High Performance Liquid Chromatography (HPLC)
- HPLC is the most widely used type of chromatography
- The reasoning behind this is its sensitivity, accurate quantitative anaylsis, its ability to separate non-volatile and thermally unstable compouns.
- HPLC can be used in most Qualitative and Quantitative analysis.
- Like all chromatographic techniques HPLC requires a Mobile Phase and a Stationary Phase.
- The Mobile Phase in HPLC drives the mixture through the stationary phase with the aid of a pump.
- The Stationary Phase in HPLC is one of the most important components of the system.
- It is a column which is packed with silica gel. The properties of the silica gel make it excellent for the separation of analytes.
- From the diagram above it is shown that the mobile phase is brought to the pump where it is sent to the column under a specific pressure.
- The sample is injected into the system before the column and is injected into the mobile phase stream.
- Once the sample is in the column the component with the greater affinity for the mobile phase will reach the detector first. it is then followed by the component which was more attracted to the stationary phase.
- The component with the greatest affinity to the stationary phase will be the last component of the mixture to reach the detector.
Chromatogram of Pain Killer containin Paracetamol and Caffeine
Once the analytes reach the detector, the information is traslated into a chromatogram which is produced by the software.
Uses of HPLC
Applications of HPLC
High Performance Liqud Chromatography has become the most widey used chromatographic technique today.