Serum creatinine assay by use of 3,5-dinitrobenzoates: a critique

Methyl-3,5-dinitrobenzoate and 3,5-dinitrobenzoyl chloride are demonstrated to react with creatinine through their conversion to 3,5-dinitrobenzoate. Analytical recovery of creatinine added to serum is improved by an "acid-supernate" modification, which is also much less susceptible to a combination of known interfering agents than is the picrate procedure. The "neutral-supernate" procedure shows negligible interference by cephalothin as compared to commonly used picrate procedures. Replacement of tetramethylammonium hydroxide by sodium hydroxide was also tested. The interrelationships between dimethyl sulfoxide concentration, recoveries of creatinine, and slopes of correlations (r greater than .992) of various dinitrobenzoate and picrate methods are critically assessed. Because of its precision, greater specificity, and other criteria, we recommend the methyl-3,5-dinitrobenzoate (or 3,5-dinitrobenzoic acid) neutral-supernate method for routine applications.