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Clinical Chemistry 33: 1129-1132, 1987;
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Clinical Chemistry, Vol 33, 1129-1132, Copyright © 1987 by American Association for Clinical Chemistry

Mechanism of interference with the Jaffe reaction for creatinine

MH Kroll, NA Roach, B Poe and RJ Elin

We investigated the mechanism of the Jaffe reaction for determination of creatinine by studying the spectrophotometric, kinetic, and equilibrium properties of the reaction of picrate with creatinine and with cyclic and aliphatic ketones. Absorbance spectra for the reaction products of picrate with all the ketones were superimposable with that of creatinine (Amax, 490 nm). Cyclic ketones not containing nitrogen had a molar absorptivity less than half that of creatinine and equilibrium constants approximately 0.01 that of creatinine. Aliphatic ketones, except for benzylacetone, had molar absorptivities similar to that of creatinine, but all of these compounds had equilibrium constants approximately a tenth or less that of creatinine. The common structure for all of the compounds reacting with picrate is the carbonyl group. The variable magnitude of interference for aliphatic and cyclic ketones is ascribable to the different rate constants, molar absorptivities, and equilibrium constants as compared with creatinine. Structures adjacent to the carbonyl group significantly affect the absorptivity and equilibrium constant, but steric hindrance is the major factor affecting the rate of reaction. We postulate that the carbonyl group is required for the Jaffe reaction, and we suggest a mechanism for the reaction.


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Copyright © 1987 by the American Association for Clinical Chemistry.