LC Columns

Q: What is the size of LC columns?

The size of an LC column varies based on its intended use. Analytical columns typically measure 4.6 mm internal diameter (ID) and 150–250 mm in length. Preparative columns have 10–50 mm ID for bulk compound isolation. Ultra High-Performance Liquid Chromatography (UHPLC) columns use smaller IDs, such as 2.1 mm, with shorter lengths for high efficiency.

Q: How do you flush an LC column?

To flush an LC column, use compatible solvents like methanol, ethanol, or acetonitrile. Select a low flow rate and flush overnight. Avoid abrupt polarity changes to maintain performance. Water can also be used but may impact column retention over time. After flushing, reconnect the column and run an analysis to ensure the method's success.

Q: How to regenerate an LC column

If flushing fails to remove contaminants, regeneration may involve backflushing or using stronger cleaning solvents. Always refer to manufacturer-specific protocols, such as those from Shimadzu, to avoid damaging the column's stationary phase.

Q: How to condition an LC column

To condition an LC column, flush it with a mobile phase or suitable solvent, running five to ten column volumes. Confirm stable pressure and a consistent baseline to ensure optimal performance and reproducibility before proceeding with analysis.

Liquid chromatography (LC) columns are invaluable additions to labs servicing a range of industries and applications. You’ll find LC columns utilized across the following industries:

Pharmaceuticals: LC columns are used to facilitate the separation and quantification of drug compounds for impurity profiling and bioanalysis.
Environmental analysis: LC columns are invaluable in the detection of contaminants in water, soil, and air, including pesticides and heavy metals after derivatization.
Food and beverage manufacturing: LC columns are used in the analysis of preservatives, flavor compounds, sweeteners, and contaminants to maintain quality control.
Chemical industry: LC columns are integrated in product-purity assessment, process monitoring, and raw material testing of chemicals.
Forensics and toxicology: LC columns are essential in the detection of drugs, poisons, and metabolites in biological samples.
Academia and clinical research: LC columns are an important component of method development, compound identification, and pharmacokinetic studies.

Column Types

Shimadzu offers a diverse lineup of LC columns tailored to analytical needs, including:

Reversed-phase (C18, C8, C4, phenyl, PFPP), industry-standard for non-polar compounds (e.g., Shim-pack Scepter, NovaCore, GIS).
Normal-phase/HILIC for polar analytes (e.g., Shim-pack Scepter HILIC, Shim-pack Arata).
Ion exchange for charged molecules (e.g., Shim-pack Bio IEX, ISC, ISA, WAX/WCX series).
Size exclusion for large biomolecules (e.g., Shim-pack Bio Diol).
Hydrophobic interaction chromatography (HIC) – for antibody-drug conjugates and protein variants (Bio HIC).
Preparative and microscale columns – high-load capacity for method development/prep (Shim-pack PREP, MC series).

FAQs

What is LC in column?

Referring to the analytical process of liquid chromatography, an LC column is a key part of many lab processes. An LC column is a tube used to separate the compounds of a sample mixture for identification. Making up the stationary phase of the analytical process of liquid chromatography, an LC column is the component where the actual separation occurs – allowing the identification and qualification of the sample’s molecular structure.

What is the size of LC columns?

The size of an LC column varies based on its intended use. Different analytical approaches require different-sized LC columns because their size impacts the pressure, resolution and sample load of each analysis. You’ll find LC columns in the following dimensions across these applications:

Analytical

4.6 mm internal diameter (ID), 150–250 mm length.

Preparative

10–50 mm internal diameter for bulk compound isolation.

Ultra High-Performance Liquid Chromatography (UHPLC)

Smaller internal diameter, e.g. 2.1 mm and shorter length for high efficiency.

How do LC columns work?

LC columns represent the stationary phase of the LC process, whereby the sample is separated to allow for the identification of all its components. It is, in fact, the way that the different compounds in the sample mixture react to this stationary phase, based on polarity, charge or size, that allows separation. And, because the components elute at different times, the equipment is able to identify and quantify the molecular structure of the sample to be mapped.

How do you flush an LC column?

Flushing your LC column is not only necessary when there are contaminants present, but doing so frequently will help prolong the instrument’s lifetime. To flush an LC column, you will need to use compatible solvents. Methanol, ethanol and acetonitrile are frequently used solvents in LC flushing, but the concentration and approach will vary based on each column’s application and dimensions. Water can also be used, but it can impact the retention of LC columns in the long term. To flush the column, turn on the pump, select a low flow rate and flush overnight. Then, reconnect the column in the normal direction, performing an analysis to confirm your method has been successful. If you need to change the solvent used for greater efficiency, do so slowly, avoiding abrupt polarity shifts to prevent carryover and maintain performance.

How to regenerate an LC column

Regenerating an LC column is required when the process of flushing has not been successful in removing contaminants. If the LC column is able, backflushing can be appropriate, where you’ll reverse the flow of the mobile phase. Otherwise, the column may benefit from the use of stronger cleaning solvents. It’s also essential to refer to Shimadzu’s manufacturer-specific protocols if regeneration is required to restore performance without damaging the stationary phase of the LC column.

How to condition an LC column

If your LC column has come from storage or is brand new, it’s important to condition it before use to prepare it for optimal analysis. LC column conditioning involves flushing with mobile phase or an appropriate solvent, running five to ten column volumes and checking for a stable baseline and pressure throughout analyses. This is an essential step in ensuring reproducibility and performance.

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