Abstract
To obtain maximal linear regression correlation coefficient (R 2) of glucose standard curve determinations, single-factor test and response surface methodology were applied to optimize conditions including reaction time, incubation temperature and phenol concentration. For investigating optimization of this method, the effects of different metal ions, amino acids and proteins, respectively, were evaluated. The contents of several polysaccharides were determined by three methods, and it was found that the absorption value at 490 nm of the reaction system (230 μL) placed in the microplate involving 150 μL sulfuric acid and 30 μL 5 % phenol solution was in linear relationship with the concentration of glucose range from 10 to 100 nmol/well at 85 °C for 20 min. Our results also suggested that the method had a relative deviation of 0.4 %, and the recovery was between 102.7 and 104.10 %.
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References
Alberto, L., Q. Anelis, and M. Sánchez. 2008. Rapid and sensitive anthrone-sulfuric acid assay in microplate format to quantify carbohydrate in biopharmaceutical products: method development and validation. Biologicals 36: 134–141.
Breuil, C., and J.N. Saddler. 1985. Comparison of the 3,5-dinitrosalicylic acid and Nelson-Somogyi methods of assaying for reducing sugars and determining cellulase activity. Enzyme and Microbial Technology 7(7): 327–332.
Dai, Y., N. Chen, S. Jia, N.N. Yuan, N. Tan, and J. Li. 2012. Purification of polysaccharide from photoautotrophic cultivation of Nostoc flagelliforme cells with sector radial flow chromatography[C]. Biomedical Engineering and Biotechnology 2012: 465–468.
Ding, X., Y.L. Hou, and W.R. Hou. 2012. Structure feature and antitumor activity of a novel polysaccharide isolated from Lactarius deliciosus gray. Carbohydrate Polymers 89: 397–402.
Dreywood, R. 1946. Qualitative test for carbohydrate material. Analytical Chemistry 18(8): 499.
Hu, Y.Z. 2000. The multivariate numerical analysis in chemistry. Beijing: Higher Education Press.
Khuri, A.I., and S. Mukhopadhyay. 2010. Response surface methodology. Wiley Interdisciplinary Reviews Computational Statistics 2: 128–149.
Laurentin, A., and C.A. Edwards. 2003. A microtiter modification of the anthrone-sulfuric acid colorimetric assay for glucose-based carbohydrates. Analytical Biochemistry 315: 143–145.
Li, H., G.Y. Zhou, J.P. Xu, J.A. Liu, H.Y. Zhang, and Y.M. Tan. 2012. Research progress on polysaccharides from Ginkgo biloba. Journal of Medicinal Plants Research 6: 171–176.
Masuko, T., A. Minami, N. Iwasaki, T. Majima, S. Nishimura, and Y.C. Lee. 2005. Carbohydrate analysis by a phenol-sulfuric acid method in microplate format. Analytical Biochemistry 339: 69–72.
Peng, H.S., J.C. Chang, C.C. Chiu, P.Y. Lai, and H.L. Lay. 2013. Optimizing strawberry fruit yield with response surface methodology in a central composite design. Crop Science 53: 786–792.
Rouau, X., and A. Surget. 1994. A rapid semi-automated method for the determination of total and water-extractable pentosans in wheat flours. Carbohydrate Polymers 24: 123–132.
Somani, B.L., J. Khanade, and R. Sinha. 1987. A modified anthrone-sulfuric acid method for the determination of fructose in the presence of certain proteins. Analytical Biochemistry 167: 327–330.
Tarley, C.R.T., G. Silveira, W.N.L. Dos-Santos, G.D. Matos, E.G.P.D. Silva, M.A. Bezerra, M. Miro, and S.L.C. Ferreira. 2009. Chemometric tools in electroanalytical chemistry: methods for optimization based on factorial design and response surface methodology. Microchemical Journal 92: 58–67.
Taylor, K.A.C.C. 1995. A modification of the phenol/sulfuric acid assay for total carbohydrates giving more comparable absorbance. Applied Biochemistry and Biotechnology 53: 207–214.
Acknowledgments
We gratefully acknowledge Professor Jianzhong Ma and Associate Professor Hui Ma for their kindly suggestions during the experimental design and data analysis.
Funding
This study was funded by Special Scientific Research Fund of Agricultural Public Welfare Profession of China (Grant Numbers 201203042, 201403048-8), Science and Technology Innovation Projects of CAAS (Grant Numbers CAAS-ASTIP-2014-LIHPS-08), Chinese National Natural Science Foundation (Grant Number 31460032), Natural Science Foundation of Gansu Province (Grant Numbers 1212RJYA008, 1308RJZA287) and the Foundation of Excellent Young Teachers of LUT (Grant Number 10-061406).
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Wang, Y., Jing, Y., Leng, F. et al. Establishment and Application of a Method for Rapid Determination of Total Sugar Content Based on Colorimetric Microplate. Sugar Tech 19, 424–431 (2017). https://doi.org/10.1007/s12355-016-0473-7
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DOI: https://doi.org/10.1007/s12355-016-0473-7