Chromatography in BIOT303 Lecture 8
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Questions and Answers

What is the primary purpose of chromatography?

Chromatography is primarily used to separate the components of a sample.

What are the two phases involved in chromatography?

The two phases are the stationary phase and the mobile phase.

Which of these can the stationary phase be?

  • Solid (correct)
  • Liquid supported on a solid (correct)
  • Gel (correct)
  • All of the above

    • What are the two possible states of the mobile phase?

    <p>Gaseous</p> Signup and view all the answers

    What is the process of chromatography?

    <p>In chromatography, a mixture to be separated is applied to a stationary phase. A pure solvent (mobile phase) is then allowed to move slowly over the stationary phase, carrying the components separately based on their solubility in the solvent.</p> Signup and view all the answers

    Chromatography is a separation method where the analyte is combined within a liquid or gaseous mobile phase and pumped through a stationary phase.

    <p>True</p> Signup and view all the answers

    In chromatography, one phase is hydrophilic and the other is lipophilic.

    <p>True</p> Signup and view all the answers

    What determines the time a component spends in the stationary phase?

    <p>The polarity of the component.</p> Signup and view all the answers

    What is the driving force for solute migration in chromatography?

    <p>The driving force is the moving fluid (mobile phase).</p> Signup and view all the answers

    What is the resulting force that contributes to separation in chromatography?

    <p>The combination of the moving fluid and the solute's affinity for the stationary phase.</p> Signup and view all the answers

    What is the specific time called when a sample component elutes from the stationary phase?

    <p>Retention time</p> Signup and view all the answers

    How are the components' signals recorded in chromatography?

    <p>The components' signals are recorded as they pass through a detector and plotted in the form of a chromatogram.</p> Signup and view all the answers

    What are the two possible states of the mobile phase in chromatography?

    <p>Liquid</p> Signup and view all the answers

    The stationary phase in chromatography can be either a solid or a liquid.

    <p>True</p> Signup and view all the answers

    What continuously exchanges solute molecules between the two phases in chromatography?

    <p>Kinetic molecular motion.</p> Signup and view all the answers

    A component that favors the moving fluid will spend more time in the stationary phase than in the mobile phase.

    <p>False</p> Signup and view all the answers

    What is the ratio of the times spent in the moving and stationary regions of a component known as?

    <p>Partition coefficient</p> Signup and view all the answers

    What is chromatography defined as?

    <p>Differential migration from a narrow initial zone.</p> Signup and view all the answers

    Based on what factor are types of chromatography categorized?

    <p>Interaction of solute with the stationary phase</p> Signup and view all the answers

    Which of these types of chromatography are categorized based on the interaction between the solute and the stationary phase?

    <p>Adsorption</p> Signup and view all the answers

    Which types of chromatography are categorized based on the physical shape?

    <p>Both A and B</p> Signup and view all the answers

    Which types of chromatography are classified based on the shape of the chromatographic bed?

    <p>Both A and B</p> Signup and view all the answers

    Which type of chromatography uses a very thin layer of adsorbent such as silica gel and alumina?

    <p>Thin Layer Chromatography</p> Signup and view all the answers

    What is the plate used in Thin Layer Chromatography known as?

    <p>Chrome plate</p> Signup and view all the answers

    Paper chromatography is an inexpensive method that uses paper sheets or strips as the stationary phase.

    <p>True</p> Signup and view all the answers

    Paper chromatography is a powerful analytical tool that uses very small quantities of material.

    <p>True</p> Signup and view all the answers

    Who discovered paper chromatography in 1943?

    <p>Synge and Martin.</p> Signup and view all the answers

    Column chromatography is a separation method where the analyte is combined within a liquid or gaseous mobile phase and pumped through a stationary phase.

    <p>True</p> Signup and view all the answers

    What does eluent refer to in chromatography?

    <p>Eluent is another name for the mobile phase.</p> Signup and view all the answers

    In column chromatography, a mobile phase moves over a stationary phase, carrying the components with higher absorptivity to a lower distance than those with lower absorptivity.

    <p>True</p> Signup and view all the answers

    Column chromatography is a 2-dimensional technique where the stationary phase is spread as a layer.

