[3] Glyoxal was first prepared and named by the German-British chemist Heinrich Debus (1824–1915) by reacting ethanol with nitric acid.

The single largest commercial source is BASF in Ludwigshafen, Germany, at around 60,000 tons per year.

Commercial bulk glyoxal is made and reported as a 40% solution in water by weight[3] (approx.

Glyoxal may be synthesized in the laboratory by oxidation of acetaldehyde with selenious acid[6] or by ozonolysis of benzene.

[11] Guanine bases in DNA can undergo non-enzymatic glycation by glyoxal to form glyoxal-guanine adducts.

[13] Coated paper and textile finishes use large amounts of glyoxal as a crosslinker for starch-based formulations.

Glyoxal solutions can also be used as a fixative for histology, that is, a method of preserving cells for examining them under a microscope.

It is estimated that, at concentrations less than 1 M, glyoxal exists predominantly as the monomer or hydrates thereof, i.e., OCHCHO, OCHCH(OH)2, or (HO)2CHCH(OH)2.

[15] Glyoxal has been observed as a trace gas in the atmosphere, e.g. as an oxidation product of hydrocarbons.