Best practice for separation columns in ion chromatography (IC) – Part 1

The high performance ion chromatography (IC) separation column is often referred to as the «heart» of the ion chromatograph. The reason for this denomination is straightforward: the column is responsible for the separation of the analytes of interest from each other as well as from interfering sample matrix ions. The unique separation capabilities of IC columns allow the determination of multiple analytes within one run. In this blog series, we will share what is required to ensure the proper operation of an IC column and how to maximize the column lifetime.

Click below to jump directly to a topic:

• Standard operating conditions
• Equilibration
• Operating limits
• Information leaflet
• Storage
• Column guards
• Pulsation absorber
• Chemical quality
• Sample preparation
• iColumn features
• Column shelf life

Standard operating conditions

To begin with, the standard operating conditions should be considered, such as the eluent (mobile phase) composition, the eluent flow rate, the column oven temperature, and the detection method. These standard conditions are specific for each individual column type and correspond to the conditions that work best with the application the column is intended for. Every analytical column sold by Metrohm is delivered with a certificate of analysis (CoA) which is recorded under standard operating conditions.

In the following table you can find the standard operating conditions for three different Metrohm IC columns.

Equilibration

Next to the standard operating conditions, the start-up parameters play an important role for the lifetime of the separation column. High mechanical and thermal stress is a frequent reason for decreased column lifetime. It is therefore recommended to slowly increase the eluent flow rate to the column and to avoid thermal and pressure shocks. For more information regarding specifics, please refer to the recommended equilibration conditions in the corresponding column leaflet.

Operating limits

Based on the different stationary phase designs that are required to achieve different selectivities and guarantee a wide range of IC applications, the chemical and physical properties of the ion exchangers can vary. Therefore, different operating limits are recommended for various column types and set according to stress tests performed during the development of a product. Before beginning to optimize an application, please refer to the respective column leaflet for the corresponding operating limits to achieve optimal results and guarantee a long column lifetime.

The chemistry of the ion exchanger defines the limits of operating temperature, eluent pH, and organic modifiers that can be present in the eluent. These values are valid for every dimension of a column type (e.g., the Metrosep C 6). Exceeding these limits can strongly affect column performance and, in the worst case, lead to irreversible damage. Tolerated flow rates and maximum pressure correlate with physical properties and of course column dimension—therefore these limits are set for every column dimension (e.g., Metrosep C 6 – 100/4.0 vs. Metrosep C 6 – 250/2.0).

Are you searching for a specific IC column for your research? Then check out our Column Finder!

Metrohm IC Column Finder

Information leaflet

In addition to the CoA, a lot of necessary valuable information can be found in the respective column leaflet. Instructions about equilibration, regeneration, operating limits, and much more can be found in the column leaflet which is provided for every IC column type offered by Metrohm.

You can download an example of a column leaflet below where the information is provided in several different languages (DE, EN, FR, and ES).

Download the Metrosep A Supp 17 leaflet

Storage

Depending on the operation conditions and the properties of the ion exchanger material, different storage conditions are recommended for different column types. These conditions (storage temperature and storage eluent) are described on the respective column leaflet and should be strictly followed.

Please note that storing a new column for long time periods as well as frequently switching between storage and operation can actually be more stressful to the column than operating it 24/7! It is therefore not recommended to stock a column for a long time without using it.

Column guards

By default, a guard column with the same material as the column should be used. However, for special applications, combining different ion exchange materials by using a different guard column can help to optimize the separation. One such example is in the case of sulfate and sulfite, as shown below.

Pulsation absorber

With the exception of the Eco IC product line, all Metrohm ion chromatography instruments are equipped with a pulsation absorber. For the Eco IC, it is strongly recommended to add the pulsation absorber as an option. As mentioned earlier, IC columns do not like repeated mechanical stress, especially those based on polyvinyl alcohol or polymethacrylate stationary phases. Therefore, the pulsation absorber is a useful tool to protect the column from possible pressure fluctuations in the system and enhance the column lifetime.

Find out more about the full line of Metrohm ion chromatography products and accessories below.

IC solutions from Metrohm

Chemical quality

In ion chromatography, the ratio between the volume of sample and the volume of eluent that flows through the column is very small, usually in the range of 1:1000. Therefore the quality of the chemical reagents used in the eluent preparation plays a crucial role for the column lifetime. To guarantee optimal performance, use chemicals denominated as «for IC» whenever possible as they are tested particularly on impurities (e.g. metals) that could harm the chromatographic column.

For dilution of the eluent components, ultrapure water is typically used in ion chromatography. To ensure good chromatographic results, the ultrapure water should have a specific resistance greater than 18 MΩ · cm and be free from particles. The ultrapure water is filtered through a 0.45 µm filter and treated with UV. Modern ultrapure water sources for laboratory use guarantee this level of water quality (Type I).

Column shelf life

How many injections are possible on a specific column? Unfortunately, it is not possible to deliver an exact answer to this question. This is because the column lifetime strongly depends on the sample matrix and elution conditions. Due to the multitude of different applications and samples that can be covered with a single IC column, it is not possible to predict the column life for every application and sample type.

During column development, several endurance tests are performed under standard conditions with a guard column using appropriate standards. Under these conditions, the column must withstand at least 2000 injections.

Your knowledge take-aways

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