The Chromatography of Steroids
1st Edition
International Series of Monographs on Pure and Applied Biology: Biochemistry
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Description
The Chromatography of Steroids details the fundamental concepts and underlying principles of laboratory techniques utilized in separating steroid mixtures. The text first covers the basic theory of chromatography, and then proceeds to tackling the chromatographic separation of steroids. Next, the selection details the techniques and apparatus employed in chromatography of steroids. Chapter IV talks about the colorimetric and radioisotopic techniques, while Chapter V deals with the structural analysis and identification of steroids by chromatography. The text also coves the typical analytical problems of steroid biochemistry. The book will be of great use to researchers who utilizes chromatographic methods in their work with steroids.
Table of Contents
Preface
Glossaries
1. Symbols and Terms
2. Description of Solvent Systems
3. The Structure and Nomenclature of the Steroids
I. The Basic Theory of Chromatography and Some General Principles
1. Introductory
2. Derivation of RF and RM Values
3. Solvent-Solute Interactions
4. Chemical Structure and Chromatographic Behavior
5. Partition Coefficients and Solvent Systems
6. The Effect of Temperature
7. Formation of Several Species of Solute
II. Chromatographic Separation of Steroids
Section 1: Descriptive
1. Introduction
2. Special Features of the Chromatography of Steroids
3. Adsorption Methods
4. Partition Chromatography
Section 2: Quantitative Treatment of Steroid Behavior on Chromatograms
1. ΔRMg Values in Typical Solvent Systems
2. ΔRMg Values in Atypical Solvent Systems
3. ΔRMg and RMr Values
4. Solvent-Solute Interactions and ΔRMg Values
III. Techniques and Apparatus
1. Transfer Operations and Volumetric Error
(a) Transfer to Adsorption Columns
(b) Transfer to Partition Columns
(c) Transfer of Material to Paper Chromatograms
2. Column Chromatography
(a) Preparation of Adsorption Columns
(b) Preparation of Partition Columns
(c) Design and Operation of Columns
(d) Multiple-Column Chromatography
3. Paper Chromatography
(a) Preparation of Sheets and Strips
(b) Impregnation of Paper with Stationary Phases
(c) No-Touch Technique for Preparing Paper Chromatograms
(d) Operation of Chromatography Tanks
(e) Ways of Running Paper Chromatograms
(f) Temperature Control
(g) General Features of Methods of Detection
(h) Special Features of Methods of Detection
(i) The Use of Dyes and Markers
(j) Elution of Material From Paper
(k) Preparative Paper Chromatography
4. Preparation of Extracts Suitable for Chromatography
(a) General
(b) Animal Tissues
(c) Blood and Plasma
(d) Sweat, Feces, Urine
IV. Quantitative Chromatography: Colorimetric and Radioisotopic Techniques
1. Introduction
2. General Procedure
3. Chromatographic Selectivity
4. Modifying Steps
5. Chromatographic Technique in Quantitative Methods
6. Direct Scanning of Paper Chromatograms
(a) Treatment of Paper Strips with Reagents
(b) Solvents and Reagents Suitable for Scanning Methods
(c) Treatment of Strips after the Reagent Has Been Applied
(d) Optical Requirements
(e) Geometrical and Planimetric Methods
(f) Electronic Recording Apparatus
(g) Fully Automatic Apparatus
7. Logistics of Quantitative Estimations Using Chromatography
8. Use of Radioactive Isotopes in Quantitative Chromatography
9. Complication and Simplicity
V. Structural Analysis and Identification of Steroids by Chromatography
1. The Basis of the Classical Method
2. The Basis of Chromatographic Identification
3. General Procedure for Structure Determination
4. Examples of Structure Determination
(a) Cortisol
(b) 2α-Me-P4-6ß, 11ß, 17α, 20ξ, 21-ol-3-One
(c) 9α-F-αP-3α, 11ß, 17α, 21-ol-20-One
(d) 18-Hydroxyöestrone
5. Systematic Investigation of Unknown Steroids
6. General Properties of Simple Functional Groups
(a) Changes By Chemical Modification —ΔRMr Values
(b) Solvent Changes — ΔRms Values
7. Special Properties of Simple and Complex Functional Groups
(a) ΔRMr on Modification
(b) ΔRMs on Change of Solvent System
8. Spectroscopic Properties, Color and Fluorescence Reactions
9. Future Developments
10. General Procedure Using ΔRMs and ΔRMg Values
VI. Some Typical Analytical Problems of Steroid Biochemistry
1. Introduction
2. Cortisol in Human Blood
3. Oestrogens in Human Urine
4. Progesterone in Blood or Urine
5. Aldosterone in Urine
(a) Direct Fluorimetry on Paper
(b) Fluorimetry in Potassium-t-Butoxide
(c) Assay of Tritium-Labeled Acetoxyl Groups
6. Testosterone in Blood or Urine
7. Difficult Separations of Non-Polar Steroids
8. Conjugates of Urinary Steroids
9. Fractionation and Estimation of Urinary Steroids
(a) Fraction 1
(b) Fraction 2
(c) Fraction 3
(d) More Complete Fractionation of Urine
(e) Δ4-3-Ketones
(f) Pregnane-3α,20α-Diol
(g) Reducing Metabolites in the Presence of Steroid Analogues
(h) Non-Alcoholic Steroid Ketones
10. Conclusion
Appendices
I. Purification of Reagents and Materials
1. Solvents
2. Adsorbents
3. Kieselguhr ("Celite")
4. Cellulose
5. Glassware
6. Reagents
II. Microchemical Reactions for Steroids
III. Methods of Detection on Paper Chromatograms
1. Selective Absorption of Light
2. Color and Fluorescence Reactions
3. Detection of Radioactive Material
IV. Calculations with RF and RM Values
How to Use the Tables
References
Name Index
Subject Index
Details
- No. of pages:
- 460
- Language:
- English
- Copyright:
- © Pergamon 1961
- Published:
- 1st January 1961
- Imprint:
- Pergamon
- eBook ISBN:
- 9781483184579
- Hardcover ISBN:
- 9780080095448
- Paperback ISBN:
- 9781483168760
About the Author

I. E. Bush
About the Editors

R. K. Callow

P. N. Campbell

S. P. Datta
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