Att 4_SOW for HOPE 1000 PFAS Analysis 6.11.26.pdf

75N94026Q00056 (UPDATED)

Statement of Work (UPDATED): Per - and Polyfluoroalkyl Substances Analysis in Health Outcomes Pregnancy Exposures 1000 Study (PFAS in HOPE 1000 Study)

June 11, 2026

This study will examine novel and legacy per- and polyfluoroalkyl substances (PFAS) in the HOPE 1000 study. HOPE 1000 is a longitudinal study of mothers and infants to understand health outcomes related to pregnancy and early life exposures which was conducted at Duke University between 2019 and 2024. Under the study design, participants are recruited prior to 24 weeks gestation (median=16 weeks) at the Duke Maternal and Fetal Medicine (MFM) clinic and complete questionnaires and provide biological specimens throughout pregnancy, including up to one plasma sample per trimester. The study is now complete and includes 477 participants for whom 903 plasma samples are stored (n=107 in the first trimester; n=389 in the second trimester; and n=407 in the third trimester). Previously, the National Institute of Environmental Health Sciences (NIEHS) funded a questionnaire that was implemented in the study to assess personal care product and dietary sources of PFAS exposure in early and late gestation. The purpose of this add-on study is to examine novel and legacy plasma PFAS concentrations in order to assess the extent of exposure and variability across the course of gestation, determine sources of exposure (based on questionnaire data collected in the cohort), and assess associations between exposure biomarkers and maternal metabolic health during pregnancy and postpartum. This work requires targeted and non-targeted analysis of PFAS congeners. Targeted analysis must be performed using liquid chromatography with high resolution mass spectrometry (LC-HRMS). Untargeted full analysis of novel PFAS, other exposures, exposomics, and metabolomics must also be performed using LC-HRMS.

To fully assess novel and legacy PFAS exposure in this study population the following is required:

I. The contractor shall perform targeted analysis of at least the PFAS, analytes identified in the Supplemental Table below including the congeners perfluoro-1-butanesulfonate (PFBS) and Trifluoroacetic acid (TFA), using liquid chromatography with tandem high resolution mass spectrometry (LC-HRMS). A list of minimum PFAS for targeted analysis and abbreviations is provided in Supplemental Table 1 below.

Average detection limit for all congeners must be between 0.01 and 0.05 ng/mL. The laboratory must measure these compounds at such detection limits in human plasma or serum samples. Additionally, the laboratory (contractor) must also participate in a nationally-recognized proficiency testing program (i.e., showing evidence of having within-range concentrations for human plasma measured on blinded samples).

II. The contractor shall perform non-targeted (also referred to as untargeted) analysis using LC-HRMS to quantitate additional PFAS features and annotate them using the best available libraries. This requires use of publicly available and in-house libraries.

The contractor must perform the analysis, clean the data using established pipelines, and annotate the features to said libraries before providing a final dataset to the investigator at NIEHS. Analysis should combine measurements across multiple modes (e.g., at least four analytic configurations) to optimize detection and identification of untargeted PFAS. All materials must be shared with investigators at NIEHS, including data scripts and tools, raw instrument files, and final data files (i.e., open source and fair data frameworks).

75N94026Q00055 (UPDATED)

III. In addition to PFAS, the non-targeted LC-HRMS data shall be annotated for other environmental exposures (exposomics) and metabolomics using the best publicly and privately available libraries. As with non-targeted PFAS data, the provider must perform the analysis, clean the data using established pipelines, and annotate the features to said libraries before providing a final dataset to the investigators at NIEHS. Analysis should combine multiple measurement strategies across multiple modes (e.g., at least four analytic configurations) to optimize detection and identification of metabolites. All materials must be shared with investigators at NIEHS, including data scripts and tools, raw instrument files, and final data files (i.e., open source and fair data frameworks). These methods combined will allow NIEHS to comprehensively assess the extent of exposure in the HOPE 1000 study population and address the specific aims of the study. These methods will be applied in the specific tasks listed below:

• Receive shipment of plasma samples (n=903), each with 250 μL, from Duke University and store at -80 degrees C until analysis;

• Analyze 903 plasma samples using untargeted LC-HRMS to estimate relative quantities of novel PFAS, other exposures, exposomics, and metabolomics, fulfilling the requirements described above;

• Analyze 903 samples using targeted analysis to quantify PFAS including PFBS and TFA, fulfilling the requirements described above;

• Re-freeze and store remaining plasma volume at -80 degrees C until analysis of all samples is complete;

• Quality control and assurance checks of collected data;

• Transfer of clean data and raw datasets to Dr. Ferguson at NIEHS;

• Shipment of remaining samples back to Duke University;

• Technical Consulting on analysis of data as needed and support for manuscripts, presentations, and other scientific materials that arise from the collected data.

Supplemental Table 1. PFAS required in targeted assay.

