CHROMtalks - Navigating Equilibration and Re-equilibration in HILIC

Equilibration and re-equilibration in hydrophilic interaction liquid chromatography (HILIC) play a pivotal role in ensuring reproducible and robust separations - yet they remain widely misunderstood. This presentation discusses the underlying mechanisms of water layer formation on polar stationary phases and explains how column chemistry, mobile phase composition, temperature, and flow rate influence equilibration dynamics. While full equilibration is essential for isocratic methods, partial equilibration proves surprisingly effective for gradient and multidimensional HILIC. Backed by recent research, this discussion offers strategies to accelerate equilibration without compromising performance - empowering analysts to optimize throughput, enhance reproducibility, and unlock HILIC’s full potential behind chromatographic retention and selectivity.