Labelled estradiol-17 beta(E2) or estrone (E1), when incubated with guinea pig liver slices, is metabolized by two main pathways. Part of each substrate is converted to estrone-3-glucuronide and estradiol-3-glucuronide. A further part of each is metabolized to estradiol-3-sulfate (E23S) and estrone-3-sulfate (E13S), which are interconverted. The latter conjugate appears to be the substrate for a 16 alpha-hydroxylase forming 16 alpha-hydroxyestrone-3-sulfate (16 alpha OHE13S). This, in turn, is further sulfurylated to yield 16 alpha-hydroxyestrone-3, 16-disulfate, accompanied by estriol-3,16-disulfate. A relatively small amount of tentatively indentified '6-hydroxyestrone disulfate accompanies these other two diconjugates. The guinea pig liver system suggests itself as a useful and relatively simple model for further study of 16 alpha-hydroxylation of E13S. The use of the latter as a natural subd E23S are present in liver, kidney, blood, gallbladder bile, intestine, uterus, and placenta after injection of labelled E2 into mature male and female guinea pigs. Some evidence has been obtained for the disulfate fraction (above) in liver and bile after injection of labelled E1.