Enzymatic Methods of Analysis

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.

Contenu

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

8. Total Change or Equilibrium Method

9. 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

2. Polarimetric Methods

3. Electrochemical Methods

4. Fluorescence Methods

5. 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

11. Amino Acid Oxidases

12. Catalase

13. Glucose Oxidase

14. Peroxidase

15. Xanthine Oxidase

    D. Dehydrogenase Enzymes

16. General

17. Lactate Dehydrogenase

18. Glucose-6-phosphate Dehydrogenase

19. Glutamate Dehydrogenase

20. a-Hydroxybutyrate Dehydrogenase

21. Alcohol and Glycerol Dehydrogenases

22. 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

7. Use of Diamine Oxidase

8. Use of Monoamine Oxidase

9. Urea Analysis with Urease

   D. Amino Acids

10. Determination of with D-amino Aids

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

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

13. Determination of Glutamic Acid

14. Determination of Alanine

15. Determination of L-aspartic Acid

16. Determination of Creatine

    E. Organic Acids

17. Acetic Acid

18. Butyric Acid and its Hydroxy Derivatives

19. DL-citric and D-isocitric Acids

20. Formic Acid

21. Fumaric Acid

22. Hyaluronic Acid

23. Lactic Acid

24. Malic Acid

25. Succinic Acid

    F. Hydroxy Compounds, Esters and Aldehydes

26. Ethanol and Aliphatic Alcohols

27. Glycerol and Dihydroxy Acetone

28. Hydroxy Pyruvate

29. Butylène Glycol

30. Sorbitol

31. Triglycérides

32. Phenols

33. Xanthine and Hypoxanthine

34. Steroid Alcohols

35. Detection and Determination of Esters

    G. Inorganic Substances

36. Peroxide

37. Ammonia

38. Nitrate

39. Hydroxylamine

40. Carbon Dioxide

41. Phosphate

42. 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

6. General

7. Flavine Mononucleotide

8. Flavine Adenine Dinucleotide

9. NAD and NADH

10. NADP and NADPH

11. Enzyme Cycling Methods for NAD and NADP

12. Adenosine-5'-triphosphate

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

14. 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

13. Anticholinesterase Compounds (Organophosphorus and Carbamate Compounds)

14. DDT, Determination by Inhibition of Carbonic Anhydrase

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

16. Assay of Pesticides with Phosphatase

    D. Determination of Other Organics

17. Sulfhydryl Binding Compounds

18. Heparin

19. Ascorbic Acid

20. Triton X-100

21. 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

5. General

6. Preparation of Gels

7. Properties of the Gel

8. Preparation of a Urea Electrode

9. Other Studies

   D. Other Methods for Immobilizing Enzymes

10. General

11. 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