SUMMARY A new chromatographic procedure for determining small amounts of pregnanediol in urine is described. The method involves (1) acid hydrolysis, (2) toluene extraction, (3) a new permanganate oxidation step, (4) chromatography on alumina columns, (5) acetylation, (6) further chromatography on alumina, and (7) colorimetry after developing a colour with sulphuric acid. The method is considered under the headings of specificity, sensitivity, accuracy and convenience. The significance of results obtained from male, proliferative phase female, and post-menopausal urine is discussed.

Peritoneal fluid was aspirated during laparoscopy from normal women at various stages of the menstrual cycle. The volume of the fluid was found to be influenced by the stage of the cycle, rising from an early proliferative mean value of 0\m=.\8ml to a mean volume of 18\m=.\7ml after ovulation and decreasing again to a mean volume of 5\m=.\4ml in the late secretory phase. The mean volume of peritoneal fluid obtained from women using the contraceptive pill was 0\m=.\8ml, whereas only negligible amounts of fluid were aspirated at laparotomy from the peritoneal cavity of men.Despite marked changes in volume, the total protein content of the peritoneal fluid ( \ m=+-\ 60% of the concentration measured in plasma) remained fairly constant throughout the cycle, although a slightly, but significantly, lower value was found in the late proliferative phase. After ovulation the concentration of oestradiol and progesterone in the peritoneal fluid was higher than in plasma in the majority of cases, suggesting that some follicular fluid had drained into the peritoneal cavity at that time. Progesterone, in contrast to oestradiol, was also found in higher concentrations in peritoneal fluid than in plasma before ovulation.These results suggest that the volume and ovarian steroid concentration of peritoneal fluid are influenced by cyclic ovarian activity. At least two mechanisms may account for these observations: changes possibly effected by oestradiol in vascular permeability leading to a change of inflow and outflow equilibrium at the level of the peritoneal membranes and ovarian\p=n-\peritoneal transfer of fluid after and possibly also before follicular rupture.

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In a group of nine perimenopausal women, aged 37-52 years, with dysfunctional uterine bleeding (DUB), serial measurements were made of urinary total oestrogen and pregnanediol excretion and of plasma gonadotrophin and steroid levels under basal conditions and during dynamic tests (oestrogen provocation and LHRH-tests). Results were compared t o those obtained in a control group of regularly menstruating women, 23 -45 years of age. Four different patterns of hypothalamic-pituitaryovarian (H.P.O.) activity were identified in the perimenopausal subjects with DUB. In two patients with a history of persistent anovulation and cystic glandular hyperplasia of the endometrium, basal plasma gonadotrophin levels were normal but there was a failure to release an adequate amount of LH in response to endogenous and exogenous oestrogen stimulation. One subject had regular ovulatory cycles but the follicular phase was shorter and circulating levels of FSH, but not of LH, were higher than in controls. A similar monotropic increase in FSH was also present in a further patient whose cycles were irregular and included an ovulatory cycle with short follicular phase, an ovulatory cycle of normal length and an anovulatory cycle. In the remaining five women follicular development was infrequent and anovulation the rule. FSH and LH levels in these women were elevated despite the presence of circulating 17Soestradiol levels in the early-mid follicular phase range. At the time of menopausal transition in one of these subjects, the decline of plasma 170-oestradiol to undetectable levels was associated with a further rise of both gonadotrophins. Conversely, following a prolonged period of follicular development with elevated urinary total oestrogen excretion in another subject, the raised gonadotrophin concentrations were suppressed and the pituitary response to LHRH was within the normal range. LHRH responses in the other four women were augmented. Oestrogen administration failed to induce a normal LH surge in three out of the five subjects. The results indicate that marked changes in the pattern of pituitary gonadotrophin secretion can be found in perimenopausal women with DUB. The observed increase in peripheral levels of FSH (with or without concomitant increase