high performance liquid chromatography, broad peaks

¿How to increase peak response in HPLC?

As we know that the high-performance liquid chromatography is a separation technique in analytical chemistry. In HPLC the system suitability parameters such as tailing factor, theoretical plate number, and resolution are most significant. The peak response is also important because the peak area is used to determine the concentration of molecules. Response factors are significant while using HPLC for quantitative analysis.

There are different types of detectors are used in HPLC such as UV, PDA. The output signal of the detector is a function of the concentration of molecules passing through the detector cell.

Here are some points that help to increase the peak response.

  • Injecting more amount of samples or higher concentrations of the sample into the HPLC injector is one of the simplest techniques to increase the peak response.
  • The wavelength of the component should be confirmed using a UV/VIS spectrophotometer since if we select a correct wavelength it can increase the peak response in HPLC.
  • Peak volume is associated with column volume therefore a smaller column will also decrease peak volume and proportionally increase the peak height. An easy technique to decrease column volume is to decrease the diameter of the column and keep other column properties the same. An additional way to decrease the peak width is to use a column with a higher theoretical plate number. This is most easily done by particle size reduction.
  • Increasing the flow rate of the system will reduce the detector response therefore the flow rate should be appropriate.


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