What is the Difference Between FPLC and HPLC?

Fast Protein Liquid Chromatography (FPLC) and High-Performance Liquid Chromatography (HPLC) are both types of liquid chromatography, but they have distinct differences in their applications, column materials, and methods. The main differences between FPLC and HPLC include:

1. Application: FPLC is used to purify large biomolecules such as proteins, nucleotides, and peptides, while HPLC is primarily used for the quantitative and qualitative analysis of small molecular weight compounds.
2. Column Materials: HPLC columns are made of silica beads with small particle sizes and high resistance to high pressures. In contrast, FPLC requires agarose or polymer material with larger particle and pore sizes.
3. Column Hardware: HPLC uses pressure-resistant stainless steel columns, while FPLC can work with glass columns due to the relatively lower pressures involved.
4. Separation Methods: HPLC primarily employs reversed phase chromatography with hydrophobic stationary phases and polar mobile phases. FPLC, on the other hand, uses a variety of methods, such as size-exclusion chromatography (SEC).
5. Monitoring: FPLC systems monitor UV absorption, pH, and conductivity, while HPLC systems typically only monitor UV absorption.
6. Software: The software for HPLC controls the instrumentation and analyzes the data, while FPLC software controls the modules and integrates the collection of purified samples.

Comparative Table: FPLC vs HPLC

Fast Protein Liquid Chromatography (FPLC) and High-Performance Liquid Chromatography (HPLC) are both types of liquid chromatography used for different purposes. Here is a table highlighting the differences between FPLC and HPLC:

Both FPLC and HPLC use pumps, detectors, valves, and software for sample separation and detection.