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    Glucose Estimation

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    ZahidKhan
    This document describes the glucose oxidase method for estimating blood glucose levels. It involves using the enzyme glucose oxidase to catalyze the oxidation of glucose into gluconic acid and hydrogen peroxide. The hydrogen peroxide then reacts with 4-aminophenazone and phenol to form a colored compound, with the intensity of color proportional to the glucose concentration. The procedure involves taking blood and standard samples, adding glucose reagent, measuring absorbance after 10 minutes, and calculating glucose concentration using the standard absorbance values. Normal blood glucose ranges provided are 70-110 mg/dl fasting and 70-180 mg/dl random.

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    Glucose Estimation

    Uploaded by

    ZahidKhan
    124 views10 pages
    This document describes the glucose oxidase method for estimating blood glucose levels. It involves using the enzyme glucose oxidase to catalyze the oxidation of glucose into gluconic acid and hydrogen peroxide. The hydrogen peroxide then reacts with 4-aminophenazone and phenol to form a colored compound, with the intensity of color proportional to the glucose concentration. The procedure involves taking blood and standard samples, adding glucose reagent, measuring absorbance after 10 minutes, and calculating glucose concentration using the standard absorbance values. Normal blood glucose ranges provided are 70-110 mg/dl fasting and 70-180 mg/dl random.

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    This document describes the glucose oxidase method for estimating blood glucose levels. It involves using the enzyme glucose oxidase to catalyze the oxidation of glucose into gluconic acid and hydrogen peroxide. The hydrogen peroxide then reacts with 4-aminophenazone and phenol to form a colored compound, with the intensity of color proportional to the glucose concentration. The procedure involves taking blood and standard samples, adding glucose reagent, measuring absorbance after 10 minutes, and calculating glucose concentration using the standard absorbance values. Normal blood glucose ranges provided are 70-110 mg/dl fasting and 70-180 mg/dl random.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    124 views10 pages

    Glucose Estimation

    Uploaded by

    ZahidKhan
    This document describes the glucose oxidase method for estimating blood glucose levels. It involves using the enzyme glucose oxidase to catalyze the oxidation of glucose into gluconic acid and hydrogen peroxide. The hydrogen peroxide then reacts with 4-aminophenazone and phenol to form a colored compound, with the intensity of color proportional to the glucose concentration. The procedure involves taking blood and standard samples, adding glucose reagent, measuring absorbance after 10 minutes, and calculating glucose concentration using the standard absorbance values. Normal blood glucose ranges provided are 70-110 mg/dl fasting and 70-180 mg/dl random.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
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    Estimation of Blood Glucose level

    Mr. Arif Ullah Khan Niazi


    Lecturer
    Bannu College of Medical Technologies Bannu
    BLOOD GLUCOSE
    There are three commonly used methods for glucose
    estimation, which are;-
    • Copper reduction method (Folin-Wu method)
    • Other reduction method such as orthotoludine
    method
    • Enzymatic methods such as
    o Hexokinase method
    o Glucose oxidase method (Recommended)
    o Glucose dehydrogenase method
    Estimation of Blood Glucose Level
    Glucose Oxidase Method (Enzymatic method)
    Apparatus:
    Spectrophotometer/Colorimeter
    Adjustable micropipette
    Blood collection kit
    Ordinary test tubes
    Centrifuge with test tubes.
    Estimation of Blood Glucose Level
    Chemical required: Blood sample, Glucose standard, Glucose
    reagent & Anti-coagulant
    1. Concentration of Glucose standard= 100 mg/dl
    2. Composition of Glucose reagent:
    I. Glucose oxidase---------------------15 KU/L
    II. Mutarotase----------------------------2 KU/L
    III. Hydrogen peroxidase-------------1.5 KU/L
    IV. Phenol------------------------------0.75 mol/L
    V. 4-aminophenazone--------------0.25 mol/L
    VI. Phosphate buffer-------------------0.1 mol/L
    Estimation of Blood Glucose Level
    Principal:
    The enzyme Glucose oxidase catalyzes the oxidation of glucose
    to Gluconic acid with the formation of Hydrogen peroxidase.
    The enzyme peroxidase breaks down hydrogen peroxidase into
    water and nascent oxygen. The nascent oxygen reacts with the
    colorless electron acceptor 4-aminophenazone which is
    oxidized to colored compound. The reaction can be followed
    photo metrically. The intensity of color produced is
    proportional to the amount of glucose in the sample:
    Glucose+H2O+O2-------------------------------Gluconic acid+H2O2
    Glucose oxidase
    Estimation of Blood Glucose Level
    Procedure:
     Take 20 micro liter of the sample in the adjustable micropipette. Pour it into a small
    test tube labeled as “UNKNOWN”.
     Take 20 micro liter of the glucose standard in another small test tube labeled as
    “STANDARD”.
     In another small test tube labeled as “BLANK” take 20 micro liters of distilled water.
     Now add 2 ml of the glucose reagent to all the three test tubes. Wait for at least 10
    minutes so that the reaction is complete at room temperature.
     Now switch on the colorimeter or spectrophotometer and do its blanking by first
    placing the “BLANK” in the path of the light.
     Use the filter/monochromator of special wavelength. The routinely used is of 546
    nm. Optical length of cuvette=1 cm. Temperature =20-25 or 37 degree centigrade.
    Note the transmittance or the absorbance of the unknown and that of the
    standard(known).
    Estimation of Blood Glucose Level
    Observation & Calculation:
    Formula : C1/C2 = A1/A2
    C1=A1/A2 × C2
    Where C1= Concentration of Unknown/Test=?
    C2= Concentration of Standard=1mg/ml.
    A1= Color absorbance of Unknown/Test.
    A2= Color absorbance of Standard.
    Estimation of Blood Glucose Level

    Observation & Calculation:

    Formula: Concentration of T × 100


    Concentration of S
    Estimation of Blood Glucose Level
    Normal Blood Glucose range:
    Venous blood (Fasting)= 70-110 mg/dl
    Venous blood (Random)= 70-180 mg/dl
    CSF = 50-80 mg/dl
    Urine= up to 25 mg/dl
    THANK YOU

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