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Glucose, quantitative determination

Glucose, quantitative determination. Hormonal regulation of blood glucose. Methods. Chemical Enzymatic. CHEMICAL METHODS. Oxidation - Reduction Methods Folin – Wu Somogyi - Nelson Condansation Methods Orto - Toluidine. Benedict reaction. Folin - Wu.

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Glucose, quantitative determination

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  1. Glucose, quantitative determination

  2. Hormonal regulation of blood glucose

  3. Methods • Chemical • Enzymatic

  4. CHEMICAL METHODS • Oxidation - Reduction Methods • Folin – Wu • Somogyi - Nelson • Condansation Methods • Orto - Toluidine

  5. Benedict reaction

  6. Folin - Wu Glucose + Cu2+>>>Gloconate+ Cu+ pH > 7 , heat Cu+ + phosphomolybdate Blue molybdeniumcomplex λ = 660 nm

  7. Somogyi - Nelson Glucose + Cu2+>>> Gloconate + Cu+ pH > 7 , heat Cu+ + arsenomolybdate>>> blue molybdeniumcomplex λ= 660 nm

  8. O - Toluidine Method o - Toluidine + Glucose>>> Glycosylamine pH < 7 , heat Schiff Base λ = 630 nm color complex NH2 CH3

  9. Interference • Bilirobin • Galactose • Mannose • Hemoglobin

  10. Enzymatic Methods • Glucose Oxidase • Hexokinase • Glucose dehydrogenase

  11. Glucose Oxidase Mutarotase α -D-Glucose β-D-Glucose O2 glucose oxidase Gluconic acid + H2O2 Peroxidase H2O2+ chromogenColor complex (o-dianisidine) + H2O (phenylamine)λ = 520 nm

  12. Glucose oxidase Inhibited by • High concentrations of uric acid, ascorbic acid, bilirubin, glutathione, creatinine. L-cysteine, L-dopa. Dopamine, methyldopa and citric acid

  13. Hexokinase method

  14. Glucose dehydrogenase method

  15. Clinical Significance • F.B.S (Fasting Blood Suger) • Fasting 12 - 14 hours • Normal Range 70 - 110 mg/dL • Decrease • Hypoglycemia • Increase • Hyperglycemia

  16. Criteria for the diagnosis of diabetes mellitus

  17. Diagnosis of Pre-Diabetes and Diabetes Mellitus

  18. Hyperglycemia • FBS > 140 mg/dL • Primary (Diabetes Mellitus) • IDDM (type I) • NIDDM (type II)

  19. Insulin Dependent Diabetes Mellitus (IDDM) • Absolute deficiency of insulin • Make up about 10% of all patients with D.M. • IDDM persent at an early age (usually before 30) • Clinical signs : • Blood glucose • Insulin (decreased) • Ketosis • Loss of body weight

  20. Noninsulin Dependent Diabetes Mellitus (NIDDM) • Deficiency of insulin receptor • Patients are commonly obese • NIDDM is the most common from of D.M. ( 80-90%) • Usually first present at an age over 40 • Clinical signs • Blood glucose • Insulin • Osmotic diuresis • Dehydration

  21. Characteristics of Type 1 and Type 2 Diabetes

  22. Response of plasma insulin to glucose stimulation

  23. Secondary hyperglycemia • Pancratectomy • Acromegaly • Cushing’s syndrome • Glucagonoma

  24. Hypoglycemia • Adult < 45 - 50 mg/dL in fasting • Infant • Preterm < 25 mg/dL • Fullterm < 35 mg/dL

  25. Causes of hypoglycemia • Insulinoma (islet cell tumors) • Glycogen Storage Diseases (GSD) • Addison’s disease

  26. OGTT Fasting 80 mg/dL 0.5 155 mg/dL 1 165 mg/dL 2 140 mg/dL 3 80 mg/dL

  27. Oral Glucose Tolerance Tests (OGTT)

  28. Screening and diagnosis of gestational diabetes mellitus

  29. OGTT curve

  30. 2hpp • 2hpp (two hours postprandial plasma glucose) • To screen for diabetes mellitus • To monitor glucose control

  31. Glycated hemoglubin • HbA1c

  32. an index of average blood glucose levels over the past 2-4 months. • People without diabetes have HbA1c levels between 4-6%. • Measure HbA1C levels every 3-6 months to monitor glycemic control

  33. Put 10 minute in boil water bath • read at 630 nm • Ct = At / Ast * Cst

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