TCA/Kreb Cycle Method

Measured Analytes

  1. Alpha ketoglutarate
  2. Citrate
  3. Fumarate
  4. Isocitrate
  5. Lactate
  6. Malate
  7. Methyl malonate
  8. Pyruvate
  9. Succinate
  10. 2-hydroxyglutarate


Liquid chromatography tandem mass spectrometry (LC-MS) was carried out with a Shimadzu Prominence HPLC system and SCIEX 4000 mass spectrometer. An aliquot of 10 µl of each sample was injected onto an Accucore 2.6 µm C18 100 x 2.1 mm column at 40oC for gradient separation. Mobile phases were: A, ddH2O with 0.1% Formic Acid; and B, Acetonitrile with 0.1% Formic Acid. Gradient schedule started at 10% B and was linearly increased to 30% B at 1 minute, 85% B at 5.5 minutes, 95% B at 5.6 minutes and held at 95% B until 6.5 minutes. At 6.5 minutes the gradient was returned to starting conditions and allowed to re-equilibrate until 8.5 minutes. Time per injection was 8.5 minutes.

Sample Processing

  1. Derivatization Method:
    Analyte derivatives for the TCA method were processed as described in Bo Tan et al [1].
  2. Extraction Protocols:
    Extraction protocols for sample processing are dependent on the matrices involved. Please see extraction methods section for more specific instructions.


[1] Anal Biochem. 2014 Nov 15;465:134-47. doi: 10.1016/j.ab.2014.07.027. Epub 2014 Aug 4.
Derivatization of the tricarboxylic acid intermediates with O-benzyl hydroxylamine for liquid chromatography-tandem mass spectrometry detection.
Tan B, Lu Z, Dong S, Zhao G, Kuo MS.


This tab might be populated with additional information like PowerPoint slides from presentations at conferences, class presentations, grant associated links, and other components that would be directly tied to the method.