Fatty Acid Ethyl Esters as Biological Markers for the Diagnosis of Fetal Alcohol Sydrome

Wei, Pei-Hsun, Natalie McCormick, Victoria Hornik-Rosinski, Hyesook Kim, and Elizabeth Roberts-Kirchhoff

Fetal alcohol syndrome can affect children born to mothers who have consumed alcohol during their pregnancy. Children with this disorder may have altered fatty acid metabolism. 13-Hydroxyoctadeca-9,11-dienoic acid (13-HODE) was converted to an ethyl ester with lipase acrylic resin and ethanol. The ethyl ester was converted to a succinate derivative via its C13 hydroxyl group. Identification of products was achieved with GC/MS. The succinate was conjugated to keyhole limpet hemocyanin (KLH), bovine serum albumin (BSA), and horseradish peroxidase (HRP). IgG was isolated from the KLH-immunized goat serum and used in immunoassays with the BSA-conjugate. The isolated IgG recognized the BSA conjugate as determined by ELISA. Western blot analysis revealed that, whereas the IgG did not bind to BSA or HRP, the IgG strongly bound to BSA- and HRP-conjugated with ethanol-esterified 13-HODE. The conjugated proteins were analyzed by immunoblot analysis after limited trypsin digestion. (Supported in part by NIAAA SBIR R43 AA014535).