Liquid Chromatography Blog

Separation Science Solutions Series

Recent Posts

Just Enough Sample Preparation

Sample preparation is an integral part to successful chromatographic analysis. Selecting the right sample preparation technique is dependent upon...

The Basics of HILIC

How can we describe the mechanism at the basis of HILIC retention? We will answer this question and give you an insight into the main interactions that govern chromatographic separation in HILIC mode.

Do you HILIC?

Are your analytes eluting close to the void volume of a reversed-phase (RP) column? Do you have problems retaining organic molecules under conventional reversed‑phase liquid chromatography (RPLC) conditions? Have you increased the initial...

HILIC & MS

The separation power of HPLC combined with the very high sensitivity and selectivity of MS detection is widely used to develop analytical methods that are required to achieve ultra-low detection and quantification limits. Examples of such...

Sample Solvent in HILIC

The sample solvent, also called dissolution solvent or injection solvent, is the solvent used to prepare the solution in which the compound to be analysed by HPLC is dissolved. The choice of the sample solvent is fundamental for a successful...

Column Equilibration in HILIC

It is matter of fact that the samples most chromatographers have to face are rather complex for sample number and compound variety. In these situations, gradient elution is largely preferred for the separation of analytes with different retention...

Column temperature in HILIC

Mostly in HILIC conditions, by increasing column temperature a decrease in the retention time is observed.

HILIC Mobile Phase: pH

The pH of the mobile phase should be selected in order to enhance the hydrophilicity of the analytes.

HILIC Mobile Phase: Salt Content

The addition of salt to HILIC mobile phases is strongly recommended to improve reproducibility and peak shape. Do you know how to solubilize the salt in mobile phases at high acetonitrile content?

HILIC Mobile Phase: Solvents

In HILIC, small variations of the elution conditions often result in differences in the retention greater than the ones observed in RPLC. Therefore, particular attention has to be paid to the selection and the preparation of the eluents. 

HILIC Stationary Phases

A common and useful classification of the HILIC stationary phases is on the basis of the presence of functional groups on the surface and their charged state: they can be divided in unbounded and bonded phases, and these ones are classified in...

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