A method for the determination of neutral steroid mono- and disulphates in human urine is described. The mono- and disulphate fractions of urinary steroids were isolated by Sephadex LH-20 chromatography. After solvolysis, the liberated steroids were fractionated on a column of silicic acid and by thin-layer chromatography and analysed by gas-liquid chromatography (GLC) as trimethyl silyi ethers on QF-1 and SE-30 liquid phases. The following neutral steroid monosulphates were determined : androsterone, epiandrosterone, etiocholanolone, 3α-hydroxy-5-androsten-17-one, dehydroepiandrosterone, 5-androstene-3β,17α-diol, 5-androstene-3β,17β-diol, 16α-hydroxyandrosterone, 16α-hydroxyepiandrosterone, 16α-hydroxyetiocholanolone, 3α,16α-dihydroxy-5-androsten-17-one, loa-hydroxydehydroepiandrosterone, 16β-hydroxydehydroepiandrosterone, 3β,17β-dihydroxy-5-androsten-16-one,7α-hydroxyandrosterone, 7β-hydroxyandrosterone, 7α-hydroxyepiandrosterone, 7β-hydroxyepiandrosterone, 7α-hydroxydehydroepiandrosterone, 7β-hydroxydehydroepiandrosterone, 5α-androstane-3α,16α,17β-triol, 5-androstene-3β,16α,17β-triol, 5-androstene3β, 16β,17α-triol, 5-androstene-3β,16β,17β-triol, 5-pregnene-3β,20α-diol, 16α-hydroxypregnenolone, 5-pregnene-3β,16α,20α-triol and 5-pregnene-3β,17α,20α-triol. In the disulphate fraction, 5-androstene-3β,17α-diol, 5-androstene-3β,17β-diol, 5-pregnene3β, 20α-diol, 21-hydroxypregnenolone and 5-pregnene-3β,17α,20α-triol were measured. The specificity of the method was established by mass spectrometric analysis of the effluent from GLC columns. The recoveries of reference steroid mono- and disulphates added to the urine were 74–101%. Dehydroepiandrosterone and loa-hydroxydehydroepiandrosterone were the most abundant compounds in the monosulphate fraction and 5-androstene-3β,17α-diol was the main compound in the disulphate fraction of neutral steroids in urine. Some figures for the daily excretion of the steroids are given.