A novel fully enzymatic method for determining glucose and 1,5-anhydro-D-glucitol in serum of one cuvette.
The aim of the study was to set up a novel fully enzymatic method for screening glucose and 1,5-anhydro-D-glucitol (1,5-AG) in one cuvette. We have determined glucose and 1,5-AG, based on glucokinase (GK) converting glucose to G6P, a compound that can be catalyzed ultimately into 6-PGA by G-6PD and its coenzyme NADP(+), and then calculated glucose concentration according to absorbance variety. Furthermore, pyranose oxidase was used to oxidize 1,5-AG with the formation of 1, 5-anhydro-fructose and H(2)O(2). Measurement was done according to Trinder's reaction principle. The mean within-run and day-to-day precision (CV) of this method for glucose was 0.88% and 1.4%, and also that for 1,5-AG was 1.05% and 1.94%, respectively. The mean recovery rate of two targets was 100.2% and 101.6%, respectively. The correlation (R(2)) between the results of 1,5-AG obtained with our proposed method (y) and those obtained with LanaAG method (x) was 0.999 (y=1.002x-0.675 micromol/l; n=86), and the correlation (R(2)) of glucose between the results obtained with our GK method (y) and those obtained with recommendatory hexokinase method (x) was 0.9999 (y=1.0043x+0.1229 mmol/l; n=86). The reference range (95%) of serological glucose and 1,5-AG was 3.7 to 5.7 mmol/l (4.70+/-0.51 mmol/l) and 83.1 to 240.7 micromol/l (161.9+/-40.2 micromol/l), respectively; and there was no difference with age and sex (P>0.05). This newly developed method was dependable and steady-going, with analysis automatization, and allows quicker and easier measurement of serum glucose and 1,5-AG in one identical reaction cuvette in-phase than previously described methods.
Jing
Feng
F
Department of Laboratory Medicine Center of Taihe Hospital, Yunyang Medical College, 442000 Shiyan, China. fengjinggood@hotmail.Com
Jun
Lu
L
Wang
Yuping
Y
Zhu
Mingan
M
Yong
Zhang
Z
Fei
Xie
X
Zhang
Jicai
J
D001786:Blood Glucose / Q000032:analysis*; D002623:Chemistry Techniques, Analytical / Q000295:instrumentation / Q000379:methods; D003847:Deoxyglucose / Q000097:blood*; D003924:Diabetes Mellitus, Type 2 / Q000097:blood / Q000378:metabolism; D005941:Glucokinase / Q000378:metabolism; D005954:Glucosephosphate Dehydrogenase / Q000378:metabolism; D006801:Humans; D008954:Models, Biological; D015203:Reproducibility of Results
D001786:Blood Glucose; C006584:1,5-anhydroglucitol; D003847:Deoxyglucose; D005954:Glucosephosphate Dehydrogenase; D005941:Glucokinase