Prapti Singh¹, Samantha Good², Karen Summers¹, Joseph A. Charbonnet², Amy E. Sparks¹, Aileen F. Keating³ 

¹ Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, University of Iowa, 200 Hawkins Drive, Iowa City, Iowa 52242, USA 

² Department of Civil, Construction, and Environmental Engineering, Iowa State University, Ames, IA, 50011, USA. 

³ Department of Animal Science, Iowa State University, Ames, IA, 50011, USA 

Introduction: Per- and polyfluoroalkyl substances (PFAS) are ubiquitous environmental toxicants associated with negative reproductive health outcomes. Our objective was to quantify PFAS in plasma and follicular fluid of female patients undergoing infertility treatment in Iowa. 

Methods: This study used plasma and follicular fluid approved for collection from the University of Iowa tissue bank (IRB #201709794). Demographic data including age, BMI, race, gravida, parity, and an infertility diagnosis of PCOS, endometriosis or unexplained infertility. Fertility outcomes included insemination method, fertilization rate, metaphase II oocytes, number of total blastocysts meeting transfer or cryopreservation criteria, live birth with IVF, number of cycles completed to achieve conception and cycle outcomes. Using solid-phase extraction and liquid chromatography-tandem mass spectrometry, we surveyed in triplicate tissue bank plasma and follicular fluid samples for 43 PFAS, including both legacy and novel species. Statistical analysis was performed with SPSS v29. Quantitative data are presented as means (±SD) and categorical data as frequencies. ANOVA (and Welch’s ANOVA as needed) was used to compare continuous data. The Chi-squared test was used to evaluate differences in categorical factors. Tukey’s HSD test was applied to make comparisons between individual groups. 

Results: There were 42 subjects included with 14 in each diagnosis group: A (PCOS), B (endometriosis) or C (unexplained infertility). Mean age (A-30 ± 2.32, B-30 ± 2.31, and C-31 ± 2.06 years; p=0.64) and BMI (A-29.74 ± 5.86, B-25.30 ± 3.25, C-25.17 ± 6.26; p=0.04) kg/m2 were similar amongst groups. Most patients (86%) identified as White and most had not had a live birth at entry to care. Live birth was similar amongst groups (A-93%, B-93%, C-86%). Of the 43 PFAS targeted, 21 species in plasma and 23 species in follicular fluid were detected above the limit of quantification. Twelve species were detected among at least 50% of patients (6:2 FTS (plasma only), L + br PFHxS, L + br PFOS, PFBA (follicular fluid only), PFBS (plasma only), PFDA, PFHpA, PFHpS, PFHxA, PFMBA (plasma only), PFNA, PFOA, PFPeS (follicular fluid only), PFUdA). Amongst the 12 PFAS, there were no differences in plasma or follicular fluid concentrations amongst the infertility groups (p-values 0.18-0.94). PFOA and L + Br PFHxS were present in practically all subjects. Five species were detected in 100% of plasma samples (L+br PFOS, PFOA, L+brPFHxS, PFNA and PFHpS), and three species were detected in 100% of follicular fluid samples (L+br PFOS, PFOA, and L+br PFHxS). PFAS concentrations were consistently higher in plasma than follicular fluid within the same patient. 

Conclusion: Legacy and novel PFAS are present in human plasma and follicular fluid samples stratified by PCOS, unexplained infertility, and endometriosis in patients seeking infertility treatment in Iowa. 

Key words: PFAS, infertility, plasma, follicular fluid