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Poll If a compound elutes at the same time as the dead time (t₀), what does this indicate?
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No interaction with the stationary phase
15 votes
88.2%
Strong interaction with the stationary phase
2 votes
11.8%
Column void volume
0 votes
0%
High resolution
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HPLC deadtime hplc
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Frequently Asked Questions: HPLC Analysis & Chromatography

High-Performance Liquid Chromatography (HPLC) is an analytical technique used to separate, identify, and quantify each component in a mixture. It relies on a pump to pass a pressurized liquid solvent containing the sample mixture through a column filled with a solid adsorbent material. Each component in the sample interacts slightly differently with the adsorbent material, causing different flow rates for the different components and leading to the separation of the components as they flow out of the column.

Column efficiency is typically measured by the number of Theoretical Plates ($N$). The most common formula is $N = 16 \times (t_r / W)^2$, where $t_r$ is the retention time and $W$ is the peak width at the base. A higher number of theoretical plates indicates a sharper peak and better analytical separation. You can calculate this instantly using our Theoretical Plates Calculator.

The ICH (International Council for Harmonisation) Q2(R1) guidelines mandate specific validation parameters for HPLC methods. These include assessing Accuracy, Precision (Repeatability and Intermediate Precision), Specificity, Detection Limit (LOD), Quantitation Limit (LOQ), Linearity, and Range. Our calculators are designed specifically to help analysts easily compute these linearity, LOD/LOQ, and %RSD parameters in compliance with ICH standards.
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