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Glucose, quantitative determination

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Presentation on theme: "Glucose, quantitative determination"— Presentation transcript:

1 Glucose, quantitative determination

2 Hormonal regulation of blood glucose

3 Methods Chemical Enzymatic

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6 CHEMICAL METHODS Oxidation - Reduction Methods Condansation Methods
Folin – Wu Somogyi - Nelson Condansation Methods Orto - Toluidine

7 Benedict reaction

8 Glucose + Cu2+ >>>Gloconate + Cu+
Folin - Wu Glucose + Cu2+ >>>Gloconate + Cu+ pH > 7 , heat Cu+ + phosphomolybdate Blue molybdenium complex λ = 660 nm

9 Glucose + Cu2+ >>> Gloconate + Cu+
Somogyi - Nelson Glucose + Cu2+ >>> Gloconate + Cu+ pH > 7 , heat Cu+ + arsenomolybdate >>> blue molybdenium complex λ = 660 nm

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

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12 Interference Bilirobin Galactose Mannose Hemoglobin

13 Enzymatic Methods Glucose Oxidase Hexokinase Glucose dehydrogenase

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

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

16 Hexokinase method

17 Glucose dehydrogenase method

18 Clinical Significance
F.B.S (Fasting Blood Suger) Fasting hours Normal Range mg/dL Decrease Hypoglycemia Increase Hyperglycemia

19 Criteria for the diagnosis of diabetes mellitus

20 Diagnosis of Pre-Diabetes and Diabetes Mellitus

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

22 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

23 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

24 Characteristics of Type 1 and Type 2 Diabetes

25 Response of plasma insulin to glucose stimulation

26 Secondary hyperglycemia
Pancratectomy Acromegaly Cushing’s syndrome Glucagonoma

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

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

29 OGTT Fasting 80 mg/dL mg/dL mg/dL mg/dL mg/dL

30 Oral Glucose Tolerance Tests (OGTT)

31 Screening and diagnosis of gestational diabetes mellitus

32 OGTT curve

33 2hpp 2hpp (two hours postprandial plasma glucose)
To screen for diabetes mellitus To monitor glucose control

34 Glycated hemoglubin HbA1c

35 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

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


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