Received on May 9, 2008
Accepted on October 24, 2008

Endocrinology and Metabolism

Steroid Profiles in Ovarian Follicular Fluid from Regularly Menstruating Women and Women after Ovarian Stimulation

¹ ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, UT, and Analytical Chemistry/Department of Physical and Analytical Chemistry, Uppsala University, Uppsala, Sweden
² Departments of Obstetrics and Gynecology, Uppsala University Hospital, Uppsala, Sweden
³ Department of Obstetrics, Gynecology and Perinatology, Donetsk State Medical University, Donetsk, Ukraine
⁴ ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, UT, and Department of Pathology, University of Utah, Salt Lake City, UT
⁵ Division of Clinical Chemistry, Department of Laboratory Medicine, Karolinska Institutet, University Hospital at Huddinge, Huddinge, Stockholm, Sweden
⁶ Analytical Chemistry/Department of Physical and Analytical Chemistry, Uppsala University, Uppsala, Sweden

^* To whom correspondence should be addressed. E-mail: kushnmm{at}aruplab.com.

BACKGROUND: Information on the concentrations of steroids in ovarian follicular fluid (FF) from regularly menstruating (RM) women has been limited because of the absence of methods for the simultaneous quantification of multiple steroids in small volumes of FF. We studied steroid profiles in FF during the early follicular phase of the menstrual cycle and after ovarian stimulation for in vitro fertilization (IVF), and compared concentrations with published values obtained by immunoassay (IA).

METHODS: We used liquid chromatography–tandem mass spectrometry (LC-MS/MS) to measure 13 steroids in 40-µL aliquots of FF samples from 21 RM women and from 5 women after ovarian stimulation for IVF. Relationships between concentrations of steroids and their ratios (representations of the enzyme activities) were evaluated within and between subgroups.

RESULTS: The concentrations of testosterone (Te), androstenedione (A4), and estradiol (E2) measured by LC-MS/MS were lower than those previously reported in studies with IAs. In RM women, androgens were the most abundant class of steroids, with A4 being the major constituent. The concentrations of 17-hydroxyprogesterone (17OHP), total androgens, and estrogens were 200- to 1000-fold greater in FF than in serum. Compared with RM women, FF samples from women undergoing ovarian stimulation had significantly higher concentrations of E2 (P = 0.021), pregnenolone (P = 0.0022), 17OHP (P = 0.0007), and cortisol (F) (P = 0.0016), and significantly higher ratios of F to cortisone (P = 0.0006), E2 to estrone (P = 0.0008), and E2 to Te (P = 0.0013).

CONCLUSIONS: The data provide the first MS-based concentration values for 13 steroids in ovarian FF from RM women, from estrogen- and androgen-dominant follicles, and from women after ovarian stimulation for IVF.