Enzymatic Methods of Analysis

International Series of Monographs in Analytical Chemistry

1st Edition - January 1, 1970

Author: George G. Guilbault

Editors: R. Belcher, H. Freiser

eBook ISBN:

9 7 8 - 1 - 4 8 3 1 - 5 0 5 4 - 3

Enzymatic Methods of Analysis attempts to cover all the aspects of modern enzymatic analysis. Enzymes possess a great potential usefulness in analytical chemistry. The specificity… Read more

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Enzymatic Methods of Analysis attempts to cover all the aspects of modern enzymatic analysis. Enzymes possess a great potential usefulness in analytical chemistry. The specificity of enzymes can solve the primary problem of most analytical chemists—the analysis of one substance in the presence of many similar compounds that interfere in the analysis. The book begins with two introductory chapters devoted to general considerations of enzymes as reagents and methods of analysis of enzymatic reactions. The next four chapters deal with methods for the assay of specific enzymes, and substrate, activator, and inhibitor analysis using enzymes. In the last two chapters the immobilization of enzymes and the automation of enzymatic reactions are discussed. In addition, a listing of all commercially available enzymes is given in an appendix. It is hoped that the information presented will prove interesting and stimulating to all individuals engaged in research and development.

Preface

Chapter 1: General Considerations

A. Principles of Enzymatic Analysis

B. Properties of Enzymes

C. Factors Influencing the Enzyme Reaction Rate

1. Effect of Enzyme Concentration

2. Effect of Substrate Concentration

3. Effect of Activators

4. Effect of Inhibitors

5. Effect of Temperature

6. Effect of pH

7. Effect of Ionic Strength

D. Determination of Concentrations

1. Total Change or Equilibrium Method

2. Kinetic Method

E. Handling Biochemical Reagents

F. Sources of Reagents

G. Enzyme Activity

References

Chapter 2: Methods of Assay

A. Chemical Methods

B. Instrumental Methods

1. Manometric Methods

2· Spectrophotometric Methods

3. Polarimetric Methods

4. Electrochemical Methods

5. Fluorescence Methods

6. Radiochemical Methods

References

Chapter 3: Determination of Enzymes

A. General

B. Hydrolytic Enzymes

1. Amylase

2. Cholinesterase

3. Cellulase

4. Chymotrypsin

5. Beta-glucosidase and Galactosidase

6. Beta-glucuronidase

7. Hyaluronidase

8. Lipase

9. Phosphatase

10. Urease

C Oxidative Enzymes

1. Amino Acid Oxidases

2. Catalase

3. Glucose Oxidase

4. Peroxidase

5. Xanthine Oxidase

D. Dehydrogenase Enzymes

1. General

2. Lactate Dehydrogenase

3. Glucose-6-phosphate Dehydrogenase

4. Glutamate Dehydrogenase

5. α-Hydroxybutyrate Dehydrogenase

6. Alcohol and Glycerol Dehydrogenases

7. Malate Dehydrogenase

E. Kinases and Transaminases

F. Other Enzymes

References

Chapter 4: Determination of Substrates

A. General

B. Carbohydrates

1. Glucose

2. Galactose

3. Fructose

4. Sucrose

5. Glycogen

6. Other Sugars

C. Amines

1. Use of Diamine Oxidase

2. Use of Monoamine Oxidase

3. Urea Analysis with Urease

D. Amino Acids

1. Determination of with D-amino Aids

2. Determination of with L-amino Acids with L-amino Acid Oxidase

3. Determination of with L-amino Acids with L-amino Acid Decarboxylases

4. Determination of Glutamic Acid

5. Determination of Alanine

6. Determination of L-aspartic Acid

7. Determination of Creatine

E. Organic Acids

1. Acetic Acid

2. Butyric Acid and its Hydroxy Derivatives

3. DL-citric and D-isocitric Acids

4. Formic Acid

5. Fumaric Acid

6. Hyaluronic Acid

7. Lactic Acid

8. Malic Acid

9. Succinic Acid

F. Hydroxy Compounds, Esters and Aldehydes

1. Ethanol and Aliphatic Alcohols

2. Glycerol and Dihydroxy Acetone

3. Hydroxy Pyruvate

4. Butylène Glycol

5. Sorbitol

6. Triglycérides

7. Phenols

8. Xanthine and Hypoxanthine

9. Steroid Alcohols

10. Detection and Determination of Esters

G. Inorganic Substances

1. Peroxide

2. Ammonia

3. Nitrate

4. Hydroxylamine

5. Carbon Dioxide

6. Phosphate

7. Pyrophosphate

References

Chapter 5: Determination of Activators and Co-enzymes

A. General

B. Determination of Inorganic Substances

1. Magnesium, Manganese and Zinc

2. Determination of Barium

3. Determination of Oxygen

4. Determination of Cyanide, Sulfide and Iodide

5. Other Substances

C. Determination of Co-enzymes

1. General

2. Flavine Mononucleotide

3. Flavine Adenine Dinucleotide

4. NAD and NADH

5. NADP and NADPH

6. Enzyme Cycling Methods for NAD and NADP

7. Adenosine-5'-triphosphate

8. Adenosine-5'-diphosphate and adenosine-5'-monophosphate

9. Creatine Phosphate

References

Chapter 6: Determination of Inhibitors

A. General

B. Determination of Inorganic Substances

1. General

2. Glucose Oxidase

3. Glucosidase

4. Hyaluronidase

5. Invertase

6. Isocitric Dehydrogenase

7. Liver Esterase

8. Peroxidase

9. Alkaline Phosphatase

10. Urease

11. Xanthine Oxidase

12. Other Enzyme Systems

C. Determination of Pesticides

1. Anticholinesterase Compounds (Organophosphorus and Carbamate Compounds)

2. DDT, Determination by Inhibition of Carbonic Anhydrase

3. Assay of Chlorinated and Other Pesticides Based on Inhibition of Lipase

4. Assay of Pesticides with Phosphatase

D. Determination of Other Organics

1. Sulfhydryl Binding Compounds

2. Heparin

3. Ascorbic Acid

4. Triton X-100

5. Other Compounds

References

Chapter 7: The Immobilized Enzyme

A. General

B. Immobilization in Starch Gel

1. General

2. Preparation of Immobilized Cholinesterase

3. Properties and Use of Immobilized Enzyme Gel

4. Automatic Monitoring of Inhibitors

C. Immobilization in Polyacrylamide Gels

1. General

2. Preparation of Gels

3. Properties of the Gel

4. Preparation of a Urea Electrode

5. Other Studies

D. Other Methods for Immobilizing Enzymes

1. General

2. Covalent Bonding to Polymeric Lattices

E. Commercial Availability of Immobilized Enzymes

F. Future Applications

References

Chapter 8: Use of Automation in Enzymic Analysis

A. General

B. Initial Slope Method

C. Fixed Concentration (Variable, Time) Methods

D. Fixed Time Method

References

Appendix

Author Index

Subject Index