GLUCOSE DETERMINATION ENZYMATIC-COLOMETRIC GLUCOSE OXIDASE-PAP METHOD (TRINDER

GLUCOSE DETERMINATION ENZYMATIC-COLOMETRIC GLUCOSE OXIDASE-PAP METHOD (TRINDER REACTION) Glucose in the blood is one of the analytes commonly tested in routine blood chemistry. It is determined either in random blood or fasting blood. It is also assayed as part of glucose tolerance. Glucose is increased in diabetes mellitus and in other secondary diseases characterized by hyperglycemia. It is decreased in various conditions of hypoglycemia such as insulinoma or functional hypoglycemia. Understanding the glucose metabolism is important to be able to correlate the abnormal levels with various disease states. Glucose is assayed in the blood, serum or plasma by various methods. The enzymatic methods are most commonly utilized both in manual techniques and automation. Early enzymatic methods for glucose determinations involved glucose oxidase to catalyze the oxidation of glucose. Keston modified this method in the early 1950’s using glucose oxidase/peroxidase enzyme system and o-dianisidine chromogen system. Since then, various alternative chromogen systems have evolved. A few colorimetric methods remain practical such as the ortho-toluidine method and neocuproine method. Proper specimen processing should be emphasized to avoid glycolysis, bacterial contamination and hemolysis as these may cause alteration in the accurate estimation of the glucose level. Glucose is also assayed in urine, spinal fluid and other body fluids. Questions for Write-up: 1. Illustrate and explain the glucose metabolism. 2. Give and describe the principles of the different methods of glucose determination. a) Copper reduction methods b) Condensation methods c) Polarographic method 3. Describe the clinical significance of abnormal glucose levels a) Hyperglycemia b) Hypoglycemia 4. Give at least 5 sources of errors that affect the accuracy of glucose assay. 5. What other specimens are analyzed for glucose? Gove the significance of the assay. 6. What are the glucose values diagnostic for diabetes mellitus? 7. Describe the following: a) Random glucose testing b) 2-hour glucose tolerance test c) O’ Sullivan glucose tolerance test d) Intravenous glucose tolerance test 8. What other tests are done to monitor diabetes mellitus? Notes: Please kindly make the answer accurate, informative and precise. Include the references.

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