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Glucose

Experiment #1. Glucose. H.AlDahshan H.AlTwaijry. - The blood glucose level is normally maintained within a narrow range under various conditions by hormones ,such as insulin, glucagon, or epinephrine.

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Glucose

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  1. Experiment #1 Glucose H.AlDahshan H.AlTwaijry

  2. -The blood glucose level is normally maintained within a narrow range under various conditions by hormones ,such as insulin, glucagon, or epinephrine. -Measurement of blood glucose is one of the most commonly performed tests in hospital chemistry laboratories.

  3. _The most frequently encountered disorder of carbohydrate metabolism is high blood glucose cause by diabetes mellitus (DM) -Patients with diabetes demonstrate an inability to produce insulin

  4. -The incidence of hypoglycemia (low blood glucose) is unknown, but, excluding patients using insulin to control their blood glucose, it is low -Clinical diagnosis should not be made on a single test result; it should integrate clinical and other lab data.

  5. Principle of the method Glucose oxidase (GOD) catalases the oxidation of glucose to gluconic acid. The formed hydrogen peroxide (H2o2), is detected by a chromogenic oxygen acceptor, phenol-aminophenazonein the presence of peroxidase (POD)

  6. COD Glucose+o2+H2o Gluconicacid +H2o2 POD H2o2+4 aminoantipyrine+PHBS RED COLOR+4H2O The intensity of the color formed is proportional to the glucose concentration in the sample

  7. Samples Serum or plasma , free of hemolysis and CSF Serum should be removed from the clot as quickly as possible Stability: Glucose is stable at 2-8 ºC for 3 days

  8. Procedure 1. Assays conditions: Wavelength……………………..510 nm (490-550) Cuvette…………………………….1 cm light path Temperature…………………….37 ºc\15-25 ºc 2 .Adjust the instrument to zero with distilled water

  9. 3. Pipette into a cuvette

  10. 4. Mix and incubate for 10 min at 37ºc or 15-20 min at room temp (15-25ºC) 5. Read the absorbance (A) of the sample and standard , against the Blank. The color is stable for at least 30 min

  11. CALCULATIONS glucose (A)Sample x 100(stdconc)= mg/dL (A)standard Conversion factor: mg/dL x 0.0555=mmol/L

  12. Reference values 60-110 mg/dL 3.33-6.10 mmol/L

  13. Interferences • Haemoglobin up to 4g/L • Bilirubin up to 20 mg/L • Creatinine up to 100mg/L • Galactose up to 1g/L Do not interfere

  14. Fasting plasma glucose (FPG) versus oral glucose tolerance test (OGTT): OGTT is performed by serial measurement of plasma glucose before and after a specific amount of glucose is given orally. Although OGTT is more sensitive than FPG, it is affected by a large number of factors that result in poor reproducibility. Therefore, an OGTT is rarely necessary for the diagnosis of DM.

  15. Whole blood versus plasma glucose: Most lab instruments measures plasma glucose, but some whole blood .Plasma glucose yields more reliable results. It is 10-15% higher than whole blood glucose, since RBC contain less water per unit volume than plasma.

  16. Capillary versus venous plasma glucose: Normally, there is little difference between capillary and venous blood glucose. However, in hyperglycemia, capillary plasma glucose may be significantly higher than venous plasma glucose.

  17. Hemoglobin A1C (glycosylated Hemoglobin): Hb A1C is a test that can be used to monitor the degree of the control of blood glucose level during the past 2 moths

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