Page 30 - Phytochemistry -1 (PG404) / Clinical Pharmacy 2nd level students ( 2019 )
P. 30
Clinical pharmacy PharmD program Third level Phytochemistry-1 (PG-504)
II. Colorimetric methods
Enzymatic Method
Estimation of glucose by enzymatic method
Principle:
• Glucose oxidase enzyme catalyses the oxidation of glucose as
follow:
• The formed H 2O 2, reacts with 4-aminoantipyrene and phenol in the
presence of peroxidase enzyme to get quitionimine which is red in
color.
• The amount of dye formed is proportional to the glucose % and can
be measured calorimetrically.
Reagents:
• Reagent 1: is a buffer solution composed of tris buffer (PH = 7.5)
92 mmol/L and phenol 0.3 mmol/L.
• Reagent 2: is composed of the enzymes glucose oxidase and
peroxidase enzymes and 4-aminoantipyrene (2.6 mmol/L).
Dissolve the content of reagent 2 bottle in the content of one bottle of
reagent 1
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