Preparative HPLC
HPLC is used to separate and refine high-purity target compounds from a mixed solution after a synthesis reaction or from natural extracts. An HPLC preparative system must offer different capabilities from a normal analysis system. It is used to fraction high-purity (and in some cases large quantities of) compounds required for subsequent evaluation, analysis, and processes in the shortest possible time. This page provides the knowledge required to select an appropriate preparative HPLC for the intended purpose and introduces various types of HPLC preparative system.
1 Setting the Fraction Volume Target
The instrument and column selection is significantly affected by the fraction volume required. Investigate preparative HPLCs by determining the minimum fraction volume required.
2 Scaling Up
• Checking the Eluent Composition
• Basic Principles of Scaling Up
• Selecting the Instrument and Column
Introduction of Preparative Systems According to Purpose
| Fractioning for structural analysis Purification of biochemicals Purification for product manufacture |
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Fractioning tens of mg Conventional HPLC (repeated fractioning operations) Fractioning hundreds of mg Semi-Preparative System Fractioning several grams Large-Scale Preparative System |
| Fractioning at high purity (Check molecular weight, if possible.) |
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Enhanced purification efficiency using ELSD or LCMS LCMS/ELSD Preparative System High separation equivalent to increasing the column length Preparative Recycle System |
| Low-cost fractioning | |
Results equivalent to multiple columns from a single column Preparative Recycle System |
| Investigating conditions and testing purity with a single system | |
Analysis and fractioning with a single system by simply switching the flow line Analytical/Preparative Automatic Switching System |
| Rapidly checking fractioning results for synthesized/purified products | |
Checking the relationship between the samples, fractionation results (chromatograms, spectra), and fraction vials at a glance Open Solution |
* Don't forget measures to handle solvent leaks.
A semi- or large-scale preparative system uses a lot of solvent, so that measures must be taken to cope with liquid leaks. For example, position a tray below the plunger head to cope with liquid leaks from the pump plunger seal. Take special care if using a flammable solvent. Recommended measures include not positioning equipment that is a potential ignition source nearby and grounding the drain tank to prevent ignition due to static electricity.
