Analytical and Bioanalytical Chemistry

, Volume 411, Issue 3, pp 787–795 | Cite as

Simultaneous determination of lactic acid and pyruvic acid in tissue and cell culture media by gas chromatography after in situ derivatization-ultrasound-assisted emulsification microextraction

  • Hua-Yin Zhang
  • Xiao-Xin Tan
  • Kai Kang
  • Wei Wang
  • Kao-Qi LianEmail author
  • Wei-Jun KangEmail author
Research Paper


Lactic acid and pyruvic acid are important metabolites in the tricarboxylic acid cycle that can reflect the cytoplasmic redox state and mitochondrial respiratory chain function. The combination of these acids is considered as a screening index for mitochondrial disorders. Due to their biological effects, a derivatization method was developed to simultaneously detect pyruvic acid and lactic acid in tissue and cell culture media using gas chromatography. In this work, the combined derivatization method with methoxyamine hydrochloride and isobutyl chloroformate was first proposed. To improve the efficiency of derivatization, in situ derivatization-ultrasound-assisted emulsification microextraction (USAEME) was used in this study. After optimizing the volume of reagents and reaction times, good linearity values were obtained from 50 to 1000 μmol/L and 1 to 100 μmol/L for lactic acid and pyruvic acid, respectively. The limits of detection (LODs) were 0.12 μmol/L for lactic acid and 0.29 μmol/L for pyruvic acid. The recoveries of the two analytes were between 93.60 and 102.80%, and the precisions were less than 6.20%. This method was successfully applied to quantify pyruvic acid and lactic acid in the animal and cellular hypoxia models which provided an auxiliary means for the diagnosis of mitochondrial diseases.

Graphical abstract


Pyruvic acid Lactic acid Derivatization USAEME Gas chromatography 


Funding information

This work has been supported by the Natural Science Foundation of Hebei Province (No. H 2018206122).

Compliance with ethical standards

All animal experiments involved in the present study were performed in accordance with Hebei Medical University Animal Experiment Center guidelines for care and use of animals and were approved by the Animal Ethics Committee of Hebei Medical University.

Conflict of interest

The authors declare that they have no conflicts of interest.

Supplementary material

216_2018_1502_MOESM1_ESM.pdf (243 kb)
ESM 1 (PDF 171 kb)


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Copyright information

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

Authors and Affiliations

  1. 1.School of Public HealthHebei Medical UniversityShijiazhuangChina
  2. 2.School of PharmacyHebei Medical UniversityShijiazhuangChina
  3. 3.Hebei Key Laboratory of Environment and Human HealthShijiazhuangChina

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