Analytical and Bioanalytical Chemistry

, Volume 410, Issue 7, pp 2011–2018 | Cite as

Simultaneous quantification of endogenous and exogenous plasma glucose by isotope dilution LC-MS/MS with indirect MRM of the derivative tag

  • Lingling Yu
  • Chao Wen
  • Xing Li
  • Shiqi Fang
  • Lichuan Yang
  • Tony Wang
  • Kaifeng HuEmail author
Research Paper


Quantification of endogenous and exogenous plasma glucose can help more comprehensively evaluate the glucose metabolic status. A ratio-based approach using isotope dilution liquid chromatography tandem mass spectrometry (ID LC-MS/MS) with indirect multiple reaction monitoring (MRM) of the derivative tag was developed to simultaneously quantify endo-/exogenous plasma glucose. Using diluted D-[13C6] glucose as tracer of exogenous glucose, 12C6/13C6 glucoses were first derivatized and then data were acquired in MRM mode. The metabolism of exogenous glucose can be tracked and the concentration ratio of endo/exo-genous glucose can be measured by calculating the endo-/exo-genous glucose concentrations from peak area ratio of specific daughter ions. Joint application of selective derivatization and MRM analysis not only improves the sensitivity but also minimizes the interference from the background of plasma, which warrants the accuracy and reproducibility. Good agreement between the theoretical and calculated concentration ratios was obtained with a linear correlation coefficient (R) of 0.9969 in the range of D-glucose from 0.5 to 20.0 mM, which covers the healthy and diabetic physiological scenarios. Satisfactory reproducibility was obtained by evaluation of the intra- and inter-day precisions with relative standard deviations (RSDs) less than 5.16%, and relative recoveries of 85.96 to 95.92% were obtained at low, medium, and high concentration, respectively. The method was successfully applied to simultaneous determination of the endo-/exogenous glucose concentration in plasma of non-diabetic and type II diabetic cynomolgus monkeys.

Graphical Abstract

The scheme of the proposed ratio-based approach using isotope dilution LC-MS/MS with indirect MRM of the derivative tag for simultaneous quantification of endogenous and exogenous plasma glucose.


Derivatization Diabetes Endogenous glucose Exogenous glucose LC-MS/MS 



This work was supported by the National key R & D program of China (No. 2017YFC0906900), the National Natural Science Foundation of China (No. 21505142), and Yunnan Provincial Science and Technology Department (No. 2012HA015).

Compliance with ethical standards

All protocols of animal experiments were approved by the Institutional Animal Care and Use Committees at Kunming Biomed International.

Conflict of interest

The authors declare that they have no conflicts of interest.

Supplementary material

216_2018_872_MOESM1_ESM.pdf (244 kb)
ESM 1 (PDF 243 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.State Key Laboratory of Phytochemistry and Plant Resources in West China, Kunming Institute of BotanyChinese Academy of SciencesKunmingChina
  2. 2.School of Chemical Science and TechnologyYunnan UniversityKunmingChina
  3. 3.University of Chinese Academy of SciencesBeijingChina
  4. 4.Kunming Biomed InternationalKunmingChina

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