Statement of Work.doc

Statement of Work (SOW) Project: Carnitine, Choline, Trimethylamine N-oxide (TMAO) and Related Analysis of Samples from the USDA MAP Study

GENERAL INFORMATION

1.0 Scope of Work: Analysis of serum samples from the USDA Meat And Pattern (MAP) Study to measure the concentration of serum choline, carnitine, trimethylamine N-oxide (TMAO) and related compounds of endogenous and microbial carnitine and choline metabolism.

2.0 Background: Previous research has shown that increased concentration of TMAO may promote atherosclerosis and increase risk for cardiovascular disease. TMAO is produced by bacteria from the precursors choline and carnitine. Dr. Steve Hazen is a world recognized physician scientist who made some of the first discoveries linking TMAO with increased risk of cardiovascular disease, with the development of analytical methods to measure these compounds, and with investigating foods, such as meats, and how their consumption might impact disease risk.

CONTRACTOR REQUIREMENTS

3.0 Technical Requirements/Tasks:

· Analyze 270 human serum samples for carnitine, choline, betaine and TMAO using LC/MS/MS methods. Analytical approach must use stable isotope dilution liquid chromatography with online LC–MS/MS on a triple quadrupole mass spectrometer. A stable isotope, such as d9(trimethyl)TMAO, should be used as an internal standard. TMAO, related compounds, and d9(trimethyl)TMAO should be monitored using electrospray ionization in positive‐ion mode with multiple reaction monitoring of parent and characteristic daughter ion transitions: m/z 76→58 and 85→66, respectively. Intraday laboratory coefficients of variation (CV) for TMAO should be <6% and interday CV should be < 10%. Published lower limit of detection (LLOD) must be at least 0.02 micromolar, the lower limit of quantitation (LLOQ) must be at least 0.05 micromolar and the upper limit of quantitation (ULOQ) should be greater than 200 micromolar. Published accuracy should be at least 97%. The analytical method should be published in the peer-reviewed scientific literature, along with data of accuracy, recovery, and linearity. Methods must be consistent with those described in Anal Biochem. 2014 Jun 15; 455: 35–40.

4.0 Government Furnished:

· Government will supply a cryovial of approximately 1 ml of serum collected in a red top tube (no gel).

· Government will ship frozen samples to contractor.

5.0 Deliverables / Schedule:

· The contractor shall provide the results in electronic spreadsheet format within 12 weeks of receiving the samples.

6.0 Data Rights:

· Data shall be furnished to the Government. Contractor shall not disclose data to others.