Preparative HPLC

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    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
    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.) 
    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 
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    * 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.

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