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A Novel Fully Enzymatic Method for Determining Glucose and 1,5-Anhydro-d-Glucitol in Serum of One Cuvette

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Abstract

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 H2O2. 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 μmol/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 μmol/l (161.9 ± 40.2 μmol/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.

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Abbreviations

1,5-AG:

1,5-anhydro-d-glucitol

PROD:

pyranose oxidase

GK:

glucokinase

G6P:

glucose 6-phosphate

HRP:

horseradish peroxidase

G-6PD:

glucose-6-phosphate dehydrogenase

HTIB:

3-hydroxy-2,4,6-triiodobenzoic acid

4-AAP:

4-aminoantipyrine

TG:

triglyceride

Hb:

hemoglobin

T bil:

total bilirubin

HK:

hexokinase

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Authors and Affiliations

  1. Department of Laboratory Medicine Center of Taihe Hospital, Yunyang Medical College, 442000, Shiyan, China

    Feng Jing, Lu Jun, Yuping Wang, Mingan Zhu, Zhang Yong, Xie Fei & Jicai Zhang

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  1. Feng Jing
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  2. Lu Jun
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  3. Yuping Wang
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  4. Mingan Zhu
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  5. Zhang Yong
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  6. Xie Fei
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  7. Jicai Zhang
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Correspondence to Feng Jing.

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Jing, F., Jun, L., Wang, Y. et al. A Novel Fully Enzymatic Method for Determining Glucose and 1,5-Anhydro-d-Glucitol in Serum of One Cuvette. Appl Biochem Biotechnol 150, 327–335 (2008). https://doi.org/10.1007/s12010-008-8173-8

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