Determination of barbituric acid, utilizing a rapid and simple colorimetric assay.

Barbituric acid is widely used in the manufacturing of plastics, textiles, polymers, and pharmaceuticals. Three assay methods are presented, which can utilize either aqueous or solid samples. The detection of as little as 10 microg of barbituric acid can be accomplished, either as an aqueous or solid sample, when using micro reaction tubes. The red-violet endpoint is easily discerned and results upon the formation of a violuric acid derivative. A Spot Test protocol is described which allows for a positive/negative indication of barbituric acid presence. The Spot Test is sensitive to as little as 18.75 microg/ml of barbituric acid. The construction of a Standard Curve for assaying multiple samples and over longer time periods is demonstrated, and is shown to be linear from 18.75 microg/ml to 2.25 mg/ml of barbituric acid. Spectrophotometer readings are made from an absorption peak appearing at 530 nm. The molar absorptivity of the violuric acid derivative is calculated to be 31.4 per mol/l per cm. The micro reaction tube assay will quantitate as little as 10 microg of barbituric acid through interpolation with controls containing a known amount of analyte. Inorganic salts such as NaCl, NaN(3), LiBr, and CaCO(3) do not interfere with endpoint determination. Many organic compounds (also pharmaceuticals) do not inhibit the reaction.