high performance liquid chromatography

The presence of unusually broad peaks is sometimes an occurring problem in liquid chromatography separation. This problem is regularly seen in isocratic separation; however, it can also occur with gradient elution. The peak broadening in HPLC is influenced by the flow rate of the mobile phase and particle size of the columns packing material. The smaller the particle with the latter, the better is the peak resolution. It is recommended that the chromatographers can try and optimize the composition and flow rate of the mobile phase and check the type of column used.

The peak broadening in high-performance liquid chromatography is due to several factors mentioned below.

  • In a very short time, several steps of gradient elution are changing from polar to non-polar.
  • Some precipitate in the sample solution.
  • Overloading of column (too much sample load)
  • Too long Retention time (RT)
  • Extra-column volume too large
  • Detector time too slow
  • Poor solubility of the sample
  • Incompletely filled sample loops
  • Saturation of the Detector
  • Contaminated column
  • Incorrect pH of the buffer or mobile phase 

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