[Technical note: proposal of a routine method of accurate measurement of urinary trans,trans-muconic acid]

A method for routine measurement of urinary trans,trans-muconic acid. A method is proposed which allows the accurate determination of urinary trans,trans-muconic acid (Ma) by HPLC with UV detection. Sample pretreatment consists of a first purification on strong anionic exchange (SAX) cartridges followed by extraction on "end capped" reversed-phase (C18-EC) ones. Purified samples are directly injected onto a chromatographic column (C18, 150 x 4.6 mm I.D., 3 microns) which is eluted with water: methanol: acetic acid (97:2:1, v/v) mixture followed by a "spike" of acetonitrile. The retention time of MA is 15 min, the coefficient of variation is lower than 3%, the limit of detection is 7 micrograms/l. The method seems suitable for accurate biological monitoring of subjects exposed to benzene.