Polarographic analysis of cholesterol and other macromolecular substances

A method for the quantitative determination of cholesterol in blood which comprises the steps of

• providing a polarographic cell including at least one electrode sensitive to hydrogen peroxide positioned behind a membrane permeable to hydrogen peroxide and impermeable to cholesterol,
• separating from said blood the component selected from the group consisting of blood plasma and blood serum,
• introducing cholesterol oxidase into said cell on the side of the membrane opposite said electrode for enzymatic reaction with cholesterol to produce hydrogen peroxide,
• providing oxygen in said cell for said enzymatic reaction,
• establishing a potential across said cell such that current is produced which is proportional to the amount of hydrogen peroxide produced,
• adding a quantity of said component for reaction with said enzyme and to effect diffusion of at least a portion of said hydrogen peroxide into said membrane and into contact with said electrode, and
• determining the current flowing across said cell as a function of the amount of hydrogen peroxide formed and as an indication of the amount of cholesterol in said blood.

A method for the quantitative determination of cholesterol in whole blood which comprises the steps of

• providing a polarographic cell including at least one electrode sensitive to hydrogen peroxide positioned behind a membrane permeable to hydrogen peroxide and impermeable to cholesterol,
• immobilizing cholesterol oxidase on the membrane on the side opposite said electrode for enzymatic reaction with cholesterol to produce hydrogen peroxide,
• providing oxygen in said cell for said enzymatic reaction,
• establishing a potential across said cell such that current is produced which is proportional to the amount of hydrogen peroxide produced,
• adding a quantity of said blood for reaction with said enzyme and to effect diffusion of at least a portion of said hydrogen peroxide into said membrane and into contact with said electrode, and
• determining the current flowing across said cell as a function of the amount of hydrogen peroxide formed and as an indication of the amount of cholesterol in said blood.