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Questions and Answers
What is the purpose of Solid-Phase Extraction (SPE) in chromatography?
What is the purpose of Solid-Phase Extraction (SPE) in chromatography?
- To analyze solid phase materials directly
- To merge different liquid samples into one
- To separate solid samples from liquid ones
- To isolate and concentrate analytes from a liquid sample (correct)
What materials can be part of the stationary phase in SPE?
What materials can be part of the stationary phase in SPE?
- Silica, polymers, and other porous materials (correct)
- Only silica particles
- Solid metals with no adsorption properties
- Various liquids with adsorption properties
What is the role of the stationary phase in SPE?
What is the role of the stationary phase in SPE?
- To dissolve the analytes completely
- To speed up the flow of the liquid sample
- To actively react with the analytes
- To selectively retain analytes based on their interactions with the solid support (correct)
After washing away interfering substances, how are the analytes of interest retrieved in SPE?
After washing away interfering substances, how are the analytes of interest retrieved in SPE?
In which fields is SPE widely used?
In which fields is SPE widely used?
Which type of adsorption chromatography relies on the specific interactions between sample components and ligands attached to the stationary phase?
Which type of adsorption chromatography relies on the specific interactions between sample components and ligands attached to the stationary phase?
What is the main principle behind ion exchange chromatography?
What is the main principle behind ion exchange chromatography?
Which chromatography technique separates components based on their ability to distribute themselves between the stationary and mobile phases?
Which chromatography technique separates components based on their ability to distribute themselves between the stationary and mobile phases?
In which type of chromatography do larger molecules elute more quickly?
In which type of chromatography do larger molecules elute more quickly?
How is elution achieved in ion exchange chromatography?
How is elution achieved in ion exchange chromatography?
Which chromatography technique involves sample components adhering to the surface of the stationary phase?
Which chromatography technique involves sample components adhering to the surface of the stationary phase?
What kind of interactions are utilized in affinity chromatography for separation?
What kind of interactions are utilized in affinity chromatography for separation?
What property is exploited in size exclusion chromatography to separate sample components?
What property is exploited in size exclusion chromatography to separate sample components?
What is the stationary phase in Thin-Layer Chromatography (TLC)?
What is the stationary phase in Thin-Layer Chromatography (TLC)?
How is the retention factor (Rf) in TLC calculated?
How is the retention factor (Rf) in TLC calculated?
What is the primary purpose of Thin-Layer Chromatography (TLC)?
What is the primary purpose of Thin-Layer Chromatography (TLC)?
What action causes the solvent to move up the plate in TLC?
What action causes the solvent to move up the plate in TLC?
What is Solid-Phase Extraction (SPE) used for?
What is Solid-Phase Extraction (SPE) used for?
In SPE, what material is typically used for the stationary phase?
In SPE, what material is typically used for the stationary phase?
During which step of SPE are unwanted components removed from the sample?
During which step of SPE are unwanted components removed from the sample?
Which industries commonly use SPE?
Which industries commonly use SPE?
Which phase in chromatography is immobile and where separation occurs?
Which phase in chromatography is immobile and where separation occurs?
Which method of chromatography involves the separation of components based on their size and shape?
Which method of chromatography involves the separation of components based on their size and shape?
In gas chromatography, the mobile phase typically used is:
In gas chromatography, the mobile phase typically used is:
What is the primary purpose of the detector in a chromatography system?
What is the primary purpose of the detector in a chromatography system?
Which type of chromatography relies on specific interactions between the sample components and the stationary phase?
Which type of chromatography relies on specific interactions between the sample components and the stationary phase?
What technique is characterized by the adsorption of sample components onto the stationary phase?
What technique is characterized by the adsorption of sample components onto the stationary phase?
Which component of chromatography is the mixture of compounds to be separated?
Which component of chromatography is the mixture of compounds to be separated?
In which type of chromatography does the separation occur through partitioning?
In which type of chromatography does the separation occur through partitioning?
What is involved in the data analysis component of chromatography?
What is involved in the data analysis component of chromatography?
In ion exchange chromatography, separation is affected by which of the following?
In ion exchange chromatography, separation is affected by which of the following?
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Study Notes
Definition and Components of Chromatography
- Chromatography is a biophysical technique used for separation, identification, and purification of mixture components for qualitative and quantitative analysis.
- Stationary phase: immobile material where separation occurs (solid, liquid, or gel).
- Mobile phase: fluid that carries the sample through the stationary phase (liquid, gas, or supercritical fluid).
- Sample: mixture of compounds to be separated.
- Column or plate: physical structure where separation takes place.
- Detector: instrument that detects separated components as they elute from the column or plate.
- Data analysis: interpretation of the chromatogram to determine identity and quantity of separated components.
Separation Mechanism
- Partitioning: separation occurs through distribution between stationary and mobile phases (gas chromatography and liquid chromatography).
- Adsorption: separation occurs through adsorption of sample components onto the stationary phase material (thin-layer chromatography and solid-phase chromatography).
- Ion exchange: separation occurs based on electrostatic interactions (ionic strength or pH of the mobile phase affects separation).
- Size exclusion: separation occurs based on size and shape of sample components (components too large to enter pores are excluded).
- Affinity: separation occurs through specific interactions between sample components and stationary phase.
Types of Chromatography
- Adsorption chromatography: relies on adsorption of sample components onto the stationary phase.
- Partition chromatography: relies on partitioning of sample components between stationary and mobile phases.
- Ion exchange chromatography: relies on electrostatic interactions between sample components and stationary phase.
- Size exclusion chromatography: relies on size and shape of sample components.
- Affinity chromatography: relies on specific interactions between sample components and stationary phase.
Thin-Layer Chromatography (TLC)
- Stationary phase: thin layer of adsorbent material (silica gel or alumina) coated on a glass or plastic plate.
- Sample components separate based on affinity for the stationary phase.
- Distance travelled by each component is measured relative to the distance travelled by the solvent (retention factor, Rf).
Solid-Phase Extraction (SPE)
- Sample preparation technique used to purify and concentrate analytes from a liquid sample.
- Stationary phase: solid material packed inside a column (silica, polymers, or other porous materials with adsorption properties).
- Sample is loaded onto the column, washed with a solvent, and then eluted with a solvent to concentrate the desired analytes.
Applications of Chromatography
- Environmental monitoring
- Drug discovery
- Food safety
- Beverage testing
- Forensic toxicology
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