    <p>False</p> Signup and view all the answers

    Chromatography is an essential part of almost any protein purification strategy.

    <p>True</p> Signup and view all the answers

    Ion exchange chromatography and size exclusion chromatography are two examples of chromatographic techniques used for protein purification and analysis.

    <p>True</p> Signup and view all the answers

    A more advanced column chromatography should be used for protein purification for specific applications.

    <p>True</p> Signup and view all the answers

    What does HPLC stand for?

    <p>High-performance liquid chromatography</p> Signup and view all the answers

    What are some key components of a chromatograph used in HPLC?

    <p>Both A and B</p> Signup and view all the answers

    Ultra High Performance Liquid Chromatography (UHPLC) is capable of high-speed analysis.

    <p>True</p> Signup and view all the answers

    Only compounds in a gaseous state can be analyzed using HPLC.

    <p>False</p> Signup and view all the answers

    What can HPLC be used for in terms of analyzing samples?

    <p>HPLC allows qualitative and quantitative analysis of the sample components.</p> Signup and view all the answers

    What is the solvent used to separate components in a liquid sample for HPLC analysis called?

    <p>Mobile phase</p> Signup and view all the answers

    In HPLC, the mobile phase is delivered to a separation column, which is otherwise known as the stationary phase.

    <p>True</p> Signup and view all the answers

    What controls the stable flow rate of the mobile phase in HPLC?

    <p>Solvent delivery pump</p> Signup and view all the answers

    What happens to the separated components downstream of the column in HPLC?

    <p>The separated compounds are detected by a detector.</p> Signup and view all the answers

    HPLC is a reliable and precise way to check product purity in pharmaceutical development.

    <p>True</p> Signup and view all the answers

    HPLC is typically the primary method used for large-scale production of drugs due to its cost-effectiveness.

    <p>False</p> Signup and view all the answers

    HPLC can be used to analyze nutrients in blood and other biological samples.

    <p>True</p> Signup and view all the answers

    HPLC is generally considered a more cost-effective alternative compared to other techniques for measuring vitamin D levels in the blood.

    <p>False</p> Signup and view all the answers

    HPLC can be used to detect drug traces in urine.

    <p>True</p> Signup and view all the answers

    HPLC can be used to identify metabolites, which are produced by a reaction to the substance, and are not typically found in the original drug.

    <p>True</p> Signup and view all the answers

    HPLC findings alone can be used as evidence in police charges.

    <p>False</p> Signup and view all the answers

    HPLC can be used to identify and quantify specific metabolites in patients with Parkinson's or heart disease.

    <p>True</p> Signup and view all the answers

    HPLC is often used to identify and quantify pesticides, preservatives, and artificial flavorings and colorants in food production.

    <p>True</p> Signup and view all the answers

    What type of substances does gas chromatography analyze?

    <p>Gas chromatography analyzes volatile substances in the gas phase.</p> Signup and view all the answers

    In gas chromatography, the mobile phase used is a chemically inert gas that serves to carry the molecules of the analyte through the heated column.

    <p>True</p> Signup and view all the answers

    Gas chromatography does utilize the mobile phase to interact with the analyte.

    <p>False</p> Signup and view all the answers

    What are the two types of stationary phases used in gas chromatography?

    <p>Both A and B</p> Signup and view all the answers

    What is a solid adsorbent stationary phase in gas chromatography called?

    <p>Gas-solid chromatography (GSC).</p> Signup and view all the answers

    What is a liquid stationary phase in gas chromatography called?

    <p>Gas-liquid chromatography (GLC).</p> Signup and view all the answers

    What is the process of vaporizing the sample in gas chromatography known as?

    <p>Sample injection.</p> Signup and view all the answers

    The detector in gas chromatography converts the amount of each component into an electrical signal.

    <p>True</p> Signup and view all the answers

    GC is primarily used for analyzing flavor and aroma compounds in various samples.

    <p>True</p> Signup and view all the answers

    GC can identify and quantify each component in a particular pharmaceutical product.

    <p>True</p> Signup and view all the answers

    GC can be used to identify and quantify specific chemicals released into the air by products.

    <p>True</p> Signup and view all the answers

    GC can be used to detect drugs from samples of bodily fluids.

