HPLC - Basics
| What does “HPLC” mean? | High Performance Liquid Chromatography, or High Pressure Liquid Chromatography |
| What does it do? | Separates a liquid sample into its components. |
| Physiology | Used for the detection of tumours and for monitoring drugs. |
| How it works |
The sample is injected into a stream of high pressure liquid (mobile phase) which sweeps the sample into a column and then to a detector. Separation takes place in the column as different components will have a different chemical affinity for the column packing. Those with least affinity will exit first. Each component elutes from the column as a narrow band or peak. The time that it takes for a peak to exit the column determines the component species and the peak height or peak area determines the quantity. |
 Chromatogram showing characteristic peaks |
|
| Units of measurement | Retention time. The time from injection until a peak of interest
emerges. Expressed in minutes. Peak height or peak area is measured and expressed in terms of concentration ie micro mole per litre |
| Typical values | Metanephrine 1 micromole per day (measured as excretion) Adrenaline – 0.01 micro mole per litre (uM/L) |
| Picture of equipment |
