|Articles|September 9, 2019

Exploring the Effect of Mobile Phase Concentration on Sensitivity of Size Exclusion-Mass Spec (SEC-MS) for Monoclonal Antibodies using a bioZen™ SEC-3 Column

In this application note we look at the impact that mobile phase concentration has on the sensitivity of SEC-MS when used to analyze intact mAb’s under non-denaturing conditions.


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Figure 2. Split injection used to measure triplicate test of MTBE, hexane, o-xylene, and 1-methylnaphthalene comparing different solvents (methanol, methylene chloride) and column insertion distances starting with 0.5 mm, 5.0 mm, 10.0 mm, 15 mm, and 20.0 mm. Methanol had the best performance at the 5.0 mm insertion distance and methylene chloride looked slightly better at the 0.5 mm. We would still recommend not going below the 5.0 mm manufacturer recommended insertion distance.
Revisiting insertion distance for methanol and methylene chloride in GC analysis and confirming that insertion depth strongly affects response and reproducibility.