    <p>True</p> Signup and view all the answers

    GC can be used to test for illegal or prohibited drug usage in legal cases and by sporting bodies.

    <p>True</p> Signup and view all the answers

    Samples analyzed by GC should be volatile and thermostable.

    <p>True</p> Signup and view all the answers

    Study Notes

    Lecture 8: Chromatography

    • The lecture was on Biological and Chemical Analytics (BIOT303) in the Faculty of Biotechnology at German International University (GIU).
    • The lecturer was Dr. Rana A. Youness, Head of Molecular Genetics Research Team (MGRT).
    • Contact information for Dr. Youness was provided.
    • Learning Objectives (LOs) were outlined. Students will be able to define chromatography, differentiate between solid and mobile phases, identify different chromatography types, understand HPLC and GC principles, and list applications in pharmaceutical and biotechnology fields.

    Chromatography

    • Chromatography is a method used for separating components in a sample based on their distribution between two phases; one stationary (solid, liquid, or gel) and one mobile.
    • The stationary phase can be packed in a column, spread as a layer, or distributed as a film.
    • The mobile phase can be gaseous or liquid.
    • Separated components are carried separately according to their solubility differences in the pure solvent.

    Chromatography Principle

    • Chromatography involves the analyte interacting with the stationary and mobile phases.
    • Polarity differences between phases result in varying retention times.
    • The amount of time spent in the mobile phase versus the stationary phase determines the separation.
    • The retention time is the specific time for each component that elutes from the stationary phase.
    • A chromatogram displays the detected signals and plots them in relation to their elution times.

    Types of Chromatography

    • Chromatography is categorized based on interaction with the stationary phase (adsorption, partition, ion exchange, size exclusion), the chromatographic bed shape (planar or column), and the physical shape of the system.
      • Examples include: Thin Layer Chromatography (TLC), Paper Chromatography, Column Chromatography, High-Performance Liquid Chromatography (HPLC), Gas Chromatography (GC), and Gas-Solid Chromatography (GSC).
    • Chromatography techniques can be two-dimensional or three-dimensional.
      • TLC uses a thin layer adsorbent (like silica gel or alumina) on a plate.
      • Paper Chromatography uses paper as the adsorbent.

    Ion Exchange Chromatography

    • Separates substances based on charge (+ or -).
    • Stationary phase contains charged groups that attract ions with opposite charges.

    Size Exclusion Chromatography

    • Separates by size.
    • Porous beads in the stationary phase trap smaller molecules.

    Partition Chromatography

    • Separates based on solubility differences.
    • The stationary phase and the mobile phase have different solubility preferences for different compounds.
    • Different compounds will migrate with differing degrees and thus become separated.

    Adsorption Chromatography

    • Separates based on the affinity of substances to the adsorbent.
    • The stationary phase has a high affinity for certain compounds, and they will spend more time interacting with and adsorbing into the stationary phase.

    HPLC (High-Performance Liquid Chromatography)

    • HPLC is used to separate compounds dissolved in a liquid, and then to analyze these separated components.
    • HPLC is a useful technique for various applications, including the analysis of nutrients in blood and the identification of drug traces in urine.

    GC (Gas Chromatography)

    • Gas chromatography is used to analyze volatile substances and their components.
    • A carrier gas in the mobile phase separates the components according to their affinities for the stationary phase.
    • The separated compounds, in turn, can be identified by the chromatogram.

    Applications

    • Applications of chromatography include pharmaceutical development (drug purity checks, identifying metabolites), medical diagnostics (measuring vitamin levels, detecting drug use), legal issues (detecting drugs or poisons), pollution monitoring, and food production (measuring pesticides or preservatives).

    Chromatography Summary

    • Chromatography is a versatile technique for separating, purifying and analyzing various compounds in different forms and phases. Different types of chromatography address different needs and chemistries.

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    Description

    This quiz covers Lecture 8 of Biological and Chemical Analytics (BIOT303), focusing on chromatography techniques. Students will explore the definition, types, principles of HPLC and GC, and applications in pharmaceutical and biotechnology fields. Test your understanding of chromatography and its significance in analytics.

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