Afraid of HILIC? Watch These Videos and Overcome Your Fear!

Hydrophilic Interaction Liquid Chromatography or HILIC is a powerful tool to analyze polar compounds that have limited retention on typical reversed-phase columns. The power of HILIC can be utilized across multiple markets including environmental, food safety, and clinical diagnostics. But to be honest, it can be challenging to implement a HILIC method and more considerations have to be taken compared to the development of a reversed-phase LC method. There are many different types of HILIC columns on the market, but no matter what type of HILIC columns you use, there are some basic principles you need to know to not only save time, but most importantly, be successful! We have made a series of 4 short videos to give you a quick look at HILIC method development.  Hopefully you will be more confident to develop your own HILIC method after watching them!

Video 1:  Understanding the HILIC Separation Method in LC

In the first video, we talk about the separation mechanisms of HILIC and a variety of ways polar analytes can interact with HILIC stationary phases. The good news is that the solvents used in HILIC methods are similar to those used in reversed-phase LC methods! For most of the LC-MS/MS analysis, HILIC methods have significant increases in instrument sensitivity because it’s operated under higher organic solvent conditions which aids in desolvation.

Video 2: Conditioning Your LC Instrument and Column for HILIC

Don’t be surprised when you see inconsistent performance (including retention time and sensitivity) from your new HILIC column upon the first few injections. This video will tell you how to condition your LC instrument and HILIC column especially when you are switching LC systems from reversed-phase to HILIC analysis. Remember that all types of HILIC columns are not built equally, and the conditioning time can vary depending on your analytes and analytical conditions.

Video 3: Buffer Choices for HILIC

Since pH plays a critical role in the performance of a HILIC method, the first step for method development is to determine a suitable pH for your mobile phases. In this video, we explain how to incorporate pH into your HILIC method development and how to fine-tune the buffer concentration in order to obtain optimal peak shapes, resolution, and sensitivity.

Video 4: Injection Solution Conditions for HILIC

During the process of method development described in videos 2 and 3, you also want to pay attention to the injection solution since it can have a great impact on the performance of your HILIC method. Matching the aqueous/organic ratio of the injection solution to the initial mobile phase conditions is the best practice for HILIC-based analysis. Therefore it is important to think ahead to determine if your sample preparation procedure and final sample solution will be compatible with your HILIC method.

So there you have it! Hope you enjoy watching these videos and don’t forget to share your HILIC method development successes with us!

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