Care of HPLC Columns

A HPLC column is the most critical component of theHPLC.system. Quality of your results will greatly depend on its performance. Though it is possible to monitor the HPLC column condition by keeping track on number of theoretical plates you are advised to look for other tell-tale signs such as :

  • 峰形状的劣化
  • 保留时间的变化
  • Sudden increase in backpressure
  • 峰之间的分辨率差
  • 鬼峰的出现
HPLC.Column
HPLC.Column

通过遵守处理,使用和存储的建议指南,可以扩展HPLC列的使用寿命

Handling

  • 避免掉落和其他机械冲击。冲击通常会导致渠道,这可以有助于尾尾
  • Operate the HPLC column within recommended pressure limits. Excessive pressure leads to same problems as mechanical shocks
  • 仅使用兼容的螺母,套管和配件。不相容的配件会导致柱螺纹泄漏和损坏
  • 永远不会拧紧结束配件
  • 永远不要让仪表用作搅拌杆,用于混合移动阶段和缓冲液。

采用

  • Install the HPLC column so that the flow direction matches the direction of the arrow
  • 使用0.45μm过滤器在使用之前,移动相应该是脱气和过滤。此外,在流动相储中使用在线滤波器。
  • 用流动相平衡柱,其与制造商储存溶剂(异丙醇)兼容。使用至少5-10卷平衡移动阶段
  • 在存储冲洗柱之前以去除任何残留的缓冲或离子配对剂。这对于防止干燥时的降解或缓冲盐沉积至关重要。
  • 三氧化硅柱的推荐pH范围为2.0 - 7.5。一些列允许从pH 1-11的范围内,但是对于安全操作和长柱的生活,您应该尽可能使用它在2.0-7.5 pH范围之间
  • 当更换流动阶段时,溶剂的选择应在极性方面是渐进的。避免变化或极端梯度。
  • 如果过度的背部压力开发,可以使用反向冲洗
  • 在注射之前通过0.45或0.2μm过滤器过滤所有样品
  • 注射器和分析柱之间的用户保护柱,以防止污染物和颗粒到达柱
  • 采用mobile phase for dissolving the sample or if it that is not possible then another solvent or a mixture of solvents of compatible polarities should be used
  • Never exceed the prescribed temperature limits and for consistency of results use a column oven

贮存

  • HPLC柱应在环境温度条件下储存,从未在冰箱或烤箱中储存
  • 存储条件取决于使用之间的时间间隔:

可以使用几小时储存到过夜 - 与上一次分析相同的溶剂。

Medium interval storage (two days to weekend ) – flush with pure water to prevent microbial or mould growth.

长期储存 - 使用不应包含超过50%的水的洗脱液。

  • 使用使用后不建议使用肥皂或洗涤剂溶液。
  • Strong acids or alkalis cannot be used as they will result in total irreversible damage.
  • During storage end plugs should be tightly fitted to prevent solvent evaporation leading to drying of stationary phase

We hope you found the guidelines useful for getting good quality results and increasing useful life of your column. Please share any experiences that you may have by leaving your comment.

Related Articles

回应

您的电子邮件地址不会被公开。必需的地方已做标记*

  1. 关于柱子的条件的情况是什么在生物分析物中使用
    because this columns must be stored between 4 and 8 °c

    谢谢
    nb : excuse my anglish it is not verry good

    1. Hi Farida, You are absolutely correct! GPC and many columns used in analysis of large molecules do ned to be stored at lower temperature.

  2. “中间间隔储存(两天至周末) - 用纯净水冲洗以防止微生物或霉菌的生长”水如何水解微生物生长?特别是室温,相反,有机溶剂防止微生物生长。

small_c_popup.png

不要被遗漏!

over 20,000 scientists read our weekly Newsletter!

Baidu