Analytical and Bioanalytical Chemistry

, Volume 411, Issue 10, pp 2045–2055 | Cite as

Quantitative characterization of glutaminolysis in human plasma using liquid chromatography-tandem mass spectrometry

  • Yunfei Hua
  • Xuping Yang
  • Ruiting Li
  • Peifang Liu
  • Peijia Liu
  • Linrui Li
  • Xia Yuan
  • Xiaoyi Hua
  • Yuan Tian
  • Zunjian ZhangEmail author
  • Yin HuangEmail author
Research Paper


Glutaminolysis is the metabolic pathway that lyses glutamine to glutamate, alanine, citrate, aspartate, and so on. As partially recruiting reaction steps from the tricarboxylic acid (TCA) cycle and the malate-aspartate shuttle, glutaminolysis takes essential place in physiological and pathological situations. We herein developed a sensitive, rapid, and reproducible liquid chromatography-tandem mass spectrometry method to determine the perturbation of glutaminolysis in human plasma by quantifying 13 involved metabolites in a single 20-min run. A pHILIC column with a gradient elution system consisting of acetonitrile-5 mM ammonium acetate was used for separation, while an electrospray ionization source (ESI) operated in negative mode with multiple reaction monitoring was employed for detection. The method was fully validated according to FDA’s guidelines, and it generally provided good results in terms of linearity (the correlation coefficient no less than 0.9911 within the range of 0.05–800 μg/mL), intra- and inter-day precision (less than 18.38%) and accuracy (relative standard deviation between 89.24 and 113.4%), with lower limits of quantification between 0.05 and 10 μg/mL. The new analytical approach was successfully applied to analyze the plasma samples from 38 healthy volunteers and 34 patients with type 2 diabetes (T2D). Based on the great sensitivity and comprehensive capacity, the targeted analysis revealed the imperceptible abnormalities in the concentrations of key intermediates, such as iso-citrate and cis-aconitate, thus allowing us to obtain a thorough understanding of glutaminolysis disorder during T2D.

Graphical abstract


Glutaminolysis LC-MS/MS Isomers Type 2 diabetes Human plasma 



The authors would like to thank the subjects for their participation.

Funding information

This study was financially supported by the National Natural Science Foundation of China (No. 81403181), the Natural Science Foundation of Heilongjiang Province of China (No. QC2016109), and the Open Project Program of the MOE Key Laboratory of Drug Quality Control and Pharmacovigilance (China Pharmaceutical University) (No. DQCP2017MS02).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants were following the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from the individual participants who provided the blood samples.

Supplementary material

216_2019_1626_MOESM1_ESM.pdf (954 kb)
ESM 1 (PDF 954 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Yunfei Hua
    • 1
  • Xuping Yang
    • 1
  • Ruiting Li
    • 1
  • Peifang Liu
    • 2
    • 3
  • Peijia Liu
    • 2
  • Linrui Li
    • 1
  • Xia Yuan
    • 1
  • Xiaoyi Hua
    • 4
  • Yuan Tian
    • 1
  • Zunjian Zhang
    • 1
    Email author
  • Yin Huang
    • 1
    • 3
    Email author
  1. 1.Key Laboratory of Drug Quality Control and PharmacovigilanceChina Pharmaceutical University, Ministry of EducationNanjingChina
  2. 2.Department of Neurology, the Second Affiliated HospitalHarbin Medical UniversityHarbinChina
  3. 3.Key Laboratory of Myocardial IschemiaHarbin Medical University, Ministry of EducationHarbinChina
  4. 4.Wuxi Center for Drug Safety ControlWuxiChina

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