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Questions and Answers
What is the main principle of HPLC?
What is the main principle of HPLC?
What is the function of the pump in an HPLC system?
What is the function of the pump in an HPLC system?
Which type of HPLC is most commonly used for separating non-polar and moderately polar compounds?
Which type of HPLC is most commonly used for separating non-polar and moderately polar compounds?
What is the main advantage of HPLC over other chromatographic techniques?
What is the main advantage of HPLC over other chromatographic techniques?
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What is the primary function of the detector in an HPLC system?
What is the primary function of the detector in an HPLC system?
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What type of HPLC is suitable for separating molecules based on their size?
What type of HPLC is suitable for separating molecules based on their size?
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Study Notes
HPLC (High-Performance Liquid Chromatography)
Principle:
- Separation of molecules based on their interactions with a stationary phase and a mobile phase
- High pressure is used to force the mobile phase through the column, resulting in faster separation and higher efficiency
Instrumentation:
- Pump: delivers the mobile phase at high pressure (up to 400 bar)
- Injector: introduces the sample into the mobile phase
- Column: contains the stationary phase, where separation occurs
- Detector: measures the absorbance or fluorescence of the separated compounds
- Data system: collects and analyzes the chromatographic data
Types of HPLC:
-
Reversed-phase HPLC: uses a non-polar stationary phase and a polar mobile phase
- Most commonly used type of HPLC
- Suitable for separating non-polar and moderately polar compounds
-
Normal-phase HPLC: uses a polar stationary phase and a non-polar mobile phase
- Suitable for separating polar compounds
-
Ion-exchange HPLC: uses a stationary phase with ionic functional groups
- Suitable for separating ions and molecules with ionic functional groups
-
Size-exclusion HPLC: uses a stationary phase with pores of specific sizes
- Suitable for separating molecules based on their size
Advantages:
- High resolution and sensitivity
- Fast separation times (typically 10-30 minutes)
- Can be used for both analytical and preparative purposes
- Can be coupled with various detection methods (e.g. MS, UV, FL)
Applications:
- Pharmaceutical analysis (e.g. drug development, quality control)
- Biotechnology (e.g. protein analysis, bioprocess monitoring)
- Environmental analysis (e.g. water and soil analysis)
- Food analysis (e.g. food safety, quality control)
- Clinical analysis (e.g. disease diagnosis, biomarker discovery)
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Description
Test your knowledge of High-Performance Liquid Chromatography, including its principles, instrumentation, types, advantages, and applications in various fields.
