The influence of incubation time on the yield of enzymatic conversion at room temperature was investigated first, using 200 ppM glucose standard solution and the assay protocol shown on the previous page. It was found that reaction yield increses linearly with stop flow time, while the blank value, although high, remains constant (A).
The influence of temperature was investigated by using 200 ppM glucose standard and incubation period of 25 seconds.(B). Since influence of temperature on peak height was small both for glucose standard (200 ppM) and reagent blank all further experiments were carried out at 25 C.
Calibration in the range of 0 to 600 ppM glucose using stop flow of 25 sec was linear with LOD 20 ppM. (C) The sampling frequency of 61 s/hr (D) could be further accelerated by decreasing stop time to 10 sec and by accelerating flowrate after peak maximum was recorded.