The Chromatography Forum

of Delaware Valley

The theme of this year’s virtual event is Advancing Chromatorgaphy in Industry . 

The event will take place over the lunchtime hour of 12:00 – 1:00 p.m. on April 13, 14, & 15, 2021.

There is no cost for the event this year due to its virtual nature so this is a perfect time to enjoy the science being shared, as well as get an inside look at the CFDV, one of the longest running chromatography organizations in the country.

11:45 am Event log-in opens
11:50 – 11:55 am Welcome message from CFDV
11:55 am – 12:00 pm Waters presentation
12:00 – 12:30 pm “Hydrophilic Interaction Chromatography in Media Analysis” Ying Zhou, Teva
12:30 – 1:00 pm “Color and Trace: Chromophores in Protein Therapeutics” Hangtian Song, BMS

“Hydrophilic Interaction Chromatography in Media Analysis”
Ying Zhou, Teva

Abstract: Hydrophilic interaction chromatography (HILIC) coupled to mass spectrometry (MS) is widely used in metabolomics research to detect polar/ionic metabolites in biological samples. Due to the complexity of sample matrix, the quantitation usually requires highly sensitive MS such as triple quadrupole with targeted MS/MS scanning for specific analyte quantitation. In this presentation, we discuss the feasibility of implementing HILIC-MS using a single quadrupole, benchtop mass detector for spent media analysis in biopharmaceutical process development environment. The discussion focuses on method development considerations for simple and fast amino acid quantitation. This HILIC-MS method is compared to a commercial, derivatization-based amino acid quantitation kit. Result indicates that this HILIC-MS method is able to detect the same trend in bioreactor daily samples without the need of derivatization, and it demonstrates better sample-to-sample consistency compared to the commercial kit.

“Color and Trace: Chromophores in Protein Therapeutics”
Hangtian Song, BMS
Abstract: Discoloration of protein therapeutics has drawn increasing attention due to safety concerns and need for improved analytics that is essential for well-controlled manufacturing processes. From past experience, many factors have been correlated to discoloration. However, two issues are still common during color investigation. One is the lack of systematic approaches. The other is, after a root cause has been identified, how to prove it is the major cause. In this study, a widely applicable workflow was developed for identifying sources that generating colors in biologics. First, excipient chromophores were identified by UV–vis spectroscopy, reversed-phase chromatography and mass spectrometry. Then, chromophores on the protein primary structure was located by peptide mapping with UV–vis photodiode array (PDA) detection and mass spectrometry. Finally, by a semi-quantitative total color scale, contribution from each source of color is estimated. As examples, the investigation of yellow discoloration of two protein therapeutic will be presented. Tryptophan oxidation and vitamin B12 were identified as the primary root causes.