Determination of aminopyrine, dipyrone and its metabolites in urine by high-performance liquid chromatography

A readily applicable and accurate isocratic high-performance liquid chromatography method for the detection of aminopyrine, dipyrone and its metabolites in urine is described. Parent drugs and four metabolites were chloroform-extracted from 1 ml of urine after addition of the internal standard isopropylaminoantipyrine and alkalinization. The organic phase was evaporated to dryness, and the residue was reconstituted in the mobile phase, which was injected onto a Spherisorb ODS 5 microns particle-size column (250 x 4.6 mm) using as mobile phase water, methanol, triethylamine, and acetic acid. The column eluent was monitored by ultraviolet absorption at 254 nm. Excellent linearity (r > 0.99) was obtained in the range 1-150 micrograms/ml urine, either for parent drugs and metabolites. This method offers a sensitive assay for aminopyrine, dipyrone (widely consumed in some countries) and its metabolites. After oral administration and collection of 24-h urine, this method allows the in vivo study of aminopyrine metabolism, which reflects liver function.