A micro-method for measurement of sterols such as cholesterol, and other macromolecular substances, is disclosed utilizing enzymes for the conversion of such substances to produce ultimately hydrogen peroxide and measurement of the generated hydrogen peroxide with a membrane covered polarographic anode. The polarographic anode is set at a voltage so as to produce current proportional to hydrogen peroxide concentration. According to the method of this invention, the macromolecular substance under analysis enters into an enzymatic reaction in a sample chamber on the side of the membrane opposite the anode and the membrane is impermeable to such substances, but senses hydrogen peroxide. The method is adapted to measure free and total blood cholesterol in a precise, rapid, sensitive and specific manner. Other substances of a macromolecular or conventional membrane impermeable nature such as high molecular weight starches or proteins, which undergo enzymatic reaction to produce ultimately hydrogen peroxide, may be analyzed by employing this polarographic technique.