THE IMPORTANCE OF CHROMATOGRAPHY IN THE ISOLATION OF PURE COMPOUND

TABLE OF CONTENT

CONTENTS                                                                         PAGE

Title Page    –        –        –        –        –        –        –        –        i

Certification         –        –        –        –        –        –        –        ii

Dedication –        –        –        –        –        –        –        –        iii

Acknowledgements        –        –        –        –        –        –        iv

Table of Contents –        –        –        –        –        –        –        vi

CHAPTER ONE

1.            INTRODUCTION       –        –        –        –        –        1

CHAPTER TWO

2.0     HISTORY OF CHROMATOGRAPHY

• Principle of Chromatography
  • Types of Chromatography
    • Column Chromatography
    • Gas chromatography
    • Liquid chromatography
    • Thin Layer chromatography
    • Size exclusion chromatography
    • Affinity chromatograophy

CHAPTER THREE

3.0     THE INTERNAL STANDARD METHOD

• Injection devices
  • Packed Column
  • Open tubular column
  • Application of chromatography in separation of compounds

CHAPTER FOUR

• SUMMARY, CONCLUSION AND REFERENCES
  •           Summary
  •           Conclusion

References

CHAPTER ONE

1. INTRODUCTION

Chromatography is based on the principle where molecules in mixture applied onto the surface or into Chromatography into the solid, and fluid stationary phase (stable phase) is separating from each other while moving with the aid of a mobile phase. The factors effective on this separation process include molecular characteristic said related to adsorption (liquid-solid), partition (liquid-solid), and affinity or differences among theirbmolecular weights (Cuatrecasas, 2004; Porath, 2006). Because of these differences, some components of the mixture stay longer in the stationary phase, and they move slowly in the chromatography system, while others pass rapidly into mobile phase, and leave the system faster (Harris, 2004).

Based on this approach three components form the basis of the chromatography technique.

• 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. Chromatography methods based on partition are very effective on separation, and identification of small molecules as amino acids, carbohydrates, and fatty acids. However, affinity chromatographies (ie. ion-exchange chromatography) are more effective in the separation of macromolecules as nucleic acids, and proteins. Paper chromatography is used in the separation of proteins, and in studies related to protein synthesis; gas-liquid chromatography is utilized in the separation of alcohol, esther, lipid, and amino groups, and observation of enzymatic interactions, while molecular-sieve chromatography is employed especially for the determination of molecular weights of proteins. Agarose-gel chromatography is used for the purification of RNA, DNA particles, and viruses (Gerberding and Byers, 2008).

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