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Questions and Answers
What is the primary function of a pump in HPLC systems?
What is the primary function of a pump in HPLC systems?
Which type of injector is more commonly used in HPLC?
Which type of injector is more commonly used in HPLC?
What is an important characteristic of HPLC detectors?
What is an important characteristic of HPLC detectors?
What is the recommended volume range for sample injections in HPLC?
What is the recommended volume range for sample injections in HPLC?
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Which factor is NOT crucial for the performance of HPLC detectors?
Which factor is NOT crucial for the performance of HPLC detectors?
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What is the primary purpose of chromatography?
What is the primary purpose of chromatography?
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Which phase of chromatography is described as the phase that moves in a definite direction?
Which phase of chromatography is described as the phase that moves in a definite direction?
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What distinguishes High Performance Liquid Chromatography (HPLC) from conventional liquid chromatography?
What distinguishes High Performance Liquid Chromatography (HPLC) from conventional liquid chromatography?
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Which of the following components is referred to as the analyte in chromatography?
Which of the following components is referred to as the analyte in chromatography?
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How long does total separation typically take in HPLC compared to conventional methods?
How long does total separation typically take in HPLC compared to conventional methods?
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What is the effect of using a smaller particle size in HPLC columns?
What is the effect of using a smaller particle size in HPLC columns?
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Which type of chromatography involves the use of a stationary phase that is solid, liquid, or a solid/liquid mixture?
Which type of chromatography involves the use of a stationary phase that is solid, liquid, or a solid/liquid mixture?
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In HPLC, what is the role of the effluent?
In HPLC, what is the role of the effluent?
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Study Notes
Chromatography
- A technique separating mixture components based on their travel time through a stationary phase using a mobile phase.
- Consists of two phases:
- Mobile phase: Liquid or gaseous, flows over or through the stationary phase.
- Stationary phase: Solid, liquid, or a solid/liquid mixture, immobilized.
Chromatography terms
- Analyte: Substance being separated during chromatography.
- Immobilized phase: Stationary phase attached to support particles or column tubing.
- Mobile phase: Moving phase, includes the sample and solvent carrying it through the column.
- Effluent: Mobile phase exiting the column.
Types of chromatography
- Paper chromatography
- Liquid chromatography
- Gas chromatography
- High performance liquid chromatography (HPLC)
High Performance Liquid Chromatography
- An advanced form of liquid chromatography.
- Uses tightly packed columns, high pressure (up to 5,000 psi) to force eluent through.
- Utilizes smaller particle size packing material for greater surface area, improving separation quality.
-
HPLC Technique:
- Liquid mobile phase separates the mixture.
- Analytes are dissolved in a solvent, then pushed through the column under high pressure (up to 400 atm).
- Mixture is separated into its components within the column.
- Separation time is often 5-10 minutes compared to hours or days for conventional methods.
-
HPLC System Components:
- Pump: Delivers the mobile phase at a stable flow rate, preventing pulsations even when mobile phase composition varies.
- Injector: Introduces the sample under pressure, minimizing band broadening and flow disturbances.
- Column: Smooth-bore stainless steel or heavy-walled glass tubing containing the packed stationary phase.
-
Detector: Monitors the elute as it leaves the column, producing an electronic signal proportional to component concentrations.
- Crucial for trace analysis
- High sensitivity
- Fast response
- Insensitive to changes in solvent type, flow rate, and temperature
Detectors Used in HPLC
- ng (nanogram) = 10^-9 g
- pg (picogram) = 10^-12 g
| Type | Principle | Detection Limit | Comments |
|---|---|---|---|
| Spectro- | Measure | ng | |
Injectors
- Two main types:
- Valve type: Connected to the column via a pipe, with valves for opening and closing, frequently used.
- Syringe type: Directly connected to the column, used for sample delivery with a syringe.
Pumps
- Flow rate usually between 0.01-10 ml/min.
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Description
Explore the fundamental concepts and types of chromatography in this quiz. Discover the roles of mobile and stationary phases, along with key terms like analyte and effluent. Test your understanding of different chromatography forms such as HPLC and paper chromatography.
