Wednesday, October 2, 2019

How to Decrease Retention Time in HPLC

The retention time is one of the most important measurements in chromatography, as it can impact on the analysis. There are several ways to increase or decrease the retention time in chromatography which depends on the nature and properties of the compound.

The high-performance liquid chromatography (HPLC) is a separation technique of analytical chemistry which used for the qualitative and quantitative determination of compound. The analytes of a sample mixture need to be separated properly and we guess the peak symmetry by the retention time.

The retention time (tR) is the time taken by an analyte to elute from the column and detected by the detector (such as a PDA, UV/VIS), which depends on various factors like, mobile phase composition, length of the column, the pH of the mobile phase and mainly flow rate, etc.

The molecules of the sample mixture have different retention times at a particular composition of the mobile phase and this is because of their affinity to the HPLC column or stationary phase.

The reverse-phase HPLC uses a polar mobile phase and the non-polar stationary phase, therefore the more polar analyte is rapidly separated from the HPLC column since the non-polar samples interact more with the stationary phase.

There are many ways to decrease the retention time in reversed-phase chromatography (RP-HPLC), you can decrease the retention by adjusting the polarity of the mobile phase, reducing the column length, increasing the column temperature, using a smaller particle size column, changing the pH value of the mobile phase, using the higher organic concentration and also mainly by the increasing flow rate of the system you can reduce the retention time in HPLC.


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