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Purification and Characterization

(1974). Recommended Methods for the Determination of Four Enzymes in Blood. Scandinavian Journal of Clinical and Laboratory Investigation: Vol. 33, No. 4, pp. 291-306.

Recommended Methods for the Determination of Four Enzymes in Blood

The metabolism of menadione (2-methyl-1,4-naphthoquinone) by isolated hepatocytes. A study of the implications of oxidative stress in intact cells.

Eine einfache enzymatische Methode zur Bestimmung des Gesamt-Cholesterins im Serum wird beschrieben. Die im Serum vorhandenen Cholcsterinester werden mittels Cholcstcrinesterase quantitativ in freies Cholestcrin und Fettsauren gespalten. Das freie Cholesterin wird in Gegenwart von Sauerstoff und Cholesterinoxydase in A-Cholestenon umgewandelt. Das bei dieser Reaktion entstehende Wasserstoffperoxyd oxydiert bei Anwesenheit von Katalase Methanol zu Formaldchyd. Dieses reagiert mit Ammoniumionen und Acetylaceton unter Bildung von 3,5-Diacctyl-l,4-dihydrolutidin, dessen Farbintcnsitat bei 405 nm gemessen wird. Richtigkeit und Prazision der Methode sind sehr gut. Die Proportionalitat ist bis 25,85 mmol Cholcsterin/1 Serum gegeben. Serumprotcine, Bilirubin, Creatinin, Hamoglobin und Pharmaka storen nicht.

[Enzymatic determination of total cholesterol in serum (author's transl)]

Comparative lipid peroxidation, antioxidant defense systems and proline content in roots of two rice cultivars differing in salt tolerance

Methoden der enzymatischen Analyse

Publisher Summary This chapter presents an overview of the principles of enzymatic analysis. The value of enzymes in analysis lies in their ability to react specifically with individual components of a mixture. The chapter highlights that the concentration of a substance which takes part in an enzymatic reaction can be determined in two ways: one, by physical, chemical, or enzymatic analysis of the product or unreacted starting material after completion of the reaction catalyzed by the enzyme; and two, from the rate of the enzyme reaction, which depends on the concentration of the substrate, cofactor, activator or inhibitor. The two methods are basically different. In the first case, the reaction should be completed as rapidly as possible. Relatively large amounts of enzyme and relatively small amounts of substrate are used. The measured values should be easily readable, not too small and not too large. In the second case, the substrate and enzyme concentrations are so arranged that the rate of the reaction, that is, the amount of substrate reacting per unit time, is not too fast and so can be measured accurately.

Principles of Enzymatic Analysis

700 authors from 29 countries describe the methods of enzymatic analysis in a way that corresponds to the various steps undertaken in laboratory routine. Designed for use in the laboratory rather than the library, the third edition takes into account recent changes in symbolism, nomenclature, and enzyme classification. The series recognizes the fact that today many more enzymes, coenzymes, and substrates are commercially available than ten years ago and also provides information on the precision of measurement, the sensitivity and limitation of a method, and on possible alternatives, such as HPLC, GC, AA, or NMR spectroscopy. In general, the methods described concern analyses that are of importance in biochemistry, and clinical chemistry. Annotations by the authors convey their personal experience by including useful hints, remarks, and experimental tricks to make the methods truly workable. A cumulative index simplifies the search for information.

Hans Ulrich Bergmeyer

Papers

Methoden der enzymatischen Analyse

Principles of Enzymatic Analysis

Lipids, amino acids and related compounds

Samples, reagents, assessment of results

Peptidases, proteinases and their inhibitors