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液相色谱柱的故障诊断与维护
作者:未知 来源:八方世纪科技有限公司 时间:2007-1-8

    This is caused by a column void forming at the head of the column or a blocked frit.  This happens over time as the column begins to fail, but can happen more quickly if the flowrate and pressure are always very high.  This causes the column bed to settle and a space to form.  Contamination or incompatible mobile phases can also cause this.
    Once upon a time there was a chromatographer new to the area of chiral separations.  He injected multiple samples onto a preparative scale, $25,000, chiral column.  These samples he dissolved in methylene chloride because he could not dissolve the in the methanol/hexane mix of the mobile phase.  After the first separation and purification, the column performance began to fall.  By the fifth sample, there was no stationary phase left.  This particular phase was soluble in methylene chloride.  The moral of the story, check the vendor specifications for the phase to ensure that the column is dying an untimely death.
    If there is only on peak that is doubled, there may be a new partially coeluting compound in the mix.  Diode array detection or MS is useful for this determination.


     Peak broadening is another common peak shape problem.
    If all of the peaks become broadened, this could be a problem with column efficiency and a loss of this efficiency, possible due to a column void.
    Large injection volume or extra column volume can lead to dispersion and peak broadening.
    The third possible cause if all peaks are broader, is high viscosity in the mobile phase.  This condition of a viscous liquid also causes dispersion and band broadening.
    If only one or some peaks are broad, it could mean that this or these peaks are from a previous injection and are very late eluting compounds from the previous analysis.
    Also, high molecular weight proteins and polymers tend to produce broad peaks.
    In a complex mix, some compounds with various isomeric forms that tautomerize or flip back and forth, will produce broad peaks.
    Amines also may give this peak shape in some cases.


     Tailing peaks have a symmetry value greater that 1.2.
    When only some peaks tail, it may be caused by a secondary retention effect such as residual silanol interactions.  A based deactivated, a highly carbon loaded, or a polymeric base material column may solve this problem.
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