What is the Difference Between FPLC and HPLC?
🆚 Go to Comparative Table 🆚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:
- 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.
- 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.
- Column Hardware: HPLC uses pressure-resistant stainless steel columns, while FPLC can work with glass columns due to the relatively lower pressures involved.
- 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).
- Monitoring: FPLC systems monitor UV absorption, pH, and conductivity, while HPLC systems typically only monitor UV absorption.
- 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:
Parameter | FPLC | HPLC |
---|---|---|
Purpose | Purification of large biomolecules (e.g., proteins, nucleotides, peptides) | Separation and analysis of small molecular weight compounds |
Column Material | Agarose or polymer material with bigger particle sizes | Silica beads with very small particle sizes |
Column Hardware | Transparent and pressure-unstable | Pressure-resistant stainless steel |
Methods | Size-exclusion chromatography (SEC), ion-exchange chromatography, affinity chromatography | Reversed-phase chromatography with hydrophobic stationary phases and polar mobile phases |
Software | Controls modules and integrates collection of purified samples | Controls instrumentation and analyzes data |
Pump Head Material | Titanium or PEEK | Stainless steel |
Flow Rate | Usually higher than HPLC | Usually lower than FPLC |
Monitoring | UV absorption, pH, and conductivity | UV absorption only |
Both FPLC and HPLC use pumps, detectors, valves, and software for sample separation and detection.
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- Distillation vs Chromatography
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- Gel Filtration vs Affinity Chromatography
- GCMS vs LCMS
- Electrophoresis vs Chromatography
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- Flow Cytometry vs FACS
- Distillation vs Extraction
- Filtration vs Centrifugation
- Decantation vs Filtration