GC and Solid Phase Extraction (SPE), for food control in a governmental institution lab.

1986 she joined Merck´s pharmaceutical R&D and QC department (Merck KGaA in Darmstadt, Germany) were she development HPLC methods for R&D and QC of pharmaceutical API´s and formulations as well as for diet convenience foods.

From 1991 on she supported customers in different positions with technical and application related inquires in Europe as a sales specialist for HPLC instruments, technical support for Chromatography and complaint management.

Since 2002 she is a Global Product Manager for Analytical Chromatography (HPLC). From 2014 she was focusing on Thin-Layer Chromatography (TLC). Since April 2018 Petra has the global responsibility for analytical HPLC columns at Merck.