Development of a sequential injection analysis system for monitoring of trehalose concentrations

Abstract

A sequential injection analysis (SIA) system was developed to on-line monitor the concentrations of trehalose in biological processes. The system is based on the enzymatic reaction of trehalose to glucose in an immobilized trehalase reactor and the subsequent reaction of glucose in the presence of glucose oxidase. The peroxide produced is detected using horseradish peroxidase with a redox indicator (ABTS, 2,2′-azino-di(3-ethylbezthiazoli-sulfonic acid-6)) and a spectrophotometer set at 435 nm. The SIA was fully automated using software written in the LabVIEWTM development environment. A number of system variablese.g. flow rate of the carrier buffer solution and volume ratio of sample to reagents, were evaluated to increase the sensitivity and performance of the SIA system. The performance of the trehalose-SIA system was linear under partially optimized operating conditions in the concentration range of 0.3 to 2.0 g/L (R2=0.999) with a sample frequency of 6 1/h. The SIA system was successfully employed to on-line monitor the concentrations of trehalose in a continuously stirred tank reactor. The on-line monitored data were in good agreement with the off-line data measured by a HPLC with a refractive index detector (n=14, R2=0.9865).