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Advantages and Molecules that Enhance Separation in Chromatography, Thesis of Pharmacy

Memorial University of NewfoundlandPharmacy

An overview of chromatography, a biophysical technique used for separating, identifying, and purifying components of a mixture. the principles of chromatography, including the role of stationary and mobile phases, and the factors that influence separation, such as molecular size, polarity, ionic charge, and binding affinity. The document also covers various types of chromatography techniques and examples of molecules used for separation. Finally, the advantages of chromatography, including the determination of retention factors and polarity of separating compounds, are discussed.

What you will learn

  • What is chromatography and how does it work?
  • What are the advantages of using chromatography for separating and identifying components of a mixture?
  • What are the three main components of chromatography?
  • What are some examples of chromatography techniques and when are they used?

Typology: Thesis

2012/2013

Uploaded on 03/17/2022

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Pan African University
Pan African University
 
 
Life and Earth Sciences Institute
Life and Earth Sciences Institute
 
 
University of Ibadan   
University of Ibadan   
Outline :
•Separation .
•Molecules that enhances separation.
•Advantages of molecules that enhances
separation.
•References .
ADVANTAGES OF MOLECULES
THAT ENHANCES SEPARATION
•Separation is an act or instance of separating
compounds present in a substance.
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Download Advantages and Molecules that Enhance Separation in Chromatography and more Thesis Pharmacy in PDF only on Docsity!

Pan African UniversityPan African University Life and Earth Sciences Institute Life and Earth Sciences Institute University of Ibadan University of Ibadan

Outline :

  • Separation.
  • Molecules that enhances separation.
  • Advantages of molecules that enhances separation.
  • References.

ADVANTAGES OF MOLECULES

THAT ENHANCES SEPARATION

  • Separation is an act or instance of separating compounds present in a substance.
  • Chromatography is an important biophysical technique that enables the separation, identification, and purification of the components of a mixture for qualitative and quantitative analysis.
  • Stationary phase: This phase is always composed of a “solid” phase or “a layer of a liquid adsorbed on the surface a solid support”.
  • Mobile phase: This phase is always composed of “liquid” or a “gaseous component.”
  • Separated molecules
  • The type of interaction between stationary phase, mobile phase, and substances contained in the mixture is the basic component effective on separation of molecules from each other. Stationary phase in chromatography, is a solid phase or a liquid phase coated on the surface of a solid phase. Mobile phase flowing over the stationary phase is a gaseous or liquid phase.
  • If mobile phase is liquid it is termed as liquid chromatography (LC), and if it is gas then it is called gas chromatography (GC).
  • Gas chromatography is applied for gases, and mixtures of volatile liquids, and solid material.

EXAMPLES OF MOLECULES USED

  • Silica resins
  • Alumina resins
  • Dextran
  • Agorose
  • Polyacrylamide
  • Cellulose
  • Anthraquinone dyes, and azo-dyes used as ligands Properties of molecules that enhances separation:

1. Polarity of molecules

  • For separations based on polarity, like is attracted to like and opposites may be repelled. That is if stationary phase used is polar {silica gel} the mobile phase should be non-polar {hexane} so as polar compound will be attracted to the stationary phase while non-polar compounds will be eluted with the mobile phase.

Examples of

chromatography

techniques based on

this property are:

  • Column chromatography
  • Paper chromatography

Examples of

chromatography

techniques based on

this property are:

  • Gas chromatography Properties of molecules that **enhances separation:
  1. Ionic charge of molecules.**
  • Molecules possessing the opposite charge as the resin will bind tightly to the resin, and molecules having the same charge as the resin will flow through the column and elute out first.
  • Example in ion- exchange chromatography. Properties of molecules that **enhances separation:
  1. Size of the molecules**

Properties of molecules that enhances separation:

5. Binding affinity of molecules.

  • If the molecule binds tightly to the enzyme and the unbound analytes will pass through in the mobile phase, and elute out of the column, leaving the substrate bound to the enzyme, which can then be detached from the stationary phase and eluted out of the column with an appropriate solvent.
  • This property is used in: affinity chromatography

Advantages

1. Helps in adsorption and solubility

  • This is a major molecular characteristic that aids separation of compounds in liquid-solid chromatography. The main principle used:
  • Differential affinities (strength of adhesion) of the various components of the analyte towards the stationary and mobile phase results in the differential separation of the components. Affinity in turn, is dictated by two properties of the molecule: ‘Adsorption’ and ‘Solubility’.

Advantages :

2. Determine retention factor (retardation factor) and polarity of separating compounds.

  • Das M, Dasgupta D. Pseudo-affinity column chromatography based rapid purification procedure for T7 RNA polymerase. Prep Biochem Biotechnol. 1998
  • Determann H. Gel chromatography gel filtration, gel permeation, molecular sieves: a laboratory handbook. Chapter 2. Materials and Methods. 2012
  • Donald PL, Lampman GM, Kritz GS, Engel RG. Introduction to organic laboratory techniques. 4th ed. Thomson Brooks/Cole;
  1. pp. 797–817. REFERENCES
  • Elmut D. Gel Chromatography, gel filtration, gel permeation, molecular sieves:a laboratory hand book. Springer-Verlag; 1969.
  • Firer MA. Efficient elution of functional proteins in affinity chromatography. J Biochem Biophys Methods.
  • Gerberding SJ, Byers CH. Preparative ion- exchange chromatography of proteins from dairy whey. J Chromatogr A. 1998
  • Harris DC. Exploring chemical analysis. 3rd ed. WH. Freeman&Co; 2004.
  • Harwood LM, Moody CJ. Experimental organic chemistry:Principles and Practice. Oxford:Blacwell Science; 1989:180–5.
  • Porath J. From gel filtration to adsorptive size exclusion. J Protein Chem. 1997

Thanks