4.7 Article

A sensitive and simple HPLC-FLD-based method for the measurement of intracellular glucose uptake

Journal

FOOD CHEMISTRY
Volume 372, Issue -, Pages -

Publisher

ELSEVIER SCI LTD
DOI: 10.1016/j.foodchem.2021.131218

Keywords

HPLC-FLD; 2-NBDG; Glucose uptake; Autofluorescence; Fluorescent natural products

Funding

  1. Guangxi Innovation-driven Development Special Foundation Project [GuiKe AA18 118049]
  2. Sichuan Science and Technology Planning Project [2020YFS0370]
  3. Macau Science and Technology Development Fund [0147/2019/A3]
  4. Research Committee of the University of Macau [MYRG2018-00239-ICMS]

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A reliable HPLC-FLD method was developed to measure intracellular glucose uptake by extracting, separating, and detecting 2-NBDG, providing a simple, sensitive, and accurate measurement. The method has been successfully used to assess the glucose uptake activity of anti-diabetic drugs and fluorescent natural products.
Glucose is a primary source of energy used in most organisms. Thus, development of reliable approaches to measure intracellular glucose uptake is an important research issue. 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino]-2-deoxy-D-glucose (2-NBDG), as a fluorescent glucose derivative, has been widely used to track intracellular glucose uptake by fluorescence imaging and measuring in mammalian cells. However, the avoid-less cross-interference of intrinsic autofluorescence background and tested fluorescent compounds limits its ability to provide trustworthy information on intracellular glucose uptake. By the extraction, separation and detection of 2-NBDG, a simple, sensitive and accurate HPLC-FLD method was established and validated for the measurement of intracellular glucose uptake in HepG2 cells. The developed method has been employed successfully to assess the glucose uptake activity of anti-diabetic drugs and fluorescent natural products. A fit-for-purpose partial validation was further performed for quantification and comparison of glucose uptake in AML12, LO2 hepatocytes, L6 myoblasts and 3T3-L1 preadipocytes.

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