Attachment 1 Statement of Work 1232SA26Q0273.pdf

Eastern Regional Research Center (ERRC), USDA ARS 600 East Mermaid Lane Wyndmoor, PA, 19038

Statement of Work – Mass Spectrometry-Based Untargeted Metabolomics Analysis

Background

Scientists from the ERRC are interested in evaluating the effect of different food components on the metabolism of the gut microbiome using in vitro and ex vivo methods.

Scope

ERRC will use this acquisition to obtain mass spectrometry-based untargeted metabolomics results to further their research goals.

Objectives

Scientists from the DFFRU at the ERRC expect to receive untargeted metabolomics results and raw data files from this analysis. Results will be analyzed within the context of the experimental design and shared within manuscripts and presentations.

Tasks or Requirements:

ERRC scientists will ship filtered supernatant overnight on dry ice for mass spectrometry-based untargeted metabolomics and data analysis.

There will be two shipments of two independent experiments. The first experiment will have 177 samples, and the second experiment will have 81 samples. The total sample number will be 258. The date of the shipments is unknown but will occur within the 12-month period of performance identified in the contract. Once samples are received the company will have 45 days to finish the metabolomics and transfer the data files. All data will be given to the location and rights, and utilization will be the sole property of USDA.

Upon delivery samples will be thawed and processed for analysis. Processed supernatants will be analyzed using three panels: primary metabolism, complex lipids, and biogenic amines. Combined these panels must be capable of providing data for over 1000 validated compounds and others of unknown chemical structure. Data will be reported with retention times, quantification masses, biochemical database identifiers, and adduct information. Metabolites can be reported as relative peak heights, but absolute quantitation should be available if desired. Metabolite annotations must be assigned a level of confidence rating (minimum 4, but ideally 5 levels), as follows:

1. Confirmed structure match based on MS, MS/MS, and RT of chemical standard

2. Probable structure based on MS/MS match in a spectral database

3. Tentative structure based on MS1 m/z database match

4. Molecular formula identification based on isotope abundance, adduct formation etc.

5. Exact mass m/z. Raw data files and methods must be available to share with ERRC.

Eastern Regional Research Center (ERRC), USDA ARS 600 East Mermaid Lane Wyndmoor, PA, 19038

Delivery:

Metabolomic results will be available within 45 days after samples are received.

Scope of Work

1. HILIC-LC-Orbitrap untargeted metabolomics acquisition

2. Sample preparation and metabolite extraction

3. Pooled QC monitoring and drift assessment

4. Feature detection and annotation (Thermo Compound Discoverer + mzCloud)

5. Normalized data matrix and QC summary report

6. Secure data delivery

Government-Furnished Property

DFFRU/ERRC scientists will send filtered supernatants from bacterial cultures for analysis.