HPLC 2018: PAT, Pot, Performance and Potential

The 47th International Symposium on High Performance Liquid Phase Separations and Related Techniques was held at the Marriott Wardman Park in Washington, DC, from July 29th to August 2nd.  The conference hosted 870 attendees from 34 countries and 203 talks.

 

New Products

While the scientific program formed the heart of the meeting, there was also activity at the booths of the exhibition hall. Several new products—including instruments, columns and consumables—were introduced. The most prominent new products in the exhibition were from Shimadzu and Activated Research Company (ARC). Shimadzu introduced the Nexera Bio, a UHPLC system designed specifically for biomolecule analysis. In the past, traditional HPLC has been unsuitable for protein analysis, due to adsorption onto stainless steel surfaces and the high salt condition in the mobile phase that causes corrosion. Nexera Bio tackles that problem by utilizing metal-free components for its wetted surface, joining other similar products on the market for these challenging samples. The new system features a carbon-coated pump head to prevent solvent contamination due to surface activity. It also has a ceramic injection needle to minimize sample carryover. Shimadzu launched Nexera Bio a week before HPLC 2018, and it is now ready to be shipped in the US.

ARC introduced a new detector for HPLC, the new Solvere Carbon Selective Detector, which utilizes flame ionization technology. Usually associated with GC detectors, ARC has modified the technology for use in HPLC. With its proprietary ActiveDiffusion technology, the detector system removes the solvent from the column effluent, and the remaining carbon analytes are then transferred into the FID for detection. The Solvere Carbon Selective Detector will be ready for shipment by October.

 

Technical Sessions

In addition to new products introduced in the exhibit hall, there were numerous technical sessions during the four-day conference. The sessions covered breakthrough technologies, applications and improvements of HPLC and related analytical techniques. Common trends observed among the sessions include HPLC applications in biomolecule analysis, online HPLC for pharmaceutical Process Analytical Technology (PAT) applications, multiple-dimension HPLC, chiral molecule analysis, performance-improvement techniques, and cannabis and drug testing applications.

One well-attended session relating to PAT, presented by Eli Lilly and Company, showcased the company’s in-house developed solution for automated process sampling, sample preparation and online chromatography. They indicated that by utilizing a mobile dilution cart with two valves between the sample preparation and process lines, PAT data management produced live chromatographic data on pharmaceutical production processes, facilitating real-time processing decisions. Other sessions presented online chromatography applications in continuous manufacturing of pharmaceuticals.

Several sessions covered many aspects of multi-dimensional HPLC. Building upon two-dimensional HPLC technology, which was was initially employed to characterize unknown peaks and identify peak purity, 4D LC/MS is now possible, as presented by Robert Kopf, PhD, from Roche. His experiment on monoclonal antibody analysis using 4D LC/MS showed improvements through online reduction and digestion of fractionated peaks, which resulted in an efficiency gain of more than 80%. Despite its powerful analyzing power, method development for multiple dimensional analysis remains costly and time consuming, due to the intricate setup and difficulty in optimizing running conditions. Bob W.J. Pirok, PhD, from the University of Amsterdam suggested the employment of artificial intelligence software to provide faster 2D HPLC method development based on data from past chromatographic data and retention patterns.

A presentation from Genentech illustrated the advantages of using size exclusion chromatography as the first chromatographic method, as it provides a useful separation with minimal chemical effects on the analytes of interest, which can then be followed by a second technique. This strategy was discussed in the context of new therapeutic modalities that are beginning to cross the boundaries of small molecule and large molecule, which require new tools and collaborative hybrid teams in order to properly characterize. While 2D separation techniques were clearly a strong focus of development, Sebastiaan Eeltink, PhD, of the Vrije Universiteit Brussel showed the current state of his lab’s ongoing research into developing microfluidic devices for 3D separations.

Enantiomeric separation of chiral molecules was another big topic at HPLC 2018, as several presenters covered the subject in the technical sessions. Erik L. Regalado, PhD, from Merck presented on the general trend towards a universal chromatographic method in the pharmaceutical industry. Ultrafast chiral chromatography is one of the most heavily researched areas in the industry, as more than half of available drugs are chiral with multiple chiral centers. Genentech has also tapped into this field with their rapid chiral compound separations with multiple heart-cutting 2D HPLC. The first dimension analyzes achiral compounds with six columns in parallel, while the second dimension further separates the chiral compounds found in the first dimension.

HPLC performance improvements were also a popular topic at the show, covering various aspects from throughput to stationary phase architecture. Fabrice Gritti, PhD, from Waters discussed the dispersion effect caused by column frits in high-throughput gradient LC and suggested a redesign of packed column frits to improve peak capacity.

Adam Socia, PhD, from Merck presented on techniques for consistent retention time in drug manufacturing. Interactions between the stationary phase and SDS (sodium dodecyl sulfate) or other excipients usually cause a shift in retention times. A suggested solution to this problem is to increase the pH of the mobile phase to minimize such interactions. Another session by Roche studied the improvement of monoclonal antibody separation using large pore superficially porous particles and ultra-high-performance hydrophobic interaction chromatography.

Last but not least, the use of HPLC in cannabis testing was covered in several sessions, as this particular application has been growing rapidly in recent years. There was a specific short course dedicated to cannabis analysis at the conference. In one of the sessions, Avinash Dalmia, PhD, from PerkinElmer presented a study of pesticide residue analysis in cannabis as regulated by California and Oregon. Among the largest cannabis markets in the US, these two states have been providing the most rigorous standards and detection limits. Analyzing pesticide residue at 100nppb to fulfill California’s state limit is possible using LC/MS coupled with an APCI source. Analysis of nicotine and illegal drugs were also covered in other sessions by utilizing chiral separations and multiple injection methods.

In 2019, two HPLC conferences are scheduled by the organizers: Milan, Italy, in June and Kyoto, Japan, in December. The following year, HPLC 2020 will be held in San Diego, California.

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