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Using the intravaginal tetrazolium method of Martin (1964), the oestrogenic activity in plasma samples (5–10ml.) taken monthly from three sows during pregnancy has been estimated.

(1) 'Free' oestrogens were extracted from the plasma with ethyl acetate. This extract was passed through a florisil column to remove corticosteroids and oestriol (Saba, 1964a). The eluate, which would contain oestradiol, oestrone and progesterone, was evaporated to dryness. The dry residue was taken up in a mixture of benzene—petroleum ether (50:50, v/v) and the phenolic oestrogens were extracted with N-NaOH. This alkaline extract was acidified and extracted with ether (Saba, 1964b).

(2) 'Bound' oestrogen. After removal of the residual ethyl acetate, the plasma was hydrolysed in 1·5N-HCI (Preedy & Aitken, 1957) and extracted with ether as described above. The residue was partitioned between benzene—petroleum ether and NaOH as under (1).

The mean recovery of oestrone added to plasma