High-performance liquid chromatographic determination of primary aromatic amines in urine after derivatization to an azo dye with 2-aminoanthracene.
A sensitive HPLC method for the determination of primary aromatic amines (anilino compounds) is described. Samples were prepared by derivatization of the substrate to an azo dye with 2-aminoanthracene (2-AA). 2-AA was found to react with the diazonium salts prepared from substituted anilines such as 4-halo, -sulfonyl, -carboxyl, -nitro or -acetyl derivatives, but not 4-hydroxy or -alkyl derivatives. In this work, three model compounds [sulfanilamide, 4-aminobenzoyl-beta-alanine and 4-aminobenzoic acid (PABA)] were used to test the linearity and accuracy of the method. Chromatographic separation was carried out using a reversed-phase column (ODS) and ultraviolet detection at 279 nm. Good linearity for the three compounds was found within the range 50-2000 ng/ml. The intra-day coefficient of variation for the three compounds (at 100, 500, 1000 ng/ml) was below 10%. Using this method, the urinary excretion of PABA and its metabolites was studied after oral administration of PABA to rats.
Hayashi
T
T
Research Laboratories, Roussel Morishita Co., Ltd, Shiga, Japan.
Amino
M
M
Uchida
G
G
Sato
M
M
D000814:Aniline Compounds / Q000737:chemistry / Q000652:urine*; D000818:Animals; D000873:Anthracenes / Q000737:chemistry*; D001391:Azo Compounds / Q000737:chemistry / Q000652:urine*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004396:Coloring Agents; D008297:Male; D051381:Rats; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet
D000814:Aniline Compounds; D000873:Anthracenes; D001391:Azo Compounds; D004396:Coloring Agents; C006593:2-anthramine