A simple automated microplate method for determining reducing sugars in food extracts and synthetic serum using cupric-neocuproine as reductant

Authors : Mustafa BENER, Esin AKYÜZ, Furkan Burak ŞEN, Kevser Sözgen BAŞKAN, Esma TÜTEM, Mustafa Reşat APAK

Pages : 794-807

View : 13 | Download : 5

Publication Date : 0000-00-00

Article Type : Research Paper

Abstract :In the present work, a simple automated microplate method based on cupric ion reduction is described for determining total reducing sugars in food extracts and synthetic serum. The reaction of Cuinsert ignore into journalissuearticles values(II);-Nc insert ignore into journalissuearticles values(cupric-neocuproine); with reducing sugars was performed in alkaline medium in microplates, and the absorbance of the formed highly colored Cuinsert ignore into journalissuearticles values(I);-Nc chelate in a plate reader at 450 nm was recorded. The proposed method was applied to reducing sugars insert ignore into journalissuearticles values(glucose, fructose, galactose, maltose, and lactose); and their linear calibration curves were constructed. The detection and quantification limits insert ignore into journalissuearticles values(LOD and LOQ); for glucose were 0.14 and 0.46 $\mu $M, respectively. Absorbances of glucose were linear within the concentration range 2.5-54.2 $\mu $M and the method showed high linearity insert ignore into journalissuearticles values(r = 0$.$9998); over a relatively broad concentration range of analyte. This automated microplate method was validated through linearity, additivity, precision insert ignore into journalissuearticles values(RSD%, 2.33-6.65);, and recovery insert ignore into journalissuearticles values(101%-103%);, revealing that the method is reliable and robust for determining reducing sugars. Total reducing sugar contents of synthetic sugar mixtures, fruit juices, milk, and synthetic serum samples were successfully determined with the proposed method. The results were compared to those of the conventional alkaline Cuinsert ignore into journalissuearticles values(II);-Nc spectrophotometric method. The proposed method offers many advantages when compared to classical methods, such as insert ignore into journalissuearticles values(sample and reagent); volume reduction insert ignore into journalissuearticles values(20-fold);, simplicity, multiple sample analysis insert ignore into journalissuearticles values(32 samples in 4 h);, and environmental friendliness.
Keywords : Reducing sugars, glucose determination, microplate reader, fruit juice, serum