Glyoxyl/Methylglyoxyl MRM Method

Measured Analytes

  1. Quinoxaline
  2. 2-methylglyoxyl
  3. 5-methylglyoxyl


Liquid chromatography tandem mass spectrometry (LC-MS) was carried out with a Shimadzu Prominence HPLC system and SCIEX 4000 mass spectrometer. An aliquot of 20 µl of each sample was injected onto an Luna 3 µm Phenyl-hexyl 200 x 2.0 mm column at 50oC for gradient separation. Mobile phases were: A, ddH2O with 0.1% Formic Acid; and B, Acetonitrile with 0.1% Formic Acid. Gradient schedule starts at 10% B and is linearly increased to 100% B at 3 minutes, then held at 100% B until 3.25 minutes, and then returned to 10% B at 3.4 at minutes and re-equilibration until 5 minutes. Time per sample injection was 5 minutes with a flow rate is 400µL/min.

glyoxyl methylglyoxyl mrm

Extraction Protocols:

Extraction protocols for sample processing are dependent on the matrices involved. Please see extraction methods section for more specific instructions.

Derivatization Protocol:

  1. Mix 400µL or Samples + 44.4µL OPD (o-phenylenediamine) Prep (100µM OPD, 100µM DPTA, 71 ng/ml 5MQ, 4.5% TCA)
  2. Vortex and Centrifuge Mixture briefly
  3. Incubate in water bath at 65°C in dark for 1 hour
  4. Centrifuge at end and add 200µL to autosampler vial

Method was adapted from the following reference:


[1] Assay of methylglyoxal and glyoxal and control of peroxidase interference
Paul J. Thornalley, Naila Rabbani
Biochemical Society TransactionsMar 20, 2014,42(2)504-510; DOI: 10.1042/BST20140009


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