High performance liquid chromatographic determination of glucose using an immobilized enzyme reactor

Abstract

Glucose was determined by using a high performance liquid chromatographic system comprised of an immobilized glucose oxidase reactor followed by amperometric detection of hydrogen peroxide. Glucose oxidase immobilized on pellicular particles of silica was packed into a short stainless steel column. The immobilized enzyme reactor incorporated with a C₁₈-bonded column. The sensitivity of this system was the same as that of chemiluminescence detection and about 5×10³-fold greater than when using differential refractive index detection. The least amount of glucose detected was 2ng (S/N=3). The uses of specific enzymatic reaction and the electrochemical detector made sample pretreatments extremely simple. This system was successfully used to determine glucose in carbonated water, sake, wine and soy sauce.