Factors Affecting Retention Time in HPLC

Retention time is the time which is spent by a molecule on the stationary phase after injecting. Retention time is one of the important parameters in chromatography because it is used to describe the chromatographic behavior of a sample component in a column. Several factors can affect the retention time (Rt value) of the molecules and can impact the analysis.

Chromatography has the peaks of separated components and peaks have the particular retention time, area, height, HPLC works on the UV or PDA detectors, it’s also showing the results in peaks. The time required to travels the component to reach the detector through a column is known as its retention time. This time is calculated from the time at which the sample is injected into the time when the chromatogram shows a maximum peak height for that analyte. Different analytes have dissimilar retention times, for a specific compound, the RT will vary it depends on the different factors.

Here are some points of the factors that influence retention time in high-performance liquid chromatography.

• Insufficient saturation of column
• Mobile phase composition
• The polarity of the mobile phase
• The viscosity of the mobile phase
• The polarity of the sample
• Type of column used
• Column length
• Column degradation
• The overloading of HPLC column
• The dimension of the column
• The flow rate of analysis
• The pH of the mobile phase
• The pH value of the sample

This means that if you are using retention time as a way of identifying compounds, then the chromatographic conditions need to be carefully controlled.

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