Kromasil^® notes

Separation of fish oil esters EPA and DHA on Kromasil C18

A nice scale-up example from column i.d. 4.6 to 21.2 mm.

Below are chromatograms illustrating the separation of fish oil esters on two Kromasil 100-10-C18 columns, 4.6 and 21.2 mm, respectively. On the two columns, both an analytical (left chromatogram) injection and an overloaded (right chromatogram) injection are performed.

The figures show a nice scalability from the 4.6 to the 21.2 mm columns. The scaling factor between the two columns is ~21 and the flow rates and loadings are scaled accordingly.

• Conditions

• Stationary phase: Kromasil 100 Å, 10 µm, C18
• Columns: 4.6 × 250 mm and 21.2 × 250 mm, respectively
• Sample: Crude containing eicosa- pentaenoic acid (EPA) and docosahexaneoic acid (DHA)
• Mobile Phase: methanol/water
• Gradient: 0-20 min 95% methanol, 20-30 min 100% methanol
• Flow rate: 1 ml/min and 21 ml/min, respectively
• Detection: UV @ 220 nm