Sodium tetrafluoro-2-(heptafluoropropoxy) propanoic acid (HFPO-DA) “GenX”

Sodium perfluoro-n-undecanoate (PFUA)

Sodium perfluoro-n-pentanoate (PFPeA)

Sodium perfluoro-n-hexanoate (PFHxA)

Sodium perfluoro-n-dodecanoate (PFDoA)

Sodium perfluoro-n-decanoate (PFDA)

Sodium perfluoro-n-butyrate (PFBA)

Sodium perfluoro-1-propanesulfonate (PFPrS)

Sodium perfluoro-1-pentanesulfonate (PFPeS)

Sodium perfluoro-1-nonanesulfonate (PFNS)

Sodium perfluoro-1-heptanesulfonate (PFHpS)

Sodium 1H,1H,2H,2H-perfluorooctanesulfonate (6:2 FTS)

Sodium 1H,1H,2H,2H-perfluorohexanesulfonate (4:2 FTS)

Sodium 1H,1H,2H,2H-perfluorododecanesulfonate (10:2 FTS)

Sodium 1H,1H,2H,2H-perfluorodecanesulfonate (8:2 FTS)

Potassium perfluorohexanesulfonate (PFHxS) (mix of isomers)

Potassium perfluoro-1-dodecanesulfonate (PFDoDS)

Potassium perfluoro-1-decanesulfonate (PFDS)

Potassium perfluoro-1-butanesulfonate (PFBS) Potassium 9-chlorohexadecafluoro-3-oxanonane-1-sulfonate (9Cl-PF3ONS) (F53B Major) Potassium 11-chloroeicosafluoro-3-oxaundecane-1-sulfonate (11Cl-PF3OUdS) (F53B Minor)

Perfluorooctanesulfonate (PFOS) (mix of isomers)

Perfluorooctanesulfonamide (PFOSA) (mix of isomers)

Perfluoro(2-ethoxyethane) sulfonic acid (PFEESA)

Perfluoro-n-tridecanoic acid (PFTrDA)

Perfluoro-n-tetradecanoic acid (PFTeDA)

Perfluoro-n-octanoic acid (PFOA)

Perfluoro-n-octadecanoic acid (PFODA)

Perfluoro-n-nonanoic acid (PFNA)

Perfluoro-n-hexadecanoic acid (PFHxDA)

Perfluoro-n-heptanoic acid (PFHpA)

Perfluoro-4-methoxybutanoic acid (PFMBA)

Perfluoro-3,6-dioxaheptanoic acid (PFDHA)

Perfluoro-3-methoxypropanoic acid (PFMPA)

N-Methylperfluorooctanesulfonamidoethanol (N-MeFOSE) (mix of isomers)

N-Methylperfluorooctanesulfonamidoacetic acid (N-MeFOSAA) (mix of isomers)

N-Methylperfluorooctanesulfonamide (N-MeFOSA) (mix of isomers)

N-Ethylperfluorooctanesulfonamidoethanol (N-EtFOSE) (mix of isomers)

N-Ethylperfluorooctanesulfonamidoacetic acid (N-EtFOSAA) (mix of isomers)

N-Ethylperfluorooctanesulfonamide (N-EtFOSA) (mix of isomers)

Dodecafluoro-3H-4,8-dioxanonanoic acid (DONA, ADONA)

Perfluoropropanoic Acid (PFPrA)

Sodium trifluoromethane sulfonate (PFMeS)

Sodum perfluoroethanesulfonate (PFEtS)

Trifluoroacetic acid (TFA)

Chloroperfluoropolyether carboxylate (MFS-N4, Cl-PFPECA (0,3) )

Chloroperfluoropolyether carboxylate (MFS-M4, Cl-PFPECA (1,2))

Chloroperfluoropolyether carboxylate (MFS-N3, Cl-PFPECA (0,2))

Chloroperfluoropolyether carboxylate (MFS-M3, Cl-PFPECA (1,1) )

Chloroperfluoropolyether carboxylate (MFS-N2, Cl-PFPECA (0,1))

Chloroperfluoropolyether carboxylate (MFS-N5, Cl-PFPECA (0,4))

8-Chloroperfluorooctylphosphonic acid (Cl-PFOPA)

6-Chloroperfluorohexylephosphonic acid (Cl-PFHxPA)

8:2diPAP

6:2diPAP

Sodium 1H, 1H, 2H, 2H-perfluorooctylphosphate (6:2PAP)

Sodium bis(perfluorooctyl)phosphinate(8:8PFPIA)

Sodium bis(perfluorohexyl)phosphinate(6:6PFPIA)

Perfluoro-4-ethylcyclohexanesulfonate

Sodium 1H, 1H, 2H, 2H-perfluorodecyltphosphate (8:2PAP)

HFPO-TrA

Sodium perfluoro-1-undecaenesulfonate (PFUdS) Sodium (1H, 1H, 2H, 2H-perfluoroocytl-1H,1H,2H,2H-perfluorodecyl)phosphate (6:2/8:2diPAP)

FOSAA

MEFBSA

HFPO-TeA