Which tips and tricks for two step purification are available?

Know your method!

For good results and a successful two step purification the methods should be well established to ensure excellent purification results. You should know the elution volume/peak of the first purification to guarantee that the first peak is stored in an appropriate vessel. A vessel that is too small will result in sample loss as some parts of the sample is transported to the waste. A vessel to big will result in sample dilution, which can affect the second purification step. An option for medium up to high sample volumes is the use of variable sample loop. These variable sample loops can be emptied completely or partially as well as filled completely or partially. This allows you to work very flexible and easily switch between different sample sizes.

Use automatic sample loading!

For sample loading by a pump, an air sensor can be used to detect the end of your sample. This protects the column from damage by running dry and supports more importantly automatic sample loading. Upon air detection, various functions can be programmed. In the case of automatic sample loading once air is detected the software will continue with column washing and elution.

Measure the pressure!

Use Pressure Control that allows you to determine the pressure difference over your column bed. The first pressure sensor measures the pressure before the column, while the second sensor measures the pressure after the column. The software PurityChrom® automatically calculates the pressure difference over the media bed. If the pressure differential value (DP) exceeds the preset limit during the run, either the run pauses or another action is applied. Thus, you can protect your valuable columns and media from overpressure.

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Based in Berlin, KNAUER has been serving the sciences since 1962. We develop and manufacture scientific instruments of superior quality for liquid chromatography systems and components, including:

Analytical HPLC / UHPLC
Preparative HPLC
Fast protein liquid chromatography (FPLC)
Multi-Column Chromatography / SMB

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