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The Isolera™ Step Gradient Explained

18 November 2014

Up to six compounds can be easily separated with an automated step-gradient optimizer embedded in Spektra software for Isolera™ flash chromatography systems. 


For many years flash chromatography has been performed using isocratic elution, linear gradients, or a combination. Often the elution methods are based on previous experience, gut-feel, or generic default gradients, while sample loading usually follows the 1% rule (1% of the cartridge media mass) resulting in less than optimal purifications. 

The patented TLC to linear gradient algorithm uses TLC data to create a linear gradient and estimate sample load for up to three compounds. But things just got even better. Software developed for Isolera™ Spektra flash chromatography systems allows optimizing the purification for up to six compounds using as few as two TLC separations to create a step gradient. Results from a proof of concept study clearly show Isolera Spektra automated TLC to step gradient reduces purification costs in several ways:

  • Increased sample loading per cartridge reduces the number of required cartridges or cartridge size required
  • Less time needed to purify therefore more productivity
  • Less solvent consumed reducing purification costs

Using the TLC to step gradient is simple and fast requiring only a minute or so to enter the data into Isolera and creating the method. The methods are scalable to larger cartridges, increasing purification savings even more.

Actual step gradient purification of 200 mg of the 5-component crude sample.

Actual step gradient purification of 200 mg of the 5-component crude sample. The separation required only 11 column volumes or 8 minutes to complete with better overall separation than the generic linear gradient.